Rushinadha Rao Kakara

BIOCHEMICAL, HEAVY METAL AND FATTY ACID PROFILING STUDIES
ON SOME SELECTED DEEP-SEA FISHES ALONG THE CONTINENTAL

SLOPE (200 TO 1200 M DEPTH) OF INDIAN EXCLUSIVE ECONOMIC ZONE
By
RUSHINADHA RAO KAKARA
Under the Supervision of
Dr. U. SREEDHAR Prof. K. SREERAMULU

INTERNAL RESEARCH DIRECTOR EXTERNAL RESEARCH DIRECTOR
FISHING TECHNOLOGY DIVISION DEPARTMENT OF ZOOLOGY,
ICAR-CIFT, VISAKHAPATNAM ANDHRA UNIVERSITY, VISAKHAPATNAM
THESIS SUBMITTED TO ANDHRA UNIVERSITY, VISAKHAPATNAM IN

PARTIAL FULFILLMENT OF THE REQUIREMENT FOR THE AWARD OF
THE DEGREE OF DOCTOR OF PHILOSOPHY IN ZOOLOGY
FEBRUARY, 2017
Dedicated To
My Beloved Parents
DECLARATION
The work incorporated in this thesis is a part of the research project
“Exploration and Assessment of demersal fishery resources along the continental
slope (200m to 1200m depth) of Indian Exclusive Economic Zone and Central Indian
Ocean” funded by Ministry of Earth Sciences, New Delhi, carried out in the
laboratory of ICAR - Central Institute of Fisheries Technology (CIFT),
Visakhapatnam. I hereby declare that this is the novel work entitled
BIOCHEMICAL, HEAVY METAL AND FATTY ACID PROFILING
STUDIES ON SOME SELECTED DEEP-SEA FISHES ALONG THE
CONTINENTAL SLOPE (200 TO 1200 M DEPTH) OF INDIAN EXCLUSIVE
ECONOMIC ZONE submitted to the Andhra University for the award of the Degree
of Doctor of Philosophy has been carried out entirely by me under the guidance of
DR. U. SREEDHAR (Internal Research Director), ICAR-Central Institute of
Fisheries Technology, Visakhapatnam and Prof. K. SREE RAMULU (External
Research Director), Department of Zoology, Andhra University, Visakhapatnam and
it has not been submitted earlier either in part time or full time to any University or
Institution for the award of any Degree
Place: Visakhapatnam,
Date: (RUSHINADHA RAO KAKARA)

Dr. U. SREEDHAR, Dt:
Principal Scientist.
CERTIFICATE
This is to certify that the thesis entitled “Biochemical, Heavy Metal and Fatty
Acid Profiling Studies on some selected deep-sea fishes along the Continental Slope (200
to 1200 m depth) of Indian Exclusive Economic Zone” is bonafide and has been carried
out by Shri. RUSHINADHA RAO KAKARA under my guidance.
(U. SREEDHAR)
Internal Guide
CERTIFICATE
This is to certify that the thesis entitled “Biochemical, Heavy Metal and Fatty
Acid Profiling Studies on some selected deep-sea fishes along the Continental Slope (200
to 1200 m depth) of Indian Exclusive Economic Zone” is bonafide and has been carried
out by Shri. RUSHINADHA RAO KAKARA under my guidance as an extramural
research scholar.
(K. SREE RAMULU)
External Guide
ACKNOWLEDGEMENTS
It is an immense pleasure to express my deep sense of gratitude to my Internal Research

Director, Dr. U. SREEDHAR, Principal Scientist, ICAR-Central Institute of Fisheries
Technology, Visakhapatnam. His dedication and keen interest above all his overwhelming attitude had
been solely and mainly responsible for completing my work. His timely advice, meticulous scrutiny,
scholarly advice and scientific approach have helped me a very great extent to accomplish this task.
I feel a huge delight to extend my sincere and heartfelt thanks to my External Research
Director, Prof. K. SREE RAMULU, Former Head & currently Board of studies Chairman,
Department of Zoology, Andhra University and Director, UGC-HRDC, Visakhapatnam, for his
motivation, excellent guidance, encouragement and co-operation throughout the study period. He
always gave me continuous scholastic supervision right from planning of research to the completion of
this thesis. It is indeed my good fortune for having the opportunity to work under him.
I am sincerely thankful to Dr. N. RAVI SANKAR, Director, ICAR-Central Institute of

Fisheries Technology for his encouragement and for providing excellent facilities to carry out my
research work.
I wish to express my sincere gratitude and respect to Prof. Ch. BHARATHI, Head and
all faculty members, Department of Zoology, Andhra University, for her support and providing
laboratory facilities to complete my research work.
I am much indebted to Dr. G. RAJESWARI, Principal Scientist and Scientist-In-

Charge, ICAR-Central Institute of Fisheries Technology, Visakhapatnam, for thier kind co-operation
and support during the course of my work.
My sincere thanks to Dr. R. RAGHU PRAKASH, Principal Scientist, ICAR-Central

Institute of Fisheries Technology, Visakhapatnam, for his support as Chief Scientist on Board and his
untiring efforts in collecting valuable resources from great depths is greatly acknowledged.
I place on record my deep sense of gratitude and indebtedness to Dr. P. VIJI, Scientist,
ICAR-Central Institute of Fisheries Technology, Visakhapatnam, for her immense help and unflagging
encouragement in carrying out this investigation.
My profound thanks to Dr. B. MADHUSUDHANA RAO, Principal Scientist, ICAR-

Central Institute of Fisheries Technology, Visakhapatnam, for his constant encouragement, co-
operation and providing laboratory facilities.
I humbly express my sincere gratitude to Dr. M.M. PRASAD, Dr. L. N.

MURTHY, Dr. R. VENKATESWARLU and Ms. JESMI DEBBARMA, ICAR-
Central Institute of Fisheries Technology, for their guidance and advice throughout the progress of
work.
I wish to record my sincere thanks to authorities of ICAR-Central Institute of
Fisheries Technology and Andhra University for providing facilities to complete my
course.
I sincerely acknowledge Ministry of Earth Sciences, New Delhi for providing

fellowship through the project “Exploration and Assessment of demersal fishery resources along the
continental slope (200m to 1200m depth) of Indian Exclusive Economic Zone and Central Indian
Ocean”. I am grateful to the authorities of Centre for Marine Living Resources and
Ecology (CMLRE), Cochin for providing facilities on board “FORV Sagar Sampada”. I
am also grateful to Chief Scientists, Crew members, Fishing Masters and Deck-hands of FORV Sagar
Sampada for their help in collecting the samples.
I am very grateful to express my special gratitude to Shri. R. UMA MAHESWARA

RAO, Co-Scholar, who has supported me a lot directly and indirectly as well as morally throughout
the work progress. I greatly acknowledge Dr. B. PREMA RAJU, Co-Scholar, for his support in
collecting and data analyzing of fishery samples onboard FORV Sagar Sampada. His valuable
suggestions and active help were the key sources of inspiration to me. I am very grateful to my Co-
scholars S. LAVANYA and D. DHANUNJAY for their invaluable service and support in
bringing out this work.
I also express my sincere gratitude to Shri. P. RADHA KRISHNA, PRASANNA

KUMAR, DHIJU DAS, BHUSANAM, PRABHAKAR and ICAR-CIFT staff members for
their assist and support during my work.
It is my pleasure to acknowledge my tenants R.S. NAIDU & R. SATYAVATHI at

Gajuwaka and I gratefully acknowledged to M. JAGANNATH, B. OM PRAKASH & K.
RAM MOHAN, members of NGO-PSS (PIOUS SCIENCE SOCIETY), for their moral
help and encouragement.
I express my warm and heartily feelings to my parents Shri. K. RAMA RAO, Smt. K.
NOOKA RATHNAM, my brother and his family Shri. K. KISHORE KUMAR & K.
VIJAYA LAKSHMI, K. BHAVESH, K. CHANDRIKA my beloved sister and her family,
Y. REVATHI, Y.V.P.R. RAO, Y. DHATRI, Y. NANDAN and my blood relation
members K. SANKARA RAO, K. VARA LAXMI, K. PARAMESWARA RAO, my
dearest sister K. RAMA for their blessings, love and inspiration for achieving the present task. I
take this opportunity to thank all my relatives for their love & prayers and I apologize for not
mentioning each one of them personally. Once again, I thank God the Divine, who continues to make
the impossible into possible through his gracious interventions and blessings.
Rushinadha Rao Kakara

PREFACE
The marine fisheries of the world can be broadly grouped under two major
categories, viz., demersal fisheries and pelagic fisheries. Demersal fisheries are cramped
to the continental shelf and continental slope and are mainly exploited by using different
types of trawls which are dragged from fishing vessels to tow along the bottom. The
fishery of organisms living freely in the water ample is determined pelagic fisheries, but
this again can be considered in two divisions, namely pelagic-neritic fisheries and
pelagic-oceanic The pelagic-oceanic fisheries with which we are concerned here
comprise the fisheries of fishes and other animals which inhabit the oceanic realm or in
other words the high seas outside the continental shelf. The wealth of marine resources
was assumed to be an unlimited gift of nature. The general concern over environmental
degradation in recent years, fishery stocks and aquatic resources has been heavily
overexploited. In many capture fisheries in the Indian Ocean, the rate of harvesting has
exceeded the natural rate of renewal, resulting in biological overfishing (Ansari et al.,
2006). FAO (1997) estimated that 69% of the world's marine stocks, for which data is
available, are in need of urgent corrective conservation and management measures: 44%
are fully to heavily exploited, 16% overexploited, 6% depleted, and 3% very slowly
recovering from overfishing. More than 69% of the stocks of demersal and pelagic fish,
crustaceans, and mollusks in various areas of the world's oceans need rehabilitation.
Global total capture fishery production in 2014 was 93.4 million tonnes, of which 81.5
million tonnes from marine waters and 11.9 million tonnes from inland waters,
continuing a positive trend that has resulted in a 37% increase in the last decade. Sixteen
countries have annual inland water catches exceeding 200 000 tonnes, and together they
represent 80% of the world total (SOFIA, 2016). Total world fishery production is
projected to expand over the period, reaching 196 million tonnes in 2025. This represents
an increase of 17% between the base period (average 2013–15) and 2025, but indicates a
slower annual growth compared with the previous decade (1.5% versus 2.5%) (SOFIA,
2016).
The potential of marine fishery resources from the Indian EEZ has been estimated
at 3.92 million tonnes and the tunas, billfishes and pelagic sharks together amount to
about, 2,46,000 tonnes, which is around 6.3% of the estimated potential. The fisheries in
India are limited to the small-scale sector with negligible inputs from the industrial
sector. During 1985-2000, the average annual production of tunas from Indian seas was
around 37,000 tonnes contributing to 3.6% of the total pelagic fish production and 1.8%
of the total marine fish production. Despite the fact that there has been a noticeable
increase in the landings of coastal fishes during the last three decades, the fish stocks
remain to be one of the least exploited pelagic resources from the EEZ of India (Pillai and
Gopakumar, 2003). The economic boom experienced by some people who ventured into
mechanized fishing has encouraged many entrepreneurs to do the same, resulting in a
sudden increase in the number of vessels. According to current estimates by the Ministry
of Agriculture, the total number of mechanized fishing crafts in the country is around
47,000 (unpublished). This has resulted in intensive non-selective fishing in order to
make the operation successful and to repay the loans obtained for the purchase of these
vessels. The use of small meshes of 20.0 µm or less in the cod end of the demersal trawl,
seines and stake nets, have resulted in the capture of juveniles of shrimp and other fishes.
The immediate need is to safeguard our resources from overexploitation and to enforce
regulations for responsible fishing (Gopakumar, 1997).

Overexploitation of marine resources is a worldwide problem. It takes place when
the maximum amount of fish that can be taken from the sea is exceeded. The
improvement in efficient fishing technology, that is becoming available to even smaller
fishing operators, is really not giving the fish in the sea much chance to escape the fishing
gear and time to reproduce. Biological overexploitation of fishery resources leads to
collapse of certain fisheries or their severe depletion (Ansari, et al., 2006). According to
Hutchings (2000), with the exception of a few species, recovery of most fishing stock
after collapse is very little. National and international efforts are, therefore, needed to
protect fish stocks under threat of overexploitation and to allow depleted stocks to
recover. Some deep-sea pelagic groups, such as the lantern fish, ridge head, marine
hatchet fish, and light fish families are sometimes termed pseudoceanic because, rather
than having an even distribution in open water, they occur in significantly higher
abundances around structural oases, notably seamounts and over continental slopes. The
phenomenon is explained by the likewise abundance of prey species which are also
attracted to the structures.
Recently, and with great sanguinity, the government implemented a policy for
deep sea resource exploitation. Experience of deep sea operations demonstrates that that
most of these vessels operate uneconomically as they do not capture enough fish of
reasonable quality. In order to that, CMLRE (Centre for Marine Living Resources and
Ecology) starts a work on deep-sea fishery resources with the help of FORV (Fisheries
Oceanographic Research Vessel) Sagar Sampada, Ministry of Earth Sciences, India.
ICAR-CIFT has taken this challenge to evaluate the Nutritional studies and Bio-
monitoring of Heavy metals in deep-sea fishery resources of Indian EEZ (Exclusive
Economic Zone).
Data collected during deep-sea fishery expeditions of FORV (Fisheries
Oceanographic Research Vessel) Sagar Sampada is depicted. The information provided
in this investigation is new and would be of a great interest and curiosity. By
encompassing the above mentioned aspects, the present thesis is divided into three
chapters.
Chapter 1 deals with the analysis of proximate composition of ten deep-sea fishes. In
this study, moisture, protein, fat and ash was observed. High moisture percentage was
observed in Alepocephalus bicolor and Narcetes erimelas in cruise 322, 332 and 338,
more amount of protein accumulation was found in Chelidoperca investigatoris Cubiceps
baxteri and Chlorophthalmus bicornis in the cruises 322, 332 and 338, almost all deep-
sea fishes comes under lean fat fish (<2%) category except Alepocephalus bicolour and
Chelidoperca investigatoris which were comes under low fat (2-4%) category.
Chapter 2 deals with monitoring of heavy metals like copper, zinc, iron, cobalt,
cadmium, lead, chromium, selenium and arsenic accumulation in the above given ten
selected deep-sea fish samples. In this study, the abundance order of metal accumulation
in cruise 322 is Fe>Zn>Cu>Cr>Co>As>Cd>Pb>Se, whereas in cruise 332 is
Fe>Zn>Cu>Co>Cr>Cd>Pb>As>Se and the order of metal content in cruise 338 is
Fe>Zn>Cu>Co>Cr>Cd>As>Pb>Se. The obtained results indicate that environmental
conditions such as the season of fish catch, temperature and the chemical composition of
water affect the physiological status of fish and the ability to accumulate metals in their
organs. The heavy metal content in deeper water fishes have not been studied extensively
and scanty literature has been found.
Chapter 3 deals with the assessment of free fatty acid profiling studies in the
given selected ten deep-sea fishes. The deep-sea fish species from the present study
contains a particularly good lipid composition for nutritional purposes. It should however
be noted that the net content of omega-3 and omega-6 is low because of the low fat
content of these fish species, and only regular consumption of such fish species could
contribute to increase the amount of omega-3 and omega-6 fatty acids in the diet. This
study demonstrates that several of the underutilized deep-sea species are able to compete
with more commercially utilized species in terms of nutritional value, and they can
definitely also compete when it comes to taste.
The results achieved during my study period were discussed and compared with
allied works.
CHAPTER-I
PROXIMATE COMPOSITION OF
DEEP-SEA FISHES
INTRODUCTION
Fisheries sector has been envisaged as a powerful engine for monetary
development and creation of employment as it stimulates enlargement of a number of
subsidiary industries. It is an imperative source of low-priced and nutritious food,
moreover it provides as an instrument of livelihood for a large section of economically
backward population of the country. Indian fisheries sector is an important factor of the
worldwide fisheries, comprising of 4.4% of the global fish production and contributes to
1.1% of the GDP and 4.7% of the agricultural GDP. The marine fisheries of the world
can be broadly grouped under two major categories, viz., demersal fisheries and pelagic
fisheries. Demersal fisheries are cramped to the continental shelf and continental slope
and are mainly exploited by using different types of trawls which are dragged from
fishing vessels to tow along the bottom. The fishery of organisms living freely in the
water ample is determined pelagic fisheries, but this again can be considered in two
divisions, namely pelagic-neritic fisheries and pelagic-oceanic fisheries. The previous
embraces the fisheries of sardines, other clupcoids, mackerel, etc., which are mainly
confined to the waters above the continental shelf. The pelagic-oceanic fisheries with
which we are concerned here comprise the fisheries of fishes and other animals which
inhabit the oceanic realm or in other words the high seas outside the continental shelf.
The wealth of marine resources was assumed to be an unlimited gift of nature. The
general concern over environmental degradation in recent years, fishery stocks and
aquatic resources has been heavily overexploited. In many capture fisheries in the Indian
Ocean, the rate of harvesting has exceeded the natural rate of renewal, resulting in
biological overfishing (Ansari et al., 2006). FAO (1997) estimated that 69% of the
world's marine stocks, for which data is available, are in need of urgent corrective
conservation and management measures: 44% are fully to heavily exploited, 16%
overexploited, 6% depleted, and 3% very slowly recovering from overfishing. More than
69% of the stocks of demersal and pelagic fish, crustaceans, and mollusks in various
areas of the world's oceans need rehabilitation. Global total capture fishery production in
2014 was 93.4 million tonnes, of which 81.5 million tonnes from marine waters and 11.9
million tonnes from inland waters, continuing a positive trend that has resulted in a 37%
increase in the last decade. Sixteen countries have annual inland water catches exceeding
200 000 tonnes, and together they represent 80% of the world total (SOFIA, 2016). Total
world fishery production is projected to expand over the period, reaching 196 million
tonnes in 2025. This represents an increase of 17% between the base period (average
2013–15) and 2025, but indicates a slower annual growth compared with the previous
decade (1.5% versus 2.5%) (SOFIA, 2016).
Deep-sea is the largest habitat on earth which covers more than 300x105 km2 and
comprises about 63% of the Earth's solid surface (Smith et al. 2008). The deep sea floor
lies between the shelf break (< 200m depth) and the bottom of the Challenger Deep (<
11000m depth). The continental slopes alone occupy 8.8 % of the world's surface,
compared to 7.5 % for the continental shelf and shallow seas (Merrett and Haedrich,
1997). The distinct habitats of the deep seafloor are diverse and include sediment-covered
slopes, abyssal plains, and ocean trenches, the pillow basalts of mid-ocean ridges, rocky
seamounts stick out above the seafloor, and submarine valleys dividing continental
slopes. Deep-sea can be categorized into four zones, the mesopelagial is uppermost and a
range of 200 to 1000m; some fisher folk use 500m as the start of the deep water habitat.
The bathypelagic goes from 1000-4000m depth zone, the abyssopelagic occupies from
4000 to 6000m and the hadal zone revealed for completeness, is the habitat of the deep
ocean trenches. For the demersal fishery, the deepwater region can be taken as the
continental slopes, starting from the shelf break and parallel to the mesopelagic and
bathypelagic, and beneath, the continental rise which extends down to the abyssal planes
at around 6000m. Complicating this picture are the existence of seabed features that may
rises thousands of metres above the surrounding areas - seamounts, or form regions of
hills or knolls. Gorden (1995) are of the view that, even though they encompass only
something like 8.8% of the ocean bottom, the slopes are among the most complex and
dynamic parts of the deep sea. Slopes are most intricate topographically than are
continental shelves. The vertical zone of the oceans and seas, which extends from the
shelf break i.e., from the outer boundary of the shelf zone, down the slope, to the oceanic
bed is called the bathyal zone. In addition to the continental slope, the slopes of islands,
seamounts and rises, including the mid-oceanic ridges, are also included in the bathyal
zone (Parin and Kotlyar, 1984). The deep sea benthic zone is further divided into an
upper bathyal zone, a middle abyssal zone and a lower hadal zone. The bathyal zone is
the environment of the continental slope and it extends down from the continental shelf to
about 3000m. It can well be defined as a broad transition zone between the sub-littoral
and the true deep sea zone. It is also referred to as archibenthic zone. Historically, the
bathyal zone is more vital for the preservation of the marine fauna (Zezina, 1985).
Deep sea creatures are also frequently equipped with a powerful sense of smell so
that chemicals released into the water can magnetize prospective mates. The shady, cold
waters of the deep are also oxygen-deprived places. Accordingly, deep-sea life entails
petite oxygen. Oxygen is transferred to the deep sea from the surface where it sinks to the
bottom when surface temperatures diminish. It was observed that oxygen is never
diminished in the deepest parts of the ocean since there are fewer animals to devour the
available oxygen. In most parts of the deep sea, the water temperature is more consistent
and stable. Pressure increases 1 atmosphere for each 10 m in depth. The deep sea varies
in depth from Zoom to more than 10000 m, therefore pressure ranges from 20atm to more
than l000atm. The deep sea creatures have adapted to pressure by developing bodies with
no surplus cavities, such as swim bladders, that would collapse under strong pressure.
The flesh and bones of deep sea marine creatures are soft and flaccid which also assist
them withstand the pressure. Deep sea creatures have developed some attractive feeding
mechanisms because of the lack of light and because food is inadequate in these zones.
Some food comes from the detritus of decomposing plants and animals from the upper
zones of the ocean. The cadavers of large animals that sink to the bottom provide
intermittent feasts for deep sea animals and are consumed swiftly by an assortment of
species. The deep sea is residence to jawless fish such as the lamprey and hagfish
(Myxinidae), which warren into the carcass quickly consuming it from the inside out.
Deep sea fish also have hefty and expandable stomachs to hold large quantities of meager
food. They don't expend energy for swimming in search of food; rather they remain in
one place and waylay their prey using amazing and clever adaptations. These ecological
characteristics of deep sea fishes can make them vulnerable to overexploitation and slow
to recover from it (Koslow et al. 1997). The species often have a slow growth rate, high
longevity, low fecundity and hence low productivity (Tracey and Horn, 1999). Catches
can be high initially in the fishery development and fishing down phases, but very low on
a long term sustainable basis, as with orange roughly in New Zealand and Australia
(Clark, 2001). The deep sea environment, being dark and cold, has generally been
regarded as a system of low energy and low productivity (Clark, 2001). The continental
slope was regarded as having little or no marketable potential (Merrett and Haedrich,
1997).
Data concerning of the Indian deep-sea fish fauna beyond 200m depth are very
scanty and the merely existing information is that the engendered from the exploratory
fishing cruises conducted by Fishery Survey of India to some extend and mostly by
FORV Sagar Sampada as part of its stock evaluating programs of deep sea fishery
species. There was information on the deep sea fishes in and around Indian waters by
former researchers during the early sixties and seventies. The bathypelagic fish,
Neoepinnula oriantalis was reported from the Konkan coast by Rao (1965). Jones and
Kumaran (1965) reported several new records of fish species from the seas around India.
A check list of fishes of the Indian EEZ based on the pelagic and bottom trawl collections
of FORV Sagar Sampada was compiled by Balachandran and Nizar (1990). The
exhaustive check list included 87 families and 242 species of both conventional as well as
non-conventional fish fauna of the Indian EEZ. Nair and Reghu (1990) reported the
distribution of Saurida spp. in the continental shelf and the upper continental slope from
the EEZ of India. Menon (1990) recorded myctophids, gonostomatids, Bregmacros, eel
larvae and juveniles of many fishes from the deep scattering layer (DSL) of Indian EEZ.
Sivakami (1990) reported the occurrence of unconventional forms like Psenopsis sp.,
Trichiurus auriga, Chlorophthalmus agassizi, Neoepinnula orientalis and Cubiceps sp.
In additional to the conventional forms especially in the deeper waters of the southwest
region, Panicker et al. (1993) reported Centrolophus sp. and Chlorophthalmus sp. as
dominant species in the depth zone 200 – 500m in the latitude 7° to 17°N, off west coast
of India.
On a global scale, fish is a chief source of food for human nutrition providing an
imperative quantity of dietary protein and lipid diet (Bouriga, et al., 2010; Ravichandran,
et al., 2011). Edible part of fish is relatively of high digestibility compared with red meat
(Pirestani, et al., 2010). It comprises essential amino acids in desirable quantities for
consumption of humans. All these properties bring the edible part of fish in the same
class as chicken protein and superior to milk, beef protein and egg albumen (Srivastava,
1959). In general, the biochemical composition of the whole body signifies the quality of
fish. Cod liver oil is rich in vitamin A and D. Fish oils have moved into the center state of
fatty acids in nutrition. It helps to prevent brain aging and Alzeimer’s disease (Whelan,
2008). Fish oil is used as an industrial chemical base in paints and linoleum.
Fish is an important source of food for mankind all over the world from the times
immemorial. Fish is very important source of animal protein in the diets of man. The
importance of fish as source of high quality, balanced and easily digestible protein,
vitamins and polyunsaturated fatty acids is well understood now. The fishes offered as a
dietary supplement to the farming pigs has considerably increased their weight and meat
yield (Ojewola and Annah, 2006). The fish and fish products consumption is
recommended as a means of preventing cardiovascular and other diseases and has greatly
increased over recent decades in many European countries (Cahu et al., 2004). Besides
this fishes are good source which possess immense antimicrobial peptide in defending
against dreadful human pathogens (Ravichandran et al., 2010).

Marine fish are known to be a very healthy food item. They are an excellent
source of protein that also brings various minerals and vitamins essential for good health
(Imad Patrick Saoud et al., 2008). Fish is a major source of food for human nutrition
gives a significant quantity of dietary protein and lipid diet in many countries. Edible part
of fish is easily digestible due to it contains long muscle fibres (Bouriga et al., 2010). The
edible part of lively fish is considered as a marker for the natural aquatic environment
(Mona et al., 2011).
Nutrients nurture the body, promote growth, maintain and repair body parts. Fish
plays a main role in human nutrition. Magnitude of fish as a source of high quality,
balanced and easily digestible protein is now well understood. Besides, it is also a well-
known source of several other nutrients and is being accepted as a healthy food.
However, endangered by diet-borne pollutants transferred along the food chain (Agrawal
and Srivastava, 2003; Jat and Kothari, 2006; Gupta and Srivastava, 2006; Ayas et al.,
2007). A healthy food should contain all the essential constituents’ viz., carbohydrates,
proteins, fats and minerals and vitamins in correct proportion. Furthermore, it has been
linked to health benefits, such as the prevention of cardiovascular diseases and some
types of cancer, including colon, breast and prostate. Understanding the composition of
food is very important to determine its nutritive value. This is particularly true for
commodities like fish as there is large variation in the composition. There are group
specific and species-specific changes in composition. The composition of the fish
depends on the season, habitat and the type of food consumed by the fish. Moisture, fat,
protein and ash which gives minerals are the main components of fish meat and the
analysis of the same is referred to as proximate composition (Love, 1970).

Biochemical composition of the fish muscle commonly indicates the quality of
fish. Therefore, proximate composition of a fish species facilitates to assess its nutritional
and edible value in terms of energy units compared to other species. Difference of
biochemical composition of fish edible part may also occur within same species
depending upon the fishing ground, fishing season, age and sex of the individual and
reproductive status. The spawning cycle and food supply are the main factors responsible
for this variation (Love, 1980). An escalating amount of evidences suggest that due to the
high content of polyunsaturated fatty acid, fish flesh and fish oil are beneficial in
reducing the serum cholesterol (Stansby, 1985). In addition to that, the special type of
fatty acid, the omega-3 polyunsaturated fatty acid, recognized as an essential nutritive
supplement to prevent a number of coronary heart diseases (Edirisinghe et al., 1998).
Lipid and moisture in the fish muscle and liver are generally quantitatively
complimentary to each other. As lipids increase, the water content decreases
correspondingly. Lipids and water together from 80% in most fishes. Protein-water
relationship is less marked. At higher concentration of lipids, the protein content at times
shows a marginal decrease. In comparison with the extensive literature dealing with
surface-living and mesopelagic (midwater) species, fewer studies have considered
biochemistry and metabolism of deep-sea species. In fact, proximate composition, energy
content and metabolic rates of a large number of pelagic crustaceans and fishes have been
studied in temperate and subtropical latitudes (Childress and Nygaard, 1973, 1974;
Bailey and Robison, 1986; Ikeda, 1988; Childress et al., 1990b; Donnely et al., 1990;
Cowles et al., 1991). Several of the cited authors revealed that mesopelagic species show
variability in proximate composition as a function of depth of occurrence and as a

function of regional productivity. Depth and productivity both affect food availability and
thus influence chemical composition. In particular, lipid and protein content (% wet
weight) both decline and as a result water concentration increases with increasing depth
of occurrence (Childress et al., 1990b). Moreover, mesopelagic crustaceans and fishes
living at greater depths have much lower metabolic rates than shallower-living pelagic
species; the ammonia excretion and oxygen consumption rates decline with increasing
depth (Donnely and Torres, 1988; Ikeda, 1988; Torres and Somero, 1988; Childress et
al., 1990a). It is worth noting that these physiological and biochemical changes have been
attributed to factors correlated to depth, aside from the possible influence of temperature
(Childress, 1971; Torres et al., 1979) or hydrostatic pressure (Teal, 1971; Meek and
Childress, 1973). Childress et al. (1990a) also found that the reduction in the metabolic
rate with depth was also related to reduction in mobility. According to the visual
interaction hypothesis of Childress et al. (1990a), the deep-sea species have a less active
prey-predator relationship than shallow-water species.
Investigations regarding the protein content of the fish and fish products have
been initiated as early as 19th century (Saha and Guha 1940). After 19th century several
worldwide investigations were made on biochemical composition of fish. Noteworthy
contributions are those of Green (1919) investigated on the biochemical changes in the
muscle tissue of king Salmon during the spawning migration. Bruce (1924) worked on
the changes in the chemical composition of Herring in relation to age and maturity.
Chanan and Saby (1932) studied on the fat metabolism of the Herring. Mc Cay and
Tunison (1936) reported on the environmental factors influence on biochemical
composition of fish. Hickling (1947) worked on the seasonal cycle in the corn fish.
George et al., (1966) on the function of lipid composition in animal body; Love (1970)
on the chemical composition of fish; Sivani (1994) investigated chemical composition in
different species of estuarine fishes. Busalmen et al., (1995) observed changes in lipids
and biochemical properties of actomyosin from pre and post-spawned hake, Merluccius
hubbsi. Stephan et al., (1996) studied the incidence of different amounts of proteins,
lipids and carbohydrates in diets on the muscle lipid composition in the turbot,
Scophthalmus maximus. Olsen and Henderson (1997) worked on muscle fatty acid
composition and oxidative stress indices of Arctic charr, Salvelinus. Jeong et al., (1998)
worked on fatty acid composition of Korean fish. Verghese and Jayasankar (1999)
studied the electrophoretic patterns of sarcoplasmic proteins in carangid fishes.
Lahnsteiner, (2000) stated Morphological, physiological and biochemical parameters
characterizing the over-ripening of rainbow trout. Rao and Rao (2002) studied the
variations in biochemical composition of Glossogobius giuris from Gosthani estuary in
relation to sex, maturity, food and feeding intensity. Nordgarden et al., (2003) stated on
seasonally changing metabolism in Atlantic salmon, Salmo salar. Gokoglu et al., (2004)
worked on Effect of cooking methods on the proximate composition and mineral contents
of rainbow trout (Oncorhynchus mykiss). Rajasree and Kurup (2005), stated some
observations on the carotenoid distribution and proximate composition in deep sea
prawns off Kerala (South India). Suman et al., (2006) studied the Species composition,
distribution, and potential yield of deep sea shrimp resources in the Western Sumatera of
the Indian Ocean EEZ of Indonesian waters. Drazen, (2007) worked on Depth-related
trends in proximate composition of demersal fishes in the eastern North Pacific.
Karuppasamy et al., (2008) studied Food of some deep sea fishes collected from the
eastern Arabian Sea. Oksuz et al., (2009) studied on A comparative study on proximate,
mineral and fatty acid compositions of deep sea water Rose Shrimp(Parapenaeus
longirostris, Lucas 1846) and golden Shrimp (Plesionika martia, A. Milne-Edwards,
1883). Suseno et al., (2010) stated Proximate, fatty acid and mineral composition of
selected deep sea fish species from Southern Java Ocean and Western Sumatra Ocean,
Indonesia. Gokhan and Karaçam, (2011) studied on Seasonal Changes in Proximate
Composition of Some Fish Species from the Black Sea. Demet and Turan (2012) stated
Proximate and Fatty Acid Composition of Some Commercially Important Fish Species
from the Sinop Region of the Black Sea. Yancey et al., (2014) worked on Marine fish
may be biochemically constrained from inhabiting the deepest ocean depths. Rao et al.,
(2016) studied on biochemical composition and heavy metal accumulation of deep-sea
crustaceans in east and west coast of Indian EEZ.
The food consumption and metabolism of deep sea fish have seldom been studied.
The main objective of this study is to provide information on the proximate composition
when utilization of new species of deep-sea fish is considered. This is because deep-sea
fishes are considered to be not only food with good source of quality protein but also
food with healthy components.

MATERIAL
&
METHODS
Sample Collection:
The fish samples were collected from deep-sea stations located in the coast of
Indian EEZ (Exclusive Economic Zone), during three deep-sea cruises (Cr-322, Cr-332
and Cr-338) of the research vessel FORV (Fisheries Oceanographic Research Vessel)
Sagar sampada. The most prominent deep-sea fishes caught in deeper waters are of
Alepocephalus bicolor, Narcetes erimelas, Talismania longifilis, Chlorophthalmus
bicornis, Lamprogrammus niger, Beryx splendens, Chelidoperca investigatoris,
Neoepinnula orientalis, Cubiceps baxteri and Psenopsis cyanea at the depth of 200 –
1200 m depth and the gear used are Expo model fish trawl and HSDT-CV (High Speed
Demersal Trawl – Crustacean version) developed by the ICAR-Central Institute of
Fisheries Technology. Sampling was done from Fisheries Oceanography Research Vessel
of the Department of Ocean Development, FORV Sagar sampada. Twenty samples of
each species of almost the same size were kept in polyethylene bags and stored in the
freezer at -20oC for further analysis (Plate 1 to 12).
Sample Preparation:
To prepare the sample for determination of proximate composition, the fishes
were washed with running tap water and the excess water was removed with the help of
blotting paper. The length and weight was measured and recorded. Non-edible portions of
fish samples (Offal, Bones and Scales) were removed immediately to avoid
decomposition. The edible portion (Muscle) was blended and aliquots weighed out for
the various analyses.

PLATE - 1
Figure1: FORV (Fisheries Oceanographic Research Vessel) Sagar sampada

PLATE - 2
Figure 2 & 3: Scanning the grounds and Fixing the Simrad ITI sensors to net
PLATE - 3
Figure 4 & 5: Monitoring the trawl and catch parameters & Hauling the net
PLATE - 4
Figure 6 & 7 : Sample collection & Scattered the samples for segregate
PLATE - 5
Figure 8 & 9 : Segregating the sample & species wise after segregation
PLATE - 6
Figure 10 & 11: Assessment of deep-sea fishes & Biology work onboard
PLATE - 7
Figure 12 & 13: Sample packing & carry for storing in -20oC
PLATE - 8
Figure 14 & 15: Alepocephalus bicolor & Narcetes erimelas

PLATE - 9
Figure 16 & 17: Talismania longifilis and Chloropthalamus bicornis

PLATE 10
Figure 18 & 19: Lamprogammus niger and Beryx spendens

PLATE 11
Figure 20 & 21: Chelidoperca investigatoris and Neoepinnula orientalis

PLATE 12
Figure 22 & 23: Cubiceps baxteri and Psenopsis cyanea

Estimation of proximate composition:
Moisture, Fat and Ash content were determined according to the AOAC
(Association of Official Analytical chemists) (2000) and Protein content was observed
using Micro- Kjeldahl by Pearson method. Five number of each sample were used to
analyse the proximate composition. All of the chemicals used in this work were high
purity GR grade.
MOISTURE:
Moisture content of all the samples was determined using Oven-dry method.
Empty Petri dishes were dried using air drying oven for 1 hour at 1000C, transferred to
the desiccators (with granular silica gel), cooled for 30 minutes, and were weighed using
an electronic balance.
Formula: Weight of the dish = w1g
Weight of dish + sample before heating = w2g
Weight of dish + sample after heating = w3g
∴Weight of sample = (w2-w1) g
Loss in moisture = (w2-w3) g
(w2-w3)
∴Percentage of moisture = ---------- × 100
(w2-w1)
Replicates of the minced samples were mixed thoroughly and ten grams of
composite fresh fillet was transferred to the dried and weighed dishes. After which the
dishes were put in an oven overnight at a temperature of 1000C. The dishes were removed
from the oven on the following day and cooled in a desiccator for about 30 minutes and
the total weight of the dishes together with dried samples recorded. The moisture content
was determined by measuring the weight of a sample before and after the water was
removed by evaporation. The loss of water was calculated as percent moisture.
PROTEIN:
Protein was estimated using Micro- Kjeldahl method (AOAC, 2000)
Principle:
The nitrogen in the compound is converted into ammonium sulphate by digesting
the compound with concentrated sulphuric acid. Ammonium sulphate thus obtained is
decomposed by alkali and the liberated ammonia is absorbed in boric acid and titrated
against standard sulphuric acid.
Procedure:
50 to 200mg of dry sample or about 1gm of wet sample is weighed into a
Kjeldahl’s digestion flask. 10ml of concentrated H2SO4 and a pinch of the digestion
mixture are added to the flask and the flask is heated on a Kjeldahl digestion rack. A
glass bead is added to the reaction mixture to prevent bumping. The mixture is digested
for 4-6 hrs or until a clear solution is obtained (greenish yellow in colour) to ensure
complete conversion of the nitrogen in the sample into ammonium sulphate. A blank is
carried out using sulphuric acid. The digested sample is transferred to a 100ml volumetric
flask and made up to the mark.
Formula:
5ml/N/70 HCL = 1mg × N2
1 × T.V × 100 × 100

Total nitrogen content of the sample = -------------------------------- mg Nitrogen
5 × 5 × wt. of the sample
Protein content = Total nitrogen (g) × 100% fresh muscle

-----------------------------------------------
16 (6.25)
5ml of the made up sample is pipetted and transferred into the chamber of
Kjeldahl distillation apparatus which has been cleaned out by steaming. 10ml of 40%
NaOH are added to the inner chamber and the liberated ammonia is steam distilled into a
receiver flask containing 10ml of 2% boric acid solution to which 2-3 drops of mixed
indicator are added. The indicator is red in boric acid and turns green when ammonia is
absorbed. The distillation is carried out for 2-5 minutes and the content of the receiver
flask (ammonia absorbed in boric acid) is titrated against standard N/70 HCL until the
colour changes to purple. The nitrogen present in the sample is calculated using the
nitrogen equivalence viz., 1ml of 0.05 N/70 HCL = 0.007g of nitrogen. The protein value
is obtained by multiplying the N content by 6.25.

FAT:
The extraction of fat from substances is often tedious and requires thorough
contact and heating with solvent. This is done in a special apparatus known as Soxhlet
Extractor. In this procedure, the apparatus is designed so that a fresh portion of solvent
comes in contact with the materials to be extracted over a relatively long period of time.
Description of the apparatus:
The apparatus consists of a flask, containing a volatile solvent and resting on
some type of heating device. The flask is connected by means of interchangeably ground
joints with a tube having a siphon arrangement and a side arm (Soxhlet Extractor). Again
the extraction tube is connected by inter-changeably ground joints with a water cooled
condenser.
Reagent: Ethyl ether or Petroleum ether.
Procedure:
2 to 3 mg of the ground moisture free material is accurately weighed into a
thimble. The mouth of the thimble is packed by absorbent cotton so as to keep the sample
in. The thimble is placed in the Soxhlet Extractor and it then connects with the pre-
weighed flask (receiver) and also the condenser. The receiver containing suitable solvent
viz., Ethyl ether or petroleum ether (B.P 40-60oC) is heated at such a rate that the ether
will drop from the condenser on to the center of the thimble at the rate of 5 to 6 drops per
second. When sufficient solvent has been thus transferred to the extracting tube to fill the
siphon it siphons back into the receiver. Thus the process is continued until the extraction
is complete, varying at times from 8 to 24 hours.

Formula:
Weight of the sample = Xg
Weight of the empty flask = w1 g
Least weight of the flask after evaporation = w2 g
∴Weight of fat collected = (w2 – w1) g
X gms of the sample contain (w2 – w1) g of fat
100 × (w2 – w1)
∴100gms contain = ---------------------
Then the flask is removed and the volatile solvent is evaporated at (60- 80oC) on a
water bath. The residue is dried in an oven maintained at 60-70oC, cooled in a desiccator
and weighed with evaporation of the ether, the weight is on the decrease and becomes
constant at last. Any further heating results in an increase in weight gradually caused by
the oxidation of the fat. The least weight of the residue gives the weight of the fat in the
sample.
ASH:
Ash is the residue remaining after food stuff is ignited until it is free from carbon,
usually at a temperature 550oC. They have an idea of the mineral content of the foodstuff
of determining the total ash, acid soluble ash and acid insoluble ash.
Formula:
Weight of the crucible = wg
Weight of the crucible + sample = w1 g
Weight of the crucible + ash = w2 g
Weight of the sample = (w1 – w) g
Weight of the ash = (w2 – w) g
(w2 – w)
∴ % of Total ash = ---------- × 100
(w1 – w)
Procedure:
A porcelain, silica or platinum crucible is ignited, cooled and weighed to constant
weight. About 3 gm. of the material is placed in the crucible and its weight determined.
The sample is carbonized by burning at low red heat. If the material contains large
amount of fat, preliminary ashing is done at low enough temperature to allow smoke off
of fat without burning. The crucible is placed in a muffle furnace at 550o±20oC until a
white ash is obtained (for 4 hours). The ash content is weighed for constant weight after
successive cooling and heating operations.

ACID INSOLUBLE ASH:
Formula:
w3 = lowest weight in g of the dish with the acid insoluble ash weight of
acid Insoluble ash
= (w3 – w) g.
(w3 – w)
∴ Acid insoluble ash, percent by weight = --------------- ×100
(w1 – w)
Dilute hydrochloric acid–1:1 prepared from concentrated hydrochloric acid. To
the ash obtained by the above procedure, add 25ml of dilute hydrochloric acid, cover with
a watch glass and heat on a water bath for 10 minutes. Cool and filter through whatsman
filter paper No.42 or its equivalent. Wash the residue with hot water until the washings
are free from chlorides as tested with silver nitrate solution, and re-turn the filter paper
and residue to the dish. Keep it in an electric oven maintained at135 ± 20oC for about 3
hours. Ignite it in a muffle furnace at 600 ± 20oC for about 1 hour. Cool in a desiccator
and weigh. Ignite the dish again for 30 minutes, cool and weigh. Repeat this process till
the difference between two successive weights note the lowest weight less than 1mg.
STATISTICAL ANALYSIS:
All the results obtained data collected in this study were presented as mean ±
standard error (SE) and analyzed by one-way ANOVA analysis using Excel to determine
the variations among the mean concentrations of moisture, protein, fat and ash of two
Tuna species viz., Euthynnus affinis and Auxis thazard . Significant between P < 0.01 and
P < 0.05 was used to indicate significant difference.

RESULTS
The biochemical composition of moisture, protein, fat and ash in body muscle of
deeper water fishes Viz., Alepocephalus bicolor, Narcetes erimelas, Talismania longifilis,
Chlorophthalmus bicornis, Lamprogrammus niger, Beryx splendens, Chelidoperca
investigatoris, Neoepinnula orientalis, Cubiceps baxteri and Psenopsis cyanea were
recorded in two years of data from three cruises – 322, 332 and 338 of FORV (Fisheries
Oceanographic Research Vessel), Sagar Sampada at the depth of 200 – 1200 m depth and
the gear used are Expo model fish trawl and HSDT-CV (High Speed Demersal Trawl –
Crustacean version) developed by the ICAR-Central Institute of Fisheries Technology.
The changes in cruise wise results of proximate composition of the muscle were endowed
in Table 1 – 4 and Graphs 1 – 4 respectively.
MOISTURE
Moisture forms the main component of proximate composition. Table 1 and
Graph 1 show the moisture values of the ten indigenous deeper water fish species. The
moisture values were in between 73.93±0.67 and 81.11±0.69 of cruise 322, the results
revealed that the moisture content was more accumulated in Alepocephalus bicolor
(81.11±0.69) followed by Narcetes erimelas (80.83±1.05) and Psenopsis cyanea
(80.03±0.81), whereas in Cruise 332 the results of moisture were ranged from 76.38±0.44
to 81.30±1.78 and the moisture was observed more in Narcetes erimelas (81.30±1.78)
followed by Alepocephalus bicolor (80.73±0.55) and Talismania longifilis (80.43±0.60),
while in Cruise 338 moisture content was accumulated in between 74.92±0.96 and
81.44±1.30, the moisture was high in Alepocephalus bicolor (81.44±1.30) followed by
Talismania longifilis (81.05±1.06) and Narcetes erimelas (80.27±2.04).

Table 1. Moisture content of deep-sea fishes from three cruises of Indian EEZ
S.No Species name Cruise 322 Cruise 332 Cruise 338
1 Alepocephalus bicolor 81.11±0.69 80.73±0.55 81.44±1.30
2 Narcetes erimelas 80.83±1.05 81.30±1.78 80.27±2.04
3 Talismania longifilis 79.34±0.42 80.43±0.60 81.05±1.06
4 Chlorophthalmus bicornis 73.93±0.67 77.51±0.72 75.91±0.54
5 Lamprogrammus niger 79.15±1.52 80.34±0.28 80.20±1.09
6 Beryx splendens 75.23±1.36 79.19±1.11 77.40±1.65
7 Chelidoperca investigatoris 75.83±1.05 76.38±0.44 74.92±0.96
8 Neoepinnula orientalis 79.55±1.82 78.39±0.60 77.46±1.43
9 Cubiceps baxteri 77.24±1.47 76.82±1.54 78.15±1.68
10 Psenopsis cyanea 80.03±0.81 79.41±0.22 80.12±1.25
Note: n=5; mean±SD
The comparison of moisture content in species wise is A bicolor, T longifilis, C
baxetri and P cyanea accumulated more moisture content in Cruise 338, N erimelas, C
bicornis, L niger, B splendens and C investigatoris having high moisture content in
Cruise 332 and N orientalis accumulated more moisture in Cruise 322 respectively. The
total mean concentration of all cruises moisture content was observed high in A bicolor
and less content found in C investigatoris. In cruise wise, there was a slight variation was
observed in overall average concentration of all deeper water fishes.
Graph 1. % of Moisture determined in deep-sea fish species from three cruises of Indian
EEZ
84
82
80
78
Moisture percentage
76
74
Cr 322
72
70 Cr 332
Cr338
Deep-sea fish samples

PROTEIN: The protein values of ten deeper water fish species were shown in Graph 2
and Table 2. The protein values were ranged in between 15.99±1.03 and 22.43±1.22 of
cruise 322, the values showed that the protein content accumulated more in Chelidoperca
investigatoris (22.43±1.22) followed by Chlorophthalmus bicornis (21.86±0.29) and
Cubiceps baxteri (21.12±1.91), while in Cruise 332, the protein content accumulated in
deep-sea fishes was from 15.79±1.08 of Narcetes erimelas to 20.85±1.00 of Cubiceps
baxteri, the protein percentage was observed more in Cubiceps baxteri (20.85±1.00)
followed by Chlorophthalmus bicornis (20.79±0.40) and Chelidoperca investigatoris
(19.45±0.26), whereas in Cruise 338, the protein content was found in between
16.08±1.05 and 21.75±0.15, and it was found more percentage in Chlorophthalmus
bicornis (21.75±0.15) followed by Chelidoperca investigatoris (21.17±0.62) and
Neoepinnula orientalis (20.57±0.38) respectively.
Graph 2. % of total nitrogen determined in deep-sea fish species from 3 cruises of Indian
EEZ
25
20
Total nitrogen content
15
Cr 322
10
Cr 332
5 Cr338
Graph 2: % of total nitrogen in deep-sea fish samples

Table 2. % of protein of deep-sea fishes from three cruises of Indian EEZ
6 Beryx splendens 20.87±0.24 18.79±0.61 19.76±0.29
Note: n=5; mean±SD
The protein percentage was compared in species wise is A bicolor, T longifilis, C
bicornis, L niger, B splendens, C investigatoris, C baxteri and P cyanea are the species
which found more protein percentage in Cruise 322, whereas N erimelas and N orientalis
having more protein content in Cruise 338. There was no higher content of protein
percentage was observed in Cruise 332 deep-sea fishes. The overall mean concentration
of all cruises protein percentage was observed more in C bicornis and lowest percentage
was found in A bicolor.
FAT
The fat values of ten deeper water fish species were shown in Graph 3 and Table
3. The fat values were observed in between 1.91±0.18 of Alepocephalus bicolor and
0.46±0.03 of Lamprogammus niger of cruise 322, whereas in cruise 332, the highest
value of fat accumulated found in Chelidoperca investigatoris (2.54±0.23) and lowest
content was observed in Chlorophthalmus bicornis (0.67±0.27), while in cruise 338, the
fat percentage was found more in Chelidoperca investigatoris (2.53±0.68) followed by
Alepocephalus bicolor (2.20±0.06) and Beryx splendens (1.97±0.42) respectively. The
overall cruise wise mean concentration of fat content was observed more in
Alepocephalus bicolor (2.12 %) followed by Chelidoperca investigatoris (2.00 %) and
Beryx splendens (1.62 %) respectively.
Graph 3. % of fat determined in deep-sea fish species from three Cruises of Indian EEZ
2.5
2 Cr 322
Fat percentage
1.5 Cr 332
1 Cr338
0.5
Graph 3: % of fat content observed in deep-sea fish samples

Table 3. Fat content of deep-sea fishes from three cruises of Indian EEZ
6 Beryx splendens 1.57±0.18 1.32±0.59 1.97±0.42
Note: n=5; mean±SD
The fat percentage was compared in species wise is Alepocephalus bicolor ,
Narcetes erimelas, Lamprogrammus niger , Chelidoperca investigatoris, Neoepinnula
orientalis and Psenopsis cyanea accumulated more amount of fat in cruise 332, whereas
Chlorophthalmus bicornis and Cubiceps baxteri contains high fat in cruise 322, while in
cruise 338 more fat content accumulated in Talismania longifilis and Beryx splendens .
The overall mean concentration of all cruises fat percentage was observed more in
Alepocephalus bicolor and lowest percentage was found in Lamprogrammus niger.
ASH: Ash results of the ten given deep-sea fish species were shown in Graph 4 and table
4. The ash values of deeper water fishes accumulated was given in cruise wise data. More
amount of ash was found in Chlorophthalmus bicornis (1.95±0.10) followed by Beryx
splendens (1.90±0.32) and Narcetes erimelas (1.83±0.12) in cruise 322, whereas in cruise
332, ash percentage found more in Lamprogrammus niger (1.45±0.53) and lowest
content was observed in Chlorophthalmus bicornis (0.69±0.10), while in cruise 338, ash
percentage observed more in Chelidoperca investigatoris (1.43±0.26) followed by Beryx
splendens (1.31±0.22) and Cubiceps baxteri (1.09±0.10). The overall cruise wise average
concentration of ash values was found more percentage in Beryx splendens (1.50 %) and
lowest ash content was observed in Psenopsis cyanea (0.73 %).

Table 4. Ash content of deep-sea fishes from three cruises of Indian EEZ
S.No Species name Cr 322 Cr 332 Cr 338
6 Beryx splendens 1.90±0.32 1.29±0.15 1.31±0.22
Note: n=5; mean±SD
Ash percentage was compared in species wise is Narcetes erimelas,
Chlorophthalmus bicornis and Beryx splendens having more amount of ash in cruise 322,
whereas Talismania longifilis, Lamprogrammus niger and Psenopsis cyanea contains

high ash percentage in cruise 332 while in cruise 338, more ash content found in
Alepocephalus bicolor, Chelidoperca investigatoris, Neoepinnula orientalis and
Cubiceps baxteri. The overall mean concentration of ash content was accumulated high
in Beryx splendens and lowest ash content found in Psenopsis cyanea.
Graph 4. % of ash determined in deep-sea fish species from three Cruises of Indian EEZ
2.5
2
Ash percentage
1.5 Cr 322
1 Cr 332
Cr338
0.5
Graph 4: % of ash content observed in deep-sea fish samples

DISCUSSION
There is an escalating exploration of deep-sea fish species throughout the
preceding years, with the intention of investigation so far under utilised fishes that can
reinstate or supplement the accessible commercial fish samples. It is therefore significant
that the novel species can contend with the traditional species concerning both taste and
nutritional values. In this study, we determined the proximate composition of 10 deep-sea
fish species which are included Alepocephalus bicolor, Narcetes erimelas, Talismania
longifilis, Chlorophthalmus bicornis, Lamprogrammus niger, Beryx splendens,
Chelidoperca investigatoris, Neoepinnula orientalis, Cubiceps baxteri and Psenopsis
cyanea from the Indian Ocean and the Arabian Sea were studied. Theses deep-sea fish
species were collected from three cruises viz., Cruise 322, Cruise 332 and Cruise 338
with the help of FORV (Fisheries Oceanographic Research Vessel) Sagar Sampada from
the depth of 200 – 1200m depth.
Proximate composition of some kind of deep-sea fishes has various percentages.
This disparity is caused by environmental variations in the midst of others, temperature,
salinity, pressure, oxygen availability and more. Deep sea fish require fat to remain body
temperature because of the environment in the deep ocean has a lower temperature than
pelagic environment or surface (Davis, 1991). In general, deep-sea fishes are smaller in
size than the pelagic fishes so that it effected to the edible part/muscle portion
(Oceanlink, 2006). Works allied with proximate composition of deep-sea fish species is
inadequate although a very few workers have been carried out on the biochemical
composition of some deep-sea fishes. Information on the proximate composition and fatty
acid distribution is important when utilization of new species of deep-sea fish is
considered. This is because deep-sea fishes are well thought-out to be not only food with
good source of quality protein but also food with healthy components. The moisture
content of the fish samples were within the acceptable limits reported by Gallagher et al., (1991).
FAO (1999) reported that moisture and lipid contents in fish muscle are inversely related and
their sum is approximately 80% with other components accounting for the remaining 20% which
partially agree with the finding of the present study.
MOISTURE:
Moisture content in deep-sea fish in contained in two forms: free water and bound
water. Free water content found in the spaces between cells and plasma that can dissolve
a wide range of vitamins, mineral salts and certain nitrogen compounds. Bound water
found with chemically bounding, psychochemical bounding and capillary bounding. The
species of the slope and open sea slopes Coryphaenoides rupestris and Anolopoma fimbria have
been found to contain greater fractions of moisture content in their muscle tissues (85 & 79%
respectively) (Krzynowek and Murphy, 1987; Crabtree, 1995). Deep-sea fish contained more
percentage of water than pelagic fishes. It was caused by having high salinity, more than
34.2%. According to Nybakken (1992), deep-sea fishes having high amount of moisture
content than pelagic fishes due to accumulation of more water in body tissues in deep-sea
fish has increased with increasing depth. Many preceding studies have been reported that
the percentage of moisture content of fresh fish is inversely related to the lipid content
(Jahncke and Gooch, 1997). The percentage of moisture, protein, fat and ash contents of
the deep-sea fishes have been tabulated in Table 1 – 4. In the study, the moisture content
found to be almost 73 to 82 % in Cruise 322, whereas in Cruise 332 moisture content
accumulated in between 76 – 82 % and in Cruise 338 moisture percentage observed in
between 74 – 82 % respectively. This is the result which is comparable to the proximate

test results for deep-sea fish species reported previously (Okland et al., 2005) and also
reported by Suseno et al., (2010). In this study, the results were similar to the proximate
composition results for deep-sea fish species reported previously (Okland et al., 2005)
There is a relation of inversion between fat and moisture percentage. Moisture
content has an inverse relation with the fish size (Hossain et al., 1999). Since the
moisture has an inverse relation with size of the fish, the deep-sea fish species in the
present study being small showed relatively higher moisture content that obtained by
Rahman et al., (1982) in small zeol fishes of Bangladesh (70.60 to 80.44%). If lower the
percentage of moisture, greater the fat and protein contents and higher the energy density
of the fish (Dempson et al., 2004). High moisture percentage augments the fish
vulnerability to microbial spoilage, oxidative degradation of polyunsaturated fatty acids
and accordingly diminishes the eminence of the fish quality for longer preservation time
(Omolara et al., 2008).
PROTEIN:
Regarding protein percentage of all the ten deep-sea fish species illustrated the
variations according to three cruise wise changes. The composition of protein of fish
affects the post harvest quality and characteristics with respect to oxidative changes in the
muscle tissues (Owaga et al., 2010). Previously many studies demeanor and showed that
deep-sea fishes are highest protein content and lowest fat content as compared to pelagic
fish (Suseno et al., 2010). However, in the present study, low amount of protein was
found in Narcetes erimelas in cruise 322 & 332 and Alepocephalus bicolor in cruise 338,
whereas high protein percentage was observed in Chelidoperca investigatoris of cruise

322, Cubiceps baxteri of cruise 332 and Chlorophthalmus bicornis of cruise 338
respectively. The protein percentage of the deep-sea fishes examined were comparable to
commercially important fish species from the Black Sea previously reported by Guner et al.
(1998), as well as to that reported by Okland et al. (2005). According to Stanby and Olcott
(1963), deep-sea fishes are rich in protein and low in fat, compared to pelagic fishes, a finding
that is confirmed in the present study.
The protein concentration of the fish species in the study was more or less
comparable to the result of Hossain et al., (1999), Kamal et al., (2007) and Kcukgulmez
et al., (2010). It therefore, seems that the encroachment in maturation is accompanied
with a greater deposition of protein in the muscle. In fishes the lowest protein was
recorded in spent or recovering fishes and highest during peak ripeness. This may
because of a greater deposition of nucleoproteins in fish (Jafri and Khawaja, 1968). The
cruise variation of protein content in the edible portion of the deep-sea fish species may be linked
with changes in the feeding activity, because the edible portion of protein losses during starvation
has been observed in N. norvegicus (Dali, 1981). Since during that period of starvation, the
abdominal muscle builds the largest contribution of protein to energy metabolism, minute
variations were observed in this tissue are sufficient to make a significant contribution to the
overall animal maintenance (Barclay et al., 1983). The difference of the protein content during
ovarian development could be the result of an augment in several protein biosynthesis, including
peptide hormones, enzymes and egg yolk proteins, which are particularly imperative in
maturation (Yehezkel et al, 2000).

FAT:
Fat has an inimitable significance to the animal body, which is mainly an energy source
in the diet. Fats also stuffing to remain various body organs in place. Moreover fats give the diet
its meticulous flavour. The percentage of fat based on the reproduction and food. The differences
in season, depending on the food availability at different time of the year, have a substantial effect
on the tissue components particularly the fat. Changes in the reproductive cycle also have a
marked effect on the body composition. Like other animals fish store fat to supply energy needed
during food scarcity and reproductive phases. Reduction of the fat content during the spawning
season has been recorded.
Deep-sea fish species were classified into four categories according to their fat levels,
lean fish (<2%), low fat (2 - 4%), medium fat (4 - 8%) and high fat (>8%) according to Dyer and
Bligh (Huss, 1988; Ackman, 1989). Moreover, it could be classified on the >5% (dwb) fat
percentage criteria for discriminating the lean (fat < 2%) percentage from fatty fishes as per
soxhlet (Owaga et al., 2010). In the present study, deep-sea fish examined were found in lean and
low fat categories and was not found in medium and high fat category. The species of lean fish in
Cruise 322 is Alepocephalus bicolor, Narcetes erimelas, Talismania longifilis,
Chlorophthalmus bicornis, Lamprogrammus niger, Beryx splendens, Chelidoperca
investigatoris, Neoepinnula orientalis, Cubiceps baxteri and Psenopsis cyanea, whereas
in cruise 332 & 338, Alepocephalus bicolor and Chelidoperca investigatoris were come
under low fat (2 – 4% category) and remaining all other deep-sea fishes were found
under lean (<2 %) category. The fat content in the species Alepocephalus bicolor was similar
to the reported previously by Ozagul and Ozagul (2007) but less amount was observed than that
reported by Guner et al. (1998). However, the observation of variation in amount of fat
from the different landing sites could be attributed to, water, temperatures, sex, age,
season of the year, food availability and salinity of the different geographical locations of
the landing sites (Stansby, 1981; Monsen, 1985; El Tay et al., 1998).
Das (2009) reported that various species illustrate different lipid levels at different
conditions (Location, Temperature, size and freezing time). Usually moisture and fat
percentages in fish species of edible part are inversely related and there sum is
approximately 80% (FAO, 1999). This inverse relationship was also well defined in this
study. Starvation alters body components. Lipid and protein are utilized as a source of
energy, diminishing progressively during starvation while moisture content increases
proportionately (Niimi, 1972).
In addition, the fat percentage of deep-sea fishes is one of the very few natural
food sources of Vitamin D and having important amounts of Vitamins A and E (α–
tocopherol) (Bhuiyan et al., 1993; Qyvind et al., 1994). Lipids are high energy nutrient
that can be utilized to partially spone (substitute for) protein in aquaculture feeds. Fat
typically comprise about 15% of fish diet and serve as transporters for fat-soluble
Vitamins. Lipids are deemed to be the most significant constituent of fish egg as a reserve
energy source (Pal et al., 2011). Bhuyan et al., (2003), Shamsan and Ansari (2010)
reported coincidence of intensive feeding with occurrence of high fat content in the
muscle of fish. Environmental factors are well known to affect the proximate analysis and
condition of White Sea bream and other fish, especially fat content (George and Bhopal,
1995; Iverson et al., 2002). Love (1957) indicated that the availability of food at different
time of the year has a considerable effect on the tissue component, particularly fat.
According to Stanby and Olcott (1963) reported that the deep-sea fishes are high in
protein and low in fat, the same result was found in this study. The low amount of fat was
found in Lamprogrammus niger of cruise 322 and 338, whereas in cruise 332 less content
of fat was observed in Chlorophthalmus bicornis. Likewise high amount of fat was
observed in Chelidoperca investigatoris of cruise 332 and 338, while in cruise 322 higher
fat was found in Alepocephalus bicolor respectively. The different fat percentage of each
fish can be prejudiced by age, seasons of year, food availability and environment such as
salinity and temperature (Stansby, 1981; Monsen, 1985). Lipids are the most essential source
of metabolic energy during the embryonic development, and their amount is usually correlated
with the size of the egg and with the time interval between spawning and hatching or larval first
feeding (Rainuzzo et al., 1997).
ASH:
Ash is an evaluated mineral content of any food including fish (Omotosho et al.,
2011). The ash percentage of the muscle tissue of deep-sea fishes was generally higher
than that of the pelagic fishes. Love (1970) reported that generally fish contains 0.5 to 5%
of ash in their edible portion. The concentrations of minerals and trace elements that
contribute for the total ash contents are known to vary in fish depending on their behavior
of feeding, increasing weight or length of the fish (Hassan, 1996), season, environment,
ecosystem and migration even within the same area (Andres et al., 2000; Canli and Atli,
2003; Abdallah and Mohamed, 2007). Total mean value of ash percentage was
found in the study is in between 0.41 % and 1.95 %. The highest ash percentage was
found in Chlorophthalmus bicornis in cruise 322, Lamprogrammus niger in cruise 332
and Chelidoperca investigatoris in cruise 338 and lowest ash content was observed in
Psenopsis cyanea of cruise 322, Chlorophthalmus bicornis of cruise 332 and
Lamprogrammus niger of cruise 338 respectively. In the study, the results of ash
percentage were relatively slightly lower than those of deep-sea fish species (Suseno et
al., 2010) with minor edible portions. This is because of pelagic fishes are consumed
whole (edible portion is 100%) unlike deep-sea fish species with edible portions of only 5
- 63% (Suseno et al., 2010). The highest total ash percentage in comparison with other
fish species is as a direct result of inclusion of skeletal muscles in the edible portion that
contains calcium and iron in high amounts; 3600 mg/100 g and 10.2 mg/100 g,
respectively (Ghelichpour and Shabanpour, 2011; Owaga et al., 2010).
Smaller sized fish species has high amount of ash percentage due to the higher
bone of muscle ratio (Daramola et al., 2007). Such fish offer minerals in their edible
forms more plentifully than large-sized fish do (Higashi, 1962). The percentage of ash
might be changes with the time of storage due to absorbance of moisture and defeat of
protein (Hassan et al., 2013). CSIR (1962) reported that the percentage of ash content of
some selected fish species in India ranged in between 1.53 and 2.60%. Only gross
measurements are made of ash. Ash concentrations increased marginally at maturity and
markedly after spawning. At maturity stage, the ash was light blue in colour. The
fluctuation of ash content made difficult to attest any relationship with the spawning
season. In general, body composition of fishes seems to depend on age, sex, season and
diet. The concentration of total ash was observed in the present studied of deep-sea fishes
in comparison with other fish species is as a direct result of inclusion of skeletal muscles
in the edible portion contains high calcium levels. The richest ash percentage is found in
the bones of the fishes than in their edible portion. As the present study was aimed
towards the evaluation of nutritive values of edible portions of the deep-sea fish species,
only the proximate percentage of the muscle part was taken into consideration.
CHAPTER-II
MONITORING OF HEAVY METAL
ACCUMULATION IN DEEP-SEA
FISHES
INTRODUCTION
Deep-sea fisheries are becoming more significant and there is a paucity of
chemical monitoring of these recent fisheries and their products. Seafood is significant in
human nutrition because of its unique nutritive value related to the presence of proteins,
fats, vitamins and minerals. At present, India is the second largest producer of fish in the
world. As the demand for fish is continuously increasing, making the required protein
available to the existing population is a challenge. With an increasing population, the
fishing pressure is also increasing in the capture sector. Most of the crafts used in Indian
waters are of 40 to 50 foot in length, having the capability of trawling only up to 200-300
m in coastal waters of Indian EEZ. The catch in coastal waters has already reached
sustainable level beyond which there may be depletion of resource in inshore areas
(Vivekanandan, et al., 2003). Hence, in the past decade, importance has been given for
exploring deep sea resources (Vivekanandan, 2001; Thirumilu & Rajan 2003; Sebastine
et al., 2011). Several exploratory surveys (Rajan et al., 2001; Karuppasamy et al., 2008)
conducted along the deep waters of both the Indian coasts indicated that deep sea
fisheries can be of much commercial and economical value.
Fishes are considered as a good indicator for heavy metal contamination in
aquatic systems. Fish is extensively consumed in several parts of the world by humans
and polluted fish may imperil the human health (Zhang et al., 2007). Metals are non-
biodegradable and consider as major environmental pollutants causing cytotoxic,
mutagenic and carcinogenic effects in animals (More et al., 2003). It is essential to
examine the lethal consequence of metals on fish since they represent an important link in
food chain and their contamination by metal causes imbalances in aquatic system (Firat
and Kargam, 2010). Marine organisms have capability of heavy metal accumulation from
different sources including soil erosion, sediments and runoff, air depositions of dust and
aerosol, and discharge of waste water (Goodwin et al., 2003). A numerous factors
including season, physicochemical properties of water, habitat, age and physiological
conditions of fish play an imperative role in accumulation of metals by fish (Kargin,
1996). Gills are directly in contact with water; therefore the concentration of metals in
gills reflects metal concentration in water where the fish lives. High level of metal
concentration accumulated in liver which represents the storage of metals from water for
detoxification (Romeo et al., 1999). Accumulation of heavy metals in within the fish
differs based on the route of metal uptake, type of heavy metal, and fish species (Begum
et al., 2009). The occurrence of higher amount of heavy metals in any part of the body
will provoke changes in biochemical metabolisms, serum biochemical changes,
histopathological changes and other induced stresses. Therefore the studies on the
accumulation of heavy metals in different organs of the fish were very much significant.
The alterations of histopathological studies in fish under the influence of heavy metals
can be used as a consistent indicator of aquatic pollution. Fishes are main source of the
human diet and it is therefore not astounding that numerous studies have been carried out
on metal pollution in different species of edible fish (Karadede and Unlu, 2000; Prudente
et al., 1997; Unlu et al., 1996 and Erdogrul and Ates, 2006). Predominantly, fish
toxicological and environmental studies have encouraged interest in the determination of
toxic elements in seafood (Waqar, 2006).
Toxic components in fish like heavy metals accumulated directly from water and
diet, and contaminant residues may ultimately reach concentrations hundreds or
thousands of times above those measured in the water, sediment and food (Goodwin et
al., 2003; Labonne et al.,2001; Osman et al., 2007). Heavy metals are general
components of marine environment that occur as a result of pollution predominantly due
to the emancipation of unprocessed wastes into rivers by many industries. The
contamination of the marine water, sediments and organisms with an extensive range of
pollutants has become a matter of great concern over the last few decades (Secrieru and
Secrieru, 2002; Ergül et al., 2008; Bat et al., 2009; Boran and Altınok, 2010). Heavy
metals are innate trace components of the marine environment, but their ranges have
amplified due to domestic, industrial, mining and agricultural activities (Bakan and
Büyükgüngör, 2000; Altas and Buyukgungor, 2007). Emancipation of heavy metals into
river or any oceanic environment can alter both the ecosystems and diversity of marine
species, due to their behavior of accumulation and toxicity (Bat, 2005; Bakan and Böke
Özkoç, 2007; Bat et al., 2009).
Marine organisms such as fish accumulate heavy metals to concentrations many
times higher than present in water or sediment (Bryan, 1976; Phillips and Rainbow, 1994;
Bat et al., 2009; Boran and Altınok, 2010). Thus, heavy metals acquired through the food
chain as a result of pollution are potential chemical hazards, threatening consumers.
Some of the heavy metals at diminutive levels such as copper and zinc are essential for
enzymatic activity and many biological processes. However, a few heavy metals like
cadmium and lead which may be introduced into the aquatic environment from
anthropogenic activities have no known essential role in living organisms, and are toxic
at even low concentrations. The essential metals also become toxic at high concentrations
(Bryan, 1976). Once in the marine environment these heavy metals can be concentrated
in fish tissues.
Bioaccumulation of heavy metals in the edible part of marine organisms has been
predicted as an indirect measure of the availability and abundance of metals in the marine
environment (Kucuksezgin et al., 2006). For this reason, monitoring seafood edible part
contamination provide an important function as an untimely warning indicator of
sediment contamination or allied water quality problems (Mansour and Sidky, 2002;
Barak and Mason, 1990) and permit us to take appropriate action to protect public health
and the environment. Manifold aspects including season, physical and chemical
properties of water can play a momentous role in accumulation of metals in various fish
tissues (Hayat et al., 2007; Romeo et al., 1999). Several studies (Ademoroti, 1996; Heath,
1987; Karthikeyan et al., 2007) have also predicted that seafood are able to accumulate
and retain heavy metals from their environment depending upon reveled concentration
and duration as well as salinity, temperature, hardness and metabolism of the animals.
Adeyeye et al. (1996) also illustrated that the metal concentration was a function of fish
species as it accumulates high in some fish species than others. Seafood is always in news
as it is asserted to be most nutritious and healthy food as well as being connected to
increasing number of food borne outbreaks across the globe. In the nutritional front, fish
accounts for 16.6 percent of the worldwide population ingestion of animal protein and
6.5% of all protein consumed (FAO, 2012). The world per capita consumption of fish and
fishery products has increased from 9.9 kg in1960s to 18.6 kg in 2010.
Seafood consumption is a momentous pathway to metal exposure in human
population living in coastal areas. Mineral intensification in the coastal zone augment the
metal load of inshore waters and aquatic animals, ensuing in major health risk from
seafood consumption (Hashmi et al., 2002). Contaminated fish consumption with heavy
metals could have hazardous effects on human health (Mai et al., 2006). Mostly seafood
have been reported to integrated these heavy metals through intake of suspended
particulates, food materials and or by constants ion exchange process of dissolved metals
across the lipophilic membranes such as the gills, absorption of dissolved metals on tissue
and membrane surfaces (Oguize, 1999; Oguize and Okosodo, 2008). As a result,
bioaccumulation of heavy metals is a major route, which augmented range of the
pollutants are transferred across food chain web generated public health problems
wherever man is involved in the food chain therefore, it is essential to continuously
resolving the bioaccumulation capacity for heavy metals by organisms specially the
edible ones, in order to evaluate potential risk to human health (Otitoloju, 2002).
The marine environment is one great connected body of water that covers more
than 70% of the earth’s surface and contains 97% of all water on the planet. It affords
food from plants and animals, energy, transportation, medicine, minerals and natural
resources. There is incredible biodiversity in the ocean; in fact, more plant and animal life
is found in the ocean than on land, almost half of the earth’s major animal groups live
only in the sea. Pollution of marine ecosystems by heavy metals is of environmental
concern worldwide. Domestic sewage, industrial effluents, combustion emissions, mining
operations and metallurgical activities are among the sources of anthropogenic metal
inputs. Even though in trace amount of heavy metals are in usual components of marine
organisms, whereas at higher levels they are potentially toxic and may interrupt the
metabolic functions of aquatic ecosystems. The heavy metals in the organs of marine
organisms are the ability to be concentrated for their toxicity and also pretense a direct
threat to both human and aquatic biota (Watling, 1983).

Heavy metals are generally institute in natural waters and some are important to
living organisms, yet they may come highly toxic when there in more concentrations
(Ibok et al., 1989). These metals also gain access into ecosystem through anthropogenic
source and get disseminated in the water body, hovering solids and sediments during the
course of their mobility (Olaijire and Imeokparia, 2000). The rate of bioaccumulation of
heavy metals in aquatic organisms depends on the ability of the organisms to digest the
metals and the concentration of such metal. Also it has to do with the concentration of the
heavy metal in the surrounding soil sediments as well as the feeding habits of the
organism. Aquatic animals (including fish) bioaccumulate trace metals in considerable
amounts and stay over a long period. Fishes have been recognized as a good accumulator
of organic and inorganic pollutants (King and Jonathan, 2003). Age of fish, fat content in
the muscle part and mode of feeding are important factors which affect the accumulation
of heavy metals in fishes. They are ultimately transferred to other animals including
humans through the food chain. Odoemelam et al. (1999) revealed high concentrations of
heavy metals such as Cd, Pb, Cu, Ni, Zn, Mn, Mg and Co in some waters within the
proximity of some industrial cities. Expulsion of industrial wastes restraining hazardous
heavy metals into water bodies may have momentous effects on fish and other aquatic
organisms, which may imperil public health through consumption of contaminated
seafood and irrigated food crops. Nwaedozie (1998) reported that zinc contamination
affects the hepatic allocation of other trace metals in fish. Zinc, copper and manganese,
which are essential elements, contend for the same site in animals. This, no doubt, would
affect tissue metal concentrations as well as certain physiological processes.

Heavy metals can be accumulated by marine organisms through a variety of
pathways, including respiration, adsorption and ingestion (Zhou et al., 2001). The levels
of heavy metal are known to increase drastically in marine environment through mainly
anthropogenic activities. This result either with the complete extinction of some non
tolerant species from these environments or they cause structural and functional damage
to the organisms by interfering with their physiological and biochemical mechanisms.
Biomonitors play an important role on in the assessment of the bio-availabilities of heavy
metals in the marine environment. Many species of marine benthic fish have been shown
to reflect ambient metal concentrations (Hornung and Ramelow, 1987; Romeo et al.,
1999; Kucuksezgin et al., 2001).
Heavy metals are innate components of the earth’s crust that cannot be tainted or
destroyed. They are metallic elements that have a relatively high density, toxic and
poisonous. According to many researchers (Blaber, 2000; Farombi, et al., 2007; Brewer,
2010), heavy metals can cause contaminations that result in a long lasting effect on the
ecological balance of the environment and the assortment of the various aquatic species
living in it. Among the aquatic species, fishes are the mostly endangered as they cannot
flight from the effects of these pollutants which has mentioned by Mansour and Sidky
(2002). Pollutants can accrue in huge quantities of heavy metals from the waters inside
their body. Even though the fish consumption can benefit its consumers and shape up
their health, it can also promote some health concerns as it can be contaminated with
hazardous metals from the pollution of their aquatic environment (Kennish, 2001;
Madhusudan et al., 2003; Kojadinovic et al., 2006). Despite the fact that most of the
heavy metal such as mercury, cadmium and lead are lethal and perilous, while there are
some heavy metals which are essential and must be taken in small amounts such as iron,
copper, manganese and zinc (Mohammad and Hossam, 2007; RinaSharlindaZabri, 2009).
Fishes can easily absorb the heavy metals which were accumulates in the muscle or
edible part of marine organisms from the surrounding areas and the ranges of heavy metal
accumulation in previous exposures can be known (Basa et al., 2003; Ashraf, 2005).
Rapid industrialization and the emancipation of potentially hazardous trace metals
into the oceanic environment has become a solemn problem (Gaspic et al. 2002).
Pollutants were accumulated in marine organisms including fish from the environment,
consequently used in marine pollution monitoring programs (Kucuksezgin et al. 2001).
Many of the researchers have been reported about metal ranges as a whole in fish, water
and sediment in a meticulous region (Usero et al. 2003; Navarro et al. 2006; Tuzen 2003;
Turkmen et al. 2005). A metal like copper is an essential element since it plays a pivotal
function in biological systems, while mercury, cadmium and lead were non-essential
metals as they were toxic even in small quantities.
The term trace metal is used to allocate the elements, which present in trace
quantities (<100 ppm) in natural biological systems. There are both essential and non-
essential trace metals. It is well known that Cu, Ni, Zn, Cr, Co, Fe and Mn are essential to
lite (lvlertz, 1981) and known as essential trace metals. The elements Al, Sb, Hg, Cd, Ge,
V, Si, Rb, Ag, Au, Pb, Br, Ti etc are believed to be attained by animal body from
environmental pollutants because of contacting of organism with the environment. These
elements are in general concentrated erratically in various organs and are called as non-
essential elements. Essential elements worked either as an electron donor system or as
ligands in complex enzymatic compounds in animals. Since indispensable elements were

used by the organisms only in small quantities, whereas their enhancement in the
organisms does not surpass the ranges which allow the enzyme system to function
without interference (Presley, 1997). Even if the heavy metal concentration is too high at
the source of supply, the homeostatic mechanism stops the function and the indispensable
heavy metals act in either acutely or chronically toxic manner. Thus, the organism may
be exaggerated even in the event of extended bioaccumulation of heavy metals.
Heavy metals in the aquatic environment and its effect on aquatic life
Trace metals are commencing in the environment from both natural sources and
as a result of human activity (Phillips et al, 1982; Martin and Scenes, 1996; Presley,
1997). A huge number of heavy metals may be contributed by corrosion of metal pipes,
smelting, refining, etc. Weathering is a natural source of dissolved and particulate trace
metals. Trace metals play an important role in the aquatic ecosystem. In marine
organisms, uptake of heavy metals was reported by many researchers (Saiki et al., 1955).
Brooks and Fiumsby (1965) studied the biogeochemistry of trace metal uptake of Ag, Cd,
Cr, Cu, Fe, Mn, Mo, Ni, Pb, V and Zn in three species of New Zealand bivalves. All the
elements analysed showed more enrichment in molluscs than in the environment.
Geological weathering of rocks produces the clays and other minerals that make up the
bulk of detrital sediments as well as the bulk of dissolved metals seawater. Another
source of metals either on land or in the sea is volcanic activity. During the 1990’s
(Badach and Heyer, 1997), several experiments data showed that cadmium in large pools
were contained in the sediments and were ingested by the benthos (Hall et al., 1996).
Some of the metals may be absorb by the phytoplanktons and as a result, a significant
concentration of mercury and cadmium were found in kidney and livers of top predators.
Metallic pollution was persistent and bioaccumulated by marine organisms with solemn
public health insinuations (Phillips and Rainbow, 1994).
The environmental impact of a metal depends less on its source than on its
behavior. Its behavior, including mobility, transport, transfer and biological uptake,
depends strongly on the chemical and physical form of the metal. The toxicity of a metal
is mainly determined by its ionic size, electron affinity, electro negativity, stability,
"solubility and its inherent capacity to adversely affect any biological activity (Wittmann,
1974). The heavy metals have high affinities for ligands containing sulphur and nitrogen,
and hence are bound easily to organic molecules such as proteins, enzymes etc
(Richardson, 1980).
Metals like Cu, Zn, and Fe are essential of indispensible metals which shows
noxious effect on human health instigates when they are present in high levels. Whereas
heavy metals like Co, Cd, Pb, Cr, Se and As are dispensable or non-essential metals and
their noxious effect on human health is eminent. As it explained formerly, these elements
may be added on the ecosystem through the activities of humans or from natural sources.
Numerous studies evaluated the concentration of these heavy metals in fish species all
over the world.
Copper (Cu)
Copper is an essential trace element for the fixation of Fe in haemoglobin and is
not a potent liver toxin except in certain cases of genetic defects resulting in the inability
to excrete copper, the primary homeostatic mechanism, for instance Wilson’s disease. It
plays a crucial role in many biological enzyme systems that catalyze oxidation/ reduction
reactions. However, if present at relatively high concentrations in the environment,
toxicity to aquatic organisms may occur. Copper under ionic forms Cu2+, Cu2OH+ and
CuOH+ is toxic to fish (Moore, 1991). High copper levels lead to an increase in the rate
of free radical formation (Gwozdzinski, 1995) teratogenicity (Stouthart et al., 1996), and
chromosomal aberrations (Fahmy, 2000). Copper in ionic form is found to be toxic and
inhibits photosynthesis and affect the growth of unicellular algae (Nielson and Anderson,
1970). Cu in excessive amounts causes haemolysis, hepatotoxic and nephrotoxic effects.
The phenomenon of green-sick oysters is caused by high content of copper in the
environmental water. The effluents from copper refineries, pesticide and fungicide
manufacturing industries bring copper to the aquatic systems. ln Taiwan, copper pollution
due to the discharges from the local copper recycling operation has been reported (Hung,
1988) and this has caused serious toxicity in green oysters (Hung et al., 1989). The
highest level of Cu in the oysters collected from the polluted area was 4400 ppm (Hung
and Han, 1991). Thus, higher level of Cu in the environment or marine organisms
adversely affects quality and fishery and can cause great economic loss.
Zinc (Zn)
The world's zinc production is rising, which means that more and more zinc ends
up in the environment. Zinc is present in large quantities in the waste water of industrial
plants, leading to the pollution of water ways. Some fish living in the Zn-contaminated
waterways can accumulate zinc in their bodies. When zinc enters the bodies of these fish,
it is able to biomagnify up the food chain. Zinc is an essential element for the life of
animal and human beings (Momtaz, 2002). It is present in many enzymes involved in
important physiological functions like protein synthesis and also it is essential for male
reproductive activity. Zinc is a metal also toxic to fish and other aquatic organisms at
higher concentrations (Pringle et al., 1968). Zinc occurs naturally in air, water and soil,
but zinc concentrations are rising unnaturally, due to human activities. Most zinc is added
during industrial activities such as mining, coal and waste combustion, and steel
processing. Zinc is a very common substance that occurs naturally. Many foodstuffs
contain certain concentrations of zinc. Zinc is a trace element that is essential for human
health. Zinc deficiency can cause birth defects. The main sources of Zn in the aquatic
systems are the effluents from factories manufacturing zinc compounds, zinc plating
wastes, galvanizing wastes, storage battery, rayon wastes, etc. Zinc has been reported to
cause the same signs of illness as does Pb, and can easily be mistakenly diagnosed as lead
poisoning, excess amount of Zn can cause system dysfunctions, cause impairment of
growth and reproduction, The clinical signs of Zn toxicosis have been reported as
vomiting, diarrhea, bloody urine, icterus (yellow mucus membrane), liver failure, kidney
failure and anemia (Duruibe et al., 2007).
Iron (Fe)
Iron is one of the most important essential plant nutrients and plays an important
role in various metabolic processes such as photosynthesis, respiration, nitrogen fixation
and detoxification of reactive oxygen species (Sunda 2002). Iron enters the aquatic
environment from weathering as well as from human activities such as burning of coke
and coal, acid mine drainage, mineral processing, sewage, iron related industries and the
corrosion of iron and steel (CCREM 1987). Generally iron exists in sea water in two
oxidation states such as iron (II) and iron (III). These oxidation states play a significant
role in the formation of soluble organic and inorganic complexes, colloids, insoluble
ferric oxyhydroxides and particle phases (Achterberg et al. 2001).
In marine system, iron is found to limit phytoplankton growth (Martin and
Fitzwater, 1988; Timmermans et al., 2001). High nutrient low chlorophyll (HNLC) areas
represent the lower limit of the Fe required by phytoplankton. In general, Fe
concentrations in these areas are low relative to other nutrients because there is little or
no atmospheric or riverine input, no upwelled water, and no contribution from
resuspended shelf sediments.
Within marine sediments, iron is generally found to have an abundance of 1-20%
(by weight) with the highest enrichment found near hydrothermal areas and
ferromanganese nodules. The surface layer of sediments are found to be areas of high
chemical activity where early diagenesis occurs, causing the oxidation of organic matter.
The resulting chemical and biologically mediated conditions mean that most particulate
iron species are potentially reducible and can be reintroduced into the overlying waters in
a more bio-available form.
Cobalt (Co)
Cobalt has been generally thought of as a scavenged-type trace element in
seawater. The deep-water concentrations of dissolved Co provide an important insight
into its geochemistry. Even though Co exhibits a scavenging tendency, at intermediate
depths in the offshore transects of both Arabian Sea and Bay of Bengal, below 500m the
concentrations increases towards bottom so that Co has a unique geochemistry, which
means that it is neither a typical nutrient-like nor a scavenged type element. Saito and
Moffett (2001) have shown that Co is organically complexed in deep sea and argued that
such complexation may inhibit the scavenging of Co as well. Such organic complexation
of Co in seawater could serve to stabilize dissolved Co, preventing its microbial co-
oxidation with Mn.
Cobalt seems to be an essential element for certain blue-green algae, nitrogen
fixing bacteria and symbiotic systems as well as for animals (Morel et al., 1994, Sunda
and Huntsman, 1995). It can be found in two oxidation states, Co2+ and Co3+. Co3+ is
found rarely in organisms as it is thermodynamically unstable in natural waters and
cannot be taken up easily. Co can also substitute other trace metals (e.g. Cu, Pb, Zn, Cd)
in a wide variety of minerals due to its similar geochemical properties, to form complexes
(EnvGovCa 2003). In addition, it can influence negatively the growth rate of, and the Ca-
uptake in aquatic herbivores (De Schamphelaere et al. 2008).
Cadmium (Cd)
Cadmium is not essential for biological function in humans. The kidney is
considered the critical target organ, for both the general population and occupationally
exposed populations. Several studies over the past decade have indicated that renal
tubular damage occurs at lower levels of cadmium body burden than previously
observed. Cadmium is an element found widely in the earth’s crust (Pinot et al., 2000).
Mineral weathering, volcanoes and forest fires are the main natural sources of Cd in soils,
and the anthropogenic source is phosphate fertilizers. Leaching from polluted soils and
industrial effluents contribute to cadmium pollution in the aquatic compartment.
Cadmium is a highly toxic metal that readily bioaccumulates in organisms, particularly

bivalves. Diet, mainly fin and shell fish, is the most important route of exposure to Cd
(Llobet et al., 1998). Cadmium affects the activity and metabolism of bone cells,
including osteoblasts and osteoclasts, and hydroxyapatite formation of the extracellular
bone matrix (Regunathan et al. 2003), although the underlying mechanisms are not fully
understood. Exposure to lower amount may cause gastrointestinal irritation, vomiting and
diarrhea. This element has been classified as carcinogenic to humans by the US
Environmental Protection Agency (EPA). It is regarded as one of the most toxic metals.
The poisoning implicated by cadmium containing food is known as ‘itai-ital’ disease in
accordance with the patient’s shrieks resulting from painful skeletal deformities. ‘ltai-itai’
disease was first reported in Jintsu River, Toyama Prefecturor, Japan (Friberg et al.,
1974). The disease was characterized by kidney malfunction, drop in the phosphate level
of the blood serum and loss of minerals from the bones. Anthropogenic sources of Cd
include the mining and minerals processing industries, Zn smelting, paint and plastic
industry, effluent from Ni/Cd batteries, urban runoff due to the elevated Cd
concentrations in phosphate fertilizers etc. Some studies have suggested that cadmium
alters calcium metabolism, contributing to osteoporosis (Jarup et al., 1998b). An
increased risk of lung cancer has been reported following inhalation exposure in
occupational settings, but there is no evidence that cadmium is a carcinogen by the oral
route of exposure (WHO Chemical Fact Sheet, 2006).
Toxicity symptoms induced by cadmium include gastrointestinal disorders,
kidney failure and hypertension. It is also reported that, intoxication with Cd in pregnant
women has been related to reduced pregnancy length and newborn weight and, recently,
to disorders of the endocrine and/or immune system in children (Schoeters et al., 2006).
Lead (Pb)
Lead is found in small amount in the earth’s crust. It can be found in all parts of
our environment. Lead is introduced into the environment by various industries such as
storage batteries, production of chemicals including paints, gasoline additives and various
metal products (eg sheet, pipes). Because of both toxicological characteristics and
dispersion in the environment, lead has been one of the most studied heavy metals. Lead
is found naturally, but the main environmental pollution sources are anthropogenic. Lead
is emitted in large amounts from municipalities, by incineration of waste products. High
levels of Pb have been found in urban runoff. Because Pb is used as an anti-knocking
agent in gasoline in many countries, elevated levels of Pb was found in urban air and by
precipitation it will be carried to the nearby water bodies. The principal source of Pb in
the marine environment appears to be the exhaust of vehicles run with leaded fuels that
reaches the sea water by a way of rain and wind blown dust (Castro and Huber, 1997).
Lead is found at high concentration in muscles and organs of fish. When accumulates in
the human body, it replaces calcium in bones (Lead and health, 1980). Lead exposure has
been mainly related to retardation of neurobehavioral and physiological problems,
particularly in children (Lidsky and Schneider, 2003; Castoldi et al., 2003). The EU
maximum residue limits permitted in fish is 0.3 µg/g for Pb, 0.1–0.3 µg/g for Cd
(Herreros et al., 2008). Data from European Food Safety Authority (EFSA) have related
exposure to Cd and Pb to effects like neurotoxicity, nephro-toxicity, carcinogenicity and
endocrine and reproductive failures in adults (Herreros et al., 2008). Moderate exposure
to Pb and Cd can also significantly reduce human semen quality and is related to many
diseases in adults and children (e.g., damage to DNA or impairment of the reproductive
function) (Telisman et al., 2000).
Chromium (Cr)
Cr is the least toxic of the heavy metals and plays a vital role in glucose tolerance.
Chromium is an important component in industrial sectors and used for the production of
ferrochrome, electroplating, pigment production and tanning. The major sources of Cr in
the marine environment are the wastes of these industries, dumping of solid wastes and
municipal wastes. Different forms of chromium are available in the aquatic environment,
but the main biologically important forms are Cr (III) and Cr (VI). Trivalent species
participates in glucose, lipid and protein metabolism (Eisler 1986), whereas hexavalent
Cr is comparatively more toxic than the trivalent form due to its oxidizing potential,
solubility and capability to cross cell membrane (Levy and Venitt 1986). The main forms
of Cr (III) are CrOH2+, Cr(OH)30 and organic and inorganic complexes, whereas
HCrO4_and CrO42-are the main forms of Cr(VI) in aquatic system (Rai et al. 1987, 1986).
Selenium (Se)
Selenium is not a true metal, but interacts with many metals in the environment. It
is an essential nutrient in small amounts but toxic in higher concentrations, damaging hair
and nails. In the environment as well as in the body, it forms an insoluble salt with
mercury, which reduces the toxicity of both mercury and selenium. The approaches by
which Se can enter an aquatic environment include atmospheric transport and deposition,
direct and indi rect riverine input, sediment resuspension and biological behavior.
Dissolved total Se was invariably undetectable in most samples. The concentration of

total dissolved Se ranged from ND to 0.020 µg/l, with average value 0.005 µg/l.
Compared to those reported for world coastal regions, the average total Se concentration
from Marsa Matrouh Beaches was lower than even Se species that was observed from
Bohai Bay, (Duan et al., 2010) due to the serious influence of anthropogenic activities
(Abdel-Moati, 1998).
Arsenic (As)
Arsenic is another cumulative poison. Large quantities of arsenical compounds
are released into the environment through mining operations and from industries
producing biocidal formulations like herbicides, pesticides, war chemicals etc. Arsenic is
also emitted during the burning of fossil fuels (Lederer and Fensterheim, 1982). Arsenic
intoxication in humans has also been reported (Nriagu, 1988). Regulatory agencies of
many countries have introduced permissible limits for As in Fish and Fish products
(Phillips et al., 1982) and in drinking water (Farmer and Johnson, 1985). Fish and
shellfish have a tendency to accumulate many folds of Arsenic in their body.
Bioaccumulation and toxicity of As has been well documented (Moore and
Ramamoorthy, 1984; Phillips, 1990). Recently in Bangladesh, high level of Arsenic
content has been observed in ground water and implicated Black foot diseases in people
residing in around 41 districts (Biswas et al., 1998). It was also found that water from
96% of tube wells of many districts of Bangladesh were not suitable for drinking. The
symptoms of As poisoning were diffused melanosis, spotted melanosis and spotted
keratoses. The ‘black foot disease’ is caused by the chronic ingestion of inorganic
arsenic.
Earlier studies have revealed that heavy metals such as copper, zinc, iron, cobalt,
cadmium, lead, chromium, selenium and arsenic were found relatively in high
concentrations in some deep-sea fish species (Nezumia aequalis, Lepidion eques and Raja
fyllae) (Mormede and Davies 2001). Some concern arose from that study, particularly in
terms of safety for human consumption. Metals, such as copper, zinc and iron are
essential metals since they play significant roles in biological systems, whereas cobalt,
cadmium, lead, chromium, selenium and arsenic are toxic, even in trace amounts. The
essential of indispensable metals can also produce lethal effects at high concentrations.
Only some metals with proven perilous nature are to be completely excluded in food for
human consumption. Metal absorption in fish is carried out via two uptake routes:
digestive tract (diet exposure) and gill surface (water exposure) (Ptashynski et al. 2002).
Metals are further transferred via blood to other target organs, such as the liver and
kidney. We selected muscles as a primary site of metal uptake and liver as tissues
specialized in metal storage and detoxification. Many other diseases like Bush Sickness,
Black foot diseases, Gena velgum diseases, Wilson’s diseases and White muscle disease
are reported as manifestations of heavy metal poisoning. It is in light of the above,
Due to the deleterious effects of metals on aquatic ecosystems, it is necessary to
monitor their bioaccumulation in key species, because this will give an indication of the
temporal and spatial extent of the process, as well as an assessment of the potential
impact on organism health (Kotze et al. 1999). Monitoring of heavy metals in marine
fishes and shellfishes presumes importance from the consumer safety point of view.
Safety of Seafoods is of paramount significant in the present global scenario. Seafood
should be free from all kinds of hazards affecting human health. When considering heavy
metal content in organisms suitable for human consumption, the most imperative factors
are its toxicity towards humans and affinity for other ligands in the enzyme or protein
matrix.
The aim of this work was to determine ranges and variations of potentially toxic
trace metals (copper, zinc, iron, cobalt, cadmium, lead, chromium, selenium and arsenic)
in the selected deep-sea fishes collected from various sampling points of the Indian EEZ
(Exclusive Economic Zone) since this seafood are an vital constituent for exporting of the
human diet in this zone, thus we wanted to estimate the health risk for these seafood
consumers.
MATERIAL
&
METHODS
Area of Investigation
The samples were collected from the west coast of India during the cruises
namely 322, 332 and 338 of the Fisheries Oceanographic Research Vessel “Sagar
Sampada” under the department of Ocean Development, Govt. of India during the period
of January 2014 to April 2015. The deep-sea fish samples were collected using Expo and
HSDT-CV (High Speed Demersal Trawl – Crustacean vessel) developed by the ICAR-
Central Institute of Fisheries Technology. Sampling was done from Fisheries
Oceanography Research Vessel of the Department of Ocean Development, FORV Sagar
sampada. Twenty samples of each species of almost the same size were kept in
polyethylene bags and stored in the freezer at -20oC, until further analysis. Table 1
showed the details of three cruises.
Cruise No. 322
The fishing was conducted along the deeper waters off west coast from Off Kochi
to Tuticorin and the area covered Lat 08°N to 11°N and Long 74°E to 78°E at the depth
of 200m – 1000m, during January 2014 in the post-monsoon season.
Cruise No. 332
Area covered during the cruise was from latitude 07° N to 10°N and longitude
75°E to 77°E, from Off Kochi to CAPE during the month of November and December
2014 in the Post-monsoon. The entire area was divided into 12 stations and sampling was
done at each station.

Cruise No. 338
This Cruise was conducted in between latitude 08°N to 13° N and longitude 73°E
to 76°E from Off Kochi to Goa during the period of 20 days in the month of April 2014
in the summer season. 8 sampling stations were done in the region.
Map showing sampling stations with reference of table 1 and 2

Table – 1: The inventory of scientific fishing operations of Cr.No. 322, 332 &338
Sl Cruise Date Name of the Latitude longitude Depth Gear CPUE
N No coast (m) (Kg/hr)
o
1 07.01.14 Ponnani 10.09.956 75.38.965 200 HSDT (CV) 22.56165
2 08.01.14 Calicut 11.04.195 74.55.430 1000 HSDT (CV) 147.6015
3 09.01.14 Kannur 11.58.355 74.16.791 1000 HSDT (CV) 109.965
4 09.01.14 Kannur 11.57.317 74.26.081 200 HSDT (CV) 1675.554
5 322 11.01.14 Kollam 8.59.618 75.55.468 200 HSDT (CV) 94.165
6 11.01.14 Kollam 8.53.593 75.27.288 1000 HSDT (CV) 440.755
7 12.01.14 Trivandrum 8.05.718 76.25.842 1000 HSDT (CV) 190.75
8 13.01.14 Trivandrum 8.21.75 76.29.800 200 HSDT (CV) 159.815
9 15.01.14 Cape 8.28.994 78.35.583 200 HSDT (CV) 165.6
10 15.01.14 Tuticorin 8.29.994 78.35.583 1000 HSDT (CV) 79.2615
11 16.01.14 Tuticorin 8.38.153 78.29.182 200 HSDT (CV) 6035.5
12 28.11.14 Cochin 9.51.849 75.29.596 950 HSDT (CV) 130.385
13 29.11.14 Cochin 9.40.357 75.39.092 200 HSDT (CV) 4021.69
14 29.11.14 Alappuzha 9.27.982 75.29.847 1000 HSDT (CV) 46.12
15 30.11.14 Alappuzha 9.27.297 75.47.053 250 HSDT (CV) 2290.01
16 332 01.12.14 Kollam 8.54.060 75.53.748 350 HSDT (CV) 221.16
17 02.12.14 Kollam 8.52.909 75.43.140 350 HSDT (CV) 224.21
18 02.12.14 Kollam 8.58.3 75.32.6 400 HSDT (CV) 75.764
19 04.12.14 Cape 7.05.797 77.26.579 250 HSDT (CV) 19.175
20 05.12.14 Cape 7.30.682 77.57.618 300 HOT 71.455
21 06.12.14 Trivandrum 8.26.550 76.25.291 300 HOT 10002.2
22 07.12.14 Trivandrum 8.27.510 76.05.536 1000 HOT 214.5
23 09.12.14 Valappadu 10.28.137 75.24.567 350 HOT 11.825
24 11.04.15 Azheekal 9.06.705 75.47.699 350 HOT 50.239
25 11.04.15 Kollam 8.50.275 75.59.370 300 HOT 5.5203
26 20.04.15 Ponnani 10.56.501 75.07.779 350 HOT 2.504
27 338 21.04.15 Ponnani 10.58.408 75.06.951 300 HOT 19.204
28 21.04.15 Calicut 11.19.613 74.50.931 200 HOT 324.7
29 22.04.15 Kannur 12.08.467 74.21.067 250 HOT 1419.6
30 22.04.15 Kazargodu 12.27.893 74.13.834 250 HOT 945.618
31 23.04.15 Mangalore 12.57.550 73.53.519 330 HOT 8.567
30112.78
Table – 2: Details of the FORV Sagar Sampada cruises undertaken for deep-sea fishery
resources investigation during study period 2014-15
S.No. Cruise Period Latitude Longitude Area Season No. of
No. stations
1 322 06.01.14 to 08°N to 11°N 74°E to 78°E Off Kochi to Post-Monsoon 11
20.01.14 Tuticorin
2 332 24.11.14 to 07° N to 10°N 75°E to 77°E Off Kochi to Post-monsoon 12
10.12.14 CAPE
3 338 10.04.15 to 08°N to 13° N 73°E to 76°E Off Kochi to Summer 8
29.04.15 Goa
Total stations 31
Analysis of Heavy metals
Reagents, chemicals and glasswares
AnalaR grade acids and reagents were used. All glasswares/plastic bottles were
washed first with water and detergent and further cleaned by soaking in 5% nitric acid for
24hr and finally rinsed with deionised water. Metal standards were prepared using pure
metals or AnalaR SOH salts.
Methods
Preparation of tissue homogenate
The fish samples were washed with potable water and the length and weight were
measured. The muscle tissue was peeled off and homogenized in a blender. The samples
were then dissected and muscle, gills and liver tissues were separately collected and kept.
They were also homogenized in a mixer. An aliquot of the homogenate was pressed
within filter paper folds so as to remove water. This was then used for digestion and
subsequent analysis. For analysis of trace metals in the whole tissue, the fish samples
were weighed and digested whole until it became clear and aliquots taken for metal
analysis.
Heavy metal standard solutions
Standard solutions
Heavy metal standards were prepared using either pure metals or AnalaR BDH
salts. In a few cases metal standards were procured from Sigma Chemicals Co. (St. Luis,
USA). A series of standard metal solutions in the optimum concentration range were
prepared by appropriate dilutions of stock metal solutions with water containing 1.5 mL
HN03/L. Stock standard solutions for Cu, Zn, Fe, Co, Cd, Pb, Cr, Se and As were
prepared from metals or metal salts as follows.
Copper Dissolve 0.1g pure copper metal in 2 ml Conc. HN03. Added 10.0 ml con.
HN03 and diluted to 1000 ml with water. 1.00 ml = 100 µg/mL Cu.
Zinc Dissolve 1gm Zn powder in 20 ml HCI (1:1) in 1L volumetric flask and

diluted to volume with deionised water. Concentration of the solution is
1000 ppm.
Iron Dissolve 0.1g iron wire in a mixture of 10 ml 1+1 HCI and 3 ml con.
HNO3. Added 5 ml conc. HNO3 and diluted to 1000 ml with water. 1.0
ml =100 μg Fe.
Cobalt Dissolve 1 gm of metal in a minimum volume of 1:1 nitric acid and dilute
to 1 litre to give 1000µg/mL Co.
Cadmium Dissolve 1.000 gm of cadmium in a minimum volume of 1:1 nitric acid.

Dilute to 1 litre to give 1000µg/mL Cd.
Lead Dissolve 0.1598 gm AnalaR grade lead nitrate, Pb (N03)2 in minimum
quantity of 1+1 HN03 added 10 ml concentrated HN03 and diluted to 1000
ml with water. 1 ml of the solution=100g µg Pb.
Chromium Dissolve 1 gm of chromium metal in 1:1 hydrochloric acid with gentle

heating. Cool and dilute to 1 litre to give 1000 µg/mL Cr.
Selenium Dissolve 1 g of selenium metal in 80 mL of 1:1 nitric acid, heat gentle to

initiate the reaction-cool if the reaction proceeds too vigorously. Cool and
dilute to 1 litre to give 1000 µg/mL Se.
Arsenic Dissolve 1.3203 gm As2O3 in a minimum volume of 20% NaOH and

neutralize with nitric acid. Dilute to 1 litre to give 1000 µg/mL As.
A series of standard metal solutions in the optimum concentration from stock
solution: The optimum concentration ranges are
Copper 1.0 ppm 2.0 ppm 4.0 ppm
Zinc 0.5 ppm 1.0 ppm 2.0 ppm
Iron 1.25 ppm 2.50 ppm 5.00 ppm
Cobalt 5.0 ppm 10.0 ppm 20.0 ppm
Cadmium 0.25 ppm 0.50 ppm 1.0 ppm
Lead 2.5 ppm 5.0 ppm 10.0 ppm
Chromium 5.0 ppm 10.0 ppm 20.0 ppm
Selenium 25.0 ppb 50.0 ppb 100.0 ppb
Arsenic 25.0 ppb 50.0 ppb 100.0 ppb

The above solutions were prepared by appropriate dilutions of stock metal
solutions with water containing 1.5 ml HNO3/L. Stock standard solutions for Heavy
metals were prepared from AnalaR metals or metal salts as follows. Working standards
for Heavy metals were prepared from commercially available AAS grade stock standard
obtained from Sigma Chemicals Co. and diluted suitably to the required level using
deionised water, acidified with nitric acid.
Glasswares and plastic wares
Sample containers were washed first with water and detergent and further cleaned
by soaking in 5% nitric acid for 24 h and finally rinsed 4-5 times with deionised water
(Milli Q Water) to prevent metal contamination.
Method of Heavy metal analysis
Pre-digestion of samples
The homogenized meat after removing the adhering water by pressing within
filter paper folds, were weighed into a Bethge’s flask and subjected to pre digestion by
adding 7ml of AH concentrated nitric acid plus 3ml of Hydrogen peroxide (H2O2) and
kept overnight. Five replicate samples were taken for analysis.
Digestion procedure for determination of Heavy metals
Heavy metals were determined in edible part by wet oxidation method (AOAC,
2000) using conc. Nitric acid and Hydrogen peroxide in the ratio 7:3 (v/v). The samples
were heated gently and cautiously at first, until the first vigorous reaction subsides,
continued heating, until the organic matter was completely destroyed, indicated by a clear
solution. If any traces of organic matter remained, as indicated by slight yellow colour,
added little 10% hydrogen peroxide and boiled to make clear. This clear solution was
cooled and made up to a known volume after filtration and kept in polythene bottles for
Flame AAS analysis.
A blank was also prepared using Hydrogen peroxide and nitric acid used for
sample digestion. The above samples were directly fed to a Flame Atomic Absorption
Spectrophotometer for heavy metal determination.
Microwave Digestion Method
The microwave-assisted sample digestion technique has become popular since
1980s and presently it is extensively used due to its safe, rapid and efficient performance
(Smith and Arsenault, 1996). It diminishes the risk of external contamination and requires
smaller quantities of acids, thus improving detection limits and the overall accuracy of
the analytical method (Valeria et al., 2003).
An aliquot of samples to be digested were placed in the clean Teflon digestion
vessel with concentrate acid mixture HNO3 : H2O2 = 7:3 (A), v/v, for the microwave acid
digestion procedure. The vessel was closed, placed into the rotor and then increasing the
microwave temperature to 120 ºC over a 15 min period and holding the temperature at
120 ºC for 15 min. The digestion was carried out with the following program (Table 3).
The vessels were cooled and carefully opened. After the digestion process, each digest
was transferred quantitatively with ultra-pure water to a 100 mL volumetric flask. These
solutions were analyzed by F-AAS (Flame-Atomic Absorption Spectrophotometry) after
corresponding dilution using external calibration standard. The standard reference
materials was digested under the same conditions.

Table-3 Digestion Programme of sample using Microwave digester
Step Power (W) Power (%) Ramp (min) Pressure (PSI) Temperature (ºC) Hold (min)
1 800 100 15 800 120 15
Determination of Heavy metals by Flame Atomic Absorption Spectrophotometer (AAS)
Heavy metals such as Cu, Zn, Fe, Co, Cd, Pb, Cr, Se and As were determined in
tissue samples by wet oxidation method using Flame Atomic Absorption
Spectrophotometer (Model GBC 902). The element to be analysed is introduced into a
flame where it becomes dissociated from its chemical bonds into an unexcited, unionized
ground state as individual atoms. The element at this state is capable of absorbing
radiation at discrete lines of narrow wavelength. When a radiation at one of this
wavelength is directed through the flame, the amount of this light absorbed as it passes
through the flame is proportional to the concentration of element being analysed. The
source of radiation is the respective hollow metal cathode lamp of the metal under
determination. The parameters of the instrument were given in Table 4.
Interferences
Interference from citric acid has been reported to suppress the absorbance by upto
50% for a citric acid level of 200μg/ml. The effect is not overcome by adjustment of
flame stiochiometry. The interference has been minimized by measuring the absorbance
in the presence of phosphoric acid. It is necessary to select an optimum burner height to
gain maximum freedom from interference. There is also some evidence that high sulphate
concentrations have a slightly depressive effect on iron determination. The use of a
nitrous oxide – acetylene flame has been found to remove all interference.
Recommended instrument parameters
Table 4. Working condition for analysis by Atomic Absorption Spectrophotometer
Heavy Wave length Lamp current Flame type Slit width Sensitivity Flame
metals (nm) (Support-Gas) (nm) (μg g-1) stiochiometry
Copper 324.7 4 mA Air-Acetylene 0.5 0.025 Oxidizing
Zinc 213.9 5 mA Air-Acetylene 1.0 0.008 Oxidizing
Iron 248.3 5 mA Air-Acetylene 0.2 0.05 Oxidizing
Cobalt 240.7 7 mA Air-Acetylene 0.2 0.07 Oxidizing
Cadmium 228.8 4 mA Air-Acetylene 0.5 0.009 Oxidizing
Lead 217.0 5 mA Air-Acetylene 1.0 0.06 Oxidizing
Chromium 425.4 7 mA Air-Acetylene 0.2 0.017 Reducing
Selenium 196.0 10 mA Air-Acetylene 1.0 0.02 Highly Reducing
Arsenic 189.0 10 mA Air-Acetylene 1.0 0.02 Reducing
Formula: Concentration-Blank × 100

Weight of the sample
STATISTICAL ANALYSIS
All the obtained resulting data collected in this study were presented as mean ±
standard deviation (SD) and analyzed by one-way ANOVA analysis using SPSS
(Scientific Package of Social Science) version 14.0 for windows, Microcal Origin pro 8
multiple range tests were used to resolve differences among treatment means of the heavy
metals like Copper, Zinc, Iron, Cobalt, Cadmium, Lead, Chromium, Selenium and
Arsenic contents of the ten deeper water fish species namely Alepocephalus bicolor,
Narcetes erimelas, Talismania longifilis, Chlorophthalmus bicornis, Lamprogrammus
niger, Beryx splendens, Chelidoperca investigatoris, Neoepinnula orientalis, Cubiceps
baxteri and Psenopsis cyanea. Correlation coefficients ‘r’ was significant between
P<0.01 and P<0.05 was used to indicate significant difference.

RESULTS
As per the best of our knowledge, this is the first report on the heavy metal
accumulation in deep sea fishes in Indian EEZ (Exclusive Economic Zone).
Concentration of heavy metals like Cu, Zn, Fe, Co, Cd, Pb, Cr, Se and As found in the
whole soft tissues of deep-sea fishes viz., A bicolor, N. erimelas, T. longifilis, C. bicornis,
L. niger, B. splendens, C. investigatoris, N. orientalis, C. baxteri and P. cyanea. The
results of heavy metal analysis indicated that its accumulation varied among the deep-sea
fin fish species. Fishes were collected from the west coast waters contained significantly
higher Fe, Zn and Cu than the other metals like Co, Cd, Pb, Cr, Se and As. Among all the
heavy metals, accumulation of Fe was the highest content followed by Zn and Cu in
overall metal accumulation and selenium was in below detectable level.
The mean concentrations of all heavy metals accumulated in the given three
cruises 322, 332 and 338. Heavy metals like copper and chromium concentration
accumulated more in cruise 322, zinc and lead content found more in cruise 332, whereas
iron, cobalt, cadmium and arsenic contents accumulated more in cruise 338, and selenium
is the metal which was observed in below detectable level in all the three cruises (Table
14) respectively. The order of metal accumulation in cruise 322 is
Fe>Zn>Cu>Cr>Co>As>Cd>Pb>Se, whereas the order of metal magnitude in cruise 332
is Fe>Zn>Cu>Co>Cr>Cd>Pb>As>Se and the order of metal content in cruise 338 is
Fe>Zn>Cu>Co>Cr>Cd>As>Pb>Se. The overall cruise wise metal concentration was
accumulated more in cruise 338 followed by 322 and 332 and the overall study
determines the order of metal accumulation is Fe>Zn>Cu>Co>Cr>Cd>As>Pb>Se, which
has shown in Table 15.

Copper content accumulated more in N orientalis (26.58±1.31) followed by C
investigatoris (23.76±0.69) and C bicornis (23.67±0.77) of cruise 322, whereas in cruise
332, copper concentration found more in N orientalis (25.49±0.42) followed by P.
cyanea (24.96±1.13) and T longifilis (24.67±0.16), while copper found high in P cyanea
(26.49±1.06) followed by N orientalis (24.65±1.19) and T longifilis (22.47±1.32) which
has shown in Table 5.
Table 6 represents the zinc content found more concentration in T longifilis
(28.41±0.82) followed by B splendens (28.31±0.32) and A bicolor (24.81±0.91) in cruise
322, while in cruise 332, zinc metal accumulated high concentration in T longifilis
(32.06±0.42) followed by N erimelas (25.71±0.06) and B splendens (24.16±0.08),
whereas in cruise 338, this metal was found highest concentration in B splendens
(31.67±1.03) and lowest concentration in C. baxteri (17.92±0.25). Iron concentration
observed more in C. baxteri (50.81±2.28) and less in L. niger (21.94±0.87) in the cruise
of 322, whereas in cruise 332, iron accumulated highest concentration in C. baxteri
(43.35±1.37) and lowest concentration in C. investigatoris (29.81±0.15), and in cruise
338, iron content observed more in B. splendens (53.61±0.74) and less concentration
accumulated in L. niger (26.43±0.28) shown in Table 7.
Cobalt had highest concentration in C. bicornis (3.05±0.06) and lowest
concentration in C. investigatoris (0.60±0.30), but this metal was in below detectable
level in B. splendens and C. baxteri in the cruise 322, accordingly cobalt found more
content in T. longifilis (2.68±0.12) and less content in C. baxteri (0.42±0.05) and this
metal was in below detectable level in C. investigatoris and N. orientalis in the cruise
332, whereas in cruise 338, cobalt found more in A. bicolor (3.16±0.06) and less in C.
baxteri (0.36±0.03) represents in Table 8. Cadmium is the toxic metal and it was
accumulated only in L. niger (0.03±0.01) and C. baxteri (0.01±0.01) and in remaining
species, this metal observed in below detectable levels in the cruise 322, correspondingly
cadmium found in a few species like N. erimelas (0.86±0.03), T. longifilis (0.35±0.01), L.
niger (0.12±0.02), C. investigatoris (0.21±0.03) and N. orientalis (0.06±0.01) and this
metal was observed in below detectable level in remaining species like A. bicolor, C.
bicornis, B. splendens, C. baxterii and P. cyanea in the cruise 332, accordingly in cruise
338, cadmium was found in some species like N. erimelas (1.05±0.02), L. niger
(0.08±0.01), C. investigatoris (0.49±0.03), C. baxterii (0.07±0.02) and P. cyanea
(1.43±0.04) and in remaining species like A. bicolor, T. longifilis, C. bicornis, B.
splendens and N. orientalis, it was in below detectable level which has shown in Table 9.
Lead is a heavy metal which was observed in below detectable levels in all the
given ten deep-sea fishes in the cruise 322 (Table 10) whereas in cruise 332, this metal
was found in a few species like T. longifilis (0.13±0.02), C. investigatoris (0.07±0.01)
and C. baxterii (0.19±0.03) and it was found in below detectable level in the remaining
species, while in cruise 338 lead accumulated only in three deep-sea fishes viz., L. niger
(0.06±0.02), C. baxterii (0.09±0.02) and P. cyanea (0.05±0.01) and in remaining species
like A. bicolor, N. erimelas, T. longifilis, C. bicornis, B. splendens, C. investigatoris and
N. orientalis, it was found in below detectable levels. Chromium is the metal
accumulated more in B. splendens (3.94±0.24) followed by N. orientalis (2.53±0.43) and
L. niger (1.54±0.09) in the cruise 322, accordingly in the cruise 332, chromium was
found more in B. splendens (1.65±0.13) followed by P. cyanea (1.57±0.27) and C.
baxterii (1.29±0.11) and this metal was observed in below detectable level in two species
viz., C. bicornis and C. investigatoris, whereas in cruise 338 chromium was found more
in N. orientalis (1.97±0.54) and less concentration was found in C. bicornis (0.53±0.11)
and in species like A. bicolor, N. erimelas, T. longifilis and C. investigatoris, chromium
was observed in below detectable levels which has been represented in Table 11.
Table 12 showed that the selenium was found in below detectable levels in all the
ten deep-sea fish species and in all the three cruises 322, 332 and 338 respectively. Table
13 represented that Arsenic, the heavy metal found in L. niger (0.05±0.02), C. baxterii
(0.08±0.03) and P. cyanea (0.11±0.03) and it was in below detectable level in the
remaining seven species in the cruise 322, correspondingly in the cruise 332, this metal
was accumulated only in few species like C. bicornis (0.07±0.02), L. niger (0.10±0.01),
C. baxterii (0.08±0.018) and P. cyanea (0.14±0.02) and in remaining species it was
observed in below detectable levels, while in cruise 338, this metal found in four species
viz., T. longifilis (0.03±0.002), L. niger (0.08±0.003), C. baxterii (0.15±0.01) and P.
cyanea (0.08±0.008) and in a few species like A. bicolor, N. erimelas, C. bicornis, B.
splendens, C. investigatoris and N. orientalis, it was found in below detectable levels
respectively.
Table 5. Copper content in ten deep-sea fishes of three cruises
1 A. bicolor 19.03±0.95 21.53±0.35 18.62±1.09
2 N. erimelas 17.46±0.54 19.86±0.29 15.31±0.64
3 T. longifilis 19.32±1.82 24.67±0.16 22.47±1.32
4 C. bicornis 23.67±0.77 18.72±0.05 20.49±0.62
5 L. niger 11.25±0.34 12.87±0.09 13.98±0.19
6 B. splendens 15.54±0.69 18.55±0.34 16.53±0.53
7 C. investigatoris 23.76±0.69 20.06±0.61 19.96±1.01
8 N. orientalis 26.58±1.31 25.49±0.42 24.65±1.19
9 C. baxterii 13.18±0.57 17.50±1.05 16.05±0.83
10 P. cyanea 23.46±0.6 24.96±1.13 26.49±1.06

Note: n=5; mean±SD
Comparison of total mean value of copper accumulation in three cruises
24
Copper Conc.
16
Cr322 Cr332 Cr338

Table 6. Zinc content in ten deep-sea fishes of three cruises
1 A. bicolor 24.81±0.91 22.19±0.35 26.83±0.24
2 N. erimelas 23.29±0.53 25.71±0.06 21.54±0.16
3 T. longifilis 28.41±0.82 32.06±0.42 25.99±0.73
4 C. bicornis 19.37±1.06 24.13±0.15 18.37±0.82
5 L. niger 16.07±1.3 21.37±0.61 19.43±0.95
6 B. splendens 28.31±0.32 24.16±0.08 31.67±1.03
8 N. orientalis 14.75±0.59 20.42±0.34 16.35±0.36
9 C. baxterii 20.60±0.78 19.57±0.59 17.92±0.25
10 P. cyanea 16.80±1.33 23.16±0.67 18.44±0.53

Note: n=5; mean±SD
Comparison of total mean value of Zinc accumulation in three cruises

40
30
Zinc Conc.
20
10
Cr322 Cr332 Cr338
Table 7. Iron content in ten deep-sea fishes of three cruises
1 A. bicolor 30.88±2.4 33.16±0.95 35.42±1.34
2 N. erimelas 36.36±1.23 31.05±1.03 39.16±0.95
3 T. longifilis 31.29±0.98 35.82±1.14 34.68±0.36
4 C. bicornis 33.80±1.21 30.43±0.29 37.86±0.49
5 L. niger 21.94±0.87 29.75±0.06 26.43±0.28
6 B. splendens 49.80±0.84 38.94±0.82 53.61±0.74
8 N. orientalis 33.57±0.89 36.91±1.25 31.43±0.26
9 C. baxterii 50.81±2.28 43.35±1.37 46.43±0.58
10 P. cyanea 40.12±1.22 37.29±1.56 36.55±0.62

Note: n=5; mean±SD
60 Comparison of total mean of Iron accumulation in three cruises
40
Iron Conc.
20
Cr322 Cr332 Cr338

Table 8. Cobalt content in ten deep-sea fishes of three cruises
1 A. bicolor 2.87±0.54 1.79±0.06 3.16±0.06
2 N. erimelas 1.19±0.09 0.82±0.03 1.64±0.11
3 T. longifilis 2.34±0.03 2.68±0.12 3.04±0.03
4 C. bicornis 3.05±0.06 2.26±0.24 2.49±0.17
5 L. niger 0.67±0.27 0.85±0.05 1.67±0.1
6 B. splendens BDL 0.94±0.07 0.88±0.06
7 C. investigatoris 0.60±0.30 BDL 1.64±0.21
8 N. orientalis 0.86±0.16 BDL 1.98±0.05
9 C. baxterii BDL 0.42±0.05 0.36±0.03
10 P. cyanea 0.63±0.08 2.54±0.03 2.53±0.07

Note: n=5; mean±SD
Comparison of total mean value of cobalt accumulation in three cruises

4
2
Cobalt Conc.
Cr322 Cr332 Cr338

Table 9. Cadmium content in ten deep-sea fishes of three cruises
1 A. bicolor BDL BDL BDL
2 N. erimelas BDL 0.86±0.03 1.05±0.02
3 T. longifilis BDL 0.35±0.01 BDL
4 C. bicornis BDL BDL BDL
5 L. niger 0.03±0.01 0.12±0.02 0.08±0.01
6 B. splendens BDL BDL BDL
7 C. investigatoris BDL 0.21±0.03 0.49±0.03
8 N. orientalis BDL 0.06±0.01 BDL
9 C. baxterii 0.01±0.01 BDL 0.07±0.02
10 P. cyanea BDL BDL 1.43±0.04

Note: n=5; mean±SD
Comparison of total mean value of Cadmium accumulation in three cruises
1.6
Cadmium Conc.
0.8
0.0
Cr322 Cr332 Cr338

Table 10. Lead content in ten deep-sea fishes of three cruises
2 N. erimelas BDL BDL BDL
3 T. longifilis BDL 0.13±0.02 BDL
5 L. niger BDL BDL 0.06±0.02
7 C. investigatoris BDL 0.07±0.01 BDL
8 N. orientalis BDL BDL BDL
9 C. baxterii BDL 0.19±0.03 0.09±0.02
10 P. cyanea BDL BDL 0.05±0.01

Note: n=5; mean±SD
Comparison of total mean value of Lead accumulation in three cruises
0.2
Lead Conc.
0.1
0.0
Cr322 Cr332 Cr338

Table 11. Chromium content in ten deep-sea fishes of three cruises
1 A. bicolor 0.14±0.02 0.08±0.03 BDL
2 N. erimelas 1.06±0.05 0.15±0.01 BDL
3 T. longifilis 0.29±0.03 0.24±0.09 BDL
4 C. bicornis 1.38±0.12 BDL 0.53±0.11
5 L. niger 1.54±0.09 0.27±0.05 0.76±0.09
6 B. splendens 3.94±0.24 1.65±0.13 1.63±0.14
7 C. investigatoris 0.78±0.10 BDL BDL
8 N. orientalis 2.53±0.43 0.32±0.05 1.97±0.54
9 C. baxterii 0.68±0.24 1.29±0.11 1.14±0.27
10 P. cyanea 0.81±0.39 1.57±0.27 1.26±0.19

Note: n=5; mean±SD
Comparison of total mean value of Chromium accumulation in three cruises
4
Chromium Conc.
Cr322 Cr332 Cr338

Table 12. Selenium content in ten deep-sea fishes of three cruises
3 T. longifilis BDL BDL BDL
5 L. niger BDL BDL BDL
7 C. investigatoris BDL BDL BDL
9 C. baxterii BDL BDL BDL
10 P. cyanea BDL BDL BDL

Note: n=5; mean±SD
Comparison of total mean value of Selenium acumulation in three cruises
2
Selenium Conc.
-2
Cr322 Cr332 Cr338

Table 13. Arsenic content in ten deep-sea fishes of three cruises
3 T. longifilis BDL BDL 0.03±0.002
4 C. bicornis BDL 0.07±0.02 BDL
5 L. niger 0.05±0.02 0.10±0.01 0.08±0.003
7 C. investigatoris BDL BDL BDL
9 C. baxterii 0.08±0.03 0.08±0.018 0.15±0.01
10 P. cyanea 0.11±0.03 0.14±0.02 0.08±0.008

Note: n=5; mean±SD
Comparison of total mean value of Arsenic accumulation in three cruises
0.16
Arsenic Conc.
0.08
0.00
Cr322 Cr332 Cr338

Table 14: Total mean concentrations of Heavy metal accumulation in three
Cruises along the Indian EEZ
Metals Cruise 322 Cruise 332 Cruise 338 Total mean conc.
Cu 26.58 25.49 26.49 26.19
Zn 28.41 32.06 31.67 30.71
Fe 50.81 43.35 53.61 49.26
Co 3.05 2.68 3.16 2.96
Cd 0.03 0.86 1.43 0.77
Pb 0.00 0.19 0.09 0.09
Cr 3.94 1.65 1.97 2.52
Se 0.00 0.00 0.00 0
As 0.11 0.14 0.15 0.13
Table 15. Order of metal concentration in cruise wise and total

mean concentration
Cruise 322 Fe>Zn>Cu>Cr>Co>As>Cd>Pb>Se
Cruise 332 Fe>Zn>Cu>Co>Cr>Cd>Pb>As>Se
Cruise 338 Fe>Zn>Cu>Co>Cr>Cd>As>Pb>Se
Total mean conc. Fe>Zn>Cu>Co>Cr>Cd>As>Pb>Se

Figure 1a. Heavy metal determination in the given ten deep-sea fish samples of Cruise 322, 332 and 338
Figure 1b. Heavy metal determination in the given ten deep-sea fish samples of Cruise 322, 332 and 338
DISCUSSION
Many outbreaks of food poisoning due to marine products containing excess
amounts of toxic metals like cadmium, lead, Cobalt Arsenic etc. have caused increasing
concern among scientists and public health authorities about the quantities of these heavy
metals in fish and other deep-sea fishes. Awareness of heavy metal concentrations in
fishes is essential in terms of management and human consumption (Rauf et al., 2009).
Anthropogenic activities constantly enhance the amount of heavy metals in environment,
especially in aquatic ecosystems. Heavy metal pollution in aquatic ecosystems is
increasing at an alarming rate worldwide due to human activities (Malik et al., 2010).
Researchers have showed that fishes accumulate heavy metals in their tissues and their
concentrations depend on many factors such as concentration and duration of exposure,
salinity, temperature and hardness of water and metabolic rate of organisms (Allen,
1995). In polluted waters, heavy metals accumulate in organisms directly through skin
and gill or indirectly via food chains (Sinha et al., 2002; Sure, 2003). Heavy metals have
toxic effects, altering physiological activities and biochemical parameters both in tissues
and blood (Nemesok and Huphes, 1988). Metals such as iron, copper, zinc etc are
essential due to their vital role in biological systems, whereas, lead, cadmium, arsenic
etc., are non-essential and toxic. The essential metals can be toxic when their
concentration in aquatic environments increases beyond the tolerable level of organisms.
The studies concerning the metals in deep-sea edible fishes from the Indian coast
are limited. Imam Khasim et al. (1993) analysed the levels of toxic metals in the tissues
of some fishes caught during the 56th cruise of FORV (Fisheries Oceanographic
Research Vessel) Sagar Sampada in the Andaman Sea. Prafulla (2002) studied the heavy
metal concentration in deep-sea fisheries from the Indian coast. Heavy metal
concentrations in some common edible fishes from Bombay and upper east coast have
been reported by Khot et al. (1985) and Prasad et al. (1990). Lakshmanan (2003) studied
the level of Cadmium in Seafood of Kerala coast. Trace metals are added to the marine
environment from both natural sources and as a result of human activity. Runoff from
land is a major source of addition of trace elements to the sea. Rivers bring in the trace
metals both as dissolved species and as adsorbed onto the suspended matter,
concentration of which may vary with the nature of rocks in catchment areas. Geologic
weathering of rocks ultimately produces the clay and other minerals that make up the
bulk of detrital sediments as well as dissolved metals in seawater. Volcanic activity either
on land or in the sea is a natural source of metals, as is diagenetic remobilization from
sediment. Many different human activities can add metals to the marine environment, like
mining, metal processing manufacturing, transportation, waste disposal etc. From, the
numerous sources of trace metals to the marine environment, it is difficult to determine
which possible source is most important. It is important to identify the source of metals
but the environmental impact of a metal depends on its behavior rather than its source.
The behaviors including mobility, transport, transfer and biological uptake, depend on the
chemical and physical form of the metal. The size of the metal species or the particle with
which it is associated is critical, as this will control its transport and settling. A given
metal will behave different as physically, chemically and biologically in each of its forms
and it will partition itself among the various possible forms in response to environmental
conditions. In this respect, it is important to note that many trace metals are reactive and
will quickly associate with particles if added to the marine environment in a dissolved
form. Data on heavy metals are very scanty for the entire Indian EEZ. The present work
is hence the first of its kind focusing on the processes affecting the temporal and spatial
distributions of heavy metals from this region. The significance of this study is in
describing the monitoring of heavy metal accumulation in deeper water fishes at the
depth of 100m – 1200 m in Indian Exclusive Economic Zone.
Copper
Copper is an essential micronutrient in living system. The principal species of Cu
(II) in seawater are hydrated Cu2+ ion, CuCO3 and Cu (OH) +. In contrast to the role of
copper as a micronutrient, the cupric ion is potentially toxic to phytoplankton at sub-
nanomolar concentrations. The mechanism of toxicity often involves competition by
copper for active sites occupied by essential metals (Sunda and Guillard, 1976). Copper
is an essential element in human nutrition and the daily dietary requirement is 2-3 mg for
an adult of 23-50 years (Anon, 1980, Gopalan et al., 1989). It has been reported by
Mckee and Wolf (1963) that Copper is toxic to man in quantities above 100 mg.
However, copper poisoning as a result of eating copper containing marine organisms is
unlikely because their taste at these levels renders them unpalatable (Imam Khasim,
1993). Human taste threshold for copper is about 5-7 ppm (Portman, 1970). The fishes
studied here form safe sources of copper in the diet. Copper is one of the most toxic
metals to fishes in higher concentrations. Copper residue in fish muscle tissue is generally
low. Ashraf and Jaffer (1998) working on 6 species from the Arabian Sea recorded
muscle levels of only 0.10-0.51 mg/kg weight. In the three species of fish analysed,
concentration of Cu in the muscle tissue ranged from 0.23 ppm in Epinephelus
diacanthus ~ to 3.5 ppm in Saurida tumbil. Gill and liver tissues accumulated more Cu
than the muscle tissue. The concentration in liver of Saurida tumbil was the highest (88
ppm), followed by that in Nemipterus japonicus (35.8 ppm). Cu concentration in the gills
was lower than that for the muscle and liver. The recommended daily allowance of Cu is
1mg. Copper has a strong tendency to form complexes with proteins and nucleic acids
and, can render these macromolecules non-functional through complex formation.
Reduced growth rates have been observed in many marine phytoplankton at cupric ion
concentrations above 10-11 M (Sunda and Hanson 1987). Copper, therefore, can play a
dual role in limiting plankton growth through nutritional deficiency or toxicity.
In near bottom waters, copper may be remobilized by interactions with sediments
(Kramer, 1982). Decomposition of partly reduced organic-rich sediments release copper
into the water column (Kremling, 1983). Enrichment of copper in bottom waters of shelf
regions in the present study may partly be due to its release from the organic-rich
sediments and partly due to its regeneration from the organism. Boyle et al., (1977) while
studying the distribution of dissolved copper in the Pacific observed a decrease from
surface maxima to top of the thermo cline, followed by an increase in bottom waters.
According to them, this unique distribution was maintained by Aeolian input to the
surface waters comparable in magnitude to the fluvial component, ubiquitous scavenging
in the surface and deep waters, and a strong bottom source. On the other hand, Bruland
(1980) observed a continuous increase in the concentration of copper from surface to
600m depth, uniform distribution between 600-1500m followed by a gradual increase up
to the bottom in the north Pacific. Though the distribution of copper in intermediate and
deep waters in both the reports were attributed to its scavenging coupled with strong
bottom source, its enrichment in surface contrary to depletion reported in the latter was
presumed to be a transient feature resulting from the advection of copper-rich near shore
surface waters into the central regions of the Pacific Ocean.
Copper (Cu) was recorded in all of the tissue samples from this study. A similar
mean values were reported by Sankar et al. (2006) in different fish species from the west
coast of India. The observed values of Cu in fish tissues were higher than those observed
by Raja et al. (2009) in fish collected from Parangipettai water, India, and Stange et al.
(1995) in Karp Farvel, Greenland and also higher than Kalay et al. (1999) reported in the
Mediterranean Sea. The mean Cu concentrations observed in commercially important
marine fish species in this study did not exceed the acceptable limit 30μg g−1 (FAO,
2008) and also suggested by the National Research Council, China and maximum Cu
content (30 mg/kg) set by FAO (1989). It is well known that Cu and Manganese are
important elements in fish, play a vital role in enzymatic processes and are essential for
the synthesis of hemoglobin. However, very high intake will cause adverse health
problems (Demirezen and Uruc, 2006; Satheeshkumar et al., 2011). Vas et al. (1993)
reported a mean copper concentration of 0.53 mg/kg in C. rupestris caught west of
Scotland, which is considerable lower than the values found in the current study, but very
low concentrations (<0.02 mg/kg) of copper in N. armatus from Rockall Trough.
Windom et al. (1987) analysed N. armatus from the Atlantic and Pacific Oceans and
reported mean copper concentrations of 0.74 and 1.09 mg/kg (on a dry weight basis),
which are less than the median value of the present study.
Zinc
Zinc is an essential trace element required by all living organisms because of its
critical role both as a structural component of proteins and as a cofactor in enzyme

catalysis (Leigh Ackland and Michalczyk 2006). Zinc is involved in numerous protein
functions such as the carbonic anhydrase and is efficiently absorbed and strongly retained
in Sepia officinalis both from the food and seawater pathways (Villanueva and
Bustamante 2006). Zn is known to be incorporated into diatom frustules (Ellwood and
Hunter, 1999) and hence covaries with Si (Bruland, 1989). Zn is also essential for various
metabolic functions including carbon fixation via carbonic anhydrase (Sunda and
Huntsman, 1992). The speciation of Zn is very sensitive to pH changes. At pH 8.2, the
principal species were Zn (OH)+, hydrated Zn2+ion and ZnC03. The distribution pattern
of Zn among the body parts clearly indicated that liver in the ten deep-sea fish species
form the major site of Zn accumulation. In general the edible muscle show relatively low
levels of Zn. Zn in the fish samples accumulated in the order Saurida tumbil >
Epinephelus diacanthus> Nemipterus japonicus. The high concentration of zinc in the
muscle of the fish samples is mainly due to the presence of a large number of fishing
vessels and trawlers which use galvanized metal coatings to prevent rusting, that
ultimately find its way into the ambient media through leaching.
Zinc was found to be the second most abundant metal in the fish species sampled
in this study. The mean value for tissue samples examined in this study was ranged from
14.75±0.59 to 28.41±0.82 in cruise 322, whereas in cruise 332, the mean values were
occurred in between 19.43±0.29 and 32.06±0.42, while in cruise 338, the mean values of
zinc varies in between 16.35±0.36 and 31.67±1.03 µg/g. Similar mean values were
reported by Raja et al. (2009) (14.1–33.5 µg/g) in Parangipettai waters, India, and Kalay
et al. (1999) (14.1–33.5 µg/g) in fish species caught from the Mediterranean Sea. Sankar
et al. (2006) reported Zn concentrations of 6µg/g in marine fish caught in Kochi waters.
The mean Zn concentration from fish samples in this study was lower than other studies
in international waters, including Kojadinovic et al. (2007) in Mozambique Channel
(41.7 µg/g), Topcuoglu et al. (2002) in the Black Sea (44.2 mg/kg) and Kumar et al.
(2011) in the Kolkata wetlands (48 µg/g). However, Kwon and Lee (1999) in Masan Bay,
Korea recorded values ranging from 6.33 to 12.9 µg/g, which is lower than the values
observed in this study. The acceptable limit of Zn in most fish is 5 µg/g or 25 mg/kg for
certain types of seafood, as suggested by the New Zealand Food Standards Code
(FSANZ, 2004). The European Union’s permissible level of Zn for human consumption
is 30 mg/kg wet weight (EU 2008). Zinc is one of the most important trace elements for
the human body because it is a key component of cells, and enzymes depend upon it as a
cofactor.
Iron
Iron is a fundamental element for many biological processes occurring in living
organisms. One of them is the interference to the thyroid metabolism and to different
enzymes in mammals and fish (Rejitha and Subhash Peter 2013). Iron is required by all
eukaryotes and most prokaryotes because it is involved in enzymatic reactions such as
oxygen metabolism, electron transfer processes, and DNA and RNA synthesis (Knaan-
Shanzer et al., 1996). The concentration of iron in unpolluted oceanic seawater was found
to range from 1.9–2.8 ng L–1 to 224–1,228 ng L–1 and higher concentrations could be
found in estuarine water (Whitehouse et al. 1998). The mean concentration of iron was
20.8 μg/g, which was found in coastal salt marshes. Ergul et al. (2010) found an iron
concentration of 0.033 ng g–1 and 25.1 μg g–1 in sea water and sediment samples,
respectively. Iron is transported to the ocean via three major pathways: fluvial (riverine)
input, atmospheric deposition, and processes occurring on the sea floor such as
hydrothermal vents, sediment resuspension, and diagenesis. Iron is initially mobilized
from the lithosphere by either mechanical action (i.e. erosion) or by thermal and chemical
reactions (i.e., leaching, anoxia, or geothermal activity). Once mobilized, several
physicochemical processes occur, altering its solubility and chemical speciation, between
the source and ocean interfaces. The iron cycling can be studied by determining fluxes of
iron in the different phases of the water column and its export from the sea. The
distribution of iron in offshore areas was not completely nutrient-like or scavenged type,
even though it exhibited a slight increase at higher depths. The deepwater concentration
has provided a picture of the geochemistry of dissolved Fe in the offshore waters of both
Arabian Sea and Bay of Bengal. The slight increasing concentration of iron with depth
suggests that it had feebly undergone a particle remineralization cycle similar to nutrients
such as nitrate (Sunda and Huntsman, 1996).
Iron (Fe) was observed in all samples and was the most abundant trace element in
tissue samples from this study. The highest Fe concentration was observed in C. baxteri
of cruise 322 (50.81±2.28) and cruise 332 (43.35±1.37), whereas in cruise 338, the
maximum concentration of Fe was found in B. splendens (53.61±0.74). High values of Fe
in the samples indicate that the environment is stressed. This study observed values
higher than permissible limits set by the FAO and the WHO. The Fe values are similar to
those reported by Kalay et al. (1999) (59.6–73.4 µg/g) in the Mediterranean Sea, except
J. elongates (240.5 µg/g), C. dussumieri (207.2 µg/g), and A. arius (112.3 µg/g). The iron
concentrations in fish samples from this study were higher than those from studies
conducted in international waters (Yilmaz, 2003; Kojadinovic et al., 2007). The high Fe
concentration observed in J. elongatus is due to their feeding habits and habitat
preference. Fe concentrations increased due to a decrease in grain size and an increased
input of organic matter and anthropogenic metals from industrial pollution. Domestic
sewage waste and hospital waste were discharged directly into the river (Satheeshkumar
and Kumar, 2011). Similar detection was reported by Zhang et al. (2007) in Yangtze
Estuary, China.
Cobalt
Cobalt is an essential component for human and other organisms since it is an
important component of Vitamin B-12. Cobalt has both natural and anthropogenic
sources. Cobalt is generally found in the form of ores. Cobalt is usually not mined alone,
and tends to be produced as a by-product of nickel and copper mining activities. Small
amount of Co are found in most rocks, soil, plants and animals. The main human sources
are coal and oil burning, industrial activities, vehicular exhausts and sewage sludge.
Cobalt is used in steel and alloys, metallurgy, electroplating, nuclear technology,
fertilizers, medicine, drier for paint, foam stabilizer in beer brewing (ATSDR 1999). The
mean concentration of cobalt detected in seawater was 0.3 μg L-1 (Huynh-Ngoc et al.
1989). The concentration of cobalt is comparatively higher in estuarine and coastal
waters than in ocean, the cobalt concentration in surface marine water is 18–300 pM,
whereas it ranged from 20 to 50 pM at depth (Donat and Bruland 1995).
Cobalt was not observed in all the given samples of three cruises. This metal was
in below detectable level in B. splendens and C. baxteri of cruise 322, and in cruise 332,
it was in below detectable level of C. investigatoris and N. orientalis. The maximum

cobalt was observed in C. bicornis (3.05±0.06), T. longifilis (2.68±0.12) and A. bicolor
(3.16±0.06) of cruise 322, 332 and 338 respectively. However, the present study showed
that the results were not similar to sivaperumal et al (2007). The results were similar to
that found by Houserova et al. (2006) and Mansouri et al. (2012b), where the rate of
bioaccumulation in skin tissue was lower than those in the liver and gills. The mucogenic
activity of the skin epithelium in fish is very high compared to gills (for accumulation)
(Paul and Banerjee, 1997). This higher mucogenesis might have a crucial role in
preventing the metal ions from entering the body, because coagulated mucus all over the
body might act as a protective ion trap (Licata et al., 2005), while the gills are the site of
active and passive exchanges between the animal and aquatic environment. The
elimination experiments started after 15 days of absorption. In the present study, the skin
and gills showed the greatest elimination of Co to sublethal concentration. Studies by
Kalay and Canli (2000), and Mansouri et al. (2012b) showed that the fish gill is a more
effective organ for metal elimination than either the liver or muscle. Turkmen et al.
(2005) have investigated Co levels in fish samples and they found lower concentration
than the present study. Turkmen et al. (2005) reported that cobalt concentration varied
between 0.73 and 1.91 mg/kg -1 in the muscles of fishes of the Iskenderun Bay and lower
Co levels in fish muscles have been reported from the Mediterranean Sea region.
Cadmium
Cadmium is not essential for biological function in humans. The kidney is
considered the critical target organ, for both the general population and occupationally
exposed populations. Several studies over the past decade have indicated that renal
tubular damage occurs at lower levels of cadmium body burden than previously
observed. Some studies have suggested that cadmium alters calcium metabolism,
contributing to osteoporosis (Jarup et al., 1998). An increased risk of lung cancer has
been reported following inhalation exposure in occupational settings, but there is no
evidence that cadmium is a carcinogen by the oral route of exposure.
Metals like cadmium is low in juvenile stages but it are accumulated with age.
The size and maturity stage of the fishes influence its accumulation levels. Cd
concentration in the muscle tissue is found to increase with the size of the fishes (Latha et
al., 2003). Concentration of Cd in the muscle ranged from below detection limit to 0.5
ppm. In the case of Cd also liver showed higher concentration. High contents of Cd
(0.213 and 0.402 ppm) were reported in squid and cuttle fish by Lakshmanan (1988) from
west coast of India. Ashajyothy and Vijayalekshmi (1999) reported levels of Cd ranging
from 0.06- 0.4 ppm in the edible muscle of fishes from Thane creek. The range in
concentration obtained in the present study correlate well with this value. A metal like
cadmium is low in juvenile stages but it is accumulated with age. The higher
concentration of Cd in liver compared to gill and muscle shows that the main route of
uptake of cadmium is through food and water. Numerous sub-acute effects have been
reported in both fresh water and marine fishes like decreased growth, morphologic
changes in the gut etc. due to Cd toxicity (Moore and Ramamoorthy, 1984). Reports
show that chronic exposure to Cd in human beings lead to renal toxicity, such as
proteiurea. A tissue residue in excess of 0.285 mg/g usually causes renal dysfunction. The
daily allowed dietary allowance for Cd per kg weight for a man is given by FAO (1975)
as 0.95 mg. The permitted tolerance limit for Cd in fish is 0.2-2 ppm (FAO, 1983). In all
the species studied from this area, the level of Cd in the edible part is below 2 ppm. This
does not pose any threat to human health. Concentration of Cd in the muscle ranged from
below detection limit to 0.36 ppm. In the case of Cd also liver showed maximum
concentration. Many of the fish samples have attained full maturity during the study
period. The higher concentration of Cd in liver compared to gill and muscle shows that
the main route of uptake of cadmium is through food and water. The permitted tolerance
limit for Cd in fish is 0.2-2 ppm (FAO, 1983). In all the species studied from this area the
level of Cd in the edible part is below 2 ppm. This does not pose any threat to human
health.
Cadmium is a serious contaminant, a highly toxic element, which is transported to
sea through air. The limit of Cd for human consumption of fish is approximately 0.5 µg g
-1 dry wt. (CSHPF, 1995). Mean concentrations of cadmium were ranged from 0.01-0.03
µg/g in cruise 322, whereas in cruise 332 the cadmium content was in between 0.06±0.01
and 0.86±0.03, and in cruise 332 the maximum cadmium concentration was observed in
P. cyanea (1.43±0.04) followed by N. erimelas (1.05±0.02). In the present study,
cadmium results were higher than maximum permissible limits recommended by FAO,
(1976). The values which obtained in cruise 338 were higher than the earlier reports
(Ashraf et al., 1991: Tariq et al., 1994 and Vijayakumar et al., 2011) but the results
which obtained from cruise 322 were lower than the permissible limits. Mendil et al.
(2010) have reported 0.11 to 0.75 µg/g cadmium in fish species and it is coincide with the
present study.
Lead
Lead is considered as a non-essential and toxic metal which also implies that it
has no known function in biochemical processes. Lead induces reduced cognitive

development and intellectual performance in children and increased blood pressure and
cardiovascular disease in adults (EC, 2001). Lead constitutes a serious health hazard to
both children and adults. The adverse toxic effect caused by lead on human was
documented by Subramanian (1988). Lead is a toxic element that has no biological role
and causes carcinogenic effects in marine biota and humans (Velusamy et al., 2014).
Radhakrishnan (1993) reported level of lead below 7 ppm in deep sea fish collected
onboard FORV (Fisheries Oceanographic ResearchVessel) Sagar Sampada. The
permitted levels of Pb in fish and fishery products in India are 5 ppm. The edible part of
the fishes studied here contains levels well below this (FAO, 1975).The maximum limit
established by European Union for crustaceans is 0.3 µg g-1 (European Union, 2006). In
the present study, lead content was below detectable level in all the crustacean species,
indicating safety for human consumption. The International Agency for Research on
Cancer (IARC, 1993) classified cadmium as a human carcinogen. According to
Lithuanian Standards of Hygiene, the Maximum Tolerable Limit of lead in fish meat is
0.4mg/kg which is same as the value adopted by the European Commission for lead in
marine fish muscle (EC, 2000) while FAO set a limit of 0.5mg/kg (FAO, 1983).
Concentrations of Lead in Engraulis encrasicholus and Merlangius merlangus fishes
in Black and Aegean Seas were found to be in the range of 0.33 to 0.93 mg/kg.
Similarly Dural et al., (2007) have also reported that the Pb contents were in the
range of 0.40 to 2.44 mg/kg in muscle and 1.41 to 3.95 mg/kg in liver tissues of fish of
Tulza Lagoon. Previous studies have demonstrated that the Pb levels were varied
with different parts of fish such as flesh, guts, and gills etc (Mansour and Sidky., 2002).
In general Pb levels were found in the order of liver> gill>flesh (Mansour and
Sidky., 2002; Kalay et al., 1999) which was comparable to the present study.
Lead is a toxic element that has no biological role and causes carcinogenic effects
in marine biota and humans. The concentration of lead in the present study varied from
0.07±0.01 to 0.19±0.03 of cruise 332 and in cruise 338 the values were found in between
0.05±0.01 and 0.06±0.02. These values were lower than the value reported by Chale
(2002) (4.9-5.30 µg g-1) and Park and Presley (1997) (1.95 to 4.79 µg g-1). For an
average adult (60 kg body weight), the provisional tolerable daily intake (PTDI) for iron,
copper and zinc were 241 µg, 48 mg and 60 mg, respectively (Joint FAO and WHO,
1999). According to WHO (1989), the maximum permissible limit of lead was 0.2
mg/kg. In the present study, the lead values in Cuddalore were higher than the
permissible limits. However, the average Pb concentration value (0.12 µg/g) observed in
the studied species was similar to values reported by Burger and Gochfeld (2005) and
lesser than reported by Sankar et al. (2006) in Kochi waters, India. Sankar et al. (2006)
reported that lead was not detected in the commercially important fish species they
studied, except for J. dussumerri (0.13 mg/kg). Sivaperumal et al. (2007) recorded Pb
concentrations in 25% of finfish, shellfish and other fishery products from the Indian
coast. However, the FDA sets a permissible Cd range of 0.001 - 0.003mg/kg. This value
is lower than the results obtained in this study (FDA, 2003). The EU’s acceptable limit
for Pb concentration is 0.5–1.0 µg/g. The Pb concentrations of several fish species were
found to be lower than the acceptable limit suggested by the European Union (EU, 2001;
EU, 2008). The fish species in this study with high concentrations of Pb require further
research, including an analysis of the number of tissue samples and maximum sample
size.
Chromium
Cr is present in the environment mainly in its inorganic forms Cr(III) and Cr(VI).
The latter is a highly toxic compound as it is soluble at each value of pH. Therefore, it is
also a quite mobile species. Cr(III) is important in the metabolism of fish, whereas Cr(VI)
has increased the maximum lifespan of females in the study of Perez – Benito (2006).
Chromium is one of the least toxic of the trace elements and the mammalian body can
tolerate 100-200 times its total body content of Cr without harmful effect. Chromium is
an essential trace element for human body for glucose tolerance (Dung 2001). Chromium
does not normally accumulate in fish and hence low concentrations of Cr were reported
even from the industrialized parts of the world (Moore and Ramamoorthy, 1984). The
permissible limits set for Cr by FEPA, (2003) are 0.05 and 0.15mg/kg respectively.
Chromium is not known to accumulate in the bodies of fish, but high concentrations of
hromium, due to the disposal of metal products in surface waters, can damage the gills of
fish that swim near the point of disposal (lenntech.com). Fish being at the higher level of
the food chain accumulate large quantities of metals and the accumulation depends upon
the intake and elimination from the body (Karadede et al., 2004). Chromium(VI) is a
danger to human health, mainly for people who work in the steel and textile industry.
People who smoke tobacco also have a higher chance of disclosure to chromium.
Chromium(VI) is known to cause various health effects. When it is a compound in
leather products, it can cause allergic reactions, such as skin rash. After breathing it in
Chromium (VI) can cause nose irritations and nosebleeds (Karadede et al., 2004).
The concentration of chromium in deep-sea fish samples varied from 0.14±0.02 to
3.94±0.24 of cruise 322, whereas in cruise 332 the mean value of chromium was found in
between 0.15±0.01 µg/g and 1.65±0.13 µg/g, while in cruise 338 the concentration of
chromium was occurred in between 0.53±0.11 µg/g and 1.97±0.54 µg/g respectively. The
Cr values from this study were not similar to those reported by Kwon and Lee (1999)
(0.18–0.25 µg/g) in Masan Bay, Korea, and higher than those measured by Sankar et al.
(2006) (0.47 µg/g) in Calicut, India, Raja et al. (2009) (0.65–0.92 µg/g) in Parangipettai
water, India and Yilmaz, 2003 (1.03–1.79 µg/g) in Iskenderun Bay, Turkey. The
concentration of Cr in fish tissues in this study was not comparable to concentrations
reported in other studies. The range of absorption values observed was higher than the Cr
contents (0.12–0.92 mg/kg wet weight) reported in fish tissue samples from New Zealand
(Winchester, 1988; Vlieg et al., 1991), Australia (0.44–0.63 mg/kg dry wt), and
California (0.27–3.0 mg/kg dry wt) waters (Moeller et al., 2003). The maximum
allowable Cr content level in fish is 12–13 mg/kg, as set by the USFDA (1993). Cr
concentration in the fish species studied met permissible level criteria for human
consumption. Cr is an important trace element and plays a critical role in glucose
metabolism (Mertz, 1969). Cr deficiency can result in impaired development and
disturbances in glucose, lipid, and protein metabolism (Calabrese et al., 1985).
Selenium (Se)
Previous studies verified the influence of feeding habits and tropic position in the
Se accumulation (Seixas et al., 2007). Watanabe et al (1997) monitoring Se and Hg
levels in fish, refers to Se intake values in fish diets ranging from 0.15 to 0.5 mg/kg (dry
weight) as necessary to adequate metabolic activity in cells and tissues. Selenium is an

essential element necessary for proper enzyme formation and function (Eisler 1985).
However, chronic exposure to significantly elevated Se levels in the diet or water can
also cause severe toxicological effects, including death. The concentration range
separating effects of Se deficiency from those of toxicity (i.e., selenosis) is very narrow
(Luoma and Presser 2000). With the exception of mortality, the two major toxicological
effects to aquatic organisms from chronic exposure are reproductive effects and
teratogenesis (i.e., malformations in developing fetus). Excessive Se contamination is
often associated with localized extinction of certain species and reduction in biodiversity.
Based on field and laboratory studies with fish and wildlife, it is apparent that elevated Se
concentrations in environmental media, including dietary components, can cause
reproductive abnormalities. These abnormalities include congenital malformations,
selective bioaccumulation by the organism, and growth retardation (Eisler 1985).
Sedimentary Se was dominated by the elemental species, making it less bioavailable than
the Se suspended in the water column (Cutter et al. 2000).
The concentration of selenium in the present study was in below detectable levels
in all the ten given deep-sea fish species in all the three cruises viz., cruise 322, 328 and
338 respectively. Skorupa (1998) carried out a study to examine the relationship between
selenium in tropical and cold waters' fish. The results indicated that fish of tropical region
waters have the minimum acute toxicity of selenium. Dietz et al. (2003) in their research
concluded that inorganic selenium is turned into organic selenium through metabolism
process and its organic forms usually have less toxicity. Hamilton (2004) in his studies
stated that the selenium rate in liver is higher than the edible tissues in fish.
Arsenic (As)
Arsenic is a naturally occurring element that is common in soils, water and
living organisms. Fish can accumulate considerable amounts of organic arsenic
from their environment, but most foods contain tiny levels of organic arsenic
and occasional consumption is not a health concern. An acute high level exposure to
arsenic can lead to vomiting, diarrhea, anemia, liver damage and death. Long term
exposure is thought to be linked to skin disease, hypertension, some forms of
diabetes and cancer (Centeno et al., 2005). Most arsenic in our diet is present in organic
form (FAO, 2011). Arsenic, a chalcophilic element, is widespread in the environment.
Although arsenic may possibly be an essential element for life (Cox 1995) and some
microorganisms are known to use arsenic for energy generation (Oremland and Stolz
2003), no firm data are available on its essentiality for biological systems (Francesconi
2005). In contrast to its possible essentiality in life, many studies have focused on its high
toxicity, which has been well known from various cases of poisoning throughout the ages
(Nriagu 2002). Arsenic has received extensive attention during the last decades due to its
relative mobility over a wide range of redox conditions, carcinogenic properties and also
its occurrence in the aquatic environment (Wang et al. 2012).
Major amounts (85–90 %) of arsenic compounds were found in edible portions of
fish (Thomson et al. 2007). In aquatic systems, arsenic can be originated from both
natural and anthropogenic sources such as volcanism, weathering, as by-product of
mining, metal refining process, burning of fossil fuels, wood preservation, herbicides,
pesticides, fungicides, cement works, electronics industries, ammunition factories,
chemistry (dyes and colors, wood preservatives, pesticides, pyrotechnics, drying agent for
cotton, oil and dissolvent recycling), pharmaceutical works (medication) (Cullen and
Reimer 1989). Arsenic is included in the priority list of dangerous substances established
by ATSDR (2007) as it shows a high ability to accumulate in lipid tissues of organisms
and can be transported easily upwards in the trophic chain. The derived compounds of
arsenic from contaminated water and/or sediments and can be possibly biomagnified
within the aquatic food web (Rahman et al. 2012). In mammals, the poisoning can be
acute (e.g. gastroenteritis) and chronic (e.g. effects on the reproductive – and the immune
system) (Norwood et al. 2007). Also in fish, chronic poisoning of As can cause problems
in the reproduction process as well as in the development of the young fish at
concentrations as low as 1 ppb (Das et al. 1980).
Arsenic is a ubiquitous element on earth with metalloid property and complex
chemistry; aquatic organisms keep and transform arsenic species inside their body after
being exposed to arsenic from food or other sources such as water and sediments
(Hasegawa et al. 2001). The maximum concentration of arsenic was found in P. cyanea
(0.11±0.03) of cruise 322, whereas in cruise 332, the concentration of arsenic found in
between 0.07±0.02 and 0.14±0.02, while in cruise 338 the arsenic content was varied
from 0.03±0.002 to 0.15±0.01. There is no information about maximum arsenic levels in
fish in the Commission Regulation (EC, 2006). Moreover, the maximum arsenic level
reported by Georgian Food Safety Rules (2001) for fish is 2.0 mg/kg wet weight. Hence,
bioaccumulation of heavy metals in fishes can be considered as an index of metal
pollution in the aquatic bodies Tawari and Ekaye (2007) and Karadede-Akin and Unlu
(2007) that might be a useful tool to study the biological role of metals present at higher
concentrations in fish (Dural et al., 2007). Tissues involved in metal metabolism

presented greater dissimilarities between minimum and maximum values with respect to
seasons.
The obtained results indicate that environmental conditions such as the season of
fish catch, temperature and the chemical composition of water affect the physiological
status of fish and the ability to accumulate metals in their organs. The heavy metal
content in deeper water fishes have not been studied extensively and scanty literature has
been found. As mentioned above research on the heavy metal accumulation of fish has
been advancing rapidly. However, additional studies are needed on the interaction
between certain metals that may alter their bioavailability.

CHAPTER-III
ASSESSMENT OF FREE FATTY
ACID PROFILING IN DEEP-SEA
FISHES
INTRODUCTION
Deep sea fishes that used to live in extreme environment are thought to have
nutritional and bioactive compounds that are beneficial for humans healthy. The human
body cannot produce all n-3 fatty acids, but two of them are linoleic acid and alpha-
linolenic acid which are widely distributed in plant oils. Fish oils contain the longer-chain
n-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), while
other marine oils, such as from seal, contain significant amounts of docosapentaenoic
acid (DPA). Fish has often been the focus of attention in both contaminants and
nutritional studies. Nutritionists (Medale et al., 2003) consider seafood to be an chief
source of high quality proteins, vitamin D, and essential fatty acids (EFAs). Unlike
vitamins, EFAs are macronutrients; these are analogous to vitamins in terms of their
significant to human health. A joint study released by the Food and Agriculture
Organization and the World Health Organization (WHO, 2003) recommends that at least
6% to 10% of our daily calorie intake be in the form of EFAs. Indispensable fatty acids
are primarily used to produce hormone-like substances that regulate a wide range of
functions in human body, including blood pressure, blood clotting, blood lipid levels, the
immune response and the inflammation response. These fatty acids are polyunsaturated
fatty acids and are the parent compounds of the omega-3 fatty acid series, respectively.
They are essential in the human diet because there is no other mechanism to synthesize
for them. A diet rich in omega-3 polyunsaturated fatty acids offer many benefits. These
fatty acids may help to reduce the risk of sudden death, abnormal heart rhythms, high
triglycerides, blood clotting as well as inflammatory and autoimmune disease
(www.fsomega 3.com). A number of studies have showed an inverse relationship
between fish and omega-3 fatty acid consumption. Declined the systolic and diastolic
blood pressure has been associated with higher intake of DHA (Rasmussen et al 2006).
Berbert et al and Covington (2004) reported that Omega-3 fatty acids are to be effective
in the treatment of hyperlipidemia, hypertension and rheumatoid arthritis. Consumption
of 1-2 serving/week of fish, especially of species higher in omega-3 fatty acids (EPA and
DHA), reduced coronary deaths by 36% and mortality by 17% (Mozafferian and Rimm,
2006).
Omega-6 fatty acids, like omega-3 fatty acids act an important role in several
biological functions. Higher intake of omega-6 fatty acids are the precursors of pro-
inflammatory eicosanoids, have been suggested to be unfavorable and the ratio of omega-
6 to omega-3 fatty acids has been suggested by some to be particularly important omega-
6 fatty acids have long been known to reduce serum total and low density lipoprotein
cholesterol, and augment in polyunsaturated fat intake, mostly as omega-6 fatty acids,
were a keystone or dietary advice during the 1960s to 1970s. Omega-6 fatty acids
diminish insulin resistance, probably by acting as ligand for peroxisome proliferation
activated receptors-gamma, and intakes have been inversely related to risk of type 2
diabetes. Sufficient intakes of both n-6 and n-3fatty acids are essential for good health
(http://www.ncbi.nlm.nih.gov/pubmed/17876199). The most important omega-6 fatty
acids is linoleic (C18:2 n-6) acid. Longer deficiency of linoleic acid in diet is fatal. All
symptoms of deficit can disappear with its inclusion into nutrition. Antioxidative and
anticarcinogenic properties are accredited to conjugated linoleic acid (CLA) and studies
and results obtained on laboratory animals are very encouraging in prevention of breast,
skin and colon tumors. The ratio of omega-6 and omega-3 fatty acids is very significant,
since these two substances act mutually in conservation of heath. Omega-3 influences
diminution of inflammatory processed whereas omega-6 (other essential fatty acids)
contributes to augment and spreading of inflammatory processes. Inadequate ratio of
these two essential fatty acids contributes to development of disease, whereas an ample
ratio of these two fatty acids contributes to maintaining and even improvement of human
health. Healthy diet should contain 2 to 4 times more omega-6 then omega-3 fatty acids.
Omega-9, or monounsaturated oleic and stearic acid, is a non essential fatty acid
produced naturally by the body whenever there is enough of either omega-3 and omega-6
essential fatty acids. This fatty acid acts a vital role in averting heart disease by
decreasing cholesterol levels. Other benefits of omega 9 are that it diminish hardening of
the arteries and develops immune function (http:/www.globalhealingcenter.com/natural-
health/benefits-of-omega-3-6-9-fattyacids). Long chain polyunsaturated fatty acids (LC-
PUFAs) represent approximately 20% of brain dry matter and its deficit is critical for
development and its function (Beltz et al, 2003). Eicosapentaenoic (EPA, 20:5w3) and
docosahexaenoic acid (DHA, 22:6w3) are the important omega-3 PUFAs, while
arachidonic acid (AA,20:4w6) is vital omega-6 PUFA, EPA and DHA are important in
treatment of arthrosclerosis, cancer, rheumatoid arthritis, psoriasis and disease of old age
such as Alzheimer’s and age related macular degeneration (Drevon et al.,1993;
Simopoulos et al., 1999). The AA and DHA are of special importance in the brain and
blood vessels, and are considered essential for pre and post natal brain and retinal
development (Crawford, 2000).
Fish oils are the major source of PUFA, and considerable evidence has indicated
that omega-3 PUFA in fish oils are actually derived via the marine food chain from
zooplankton consuming omega-3 PUFA-synthesizing microalgae (Yongmanitchai and

Ward, 1989). Docosahexaenoic acid is very important for pregnant or breastfeeding
women and infants. The supplement is claimed to help with fetal and infant brain and eye
development. There may also be cardiovascular benefits of docosahexaenoic acid.
Studies show that fish oil may present cardiovascular disease and help lower
triglycerides. Researches recommended that the fish oil (which contains docosahexaenoic
acid along with other active components) may prevent cardiovascular disease (including
heart attacks and strokes). In people who have already had a heart attack, fish oil may
decrease the risk of another attack and the risk of death
(http://heart_diseasemedtv.com/dha/benefits_of_dha_p2.html). Docosahexaenoic acid is
the most abundant omega-3 fatty acid (PUFAs) in the brain and retina. It comprises 40%
of the PUFAs in the brain and 60% of the PUFAs in the retina. Half of the weight of the
neuron’s plasma membrane is composed of docosahexaenoic acid (Singh, 2005). Rist et
al (2007) stated that 50% more linoleic acid was found in mother’s milk which is a
significant for children’s health. A important thing is to commence through food
sufficient amount of linoleic acid since deficiency will be expressed in the following
systems: changes in skin similar to eczema, hair loss, liver degeneration, changes in
behavior, inclination to infections, wounds heal very slowly, male sterility, abortions in
women, heart and circulation disorders and slow growth (Yoneyama et al, 2007).
Recommendations of the International Society for Study of Fatty Acids and Lipids
(ISSFAL) suggest an adequate intake of omega-3 PUFAs to be 0.65 g of
docosahexaenoic acid plus EPA per day and 1.0 g of ALA per day. The ratio of omega-6
to omega-3 in the diet should be 4 to 1 (Nettleton 1995). A number of research papers
recommend consumption of omega-3 PUFA (Soltan and Gibson 2008).

The fatty acid profiles in Gulf of Mexico fishes studied by Lytle and Lytle (1988).
This study revealed that there was a slight decline in concentration of most fatty acid
from winter to spring though decline is lower than 10%. However this study showed no
significant differences between the seasons of omega-3/omega-6 ratio,
unsaturated/saturated ratio and EPA+DHA/omega-3%.
At different places in the edible fillets of deep-sea fish blue mackerel (Scomber
australasicus) Body and Vlieg (1989) identified the distribution of lipid classes and
ecosapentaenoic (20:5) and docosahexaenoic (22:6). Dark muscle contained 20% lipid
whereas white muscle had only 4% lipid. However, white muscle lipid was relatively
much higher in phospholipids 19% and 10% in dark muscle lipid.
Fatty acid compositions and lipid levels of five common several of marine fishes
from temperate Australian waters (Armstrong et al 1991). The total lipid content of each
of the five Australian species was significantly lower as compared to northern
hemisphere counterparts. Omega-3/Omega-6 ratio and the levels of arachidonic acid
(AA) and docosahexaenoic acid (DHA) in the lipids were comparable for the two, but the
Australian fishes had lower levels of eicosapentaenoic acid (EPA). Oils from Australian
fishes were shown to have nutritional properties as beneficial as their northern
hemisphere counterparts.
Krzynowek et al (1992) analyzed marine sardines harvested in June, July,
October and November for proximate composition and fatty acids either as raw dressed,
whole fish, steam precooked and after canning in soy oil, menhaden oil or in liquid
exuded after steam precooking (cookout liquid). They found fat content to range from 5%
for juvenile herring sampled in June and October to 11% in maturing herring harvested
prior to the spawning season in July. All canning liquids contributed to elevated fat
content in the finished product. Nutritional composition of the raw product was
unaffected by heat processes but was altered significantly by canning solutions. The
amounts of EPA and DHA in samples were primarily a function of the amount of fat. The
fat content of sardines packed in canning solutions increased significantly with no change
in n-3/n-6 ratio. The juvenile sardines with significantly less fat than the mature juveniles
had significantly less EPA and DHA/serving than mature juveniles of all product forms.
Commercially packed sardines in soy oil appeared to be a good source of omega-3 fatty
acids while contributing moderate amounts of cholesterol. Canning in menhaden oil more
than doubled the amount of EPA and DHA in the sardines, but the final product had
slightly elevated levels of cholesterol. Steam loss liquid appeared to be the best canning
solution contributing substantially to the amounts of EPA and DHA while maintaining
moderate levels of cholesterol.
Gutierrez and Silva (1993) examined total fat and fatty acid composition of nine
marine fish and seven freshwater commercially important in Brazil were determined.
Palmitic acid was the primary saturated fatty acid in both freshwater fish the total C-16
fatty acids were higher than in marine fish. Oleic acid was the most abundant mono-
saturated fatty acid and it was found in higher levels in freshwater fish from Brazil
examined were a poor source of EPA and DHA.
Tornaritis et al (1994) reported the fatty acids and the total fat content of eight
species of Mediterranean fish (Scamber japonicus, Boops boops, Meillus barbatus,
Meillus surm uletus, Merluccius merluccius, Pagelius erythrinus, Pagrus pagrus, Sparus
aurata) normally devoured in this area were analyzed. The Gas Chromatography of the
fatty acids showed the ω-3 to range from 12.6 to 28.3% of the observed acids. The ω-6/ω-
3 ratio of fatty acids was found to be 0.2-0.7, and saturated fatty acids range from 28 to
33.2%. These results revealed that the eight species of fish studied from the
Mediterranean Sea are a good source of omega-3 fatty acids.
The seasonal changes in lipid composition of sardine (Sardina pilchardus)
(Bandarra et al., 1997). They reported that sardine, a small pelagic fish have important
nutritional characteristics because of their high level of omega-3 fatty acids. Their study
revealed that fat content of sardines showed important seasonal changes during the year
which was typical of pelagic species. The minimum value was reached in March-April
which coincided with the period of spawning and the highest percentage was found in
September-October after a heavy feeding period. Polar lipids were dominant in the lean
period and phosphatidyl choline was the main phospholipid followed by phosphatidyl
ethanolamine which attained the highest level in winter. According to the total levels of
DHA and EPA in mg/100g edible flesh, the best period for consumption of sardines
would be September – October when it is tastier and more abundant.
Fatty acid composition of two marine fish oil namely Eusphyra blochii and
Carcharhinus bleekeri. Twenty five individual fatty acids from the marine fish oil were
analysed among those the Palmitic acid was a major saturated fatty acid while Stearic
acid was the other major constituent. Among unsaturated fatty acid monoenoic e.g. oleic
and palmitoleic acids were the major constituent and traces of dienoic and trienoic fatty
acids were also found (Saify et al., 2000).

Osman et al (2001) reported that the total fat, fatty acid composition and
cholesterol content of selected marine fish in Malaysian waters. The composition of fatty
acids showed that n-3 PUFAs (29.7 - 48.4%) were the highest followed by other PUFA
(27.7-40.0%), n-6 PUFAs (11.0-20.0%), saturated fatty acid (3.63-11.4%) and finally
monounsaturated fatty acid (1.37-9.12%). Most of the fish contained <6 percent lipid by
weight and total cholesterol was 37.1 - 49.1 mg/100g. The fish had a higher n-3/n-6 ratio
(2.16-4.14) than the standard menhaden oil (2.03) except for Four Finger Threadfin
(1.50), Indian Mackerel (1.67) and Striped Sea Catfish (1.78). The PUFA/saturated ratio
ranged from 5.49-25.2.
Although, all fishes contain EPA and DHA, the abundance of fatty acids vary
among species and within a species according to environmental variables such as diet and
whether fish are wild or farm raised. Farm raised catfish tend to have less EPA and DHA
than do the wild catfish. Farm raised salmon and trout, however, contain similar amounts
as compared to their wild counterparts (http://www.nalusda.gov/fnic/foodcomp).
According to USDA Nutrient Data Laboratory (assessed October 3, 2002), fishes are
major source of EPA and DHA.
Anchovy (Engraulis encrasicholus) and horse mackerel (Trachurus trachurus)
are the two fish species and three commercial fish products were compared by gas
chromatography mass spectrometry (GC-MS) according to their EPA and DHA content
(Kartal et al., 2003). PUFA and ω-3 were detected in abundant amounts, especially in the
anchovy oil. Anchovy oil can be used in the form of gelatin capsules as dietary
supplements in Turkey.
Chakraborty et al (2004) observed that Bhola Bhetki (Nibea soldado) for lipid
composition of its specific organs. This study revealed that the fish had much higher lipid
percentage in brain in comparison with its other organs while the muscle and digestive
tract had almost similar amount of total lipids. The cholesterol content in various organs
was high whereas phospholipid content was low. Fatty acid composition of eye and
digestive tract were richer in variety than those of muscle and brain. The total saturated
and PUFAs were more or less equal in muscle and brain lipid. The total MUFAs were
more or less same in the muscle, brain and eye. The content of EPA was very high in all
the organs. Arachidonic acid was also present in considerable amounts in brain, eye and
digestive tract. They suggested that the variation of cholesterol, phospholipid and fatty
acid composition in fish organs would be beneficial for human consumption from health
point of view.
Stokens et al (2004) examined the lipid class and fatty acid composition in eyes
and brain from teleosts and elasmobranches. This study reported that the lean species
contained high ratios of DHA versus EPA, and high ratio of omega-3 fatty acids versus
omega-6 fatty acids, while these ratios were significantly lower for the fatty fish species.
Analysis of the fatty acid composition of brains revealed that the deep-sea
elasmobranches contained a level of arachidonic acid (AA) that was higher than their
EPA and fivefold higher than in the brains of teleosts.
Results obtained from the comparative study of body composition of different fish
species from brackish water pond did not show any adverse effect of salinity on fish
species (Ali et al., 2005). Minimum amount of water content and maximum amounts of
lipids, organic content and condition factor were observed in Cyprinus carpio indicating
that Cyprinus carpio show overall better growth in brackish water as compared to other
species. The mean values of body constituents, except for protein content differed
significant (p<0.05) among various fish species. Chakraborty et al (2005) studied the
fatty acid profiles of Pomfret fish (Pampus argenteus) organs. This pomfret fish showed
accumulation of high fat percentage in brain than in other organs. The lipid composition
including fatty acids in the different organs of pomfret fish varied significantly. The
protein, fatty acid, nutritional value in muscle of fish species Mora moro (Risso, 1810)
inhabiting deep Mediterranean waters (Rossano et al., 2005). The major fatty acids were
docosahexaenoic acid (C22:6 n-3), palmitic acid (C16:0), oleic acid (C18:0 n-9),
arachidonic acid (AA; C20:4 n-6) and EPA (C20:5 n-3). The PUFA composition was
higher than that of both saturated and monounsaturated fatty acids but the ratio of
PUFA/SFA was lower than the value reported in other studies. Varljen et al (2005)
examined fatty acid composition of the lipid fractions of 2-banded sea bream (Diplodus
vulgaris). The total lipid content was 1.1±0.3% in the edible part and 4.5±1.0% in the
liver. The major fatty acids found were saturates Palmitic (20.8-43.4%) and Stearic acids
(6.3-33.7%); monosaturates - oleic (10.0-33.6%) and palmitoleic acids (2.1-11.0%); and
polyunsaturates - arachidonic (1.5-8.6%), docosatrienoic (0.8-11.6%), eicosapentaenoic
(1.1-5.5%) and docosahexaenoic acids (1.5-9.9%). The liver contained more n-3 PUFA
than the muscle or meat part of the fish. No significance was observed in between the
relative amounts of EPA in phosphatidylethanolamine of muscle and liver tissue. The
relative ratio of DHA was significantly higher in liver triacylglycerols (P<0.0005) and
phosphatidylcholine (P<0.05) compared with the flesh.

Fatty acid composition of three marine fishes of the Bay of Bengal was studied by
Bhuiyan et al (2006). The three fishes namely stingray, anchovy and eel. The SFA
contents were 52.95, 45.28 and 52.29% and the unsaturated fatty acid contents were
43.97, 54.72 and 33.39% in stingray, anchovy and eel, respectively. The highest PUFA
content was found in anchovy (13.65%) and the lowest was in stingray (8.06%). The w-3
fatty acid contents were 4.58, 3.65 and 9.91% and w-6 fatty acid contents were 3.48,
10.00 and 5.50% in stingray, anchovy and eel. Two EFA lionleic acid (4.12%) and
arachidonic acid (5.32%) contents were found higher in anchovy whereas another EFA
linolenic acid (8.98%) found higher in eel. Another important PUFA EPA (2.12%) found
higher in stingray. Shamsudin and Salimon (2006) estimated the total lipids and fatty acid
compositions in aji-aji fish (Seriola nigrofasciata).The result showed that aji-aji fish
content about 18.3% lipid (wt/wt). The lipid consists of high saturated fatty acid
(45.8±1.8%) compared to MUFA (32.4±0.5%) and omega-6 PUFA (1.0±0.2%) and
omega -3 PUFA (9.8±0.5%). Aji-aji fish oil shows relatively high in omega-3 PUFA
content comparable with other local fish oils. The EPA and DHA are the major PUFA
content which are 1.1±0.1% and 8.0±1.0% respectively. However its omega-3 PUFA is
quite low as compared to commercial menhaden fish oil.
Mnari et al (2007) analyzed the fatty acids in dorsal and ventral muscles and liver
of Tunisian wild and farmed gilthead sea bream, Sparus aurata. They statement the
quantity of fish lipid was higher in farmed fish than in wild fish in all examined samples
and the highest level of all was observed in liver. The palmitic acid (16:0) and oleic acid
(18:1 omega-9) were the primary saturated and monounsaturated fatty acids. The farmed
fish contained a higher level of n-3 PUFAs particularly DHA and EPA and omega-
3/omega-6 ratio whereas wild fish contained a higher level of omega-6 PUFA.
Arachidonic acid (AA) was primary omega -6 PUFA in wild fish whereas in farmed fish,
linoleic acid was major omega -6 PUFA. Farmed fish were characterized by higher
omega-3/ omega -6 ratio for all samples due to abundance of omega-3 PUFA, particularly
DHA. The estimation of fatty acid and proximate composition in the flesh of thornback
ray (Raja clavata) was determined by Turan et al (2007). The total SFA, MUFA and
PUFA content were 48.27%, 13.98% and 24.10% of total fatty acids. Palmitic acid
(26.45%) and stearic acid (10.62%) were the dominant saturated fatty acids. The major
unsaturated fatty acids were determined as DHA (12.21%), oleic acid (8.88%) and
linoleic acid (3.43%). The results showed that thornback rays are excellent source of
protein.
The effects of seasonal variations on the proximate compositions and fatty acid
profiles of chub mackerel (Scomber japonicus) and horse mackerel (Trachurus
trachurus) captured in the north-eastern Mediterranean Sea which was investigated by
Celik (2008). The fatty acid composition of both species was lower in winter than in
autumn and spring. In all seasons, the major fatty acids in both species were observed to
be palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1 n-9), eicosapentaenoic acid
(20:5 n-3) and docosahexaenoic acid (20:6 n-3). Chub mackerel and horse mackerel
showed the seasonal fluctuations in their fatty acid contents. The fatty acid profile of the
two species had a higher degree of unsaturation during winter. The levels of EPA in chub
mackerel in winter, spring and autumn were 5.96%, 4.86% and 4.33%, respectively,
while those of DHA were 24.94%, 18.75% and 17.12%, respectively. The levels of EPA
in horse mackerel in winter, spring and autumn were 5.42%, 5.03% and 4.86%, while
those of DHA were 14.96%, 13.31% and 11.10%, respectively. The PUFA
(polyunsaturated fatty acids) values and omega-3/ omega-6 ratios in the two species were
highest in winter. According to this study, the chub mackerel and horse mackerel
captured in the north-eastern Mediterranean Sea, which are among the most imperative
fish in Turkey and of international commercial value, are a excellent source of nutrition
for human consumption in terms of their proximate composition and fatty acids. The
quality characteristics of sea bass (Dicentrarchus labrax) intensively reared and from
lagoon as affected by growth conditions and the aquatic environment (Orban et al.,
2008). Fish from either productive system showed comparably high total PUFA levels
and n-3/n-6 ratio values. Differences in the mineral content occurred among the fishes,
with the levels of mercury and selenium significantly higher in those from the lagoon.
Farmed fish had a total lipid content (10.57±0.17 g/100g) significantly higher (P<less or
= >0.001) than that of wild fish (1.78±0.01 g/100g). Meynier et al (2008) indomitable
that the proximate composition, energy content and fatty acid composition of marine
species from Campbell Plateau, New Zealand. The variation of energy content between
fish species was not significant, but their fatty acid and protein contents varied
significantly. Fatty acid signatures eminent the species analysed and at a broader scale,
the type of habitat. In general, the diet inference from Fatty acid trophic markers was
consistent with reported diets from stomach contents.
Huynh and Kitts (2009) determined the quality of nutritional of Pacific Northwest
coastal water fish species from fatty acid signatures. They determined that Pacific fish
species have relatively high proportion of omega -3 highly unsaturated fatty acids (omega
-3 HUFAs), of which more than 80% was reported by both EPA and DHA with species
specific and lower proportions of oleic acid and palmitic acid also dominating. The
MUFA contents of fish were lower (P<0.05) in the lipids of lean and low fat fish
compared to fattier fish. In contrast, higher (P<0.05) proportions of PUFA accumulated
in the low-fat species with DHA contents ranging from 18-29% in the low fat fish and
from 8-10% in fattier fish such as herring and capelin. Expressing the same fatty acid
content data in terms of absolute amount of fatty acids (e.g. FA g/100g wet tissue)
showed that both EPA and DHA contents in the flesh of pollock and hake were indeed
many fold lower than those found in fatty fish, such as herring. According to the authors,
their findings confirmed that it was important that both the total lipid content and the
fatty acid composition of Pacific fish food sources be considered when making
evaluations on the nutritional quality. Oksuz et al (2009) reported biochemical
composition, fatty acids (FA) and elements content of two shrimp species, deep seawater
rose shrimp (Parapenaeus longirostris) and red shrimp (Plesionika martia). The quantity
of fatty acid in P. longirostris and P. martia was found as 1.1 and 2.61%. Percentage of
lipid in both shrimp was lower than that of marine fish. A fatty acid profile of these two
shrimp species was similar with that of marine fish. The amounts of PUFA’s in both
shrimp species were found higher than of SFA and MUFA. Levels of DHA in P.
longirostris was significantly (p<0.05) higher than that of P .martia. Tang et al (2009)
predicted the fatty acid profiles of edible part from large yellow croaker (Pseudosciaena
croceal) of various age. Crude protein, fat, moisture and ash content showed that
significant difference was not observed in between the two age groups. The content of
PUFAs and DHA were significantly higher and EPA content was significantly lower in
the lower two year old large yellow croaker than in the one year old (P<0.05). No
significant differences were observed in the contents of total SFAs and MUFAs or the
ratio of n-3/n- 6 fatty acids among the large yellow croakers of the two age groups.
Estimation of fatty acid and amino acid profiles in three silverside populations
caught in Tunisian waters, open sea, lagoon and island coasts (Bouriga et al., 2010).
Saturated fatty acids reached in total lipids 43.54%, 39.96% in marine and 33.64% in
insulkar silverside. EPA, DHA and linoleic acid were the prominent fatty acids in A.
lagunae (lagoon). The n-3/n-6 index showed a significant level indicating a tendency to
accumulate n-3 fatty acids in A. boyeri and A. lagunae and n-6 fatty acids in Atherina sp.
Proximate content, fatty acid and mineral compositions were determined for the ten
species of deep-sea fish from Southern Java Ocean and Western Sumatra Ocean,
Indonesia (Suseno et al., 2010). The proximate composition was found to be 23.0-24.8 %
protein, 1.9-4.1% fat , 0-1.75 % carbohydrate, 1.7-2.4 % ash and 70.1-72.1% water,
whereas the fatty acid compositions consisted of 0.86 - 49.63 % saturated fatty acids
(SFA), 0.2 - 50.09 % monounsaturated fatty acid (MUFA) and 2.85 % - 46.32 %
polyunsaturated fatty acids (PUFAs). Among them, those occurring in the highest
proportions were myristic acid (C14:0, 0.12-7.59%), palmitic acid (C16:0, 0.02–20.5%),
stearic acid (C18:0, 0.42–49.19), oleic acid (C18:1, 0.29–50.09 %), linoleic acid (C18:2,
0.23–44.91%), eicosapentaenoic acid (EPA, C20:5n3, 0.41– 4.61%) docosahexaenoic
acid (DHA, C22:6n3, 0.28–3.44%). The rest of the microelements, Cd, Hg, and Pb were
all present in amounts below toxic levels. Kuçukgulmez et al (2010) examined the
seasonal variation of the biochemical and fatty acid compositions in the round herring
(Etrumeus teres) and tub gurnard (Chelidonichthys lucernus) caught from the North-
Eastern Mediterranean Sea. Biochemical compositions and fatty acid contents of both
fish species showed statistically significant seasonal variations (P < 0.05). While the
protein contents were at its highest in winter, the lipid contents were at its highest in
autumn. The main fatty acids were C16:0 (palmitic acid), C18:0 (stearic acid), C16:1 n-7
(palmiteoleic acid), 18:1 n-9c (oleic acid), C20:5 n-3 (eicosapentaenoic acid) and C22:6
n-3 (docosahexaenoic acid). The total polyunsaturated fatty acid in round herring was
highest in winter, whereas that in tub gurnard was highest in spring.
The edible portion of Diaphus watasei, a bentho-pelagic fish collected off Quilon,
south-west coast of India for determination of proximate composition and fatty acid
profile (Manju et al., 2011). The fleshy part of the fish was found to contain 15.62%
protein, 11.71% fat, 0.47% minerals, 0.28% soluble carbohydrate and 0.01% crude fibre.
The dry matter in the fish was found to be about 28%. Monounsaturated fatty acids
(MUFA) were found to have the highest (36.7%) share among total fatty acids followed
by saturated fatty acids (SFA) (33.3%) and polyunsaturated fatty acids (PUFA) (25.5%).
The abundant fatty acids were found to be oleic acid, palmitic acid, docosahexaenoic
acid, stearic acid, myristic acid, linolenic acid, eicosapentaenoic acid and palmitoleic
acid. The most predominant fatty acid was recorded as oleic acid which contributed 32%
to the total fatty acids. Docosahexaenoic acid formed the single largest component of
PUFA (9.33%) followed by γ linolenic acid (3.97%) and eicosapentaenoic acid (3.83%).
The omega -3 PUFA contributed about 70% of the total PUFAs. The most important
omega-3 PUFAs were EPA and DHA that contributed 73% to the total omega-3
polyunsaturated fatty acids.
Deep-sea fishes (myctophids) can be utilized for the production of commercial
fishery products like fish meal, fish oil, fish silage, surimi, seasoning products, feed for
cultured fish, nutrient resource in the formulation of poultry feed as well as crop
fertilizers and products like lubricating oil, cosmetics and wax (Rajamoorthy et al.,
2013). This study have shown significantly higher fat content in Diaphus watasei
(15.13%), a myctophid species commonly seen in the discards of deep sea shrimp
trawlers operating off south-west coast of India. They had also reported higher protein
content in myctophid fishes viz., D. watasei (21.40%) and Myctophum obtusirostre
(22.64%) respectively. Determination of fatty acid and steroid content of escolar
(Lepidocibium plavobrunneum) and deep-sea lobster (Linuparus somniosus), the fatty
acid analysis was carried out using methods of gas chromatography (GC) (Sugeng et al.,
2013). Oleic acid (C18:1, 35.16%) was dominant monounsaturated fatty acid (MUFA) in
escolar flesh. The highest fatty acid composition of deep-sea lobster can be found in
monounsaturated fatty acids (MUFA) and saturated fatty acids (SFA). Deep-sea lobster’s
flesh and viscera contained oleic acid (5.72% and 12.40%) as major monounsaturated
fatty acids. The dominant saturated fatty acid of lobster’s flesh and viscera was palmitic
acid (5.06% and 12.02%). The result of Liebermann-Burchard analysis showed that deep-
sea lobster contained more steroids than escolar. Based on the results, the deep-sea fishes
have potential as a natural source of aphrodisiac.
Fernandez et al (2014) examined the comparison of Nutritional Characteristics of
Myctophid Fishes (Diaphus effulgens and D. hudsoni) with Common Indian Food Fishes.
Biochemical composition shows myctophid fishes having comparable quantity of protein
and fat with food fishes and in D. effulgens, the protein content was higher than in S.
longiceps. Analysis of minerals (calcium, sodium and potassium) and trace elements
(copper, iron, manganese, zinc and cadmium) shows that myctophid fishes have
comparable quantities of sodium, potassium and calcium with the food fishes studied. D.
hudsoni contained higher amount of calcium than D. effulgens, but sodium, potassium
and barium were observed in significantly higher level in the latter. Proximate
composition indicates that protein and lipid levels in these myctophid fishes are
comparable to that of the selected food fishes. These myctophids contained omega-3
polyunsaturated fatty acids (omega-3 PUFA) and essential amino acids in significant
proportions.
Ramesh et al (2015) evaluated the seasonal changes in nutritive value of ribbon
fish, in relation to size, total lipid and fatty acid profiles of different sizes of ribbon fish,
Lepturacanthus savala captured in different seasons were studied. Total lipid content was
significantly (p<0.05) high in large sized ribbon fish captured during winter and lowest in
medium size groups during post-monsoon season. A total of 37 fatty acids were identified
by Gas Chromatography Mass Spectrometry encompassing saturated, mono unsaturated,
n-6, n-3 poly unsaturated and odd and branched chain fatty acids. The highest content of
eicosapentaenoic acid (9.42%) and docosahexaenoic acid (33.16%) were found in the
post-monsoon season. The high levels of omega-3 PUFA (12.37 – 43.14%) and favorable
omega -3/ omega-6 PUFA ratio of 1.79 to 5.04 revealed nutritional significance of ribbon
fish in human diet. The difference between small, medium and large groups of ribbon
fishes of the respective season based on fatty acid composition was clearly established by
principal component analysis.
The omega-3 fatty acids [docosahexaenoic acid (DHA) and eicosapentaenoic acid
(EPA)] profile of four myctophid species of fishes (Benthosema pterotum, Benthosema
fibulatum, Diaphus jenseni, Myctophum spinosum) caught from Arabian Sea was
compared with commerson’s anchovy (Stolephorus commersonii) to evaluate their
nutritional quality for human health benefits (Navaneethan et al., 2016). The protein
content of myctophid fishes ranged between 19.36% for Myctophum spinosum to 13.74%
for Benthosema pterotum. Total fat (3.64%) and moisture content (81.03%) were high in
Benthosema pterotum (3.64%). Stolephorus commersonii was found to have higher ash
content (3.23%). Myctophid omega-3 PUFA profile is comparable to Commenrson’s
anchovy fatty acid composition. Presence of higher DHA content in M. spinosum
(20.2%), B. pterotum (17.48%) was comparable to edible S. commersonii (16.9%)
emphasizes that these underutilized non-edible myctophids may be considered as a
potential nutrient resource of omega-3 PUFA especially DHA for the formulation of
nutraceuticals, cosmetics and animal feeds. The biochemical composition of deep sea
squid species especially Mastigoteuthis flammea available in Arabian Sea is scanty
(Jayarani et al., 2016). This study has investigated the proximate composition, amino acid
composition and fatty acid profiling of M. flammea caught from Arabian Sea. The
moisture, crude protein, crude lipid and ash contents of the squid were 60.20 g/100g,
23.13g/100 g, 17.05g/100 g, and 1.61g/100 g respectively. The squid species was found
to contain the essential and non-essential amino acids in a balanced proportion, which is
an essential criterion for attenuating protein deficiency related malnutrition disorders.
Interestingly, its body oil was found to comprise of healthcare important n-3
polyunsaturated fatty acids (omega -3 PUFA) such as eicosapentaenoic acid (EPA) and
docosahexaenoic acid (DHA) in considerable quantities. The results of the present study
suggest that the deep sea squid M. flammea can be considered as a potential resource of
nutrients required for the normal maintenance of human health. Determination of nutrient
profiling of two deep sea fish species (Neoepinnula orientalis and Chlorophthalmus
corniger) was compared with a well known edible brackish water fish (Scatophagus
argus) Vijayan et al., 2016). The deep sea fish of interest (N. orientalis and C. corniger)
were observed to possess relatively similar quantities of crude protein (18.6 ± 0.9 and
19.4 ± 0.9% respectively) as that of S. argus (20.4 ± 0.8%). The fat content of N.
orientalis is commensurate with that of spotted scat, whereas C. corniger was found to
contain very high fat content (14.6 ± 0.7%). Though their amino acid composition display
slight variation with that of the brackish water fish, the deep sea fish were analyzed to
contain significant amount of the essential amino acids viz. lysine, phenyl alanine,
histidine, as well as the non essential amino acids aspartate, arginine, serine, glutamate,
proline, glycine, alanine. The studies have also conceded that the fish from deep waters
are the comparable sources of minerals, with those of the brackish water fish. Among the
three fish of interest N. orientalis was noticed to be the richest source of sodium (5746 ±
27 mg/kg), potassium (3438 ± 19 mg/kg), calcium (4247 ± 16 mg/kg) and magnesium
(2253 ± 21 mg/kg). Meanwhile, C. corniger is having highest levels of iron (120 ± 1.5
mg/kg) and zinc (135 ± 2.8 mg/kg), whereas S. argus was found to have the highest
levels of manganese (35.8 ± 2.8 mg/kg) and nickel (10.1 ± 0.8 mg/kg). The level of
cadmium in N. orientalis (0.75 ± 0.01 mg/kg) was demonstrated to be slightly higher than
the prescribed limit.
The food consumption and metabolism of deep sea fish have seldom been studied.
Information on the proximate content and fatty acid distribution is important when
utilization of new species of deep-sea fish is considered. This is because deep-sea fishes
are considered to be not only food with good source of quality protein but also food with
healthy components. Polyunsaturated fatty acids (FAs) such as eicosapentaenoic acid
(EPA, 20:5 (n-3)) and docosahexaenoic (DHA, 22:6 (n-3)) have been recommended for
human health and fish fecundity; particularly in DHA, has a therapeutic effect on human
physiology.
MATERIAL
&
METHODS
DETERMINATION OF FATTY ACID PROFILE BY GAS
CHROMATOGRAPHY
Lipid Extraction
The wet muscle was homogenized with 2:1 mixture of chloroform and methanol. The
chloroform-methanol mixture extracts the total lipid from the tissue in to a single phase of
solvent. Disturbing the equilibrium between chloroform and methanol separates the
chloroform soluble fat.
Procedure
Five gram of meat was triturated with 15 volumes of chloroform - methanol
mixture in a mortar and pestle. The mixture was filtered to remove the residue. The
extract was measured and taken in a separating funnel, to which 20% volume of water
was added and mixed for phase separation. The lower layer is collected after passing
through anhydrous sodium sulphate to absorb moisture. The solution was concentrated to
10 ml in a vacuum flash evaporator and kept under nitrogen pending analysis. One
milliliter of the concentrate was taken in a pre-weighed test tube and solvent was
evaporated. The test tube was cooled in a desiccator and weighed.
Calculation
W2 x V1 x 100
Fat content (g/100g meat):
V2 x W1
Where:
 V1: total volume of extract
 V2: Volume of extract taken for drying
 W2: weight of dried lipid
 W1: weight of sample for fat extraction
Preparation of Fatty acid Methyl esters (FAMEs)
One millilitre of the above chloroform extract was taken in a 30 ml screw capped
test tube and solvent was evaporated. To the lipids 1.5 ml of 0.5 M methanolic NaOH
was added and flushed with nitrogen gas. The lid was tightly closed and heated at 100 0C
for 5 min in a hot air oven for saponification. The tubes were cooled and 2 ml of Boron
trifluoride methanol was added and flushed with nitrogen gas. The lid was tightly closed
with the cap and heated at 100 0C for 30 min in a hot air oven for transesterification. The
tubes were cooled and 1 ml of hexane was added and thoroughly mixed. To this 5 ml of
saturated sodium chloride solution was added and thoroughly mixed. The cap was
loosened and allowed to stand for 5 min for phase separation to take place. Carefully
using a micropipette, about 0.8 ml of upper hexane phase was removed and transferred to
a glass vial. The extraction was repeated one more time with 1 ml of hexane and the
extracts were pooled. One microlitre of the prepared FAME was injected for fatty acid
profiling.
Analysis of the fatty acid methyl esters was performed in a Varian CP 3800 gas
chromatograph with a split/splitless injector, a column oven and a flame ionization
detector. A capillary column of 100 meter length was used for separation. The system
was calibrated with a 37 component FAME mix if analytical standard (Supelco).

Gas chromatography conditions:
Slit-1:5 to 1:50, Injector temp-260 0C, Oven temp- 140 0C for 5 min, ramp rate –
2.2 0C/min up to 200 0C, ramp rate -20C/min up to 240 0C, Detector temp- 270 0C, Carrier
gas flow- 3.5 ml/min N2 .The program took 60 min for complete separation of all fatty
acid methyl esters.
Samples were identified using retention time by comparing with respective
standards. Area of each component was obtained from the computer-generated data and
concentration was calculated using the software by external standard.

RESULTS
Three major groups of fatty acids viz., polyunsaturated fatty acids (PUFAs),
monounsaturated fatty acids (MUFAs) and saturated fatty acids (SFAs) have been
identified in the muscle of deep-sea fish species viz., Psenopsis cyanea, Bembrops
caudimacula, Neoepinnula orientalis, Nemipterus japonicus, Chascanopsetta lugubris,
Cubiceps baxteri and Lamprogrammus niger. The PUFAs included omega-3 (n-3)
omega-6 (n-6) and omega-9 (n-9) fatty acids. The major n-3 fatty acids were linolenic
acid (Len; C18:3 n-3), eicosapentaenoic acid (EPA; C20:5 n-3), docosapentaenoic acid
(DPA; C22:5 n-3) and docosahexaenoic acid (DHA; C22:6 n-3), whereas the major n-6
fatty acids included linoleic (Lin; C18:2 n-6) , arachidonic acid (AA; C20:4 n-6) and
eicosadienoic acid ( EDA; C20:2 n-6). The only n-9 fatty acid was eicosaenoic acid (C
20:1 n-9). Palmitoleic acid (C16:1 n-7) and oleic acid (C18:1 n-9) were the major. The
SFAs included capric acid (C10:0), undecyclic acid (C11:0), lauric acid (C12:0), myristic
acid (C14:0), pentadecanoic acid (C15:0), palmitic acid (C16:0), margaric acid (C 17:0),
stearic acid (C18:0), and eicosanoic acid (C20:0).
Saturated Fatty Acids (SFA):
The total mean value of saturated fatty acid percentage in the following samples
was Psenopsis cyanea (37.11%), Bembrops caudimacula (30.88%), Neoepinnula
orientalis (12.67%), Nemipterus japonicus (34.61%), Chascanopsetta lugubris (33.88%),
Cubiceps baxteri (42.02%) and Lamprogrammus niger (26.34%). Among these samples,
saturated fatty acid accumulated more percentage in Cubiceps baxteri followed by
Psenopsis cyanea and Nemipterus japonicus while less quantity of saturated fatty acid
was found in Neoepinnula orientalis. Among saturated fatty acids, palmitic acid was the
most prominent fatty acid followed by stearic acid and myristic acid in the samples of
Psenopsis Cyanea, Bembrops caudimacula, Nemipterus japonicus, Chascanopsetta lugubris and

Lamprogrammus niger, whereas the samples of Neoepinnula orientalis and Cubiceps baxteri
palmitic acid was the highest amount of fatty acid followed by myristic acid and stearic acid
(Table 1) respectively.
Mono-Unsaturated Fatty Acids (MUFA):
The total mean value of mono-unsaturated fatty acid percentage in the following
samples was Psenopsis cyanea (35.07%), Bembrops caudimacula (18.66%), Neoepinnula
Cubiceps baxteri (26.45%) and Lamprogrammus niger (24.89%). Among these,
Neoepinnula orientalis accumulated highest percentage of mono-unsaturated fatty acids
followed by Psenopsis cyanea and Cubiceps baxteri, while least quantity of MUFA was
found in Nemipterus japonicus Octadec-9-enoic acid – (C18:1) was the dominant MUFA
in the given deep-sea fish species. The percentage of Octadec-9-enoic–(C18:1) fatty acid
in the deep-sea fish species were Psenopsis cyanea (27.49%), Bembrops caudimacula
(10.40%), Neoepinnula orientalis (48.38%), Nemipterus japonicus (9.82%),
Chascanopsetta lugubris (19.76%), Cubiceps baxteri (22.02%) and Lamprogrammus
niger (14.04%), while Neoepinnula orientalis had more percentage and Nemipterus
japonicus had least percentage of Octadec-9-enoic acid in mono unsaturated fatty acids.
The percentage of Cis-9-Hexadecenoic (C16:1) fatty acid in the given seven samples was
Psenopsis cyanea (3.79%), Bembrops caudimacula (3.48%), Neoepinnula orientalis
(7.42%), Nemipterus japonicus (5.24%), Chascanopsetta lugubris (3.52%), Cubiceps
baxteri (0.49%) and Lamprogrammus niger (2.02%). Neoepinnula orientalis and
Nemipterus japonicus accumulated more quantity of Cis-9-Hexadecenoic fatty acid
(Table 1).
Poly-Unsaturated Fatty Acids (PUFA):
The overall mean concentrations of poly-unsaturated fatty acids in the following
sample were Psenopsis cyanea (27.81%), Bembrops caudimacula (50.47%), Neoepinnula
Cubiceps baxteri (31.53%) and Lamprogrammus niger (48.77%). In these deep-sea fish
species, Bembrops caudimacula accumulated more quantity followed by
Lamprogrammus niger and Nemipterus japonicus whereas less percentage of poly-
unsaturated fatty acid was observed in Cubiceps baxteri respectively. Docosa-4, 7, 10,
13, 16, 19-hexaenoic acid (DHA) - C22:6 was the most significant fatty acid in poly
unsaturated fatty acid and DHA found in more quantity in all the given samples. Eicosa-
5, 8, 11, 14, 17-pentaenoic acid (EPA) was found more concentration after the DHA in
Psenopsis cyanea, Neoepinnula orientalis and Cubiceps baxteri whereas eicosa-
5,8,11,14-tetraenoic acid ETA found more percentage after the DHA in Bembrops
caudimacula, Nemipterus japonicus, Chascanopsetta lugubris and Lamprogrammus
niger respectively. Among the seven deep-sea fish samples, the total percentage of
Eicosa-5, 8, 11, 14, 17-pentaenoic acid (EPA) was found more in Bembrops caudimacula
(6.84%) followed by Neoepinnula orientalis and Nemipterus japonicus, while eicosa-
5,8,11,14-tetraenoic acid (ETA) found more percentage in Bembrops caudimacula
followed by Lamprogrammus niger and Nemipterus japonicus (Table 1) respectively.
The total percentage of mean value of fatty acids in all the given deep-sea fishes,
more quantity of saturated fatty acids (SFA) observed in Psenopsis cyanea and Cubiceps
baxteri, while more amount of mono unsaturated fatty acids (MUFA) was found only in
Neoepinnula orientalis, whereas high percentage of poly unsaturated fatty acids (PUFA)
was found in Bembrops caudimacula, Nemipterus japonicus, Chascanopsetta lugubris,
and Lamprogrammus niger (Table 2 & Graph 1) and in overall percentage of fatty acids,
poly unsaturated fatty acids (40.095%) accumulated more in deep-sea fishes followed by
SFA (31.072%) and mono unsaturated fatty acids (28.83%). Gas chromatograms of fatty
acid standard with nitrogen as a carrier gas of fatty acid profiling pictures were shown in
figure 1 to 7 respectively.
Table 2: saturated and unsaturated fatty acid composition in deep-sea fishes
P. B. N. N. C. C. L.
Cyanea Caudimacula Orientalis japonicus lugubris baxteri niger
SFA (%) 37.11 30.88 12.67 34.61 33.88 42.02 26.34
MUFA (%) 35.07 18.66 55.80 17.66 23.28 26.45 24.89
PUFA (%) 27.81 50.47 31.53 47.71 42.85 31.53 48.77
60
50
40 SFA
30
MUFA
20
10 PUFA
0
Graph1: composition of saturated and unsaturated fatty acids in deep-

sea fishes
Table 1: Free fatty acid profiling in deep-sea fishes along the Indian EEZ and Central Indian Ocean
Psenopsis Bembrops Neoepinnula Nemipterus Chascanopsetta Cubiceps Lamprogrammus
Fatty acid (%) Cyanea caudimacula orientalis japonicus lugubris baxteri niger
C4:0 0 0 0 0 0 0 0
C6:0 0 0 0 0 0 0 0
C8:0 0 0 0 0 0 0 0.6
C10:0 0 0.18 0 0 0 0 0
C11:0 0 0 0 0 0 0 0
C12:0 0.04 0.26 0 0 0 0.07 0.89
C13:0 0.06 0 0 0 0 0.11 0
C14:0 3.32 1.64 2.42 3.72 2.9 5.58 0.82
C15:0 0.81 0.52 0 1.38 0 1.07 0
C16:0 22.69 19.2 8.71 20.83 23.66 26.26 17.48
C17:0 0.98 0.81 0 1.25 0 1.05 0
C18:0 7.51 6.94 1.54 6.58 7.32 5.46 4.97
C20:0 0.94 0.27 0 0.56 0 1.4 0
C21:0 0.18 0.63 0 0 0 0.3 0.91
C22:0 0.49 0 0 0.29 0 0.59 0

C23:0 0.09 0.43 0 0 0 0.13 0.67
C24:0 0 0 0 0 0 0 0
SFA 37.11 30.88 12.67 34.61 33.88 42.02 26.34
C14:1 0.11 1.91 0 0.53 0 0.07 3.19
C15:1 0 0 0 0 0 0 0
C16:1 3.79 3.48 7.42 5.24 3.52 0.49 2.02
C17:1 0.43 2.09 0 0.7 0 0.07 3.97
C18:1 trans 9 0.22 0 0 0 0 0.2 0
C18:1 cis 9 27.49 10.4 48.38 9.82 19.76 22.02 14.04
C20:1 cis 11 1.89 0.5 0 0.38 0 2.28 1.67
C22:1 cis 13 1.05 0 0 0.45 0 1.26 0
C24:1 cis 15 0.09 0.28 0 0.54 0 0.06 0
MUFA 35.07 18.66 55.8 17.66 23.28 26.45 24.89
C18:2 trans
9,12 0.1 0.31 0 0.4 0 0.21 0
C18:2 cis 9,12 0.85 1.05 1.3 2.17 0 1.12 1.34
C20:2 cis
11,14 0.29 0.38 0 0.69 0 0.39 0
C22:2 cis
13,16 0.09 0.35 0 0.5 0 0.06 0
C18:3 cis
6,9,12 gamma 0.07 0 0 0.35 0 0.44 0
C18:3 cis
6,9,12,15 alpha 0.44 0.23 0 0.88 0 1.35 0
C20:3 cis
8,11,14 0.16 0.28 0 0.34 0 0.15 0
C20:3 cis
11,14,17 0.64 0 0 0 0 0.61 0
C20:4 cis
5,8,11,14 ETA 3.56 8.69 4.09 7.88 7.86 2.36 8.11
C20:5 cis
5,8,11,14,17
EPA 5.04 6.84 5.44 5.12 4.93 4.07 3.51
C22:6 cis
4,7,10,13,16,19
DHA 16.57 32.34 20.7 29.38 30.06 20.77 35.81
PUFA 27.81 50.47 31.54 47.71 42.85 31.53 48.77

Figure 1: Fatty acid profile of Psenopsis Cyanea
Figure 2: Fatty acid profile of Bembrops caudimacula
Figure 3: Fatty acid profile of Neoepinnula orientalis
Figure 4: Fatty acid profile of Nemipterus japonicus
Figure 5: Fatty acid profile of Chascanopsetta lugubris
Figure 6: Fatty acid profile of Cubiceps baxteri
Figure 7: Fatty acid profile of Lamprogrammus niger
DISCUSSION
The main health benefits of seafood are attributed to the polyunsaturated fatty
acid content in marine lipids. The most important among PUFA are Eicosapentanoic acid
(EPA) and Docosahexanoic acid (DHA). Although the rate of denovo synthesis is very
low by fish, they accumulate EPA and DHA by consuming phytoplankton rich in these
fatty acids. The total amount of EPA and DHA in cultured fresh water fish which are
generally fed vegetable oil based diets is negligible while marine fish contain up to 25 %
out of total fatty acids. Consumption of EPA and DHA are linked to improvement in
cardiovascular functions, immunity, fetal development, cognitive function, and
inflammatory disorders. Hence, consumption of marine lipids has long term health
benefits. Nutritional studies of some potential demersal resources are of paramount
importance in the present context of diversification of fishing effort from the coastal
waters to the deeper waters. The output from the project will be in the form of precise
knowledge on the magnitude and abundance of new fishery resources from continental
slope. Demersal fish feed in deep water or on the seabed. Their meat tends to be white
and relatively low in fats. Nutritionally, the demersal group provides mainly protein.
Demersal fish in general do not have a lot of lipid in their flesh, generally lipid stores are
found in the liver. The high levels of lipid found in the liver of the Gadidae family
(including cod, coley and haddock) is exploited to produce cod liver oil, a rich source of
the Omega 3 long chain polyunsaturates as well as vitamins A and D. Information on the
nutritional profiling of demersal fish and shellfish species is useful to help consumers in
choosing fish and shellfish based on their nutrient values, besides to complement
information in food composition database.

Fatty acids may be classified as saturated (no double bonds between individual
carbon atoms) fatty acids and mono- or polyunsaturated fatty acids according to the
number of double bonds in the chain. The most common saturated fatty acids in human
food are: lauric, myristic, palmitic, and stearic acids (ranging from 12 to 18 carbon
atoms). Unsaturated fatty acids are classified into two main categories: polyunsaturated
fatty acids, represented by omega-6 fats (with main representatives being linoleic and
arachidonic acids) and omega-3 fats (with main representatives being linolenic,
eicosapentaenoic (EPA), and docosahexaenoic (DHA) acids) or monounsaturated fatty
acids, represented by the omega-9 (omega-9 – oleic) fats. Omega-3 and omega-6 fatty
acids are considered essential, as they are not synthesized by the human body. The
linoleic acid (18:2 omega-3) is the precursor of the other polyunsaturated fatty acids in
omega-6 fats, which has soy, corn, and sunflower vegetable oils as their main food
sources. In the omega-3 family, the linolenic acid (18:3 omega-3) is found in a number of
vegetables, such as canola and linseed, whereas EPA (20:5 omega-3) and DHA (22:6
omega-3) are found in deep sea cold water fish (mackerel, sardines, salmon, herring). On
the other hand, oleic acid (18:1 omega-9) can be synthesized by the body and its main
diet sources are olive oil, canola oil, olives, avocado and nuts (peanuts, chestnuts,
walnuts, almonds) (Bhangle and Kolasinski, 2011).
Deep-sea fishes are considered to be not only food with good source of quality
protein but also food with healthy components. Several kind of deep-sea fish already be
important food and often looked in the markets. Deep sea is part of the marine
environment that lies below the depth that can be illuminated by sunlight in the open sea
and deeper than the continental shelf (> 200 m) (Nybakken, 1992). The study of Suseno
et al. (2007) showed that several deep sea species from western Sumatra Ocean such as
Dietmoides pauciradiatus, Benthodesmus tenuis, Beryx splendens, Hoplothethus
crassipinus, Hoplothethus sp., Ophidiidae, Ostracoberyu dorygenis, and Godamus colleti
had positive results for the presence of steroid compounds. Fish and seafood provide
some of the leanest sources of protein and are one of the only natural sources of the
omega 3 fatty acids - EPA and DHA. The American Heart Association recommends that
everybody gets at least 2 servings of oily fish each week because research has proved
time and again that both EPA and DHA support a healthy cardiovascular system.
Fatty acids play an important role in the life and death of cardiac cells because
they are essential fuels for mechanical and electrical activities of the heart (Honore et al
1994; Reiffel and McDonald 2006; Landmark and Alm 2006). The fatty acid composition
of fish tissue can be affected by diet, size, age, reproductive cycle, salinity, temperature,
season and geographical location (Lucia et al., 2003). The fatty acid composition of
different fish from the same species can vary because of diet, location, gender and
environmental conditions (Gruger, 1967). The demersal fish contained more MUFA than
pelagic fish, while pelagic fish contained more PUFA than demersal fish (Kusumo,
1997). Consumption of fish rich in n-3 fatty acids is suggested to reduce cholesterol
cancer risk through inhibition of the arachidonic acid cascade that plays a role in
inflammation and has been linked to carcinogenesis (Roynette et al., 2004). Deficit in
omega-3 fatty acids can result in neurological abnormalities and retardation on growth,
weakness, reduced learning ability, disturbances in motor coordination, changes in
behavior, high level of triglycerides, high blood pressure, oedema, dry skin, mental
retardation, immune dysfunction etc. all symptoms of deficit can disappear with the
return of omega-3 into nutrition. The distinctive fatty acid composition found in the
edible part (large amount of MUFA), can be due to the metabolic functions of this organ,
namely as storage of energy, since the MUFA, as well as the SFA, are quantitatively
more important than PUFA for the supply of metabolic energy (Sargent, 1995). The
predominance of unsaturated fatty acids and particularly the high levels of highly
unsaturated fatty acids agree, in general, with the known fatty acid composition of marine
fishes (Gruger et al. 1964; Sargent et al., 1973; Yamada and Hayashi, 1975; Body, 1983;
Henderson and Almatar, 1989; Bakes et al., 1995).
Polyunasaturated fatty acids (PUFAs) are important structural components of cell
membranes, and are useful in growth and development of human beings (Chakraborty et
al., 2010). PUFAs, especially of longer chain length, viz., EPA and DHA were reported
to be found abundant in marine fish, and have beneficial properties to the prevention of
atherosclerosis and other diseases. A comparison of the fatty acid composition and the
depth of fish habitat have no correlation related to the profile of saturated fatty acid
(SFA), monounsaturated acid (MUFA) and polyunsaturated acid (PUFA). These are more
related to the food chain while deep sea fish moved vertical mobile at night for look far
distance a feed for example plankton and phytoplankton. In generally they showed
similar fatty acid composition with depth difference. A comparison of omega-3 fatty acid
content of various foods has clearly demonstrated that dietary sources of EPA and DHA
are almost entirely confined to fish (Givens et al 2000). Thus, fish represents a virtually
unique source of n-3 HUFAs. Consumption of fish has been increasing worldwide. In
USA, per capita consumption of fish has increased from an average of 4.5 kg in 1960 to
about 7.4 kg in 2005 as per the data provided by the National Marine Fisheries Service,
U.S. Department of Commerce (2006). The annual per capita consumption of fish in
India is still very low, although it has steadily risen from 2.9 kg before 1981 to 4.7 kg
after 2000 (Venkiteswaran 2007). The highest value of PUFA was found at
Neoscophelus porosus while the highest of SFA was Lamprogrammus niger.
Ostracoberyx dorygeny had the highest concentration of oleic acid (443.730 mg/gr) while
the lowest oleic acid was found at Seriola sp. (66.174 mg/g). Eskimos diet has a large
amount of seafood, which is a source of polyunsaturated fatty acids (Kronmann and
Green, 1980). Thereafter, several studies were conducted to find the role of these lipids in
many chronic diseases in which inflammation plays a major pathogenetic role.
Very modest literature is available regarding the biochemical compositions of
deep sea creatures belonging to the Indian EEZ and hence it was decided to analyse the
resources obtained during cruises on the FORV Sagar Sampada with appreciable lipid
content of deep-sea fishes and thereby study their fatty acid profiles. A 2007 study of
nearly 12,000 pregnant women found that children born to mothers who ate more than
340 grams of seafood per week during pregnancy scored six points higher on tests of
verbal IQ than kids born to mothers who had other foods on the menu. Fish oil appears to
have anti-inflammatory properties, and has been researched as a treatment for many
conditions including inflammatory bowel disease and rheumatoid arthritis. It also has
some preventive effect for Parkinson’s disease (Stark et al., 2015). The omega-9 fatty
acids are effective in decreasing low density lipoproteins cholesterol blood and also for
increasing high density lipoproteins cholesterol blood. They also have better capability
for increasing high density lipoproteins than omega-3 and omega-6nand also the
capability to impede the production of eicosanoid structure (stimulants causing cancer in

animal tests) (Pranoto, 2006). The fatty acid composition of deep-sea fish examined
were similar to that reported previously (Hayashi and Kishimura, 2003). Deep sea fish is
composed mainly of monoenoic fatty acid, which is important for industrial use in food
processing of hydrogenated fish oil, and are raw materials of margarine, fat spread or
shortening.
The fatty acid composition of the species investigated is summarized in the Table
1. The results demonstrated that a significant part of the fatty acid in the muscles was
saturated acid (SFA), monounsaturated fatty acid (MUFA) and polyunsaturated fatty
acids (PUFA). The content of SFA varied from 12.67% to 42.02% of the total fat content
among the different species observed, the range of MUFA content was between 17.66%
and 55.80% and PUFA varied from 27.81% to 50.47%. Deep sea fish is composed
mainly of monoenoic fatty acid, which is important for industrial use in food processing
of hydrogenated fish oil, and are raw materials of margarine, fat spread or shortening.
According to Okland et al. (2005) the deep-sea teleosts and elasmobranchs contain lipids
that consisted mainly of polyunsaturated fatty acid (PUFA) and docosahexaenoic acid
(DHA). In his studies on fatty acid profiles and fat contents of commercially important
seawater and freshwater fish species Ozagul et al. (2007) showed that the proportion of
n3 PUFAs of seawater fish were higher than that of fresh water. The concentration of ω3
PUFAs differs among species and can be influenced by a number of factors. Due to their
molecular similarity, linoleic acid and - linolenic acid compete for the same enzymes to
synthesize derivatives with 20 carbon atoms: Arachadonic acid (AA) (20:4 omega-6),
eicosapentaenoic acid (EPA) (20:5 omega-3) e eicosatrienoic acid (ETA) (20:3 omega-9).
The -linolenic acid has a 22-carbon atom derivative, the docosahexaenoic acid (DHA)
(22:6 omega- 3) ((Bhangle and Kolasinski, 2011; Bistrian, 2003; Leonard et al.,
2004).Thus, the ratio between daily intake of omega-6 and omega-3 fatty-acid sources
assumes great importance in human nutrition (Bistrian, 2003).
The positive health benefits of fish and omega-3 fatty acids are well established
(Flick and Martin, 1992; Stone, 1996; Kris-Etherton et al., 2002; Mozaffarian and Rimm,
2006). EPA and DHA are nutritionally important n-3 fatty acids. Observational studies,
meta-analyses, intervention trials and systematic reviews suggest that intake of EPA and
DHA prevents fatal cardiovascular diseases (Brouwer, 2008). EPA acts as a precursor for
prostaglandin-3, thromboxane-3 and leukotriene-5 groups. DHA is a major fatty acid in
brain phospholipids and in the retina. The fish species cannot be compared to fat fish
species when it comes to total amount of n-3 FAs.
Previous studies have also demonstrated a beneficial role of lean fish, as well as
fatty fish consumption, in the prevention of cardiovascular diseases (Kromhout, 2001).
The bony species, roughhead grenadier and mora contained almost five times more EPA
than the cartilaginous species. DHA accounted alone for about 30–40% of total fat
content, which is considered to be particularly high. However, it has previously been
demonstrated such high ratio of DHA in the muscle of the tuna Euthynnus (Katsuwonus)
pelamis (Saito et al., 1997). Arachidonic acid (20:4n-6) was the dominant n-6 fatty acid
in muscle from all the deep-sea fish species examined. The content of arachidonic acid
was more than twice as high as the level found in muscle in the upper strata, cod and
piked dogfish (Hege et al., 2005).

In India, deep-sea fisheries sector is not well developed probably due to lack of
awareness among fisher folk, poor governmental support together with short fall of
expertise. The current level of commercial exploitation of deep-sea resources is limited
only to deep-sea fishes (Vivekanandan, 2011). However, unfamiliar appearance and rapid
discoloration due to melanosis made less market preference for these species. It is
essential to assess the nutritive value of these resources as many of them are thrown back
to the sea because of the non availability of information pertaining to its biochemical
composition. Information about composition of deep-sea edible part might raise their
value as table food. A very few literature is available for either fin fish or shell fish from
deep waters off the Indian EEZ. The studied deep-sea fish species also contains
significant concentration of Eicosapentaenoic acid (EPA), another important omega -3
PUFA, essentially required for the regulation of neuronal and cardiovascular functions.
omega -3 PUFA are well known to exert neuroprotective properties and to represent
potential nutritional and therapeutic interventions in the treatment for variety of
neurodegenerative and neurological diseases and disorders such as Alzheimer's Disease,
Brain Cancer, Degenerative Nerve Diseases, Epilepsy, Genetic Brain Disorders
Encephalitis, Head and Brain Malformations, Hydrocephalus, Stroke, Parkinson's
Disease, Multiple Sclerosis, Amyotrophic Lateral Sclerosis etc. Other potential beneficial
actions of EPA and DHA include regulation of inflammation, transcription, and cell
membrane properties. It is noticed that the other fatty acids like palmitic acid, stearic acid
and eicosenoic acid form lesser part in composition of fatty acids as compared to omega -
3.Nutritional lipid quality indexes such as total omega-3/ omega-6, DHA and EPA
consumption of the selected deep-sea fish species are advisable in a balanced diet.
The deep-sea fish species from the present study contains a particularly good lipid
composition for nutritional purposes. It should however be noted that the net content of
n-3 and n-6 is low because of the low lipid content of these fish species, and only regular
consumption of such fish species could contribute to increase the amount of omega-3 and
omega-6 fatty acids in the diet. This study demonstrates that several of the underutilized
deep-sea species are able to compete with more commercially utilized species in terms of
nutritional value, and they can definitely also compete when it comes to taste.
SUMMARY
&
CONCLUSION
The wealth of marine resources was assumed to be an unlimited gift of nature.
Deep sea fishes that used to live in extreme environment are thought to have nutritional
and bioactive compounds that are beneficial for human health. Deep-sea fishes
(myctophids) can be utilized for the production of commercial fishery products like fish
meal, fish oil, fish silage, surimi, seasoning products, feed for cultured fish, nutrient
resource in the formulation of poultry feed as well as crop fertilizers and products like
lubricating oil, cosmetics and wax. Deep-water fisheries are becoming more and more
important, and there is a paucity of safety monitoring of the ecosystem. The food
consumption and metabolism of deep-sea fish was not much studied. Some deep-sea
pelagic groups, such as the lantern fish, ridge head, marine hatchet fish, and light fish
families are sometimes termed pseudoceanic because, rather than having an even
distribution in open water, they occur in significantly higher abundances around
structural oasis, notably seamounts and over continental slopes. The phenomenon is
explained by the likewise abundance of prey species which are also attracted to the
structures.
Information on the proximate composition will be important when utilization of
new species of deeper water fishes is considered. This is because deeper water fishes are
considered to be not only food with good source of quality protein but also food with
healthy components. The present study concerning to proximate composition of cruise
wise variations of moisture, protein, fat and ash in the edible part of deeper waters are of
Alepocephalus bicolor, Narcetes erimelas, Talismania longifilis, Chlorophthalmus
bicornis, Lamprogrammus niger, Beryx splendens, Chelidoperca investigatoris,
Neoepinnula orientalis, Cubiceps baxteri and Psenopsis cyanea along the continental
slope (200m – 1200 m depth) of Indian EEZ (Exclusive Economic Zone). Proximate
composition of fish species greatly varies due to physiological reasons and changes in
environmental conditions, i.e., spawning, migration, and starvation or heavy feeding.
This overview will explain the percentages of moisture, protein, fat and ash compositions
of selected deep-sea fishes caught from the Indian EEZ (Exclusive Economic Zone).
The present study provides information on proximate composition of few deep
sea fishes. The results indicated that the fish resources analysed contain significant
protein content, and hence can be exploited commercially for meeting protein
requirements. The findings of the study are important in the perspective of exploring deep
sea resources as edible seafood. In a fast growing country like India, exploitation of deep
sea resources within the judicious management practices is necessary to overcome
resource crunch to ensure food and nutritional security.
The presence of heavy metals in the aquatic environment is a major concern
because of their toxicity and threat to plant and animal life disturbing the natural
ecological balance. The specific problem associated with the heavy metals in the
environment is their accumulation through food chain and persistence in nature. In fact,
heavy metal pollution and its management has been a major global concern for
environmentalists due to their non-biodegradable and hazardous nature. Fish absorb
metals through ingestion of water or contaminated food. Heavy metals have been shown
to undergo bioaccumulation in the tissue of aquatic organisms. On consumption of fish
and other aquatic organisms these metals are transferred to man. Uptake of heavy metals
through food chain in human being may cause various physiological disorders like
hypertension, sporadic fever, nausea, renal damage, cramps etc. Other toxic responses
include irritability, less appetite, damage to nervous system and kidneys, anemia and
gastrointestinal problems. The present study pertaining to monitoring of heavy metals
like copper, zinc, iron, cobalt, cadmium, lead, chromium, selenium and arsenic
accumulation in the given above selected ten deeper water fishes
The current study results represent that consumption of these deep sea fishes do
not pose threat of heavy metals to consumers as their levels were in below detectable
levels/negligible. The nominal differences noticed in the accumulation of metals between
the deeper water fishes could be attributed to the variability of feeding habits, ecological
needs, metabolism, age and size of the species and their habitats. The concentrations of
heavy metals analyzed were well within the regulatory limits indicating that the samples
off deep waters do not pose any threat to the consumers. Therefore the results of this
study provide new information on the concentration of heavy metals in deeper water
fishes from Indian EEZ (Exclusive Economic Zone) which could serve as a useful tool
for future assessment programme can be compared.
In this study, the fatty acid profile shows that the edible part of deeper water
fishes contains more percentage of unsaturated fatty acids than saturated fatty acids. In P.
Cynea, saturated fatty acids (37.11 %) form the major fatty acids followed by
monounsaturated fatty acids (35.07%) and polyunsaturated fatty acids (27.81 %),
whereas in B. caudimacula, N. japonicas, C. lugubris and L. niger accumulated more
percentage of polyunsaturated fatty acids (50.47%, 47.47%, 42.85% and 48.77%)
followed by saturated fatty acids (30.88%, 34.61%, 33.88% and 26.34%) and
monounsaturated fatty acids (18.66%, 17.66%, 23.28% and 24.89%), while in N.
Orientalis forms more content of monounsaturated fatty acids (55.80 %) followed by

polyunsaturated fatty acids (31.53 %) and saturated fatty acids (12.67 %) and in C.
baxteri contains more percentage of saturated fatty acids (42.02 %) followed by
polyunsaturated fatty acids (31.53 %) and monounsaturated fatty acids (26.45 %)
respectively. The most abundant fatty acids were oleic acid, palmitic acid,
docosahexaenoic acid, eicosapentaenoic acid, stearic acid, palmitoleic acid; of the total
fatty acids. Palmitic acid (C16:0) was found to be the most prominent fatty acid. SFA has
the principal component as palmitic acid. Oleic acid (C18: 1n9) is the major component
of MUFA. Docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and ɣ linolenic
acid (GLA) constitutes the principal components of PUFA.
Present study shows that deep sea fishes is a good source of protein, fat and
PUFA and hence it could will be a potential source of alternative low cost protein and fat
for future. At present, there is no demand for the resource and so used in the fish meal
industry. More studies are required for the development of value added products.
Research is required on post harvest technologies, like processing, marketing and
utilization for better profit from this sector. Their enormous biomass may make them
suitable for much greater commercial exploitation in future.
Ultimately, the conclusion have also been afforded for easy perceptive of the
current study. The findings accomplish during the research study period were
meticulously discussed and compared with related works done in India and abroad.
Especially, the nutritional awareness aspects on presence of healthcare important omega
– 3 polyunsaturated fatty acids in deep-sea fishes is relatively low due to non-availability
of database on the biochemical composition, monitoring of heavy metals and nutritional
profiling studies in deeper water fishes available in Indian waters.

The present study accomplish the generation of proper information on the
proximate composition, heavy metals and free fatty acid profiling studies of Indian EEZ
(Exclusive Economic Zone) and will be more important when utilization of new species
of deeper water fishes for human healthcare benefits especially in pregnant women,
young children and aged population. This is because deeper water fishes are considered
to be not only food with good source of quality protein but also food with healthy
components.
REFERENCES
Abdallah. and Mohamed, M.A. (2007) Speciation of trace metals in coastal sediments of
El- Max Bay south Mediterranean Sea-west of Alexandria, (Egypt). Environ.
Monitor. Assess. 132: 111-113.
Abdel-Moati, M.A.R. (1998) Speciation of selenium in a Nile Delta lagoon and SE
Mediterranean Sea mixing zone. Estuarine. Coastal. Shelf. Sci. 46: 621-628.
Achterberg, E.P., Holland, T.W. Bowie, A.R. Mantoura, R.F.C. and Worsfold. P.J.
(2001) Determination of iron in seawater. Anal. Chim. Acta. 442:1-14.
Ackman, R.G. (1989) Nutritional composition of fats in sea foods. Prog. Food. Nutr. Sci.
13: 161-289.
Ademoroti, C.M.A. (1996) Environmental Chemistry and Toxicology, Foludex Press
Ltd., Ibadan. p. 171-204.
Adeyeye, E.I., Akinyugha, R.J., Febosi, M.E. and Tenabe, V.O. (1996) Determination of
some metals in Clarias gariepinus (Cuvier and Valenciemes), Cyprinus carpio (L)
and Oreochromis niloticus (L) fishes in a polyculture fresh water pond and their
environment. Aquacut. 47: 205-214.
Agrawal, M. and Srivastava N. (2003) Effect of chronic zinc exposure on the thyroid
gland activity of a fresh water fish Channa punctatus (Bloch). J. Ecophysio.
Occup. Hlth. 3: 273-278.
Ali, M., Iqbal, F., Salem, A., Iram, S. and Athar, M. (2005) Comparative study of body
composition of different fish species from brackish water pond. Int. J. Environ.
Sci. Tech. 22: 77-80.

Altas, L. and Buyukgungor, H. (2007) Heavy metal pollution in the Black Sea shore and
offshore of Turkey. Environ. Geol. 52(3): 469-476. doi 10.1007/s00254-006-
0480-1.
Andres, S., Ribeyre, F., Toureneq, J.N. and Boudou. (2000) Interspecific comparison of
cadmium and Zinc contamination in the organs of four fish species along a
polymetallic pollution gradient (Lot River, France). Sci. Total. Environ. 284: 11-
25.
Ansari, Z.A., Achuthankutty, C.T., Dalal, S.G. (2006) Research Paper entitled Ecosystem
based Fishery Management for Sustainable Coastal Fisheries in Goa West Coast
of India. National Institute of Oceanography, Dona Paula, Goa, India.
Armstrong, S.G., Leach, D.N. and Wyllie, S.G. (1991) Nutritional Evaluation of Lipids in
Fish from Temperate Australian Waters. J. Food. Sci. 56: 1111-1112.
Asha, J.K. and Vijayalakshmi, R.N. (1999) Concentration of metals in fishes from Thane
and Bassein creeks of Bombay, India. Indian. J. Mar. Sci. 28: 39-44.
Ashraf, W. (2005) Accumulation of heavy metals in kidney and heart tissues of
Epinephelus microdon fish from the Arabian Gulf. Environ. Monit. Assess. 101:
311-315
ATSDR (Agency for Toxic Substances and Disease Registry) (2007) Agency of toxic
substances and disease registry. http://www.atsdr.cdc.gov/. Accessed May 2013.

Ayas, Zafer, Guler, E., Sedat, V.Y. and Murat, O. (2007) Heavy metal accumulation in
water, sediments and fishes of Nallihan Bird Paradise, Turkey. J. Environ. Biol.
28: 545-549.
Badach, G. and Heyer, K. (1997) Cd budget for the German bight in the North Sea. Mar.
Pollution. Bull. 84: 875-881.
Bailey, T.G. and Robison, B.H. (1986) Food availability as a selective factor on the
chemical composition of mid water fishes in the eastern North Pacific. Mar. Bio.
91: 131-141.
Bakan, G. and Boke, O.H. (2007) An ecological risk assessment of the impact of heavy
metals in surface sediments on biota from the mid-Black Sea coast of Turkey. Int.
J. Environ. Studies. 64: 45-57. doi: 10.1080/00207230601125069.
Bakan, G. and Buyukgungor, H. (2000) The Black Sea. Marine. Poll. Bull. 41: 24-43.
doi: 10.1016/S0025-326X(00)00100-4.
Bakes, M.J., Elliot, N.G., Green, G.J. and Nichols, P.D. (1995) - Variation in lipid
composition of some deep-sea fish (Teleostei: Oreosomatidae and
Trachichthydae). Comp. Biochem. Physiol. 111B: 633-642.
Balachandran, K. and Nizar. A.M. (1990) A check list of fishes of the Exclusive
Economic Zone of India collected during the research cruises of FORV Sagar
Sampada. In: Proc. First Workshop Scient. Resul. FORV Sagar Sampada, 1989.
305-324.
Balchand, A.N., Nair, S.M. and Nambisan, P.N.K. (1990) A survey of the protein content
in estuarine waters. Toxical Environ. Chem. 27: 285-296.
Bandarra, N.M., Batista, I., Nunes, M.L., Empis, J.M. and Christie, W.W. (1997)
Seasonal changes in lipid composition of Sardine (Sardina pilchardus). J. Food.
Sci. 62: 40-42.
Bande, V.N., Menon N.G. and Balachandran, K. (1990) Studies on the distribution and
abundance of Bull's eye (Priacanthus spp.) in the EEZ of India. In: Proc. First
Workshop Scient. Resul. FORV Sagar Sampada, 1989. 233-239.
Barak, N. and Mason, C. (1990) A Survey of Heavy Metal Levels in Eels (Anguilla
anguilla) from Some Rivers in East Anglia, England - The Use of Eels as
Pollution Indicators. Int. Rev. Gesamten. Hydrobiol. 75: 827-833.
Barclay. M.C., Dali. W. and Smith. D.M. (1983) Changes in lipid and protein during
starvation and the moulting cycle in the tiger prawn. Penaeus esculentus Haswell.
J. Exp. Mar. Biol. Ecol. 68: 229-244.
Basa., Siraj, P. and Usha Rani, A. (2003) Cadmium induced antioxidant defense
mechanism in freshwater teleost Oreochromismossambicus (Tilapia).
Eco.Toxicol. Environ. Saf. 56: 218-221
Bat, L. (2005) A Review of Sediment Toxicity Bioassays Using the Amphipods and
Polychaetes. Turkish. J. Fish. Aquat. Sci. 5: 119-139.

Bat, L., Gokkurt, O., Sezgin, M., Ustun, F. and Sahin, F. (2009) Evaluation of the Black
Sea Land Based Sources of Pollution the Coastal Region of Turkey. The Open
Marine Biology Journal. 3: 112-124. doi: 10.2174/18744508009030100112.
Begum, A., Harikrishna, S. and Khan, I. (2009) Analysis of heavy metals, sediments and
fish samples of madivala lakes of Bangalore, Karnataka. Int. J. Chem. Tech. Res.
1: 245-249.
Beltz, B.S., Tlusty, M.F., Benton, J.L. and Sandeman, D.C. (2007) Omega-3 fatty acids
upgregulate adult neurogenesis. Neuroscience Letters. 415: 154-158.
Berbert, A.A., Kondo, C.R., Almendra, C.L., Matsuo, T. and Dichi, S. (2005)
Supplementation of fish oil and olive oil in patients with rheumatoid arthritis.
Nutr. 21: 131-136.
Bhangle, S. and Kolasinski, S.L. (2011) Fish oil in rheumatic diseases. Rheum. Dis. Clin.
N. Am. 37:77-84.
Bhuiyan, A.K.M., Ratnayake, W.M.N. and Ackman, R.G. (1993) Nutritional composition
of raw and smoked Atlantic mackerel (Scomber scombrus): oil and water soluble
vitamins. J. Food. Comp. Anal. 6: 172-184.
Bhuiyan, H.R., Nath, K.K., Seal, P. and Hossain, M.E. (2006) Fatty Acid Composition of
Three Marine Fishes of the Bay of Bengal Bangladesh, Journal of Science and
Industrial Research, Karachi. 41: 47-54.

Bhuyan, H.R., Chowdhury, M.B., Nath, K.K.., Seal, P. and Hag, M.A. (2003) Studies on
the biochemical parameters of Cynoglossids in the Kutuboha Channel,
Bangladesh. Bang. J. Sci. Ind. Res. 38: 91-96.
Birader, R.S. (1989) Course Manual Fisheries Statistics. Central Institute of Fisheries
Education. 110-113.
Bistrian, B.R. (2003) Clinical aspects of essential fatty acid metabolism: Jonathan Rhoads
lecture. J. Parenter. Enteral. Nutr. 27:168-175.
Biswas, B.K., Dhar, R.K., Samanta, G., Mandal, B.K., Chakraborti, D., Farak, I., Islam,
K.S., Chowdhury, M.M., Islam A. and Bay, S. (1998) Detailed study report of
Samta, one of the arsenic affected villages of Jessore District, Bangladesh. Curr.
Sci. 74: 134-145.
Blaber, S.J.M. (2000) Tropical Estuarine Fishes: Ecology, Exploitation and Conservation.
1stEdn Blackwell Sci., Oxford.
Body, D.R. (1983) The nature and fatty acid composition of the oils from deep-sea fish
species from New Zealand waters. J. Sci. Food. Agricult. 34: 388-392.
Body, D.R. and Vlieg, P. (1989) Distribution of the lipid classes and eicosapentaenoic
(20:5) and docosahexaenoic (22:6) acids in different sites in blue mackerel
(Scomber australasicus). fillets. J. Food. Sci. 54: 569-572.
Boran, M. and Altinok, I. (2010) A review of heavy metals in water, sediment and living
organisms in the Black Sea. Turkish. J. Fish. Aquat. Sci. 10: 565-572. doi:
10.4194/trjfas.2010.0418.
Borlongon, I.G. (1992) Dietary requirement of milk fish, Chanos chanos (Forskol)
Juveniles for total aromatic amino acids. Aquacult. 102: 309-317.
Bouriga, N., Selmi, S., Faure, E. and Trabelsi, M. (2010) Biochemical composition of
three Tunisian silverside (fish) populations caught in open sea, lagoon and island
coasts. Afr. J. Biotechnol. 9: 4114-4119.
Brewer, G.J. (2010) Copper toxicity in the general population. Clin. Neurophysiol. 121:
459-460
Brooks, B.R. and Fiumsby, M.G. (1965) The biogeochemistry of trace element uptake by
some New Zealand bivalves. Limnol. Ocean. Org. 12: 521-527.
Brouwer, I.A. (2008) Fish, omega-3 fatty acids and heart disease. In : Borresen, T. (Ed.),
Improving seafood products for the consumer. CRC Press, Woodhead Publishing
Limited, England, p 165.
Bruce, J.R. (1924) Changes in the chemical composition of the tissue of the herring in
relation to age and maturity. Biochemic. J. 18: 469-485.
Bryan, G.W. (1976) Some aspects of heavy metal tolerance in aquatic organisms. In:
A.P.M. Lockwood (Ed.), Effects of Pollutants on Aquatic organisms. Cambridge
University Press. UK: 7-34.
Burger, J. and Gochfeld, M. (2005) Heavy metals in commercial fish in New Jersey.
Environ. Res. 99: 403-413.

Busalmen, J.P., Roura, S.I., Roldan, H. and Crupkin, M. (1995) Changes in the lipids and
biochemical properties of actomyosin from Preand Post-spawned hake
(Merluccicus hubbsi Marini). Comp. Biochem. Physiol. 112: 743-748.
Cahu, C., Salen, P. and De Lorgeril, M. (2004) Farmed and wild fish in the prevention of
cardiovascular diseases: Assessing possible differences in lipid nutritional values.
Nutr. Metab. Cardiovasc. Dis. 14: 34-41.
Calabrese, E.J., Canada, A.T. and Sacco, C. (1985) Trace elements and public health.
Annu. Rev. Pub. Health. 6: 131-146.
Canli, M. and Atli, G. (2003) The relationship between heavy metal (Cd, Cr, Cu, Fe, Pb
and Zn) levels and size of six Mediterranean fish species. Environ. Pollut. 121:
129-136.
Castoldi, F., Coccini,T. and Manzo,L. (2003) Neurotoxic and molecular effects of
methylmercury in humans. Rev. Environ. Hlth. 18: 5-24.
Castro, P. and Huber M. (1997) Marine Biology. 2 ed. WCB/McGraw Hill.
CCREM (Canadian Council of Resource and Environment Ministers). (1987) Canadian
water quality guidelines. Prepared by the Task Force on Water Quality
Guidelines.
Celik, M. (2008) Seasonal changes in the proximate chemical compositions and fatty
acids of chub mackerel (Scomber japonicus) and horse mackerel (Trachurus
trachurus) from the north eastern Mediterranean Sea. Int. J. Food Sci. Technol.
43: 933-938.
Centeno, R., Collados, M., and Trujillo B.J. (2005) ApJ, submitted Collados, M. 2003,
Proc. SPIE, 4843, 55.
Chakraborty, K., Vijayagopal, P., Chakraborty, R.D. and Vijayan, K.K. (2010)
Enrichment of eicosapentaenoic acid concentrates from sardine oil by bacterial
(Bacillus circulans) lipase isolated from seaweed Turbinaria conoides. Food.
Chem. 120: 433-442.
Chakraborty, S., Ghosh, S. and Bhattacharyya, D.K. (2004) Lipid profiles of Bhola bhetki
(Nibea soldado) organs. J. Oleo. Sci. 53: 367-370.
Chakraborty. S., Ghosh, S. and Bhattacharyya, D.K. (2005) Lipid profiles of pomfret fish
(Pampus argenteus) organs. J. Oleo. Sci. 54: 85-88.
Chale, F.M.M. (2002) Trace metal concentrations in water, sediments and fish tissue
from lake Tanganyika. Sci. Total. Environ. 299: 115-121.
Chanan, H.J. and Saby, M.K.E.I. (1932) Fat metabolism of the herring. I. Preliminary
survey. Biochemic. J. 26: 2021-2034.
Chandrasekhara rao, A. and Krishnan, L. (2011) Biochemical composition and changes
in biological indices associated with maturation of the ovary in the spiny cheek
grouper Epinephelus diacanthus (Valenciennes, 1828). Indian. J. Fish. 58: 45-52.
Chezhian, A., Kabilan, N., Kumar, S.T., Senthamilselvan, D. and Sivakumari, K. (2010)
Impact of common mixed Effluent of spicot industrial Estate on histopathological
and biochemical changes in Estuarine fish Lates calcarifer. Curr. Res. J. Boil.Sci.
2: 201-209.
Childress, J.J. (1971) Respiratory rate and depth of occurrence of midwater animals.
Limnol. Oceanogr. 16: 104-106.
Childress, J.J. and Nygaard, M.H. (1973) The chemical composition of midwater fishes
as a function of depth of occurrence off Southern California. Deep. Sea. Res. 20:
1093-1109.
Childress, J.J. and Nygaard, M.H. (1974) The chemical composition and relative
buoyancy of midwater crustaceans as a function of depth off Southern California.
Mar. Bio. 27: 225-238.
Childress, J.J., Cowles, D.L., Favuzzi, J. and Mickel, T.J. (1990a) Metabolic rates of
benthic deep-sea decapod crustaceans decline with increasing depth primarily due
to the decline in temperature. Deep. Sea. Res. 37: 929-949.
Childress, J.J., Price, M.H., Favuzzi, J.A. and Cowles, D.L. (1990b) Chemical
composition of midwater fishes as a function of depth occurrence off the
Hawaiian Islands: food availability as a selective factor. Mar. Bio. 105: 235-246.
Clark, M.R. (2001) Are deepwater fisheries sustainable. The example of orange roughy.
Fish. Res. 51: 123-135.
Commission of the European Communities (2001) Commission Regulation (EC) No.
221/2002 of 6 February 2002 amending regulation (EC) NO. 466/2002 setting
maximum levels for certain contaminants in foodstuffs. Official Journal of the
European Commucities, Brussels, 6 February 2002.
Covington, M.B. (2004) Omega-3 fatty acid. Am. Fam. Physician. 70: 133-140.
Cowles, D.L., Childress, J.J. and Wells, M.E. (1991) Metabolic rates of midwater
crustaceans as a function of depth occurrence off the Hawaiian Islands: food
availability as a selective factor. Mar. Bio. 110: 75-83.
Cox, P.A. (1995) The Elements on Earth: Inorganic Chemistry in the Environment.
Oxford University Press, Oxford, pp 287.
Crabtree, R.E. (1995) Chemical composition and energy content of deep sea demersal
fishes from tropical and temperate regions of the western north atlantic. Bull. Mar.
Sci. 56: 434-449.
Crawford, M. (2000) Placental delivery of arachidonic and docosahexaenoic acids:
implications for the lipid nutrition of preterm infants. Am. J. Clin. Nutr. 71:275-
284.
CSHPF Conseil Superieur d’Hygiene Publique de France. (1995) Plomb, cadmium et
mercure dans l’alimentation:´Evaluation et gestion du risque. Minist‘ere du
Travail et des Affaires Sociales,´editeur. Paris: Lavoisier Tec. Doc.
CSIR (Council of Scientific and Industrial Research). (1962) Fish and Fisheries Raw
Materials, India. 4: 132p.
Cullen, W.R. and Reimer, K.J. (1989) Arsenic speciation in the environment. Chem. Rev.
89:713-764.
Cutter, S.L., Mitchell, J.T. and Scott. M.S. (2000) Revealing the Vulnerability of People
and Places: A Case Study of Georgetown County, South Carolina. Ann. Assoc.
Am. Geogr. 90:713-737.

Dali, W. (1981) Lipid absorption and utilization in the Norwegian lobster Nephrops
norvegicus (L.). J. Exp. Mar. Biol. Ecol. 50: 33-45.
Daramola, J.A., Fasakin, E.O. and Adeparusi, E.O. (2007) Changes in physicochemical
and sensory characteristics of smoke-dried fish species stored at ambient
temperature. Afr. J. Food Agric. Nutr. Dev. 7: 169-183.
Das, K.K., Dastidar, S.G., Chakrabarty, S. and Banerjee, S.K. (1980) Toxicity of
mercury: a comparative study in air – breathing and non air – breathing fish.
Hydrobiologia. 68: 225-229.
Das, K.P. (2009) Effect of ambient temperature, icing and freezing on nutrient
composition of Rohu (Labeo rohita), Grass carp (Ctenopharyngodon idella) and
Tilapia (Oreochromis mossambica). B.Sc Thesis, Fisheries and Marine Resources
Technology Discipline, Khulna University, Khulna, Bangladesh, 20-30.
Davis, R.A. (1991) Oceanography, An Introduction to the Marine Environment Ed ke-2.
Wm. C. Brown Publishers, USA.
De Schamphelaere, K.A.C., Koene, J.M., Heijerick, D.G. and Janssen, C.R. (2008)
Reduction of growth and haemolymph Ca levels in the freshwater snail
Lymnaea stagnalis chronically exposed to cobalt. Ecotoxicol. Environ.
Saf. 71: 65-70.
Deepu, A.V., Divya V.H. and Kurup, B.M. (2007) Catch and biology of Alepocephalus
bicolour (Alcock, 1891) from the southwest coast of India. J. Mar. Biol. Ass.
India. 49: 239-242.

Demet K. and Hulya T. (2012) Proximate and Fatty Acid Composition of Some
Commercially Important Fish Species from the Sinop Region of the Black Sea.
Lipids. 47:635-641. DOI 10.1007/s11745-012-3658-1
Demirezen, D. and Uruc, K. (2006) Comparative study of trace elements in certain fish,
meat and meat products. Meat. Sci. 74: 255-260.
Dempson, J.B., Schwarz, C.J., Shears, M. and Furey, G. (2004) J. Fish. Biol. 64: 1257-
1271.
Dietz, C., Sanz, J., Ximenez, P., Madrid, Y. and Camara, C. (2003) Volatile organo-
selenium speciation in biological matter by solid phase microextraction -
moderate temperature multicapillary gas chromatography with microwave
induced plasma atomic emission spectrometry detection. Analytica Chimica Acta:
International Journal Devoted to All Branches of Analytical Chemistry. 501: 157-
167. DOI: 10.1016/j.aca.2003.09.027.
Divya, T., Hashim M. and Jayaprakash, A.A. (2007) Distribution and abundance of deep
sea eel, Gravialiceps taeniola along the continental slope off Indian EEZ. J. Mar.
Biol. Ass. India. 49: 81-85.
Donga, C.D., Chena, C.W. and Liub, L.L. (2004) Seasonal variation in the composition
and concentration of butyltin compounds in marine fish of Taiwan. Environ.
Pollut. 131: 509- 514.
Donnelly, J., Toeres, J.J., Hopkins, T.L. and Lancraft, T.M. (1990) Proximate
composition of Antarctic mesopelagic fishes. Mar. Biol. 106: 13-23.

Donnely, J. and Torres, J.J. (1988) Oxygen consumption of midwater fishes and
crustaceans from the eastern Gulf of Mexico. Mar. Biol. 97: 483-494.
Douglas, R., Tocher, E., Bendiksen, A., Patrick., C.J. and Gordon B.J. (2008) The role of
phospholipids in nutrition and metabolism of teleost fish. J. Aqacult. 280: 21-34.
Drazen, J.C. (2007) Depth-related trends in proximate composition of demersal fishes in
the eastern North Pacific. Deep. Sea. Res. I. 54: 203-219.
Drevon, C.A., Baksaas, I. and Krokan, H.E. (1993) (eds), Omega-3 fatty acids:
Metabolism and Biological Effects, Birkhauser Verlag Basel. p. 389.
Duan, L., Song, J., Li, X., Yuan, H. and Xu, S. (2010) Distribution of selenium and its
relationship to the eco-environment in Bohai Bay seawater. Mar. Chem. 121: 87-
99.
Dung, L.Q. (2001) Effects of some heavy metals in aquatic environment on the growth of
the grass shrimp (Penaeus monodon). M. Sc. Thesis.
Dural, M., Goksu, M.Z.L. and Ozak, A.A. (2007) Investigation of heavy metal levels in
economically important fish species captured from the Tuzla Lagoon. Food.
Chem. 102: 415-421.
Duruibe, J.O., Ogwuegbu, M.C. and Egwurugwu, J.N. (2007) Heavy metal pollution and
human biotoxic effects. Int. J. Phys. Sci. 2: 112-118.
EC (COMMISSION REGULATION) (2000) Setting maximum levels for certain
contaminant in foodstuffs. EC No 104/2000, Brussels, 2-28.

EC (COMMISSION REGULATION) (2006) Setting maximum levels for certain
contaminants in foodstuffs, No 1881/2006 of 19 December 2006.
Edirisinghe, E.M.R.K.B., Perera, W.M.K., Tayasooriya, S.P. and Bamunuarachchi, A.
(1998) Health related fatty acids in some pelagic fishes in Sri Lanka. Sri Lanka. J.
Aquat. Sci. 3: 97-107.
Ehrlich, K.F. (1974) Chemical changes during growth and starvation of larval
Pleuronectes platessa. Mar. Biol. 24: 34-48.
Eisler, R. (1986) Chromium hazards to fish, wildlife, and invertebrates: A synoptic
review. Biol. Report. 85: 1-38.
Eisler, R. (1985) Selenium hazards to fish, wildlife and invertebrates: a synoptic review.
U.S. Fish and Wildlife Service. Biol. Rep. 85: 1-5.
El-Tay, N.O.A., Abdeltif, E.M. and Ali, M.E. (1998) Chemical composition and quality
grading of three commercial fishes from the Nile. In: Proceedings of FAO Expert
Consultation on Fish Technology in Africa (Kisumu, Kenya). Report no. 574
FAO (Rome): 215-218.
Elvingson. and Januja, L.O. (1992) Determination of fat, protein and dry matter content
of fish by mid-infra red transmission spectroscopy. Aquacult. Fish. Manage. 23:
209-220.
EnvGovCa. (2003) Technical report – water quality guidelines for cobalt.
http://www.env.gov.bc.ca/wat/wq/BCguidelines/cobalt/cobalt_tech.pdf. Accessed
June 2013.
Erdogrul, O. and Ates, D.A. (2006) Determination Cadmium and Copper in fish samples
from Sir and Menzelet Dam Lake Kahramanmaras, Turkey. Environ. Monit.
Assess. 117: 281-290
Ergul, H.A., Topcuoglu, S., Olmez, E. and Kirbasoglu, C. (2008) Heavy metals in sinking
particles and bottom sediments from the eastern Turkish coast of the Black Sea.
Estuar. Coast. Shelf. Sci. 78: 396-402. doi:10.1016/j.ecss.2008.01.006.
EU (European Union). (2001) Commission Regulation as regards heavy metals. Directive
2001/22/EC, No: 466/2001.
EU (European Union). (2008) Commission Regulation (EC) No. 629/2008. Setting
maximum levels for certain contaminants in foodstuffs. Official Journal of the
European Union. p 173.
Fahmy, M.A. (2000) Potential genotoxicity in copper sulfate treated mice. Cytologia. 65:
235-242.
FAO (Food and Agricultural Organisation). (1975) Toxicity of heavy metals in the
environment" 16th Report of Joint FAO/ WHO Expert Committee on Food
additives cited from Part 1 (Frederick. W. Oehme, Ed.) 7, Marcee Dekkering,
New York, 1978, p 7.
FAO (Food and Agricultural Organisation). (1976) List of maximum levels
recommended for contaminants by the Joint FAO/WHO Codex Alimentarius
Commission. Second Series. CAC/ FAL, Rome, 3: 1-8.
FAO (Food and Agricultural Organisation). (1983) Circular No. 764, FAO, Rome.
FAO (Food and Agricultural Organisation). (1989) Evaluation of certain food additives
and the contaminants mercury, lead and cadmium. WHO Technical Report, Series
No. 505.
FAO (Food and Agricultural Organisation). (2008) Statistics division, food security
statistics, food consumption. . Underwood, E. J. (1977). Trace elements in human
and animal nutrition pp. 258–270 (4th ed.). New York: Academic Press.
FAO (Food and Agricultural Organisation). (2011) FAO/WHO Expert Committee on
Food Additives (JEFCA): Safety evaluation of certain contaminants in food.
http://whqlibdoc.who.int/publications/2011/9789241660631_eng.pdf. Accessed
Aug. 21, 2014.
FAO (Food and Agricultural Organisation). (2012) The State of World Fisheries and
Aquaculture 2012, FAO Fisheries and Aquaculture Department. Rome, FAO.
209 p. Available at http://www.fao.org/docrep/016/i2727e/i2727e.pdf
FAO (Food and Agriculture Organization). (1997) The State of World Fisheries and
Aquaculture 1996 Rome. p 43.
FAO (Food and Agriculture Organization). (1999) World production of fish, crustaceans
and mollusks by major fishing areas. Fisheries Information Data and Statistics
unit (FIDI), Fisheries Department, FAO Rome. 33p.
Farmer, J.G. and Johnson, L.R. (1985) The arsenic content of bottled mineral waters.
Environ. Geochem. Health. 7: 124-126.

Farombi, E.O., Adelowo, O.A. and Ajimoko, Y.R. (2007) Biomarkers of oxidative stress
and heavy metal levels as indicators of environmental pollution in African Cat
fish (Clariasgariepinus) from Nigeria ogun river. Int. J. Environ. Res. Public
Health. 4(2): 158-165
FDA (Food and Drug Administration). (2003) FDA Consumer Advisory. Food and Drug
Administration, Washington, DC. Available from:
<http://www.fda.gov/bbs/topics/ANSWERS/2000/advisory.html>.
FEPA (Federal Environmental Protection Agency). (2003) Guidelines and standards for
environmental pollution control in Nigeria, p 238.
Fernandez, T.J., Pradeep, K., Anandan, R., Zynudheen, A.A. and Sankar. T.V. (2014)
Comparison of nutritional characteristics of myctophid fishes (Diaphus effulgens
and D. hudsoni) with common Indian food fishes. Fish. Technol. 51: 173-178.
Firat, O. and Kargm, F. (2010) Biochemical alterations induced by Zn and Cd
individually or in combination in the serum of Oreochroms niloticu. Fish. Physiol.
Biochem. 36: 647-653.
Flick, J.R. and Martin, R.K. (1992) The implications of omega 3 fatty acids in human
health. In: Advances in seafood biochemistry composition and quality. Technomic
Publishing Co, Inc., Basel, Switzerland. p. 69-91.
Francesconi, K.A. (2005) Current perspectives in arsenic environmental and biological
research. Environ. Chem. 2:141-145.

Friberg, L., Piscator, M., Nordberg, G.F. and Kjellstrom, T. (1974) Cadmium in the
Environment. 2nd edition. CRC Press, Cleveland, OH, USA.
FSANZ (2004) Food Standards Code. Food Safety Standard 1.4.1. Food Standards
Australia, New Zealand.
Gallagher, M.L., Harrel, M.L. and Rulifson, R.A. (1991) Variation in lipid and fatty acid
contents of Atlantic Croaker, Stiped Mullet and Summer Flounder. T. Am. Fish.
Soc. 120: 614-619.
Gaspic, Z.K., Zvonaric, T., Vrgoc, N., Odzak, N. Baric, A. (2002) Cadmium and lead in
selected tissues of two commercially important fish species from the Adriatic Sea.
Water. Res. 36:5023–5028
George, M.G., Qasim, S.Z. and Siddiqui, A.Q. (1966) A limnological survey of the river
Kali with reference to fish mortality. Environ. Hlth. 8: 262-269.
George, R. and Bhopal, R. (1995) Fat composition of free living and farmed sea species:
implications for human diet and sea-farming techniques. Bri. Food. J. 97: 19-22.
Georgian Food Safety Rules (2001) Fish, other river/sea products and products made
from them.SanPiN- 2.3.2.560-96. The Minister's Decree 16/08/2001 N301/n for
Health, Labour and Social Affairs.
Ghelichpour, M. and Shabanpour, B. (2011) The investigation of proximate composition
and protein solubility in processed mullet fillets. Int. Food. Res. J. 18: 1343-1347.
Givens, D.I., Cottrill, B.R., Davies, M., Lee, P.A., Mansbridge, R.J. and Moss, A.R.
(2000) Sources of n-3 polyunsaturated fatty acids additional to fish oil for
livestock diets – A review. Nutr. Abstr. Rev. 70: 54-59.
Gokhan, B. and Hikmet, K. (2011) Seasonal Changes in Proximate Composition of Some
Fish Species from the Black Sea. Turkish. J. Fish. Aquat. Sci. 11: 01-05.
Gokoglu, N., Yerlikaya, P. and Cengiz, E. (2004) Effect of cooking methods on the
proximate composition and mineral contents of rainbow trout (Oncorhynchus
mykiss). Food. Chem. 84: 19-22.
Goodwin, T.H., Young, A.R., Holmes, M.G.R., Old, G.H., Hewitt, N., Leeks, G.J.L.,
Packman, J.C. and Smith, B.P.G. (2003) The temporal and spatial variability of
sediment transport and yields within the Bradford Beck catchment, West
Yorkshire. Sci. Total. Environ. 31: 475-494.
Gopalakrishnan, K., Varghese, B.C., Ali, D.M. and Pandian, P. (1988) Deep sea fin fish
resources of Indian Exclusive Economic Zone (beyond 150m). Proc. Nat.
Workshop Fish. Resour. Data Fish Indus., (FSI, Bombay).
Gordon, A.L. (1985) Indian-Atlantic Transfer of Thermocline Water at Agulhas
Retroflection. Science. 227: 1030-1033.
Gruger, E.H. (1967) Fatty acid composition. In: M. E. Stansby, Ed, Wesport, CT: AVI
Publishing Co. p. 3.
Gruger, E.H., Nelson, R.W. and Stansby, M.E. (1964) Fatty acid composition of oils
from 21 species of marine fish, freshwater fish and shellfish. J. Am. Oil. Chem.
Soc. 41: 662- 667.
Guner, S., Dincer, B., Alemdag, N., Colak, A. and Tufekci, M. (1998) Proximate
composition and selected mineral content of commercially important fish species
from the Black Sea. J. Sci. Food. Agricul. 78: 337-342.
Gupta, P. and Srivastava, N. (2006) Effects of sub lethal concentrations of zinc on
histological changes and bioaccumulation of zinc by kidney of fish, Channa
punctatus (Bloch). J. Environ. Biol. 27: 211-215
Gutierrez. L.E. and Silva, R.C.M. (1993) Fatty acid composition of commercially
important fish from Brazil. Sci Agric Piracicaha 50: 478-483.
Gwozdzinski, K. (1995) Structural changes in erythrocyte components induced by copper
and mercury. Radiat. Phys. Chem. 45: 877-882.
Hall, J.A, Frid, C.L.J. and Proud foot, H.K. (1996) Effects of metal contamination on
macrobenthos of two North Sea estuaries. ICES J. Mar. Sci. 53: 1014-1028.
Hamilton, S.J. (2004) Review of selenium toxicity in the aquatic food chain. Sci. Total.
Environ. 326: 1-31. DOI: 10.1016/j.scitotenv.2004.01.019.
Hasegawa, H., Sohrin, Y., Seki, K., Sato, M., Norisuye, K., Naito, K. and Matsui, M.
(2001) Biosynthesis and release of methylarsenic compounds during the growth
of freshwater algae. Chemosphere. 43: 265-272.

Hashim M.M., Divya, T., Jayaprakash, A. and Ganga, U. (2009) Diversity of deep-sea
finfish resources of the Indian continental slope. Book of Abstracts, MECOS,
Kochi, February, 19-21.
Hashmi, M.I., Mustafa, S. and Tariq, S.A. (2002) Heavy metal concentrations in water
and tiger prawn (Penaeus monodon) from grow-out farms in Sabah, North
Borneo. Food. Chem. 79: 151-156.
Hassan, M. (1996) Influence of pond fertilization with broiler dropping on the growth
performance and quality of major carps. Ph.D. Thesis, University of Agriculture,
Faisalabad.
Hassan, M.N., Rahman, M., Hossain, M.M., Nowsad, A.A.K.M. and Hossain, M.B.
(2013) Post-Harvest Loss and Shelf Life of Traditionally Smoked Shrimp
Products Produced in Bangladesh. World J. of Fish. and Marine. Sci. 5: 14-19.
Hayashi, K. and Kishimura, H. (2003) Amount of squalene and fatty acid Composition of
Triacylglycerol and Phospolipids in flesh and liver lipids of Some Deep-sea
Teleost Fish, Morid Cods and Whiptails. J. Oleo. Sci. 52: 339-345.
Hayat, S., Javed, M. and Razzaq, S. (2007) Growth Performance of Metal Stressed Major
Carps viz. Catla Catla, Labeo Rohita and Cirrhina Mrigala Reared under Semi-
Intensive Culture System. Pak. Vet. J. 27: 8-12.
Heath, A.G. (1987) Water Pollution and Physiology. CRC press Inc., Boca Raton. p 145.
Hege, M.W.A., Iren, S., Stoknes, Jannicke, F., Remme, Margareth, K. and Marianne, S.
(2005) Proximate composition, fatty acid and lipid class composition of the
muscle from deep-sea teleosts and elasmobranchs. Comp. Biochem. Physiol. B-
Biochem. Mol. Biol. 3: 437 – 443.
Henderson, R.J. and Almatar, S.M. (1989) Seasonal changes in the lipid composition of
herring (Clupea harengus) in relation to gonad maturation. J. Mar. Biol. Ass. U.K.
69: 323-334.
Herreros, M., Inigo-Nunez, S., Sanchez-Perez, E., Encinas, T. and Gonzalez-Bulnes, A.
(2008) Contribution of fish consumption to heavy metals exposure in women of
childbearing age from a Mediterranean country (Spain). Food. Chem. Toxicol. 46:
1591-1595.
Hickling, C.F. (1947) The seasonal cycle in the corn fish Pilchard, Sardina pilcharda
Wolf. J. Mar. Biol. Ass. U.K. 21: 115-138.
Higashi, H. (1962) Relationship between processing techniques and the amount of
vitamins and minerals in processed fish. In Fish to Nutrition ed. R. Heenan R.
Krauzer. London Publ. New (Bks) Ltd. London. 125-131.
Ho, B.T. and Paul, D.R. (2009) Fatty acid profile of Tra Catfish (Pangasius
hypophthalmus) compared to Atlantic Salmon (Salmo solar) and Asian Seabass
(Lates calcarifer). Int. Food. Res. J. 16: 501-506.
Honore, E., Barhanin, J., Attali, B., Lesage, F. and Lazdunski, M. (1994) External
blockade of the major cardiac delayed- rectifier K+ channel (kv1.5) by
polyunsaturated fatty acids. Proc. Natl. Acad. Sci. USA. 915: 1937-1941.
Hornung, H. and Ramelow, G. (1987) Distribution of Cd, Cr, Cu and Zn in Eastern
Mediterranean fish. Mar. Pollut. Bull. 18: 45-49.
Hossain, M.A., Afsana, K. and Shah, A.K.M.A. (1999) Nutritional value of some Small
Indigenous Fish Species (SIS) of Bangladesh. Bangladesh. J. Fish. Res. 3: 77-85.
Houserova, P., Kuban, V., Spurny, P. and Habarta P. (2006) Determination of total
mercury and mercury species in fish and aquatic ecosystems of Moravian rivers.
Veter. Med. 3: 101-110.
http:// heart_disease. Emedtv. Com/dha/benefits_of_dha_p2.html.
http://www.fsomega.3.com.
http://www.globalhealingcenter.com/natural_health/benefits_of_omega_3_6_9_fatty_aci
ds/.
http://www.lenntech.com/periodic/elements/cr.htm
http://www.ncbi.nlm.nih.gov/pubmed/17876199.
https://www.nal.usda.gov/fnic/food-composition.
Hung, T.C. (1988) Heavy metal pollution and marine ecosystem as a case study in
Taiwan. ln: Hazardous waste detection, control, treatment. Edited by R. Abbou
Elsevier. Amsterdam. p. 869-877.
Hung, T.C. and Han, B.C. (1991) Green oysters: Bioavailable copper and copper
assimilatlve capacity in seawater. ln: Environmental protection of coastal

ecosystem. Edited by D. F. Hwang, T. Yoshida, C. C. Chen, M. Y. Wu and S. S.
Jeng. p. 221-229.
Hung, T.C., Han, B.C. and Wu, S.T. (1989) Green oyster species and forms of copper in
the charting coastal water. Acta. Oceanogr. Taiwan. 23: 33-42.
Huss, H.H. (1988) Fresh fish-quality and quality changes. FAO/ DANIDA (Rome). 15-
75.
Huynh, M.D. and Kitts, D.D. (2009) Evaluating nutritional quality of pacific fish species
from fatty acid signatures. Food. Chem. 114: 912-918.
IARC. (1993) Cadmium and cadmium compounds. In Beryllium, Cadmium, Mercury and
Exposures in the Glass Manufacturing Industry. IARC Monographs on the
Evaluation of Carcinogenic Risk of Chemicals to Humans, vol. 58. Lyon, France:
International Agency for Research on Cancer. p. 119-239.
Ibok, U.J., Udosen, E.D. and Udoidiong, O.M. 1989. Heavy metals in fishes from some
streams in Ikot Ekpene area of Nigeria. Nig. J. Tech. Res., 1: 61-68.
Ikeda, T. (1988) Metabolism and chemical composition of crustaceans from the Antarctic
mesopelagic zone. Deep. Sea. Res. 35: 1991-2002.
Imad, P.S, Malek, B., Joly, G. and Nada, L. (2008) seasonal evaluation of nutritional
benefits of two fish species in the eastern Mediterranean Sea. Int. J. Food. Sci.
Tech. 43: 538-542.

Imelda, J., Shoji, J., Boby, I., Syda Rao, G., Sobhana, K.S., Prema, D. and Molly, V.
(2010) A pilot study on culture of Asian seabass Lates calcarifer (Bloch) in open
sea cage at Munambam, Cochin coast, India. Indian. J. Fish. 57: 29-33.
Iverson, S.J., Frost, K.J. and Lang, S.L.C. (2002) Fat content and fatty acid composition
of forage fish and invertebrates in Prince William Sound, Alaska: factors
contributing to among and within species variability. Marine Ecol. Progress.
Series. 241: 161-181.
Jafri, A.K. and Khawaja, D.K. (1968) Seasonal changes in the biochemical composition
of the freshwater murrel Ophiocephalus punctatus (Bloch). Hydrobiologia. 32:
206-213.
Jahncke, M.L. and Gooch, J.A. (1997) Sensory and chemical characteristics of selected
Gulf of Mexico coastal herring species. J. Food. Sci. 62: 626-631.
James, P.S.B.R. and Pillai, V.N. (1990) Fishable concentrations of fishes and crustacean
in the offshore and deep sea areas of the Indian Exclusive Economic Zone based
on observations made onboard FORV Sagar Sampada. In: Proc. First Workshop
Scient. Resul. FORV Sagar Sampada, 1989, 201-213.
Jarup, L., Berglund, M., Elinder, C., Nordberg, G. and Vahter, M. (1998) Health effects
of cadmium exposure-a review of literature and a risk estimate. Scand. J. Work.
Environ. Hlth. 24: 1-52.
Jat, D. and Kothari, S. (2006) Distribution of zinc in the gills of experimentally exposed
cat fish: Histological and histochemical study. Asian J. Exp. Sci. 20: 165-170
Jayaprakash, A.A., Kurup, B.M., Sreedhar, U., Venu, S., Divya, T., Hashim, M., Anish,
V.P., Paul, T. and Siva, S. (2006) Distribution, diversity, length-weight
relationship and recruitment pattern of deep-sea fin fishes and shellfishes in the
shelf break area of the southwest Indian EEZ. J. Mar. Bio. Ass. India. 48: 56-67.
Jayarani, R., Vijayan Divya, K., Suseela M., Sankar, T.V., Mohanthy, B.P. and Anandan,
R. (2016) Importance of deep sea squid Mastigoteuthis flammea in healthcare as a
rich source of ω -3 polyunsaturated fatty acids, especially eicosapentaenoic and
docosahexaenoic acids. Int. J. Fish. Aquat. Stud. 4: 620-623.
Jeong, B.Y., Choi, B.D., Moon, S.K. and Lee, J.S. (1998) Fatty acid composition of 72
spsecies of Korean fish. J. fish. Sci. Technol. 1: 129-146.
John, M.E. and Sudarsan, D. (1988) Assessment of the Stock of Big eye Snappers (Fam.
Priacanthidae) in the Indian seas with a note on their pattern of abundance. Symp.
Tropical Marine Living Resources, MBAI, 12-16 Jan. 1988, Cochin (Abstract
No.232).
Joint FAO and WHO, (1999) Expert Committee on Food Additives. Summery and
Conclusions, 53rd Meeting, Rome, 1-10.
Jones, S. and Kumaran, M. (1965) New records of fishes from the seas around India -
Part II. J. Mar. Bio. Ass. India. 7:108-123.
Jong-Bang, E., Hee, J., Chung. and Hearnsberger, J.O. (1994) Chemical Composition and
Microflora of Channel Catfish (Ictalurus punctatus) Roe and Swim Bladder. J.
Agric. Food. Chem. 42: 714-717.

Joseph, K..M. (1984) Salient observations on the results of fishery resources survey
during 1983-84. Bull. Fish. Surv. India. 13: 1-11.
Joseph, K.M and John, M.E. (1986) Potential marine fishery resources. Paper presented
in the seminar on Potential Marine Fishery Resources. April 23, 1986. Cochin.
Kalay, M. and Canli M. (2000) Elimination of essential (Cu, Zn) and non-essential (Cd,
Pb) metals from tissues of a freshwater fish Tilapia zillii. Turk. J. Zool. 24: 429-
436.
Kalay, M., Ay, O. and Canli, M. (1999) Heavy metal concentrations in fish tissues from
the Northeast Mediterrenean Sea. Bull. Environ. Contam. Toxicol. 63: 673-681.
Kalpana, D.S. (1980) Seasonal variation in the chemical composition of the cat fish
Heteropneustes fossilus (Bloch) Proc. Indian. Acad. Sci. 89: 191-196.
Kamal, D., Khan, A.N., Rahman, M.A. and Ahamed, F. (2007) Biochemical composition
of some small Indigenous freshwater fishes from the River Mouri, Khulna,
Bangladesh. Pak. J. Bio. Sci. 10: 1559-1561.
Karadede, H. and Unlu, E. (2000) Concentrations of some heavy metals in water,
sediment and fish species from the Atatu¨rk Dam Lake (Euphrates), Turkey.
Chemosphere. 41:1371–1376.
Karadede, H.I., Oymak, S.A., Acenla, E. (2004) Heavy metals in mullet, Liza abu and
catfish, Silurus triostegus, from the AtatÃrk Dam Lake (Euphrates), Turkey.
Environ. Inter. 30:183-188.

Karadede-Akin, H. and Unlu, E. (2007) Heavy metal concentrations in water, sediment
and fish and some benthic organisms from Tigris River, Turkey. Environ. Monit.
Assess. 131: 323-337.
Kargin, F. (1996) Seasonal changes in levels of heavy metals in tissues of Mullus
barbatus and Sparus aurata collected from Iskenderum Gulf (Turkey). Water. Air.
Soil. Poll. 90: 557-562.
Kartal, M., Kurucu, S., Aslan, S., Ozbay, O., Ceyhan, T., Sayar, E. and Cevheroglu.
(2003) Comparison of omega-3 fatty acids by GC-MS in frequently consumed
fish and fish oil preparations on the Turkish market. J. Pharm. Sci. 28: 201-205.
Karthikeyan, S., Palaniappan, P.L.R.M. and Sabhanayakan, S. (2007) Influence of pH
and water hardness upon Nickel accumulation in edible fish Cirrhinus mrigala. J.
Environ. Biol. 28: 484-492.
Karuppasamy, P.K., Balachandran, K., George, S., Balu, S., Vimala, P. and Menon, N.G.
(2008) Food of some deep sea fishes collected from the eastern Arabian Sea. J.
Mar. Biol. Ass. India. 50: 134-138.
Kcukgulmez, A., Celik, M., Ersoy, B. and Yanar, Y. (2010) Effects of season on
proximate and fatty acid compositions of two Mediterranean fish – the round
herring (Etrumeus teres) and tub gurnard (Chelidonichthys lucernus). Int. J. Food.
Sci. and Tech. 45: 1056-1060.

Kemp, K.M., Fraser, K.P.P., Collins, M.A. and Priede, I.G. (2008) Seasonal variation in
the white muscle biochemical composition of deep-sea macrourids in the North-
east Atlantic Mar. Biol. 155: 37-49.
Kennish, M.J., (2001) Coastal Salt Marsh Systems in the U.S. A Review of
Anthropogenic Impacts. J. Coast. Res. 17: 731-748
Khan, F. M., Zacharia, P.U., Nandakumaran, K., Mohan, S., Arputharaj, M.R., Nagaraja,
D. and Ramakrishnan, P. (1996) Catch, abundance and some aspects of biology of
deep-sea fish in the southeastern Arabian Sea. In: Proc. Asian. Fish. Sci. 617-629.
King, R.P. and Jonathan, G.E. (2003) Aquatic environmental perturbations and
monitoring, African experience, USA.
Knaan-Shanzer, S., Valerio, D. and Van Beusechem, V.W. (1996) Cell cycle state,
response to hemopoietic growth factors and retroviral vector-mediated
transduction of human hemopoietic stem cells. Gene. Ther. 3: 323-333.
Kojadinovic, J., Potier, M. and Le Corre, M. (2006) Mercury content in commercial
pelagic fish and its risk assessment in the Western Indian Ocean. Sci. Total.
Environ. 366: 688-700.
Kojadinovic, J., Potier, M., Le Corre, M., Cosson, R.P. and Bustamante, P. (2007)
Bioaccumulation of trace elements in pelagic fish from the Western Indian Ocean.
Environ. Pollut. 146: 548-566.
Koslow, J.A., Bax, N.J., Bulman, C.M., Kloser, R.J., Smith, A.D.M. and Williams, A.
(1997) Managing the fishdown of the Australian orange roughy resource. In:
Hancock, D.A., Smith, D.C., Grant, A., Beumer, J.P., (Eds.), Developing and
sustaining world fisheries resources: the state of science and management. CSIRO
Australia, 558-562.
Kotze, P., Preez, H.H., and van Vuren, J.H.J. (1999) Bioaccumulation of copper and zinc
in Oreochromis mossambicus and Clarias gariepinus, from the Olifants river,
Mpumalanga, South Africa. Water SA. 25: 99-100.
Kozlova, A.U.T.A. and Khotimchenko, S.V. (1998) Lipids and fatty acids of two pelagic
cottoid fishes (Comephorus species) endemic to Lake Baikal. J. Aqacul. 126: 477-
485.
Kris-Etherton, P.M., Harris, W.S. and Appel, L.J. (2002) Fish consumption, fish oil,
omega-3 fatty acids, and cardiovascular disease. Circulation. 106: 2747-2757.
Kromhout, D. (2001) Diet and cardiovascular diseases. J. Nutr. Health Aging 5: 3144-
3149.
Kronmann, N. and Green, A. (1980) Epidemiological studies in the Upernavik district,
Greenland: incidence of some chronic diseases 1950-1974. Acta. Med. Scand.
208: 401–406.
Krzynowek J., Uljua, D.S., Panunzio, L.J. and Maney, R.S. (1992) Factors Affecting Fat,
Cholesterol and Omega-3 Fatty Acids in Maine Sardines. J. Food. Sci. 57: 63-65.
Krzynowek, J. and Murphy, J. (1987) Proximate composition, energy, fatty acid, sodium
and cholesterol content of finfish, shellfish and their products. NOAA Technical
Report NMFS 55: 1-50.

Kucukgulmez, A., Celik, M., Ersoy, B. and Yanar, Y. (2010) Effect of season on
proximate and fatty acid compositions of two Mediterranean fish- the round
herring (Etrumeus teres) and tub gurnard (Chelidonichthys lucernus). Intr. J.
Food. Sci. Tech. 45: 1056-1060.
Kucuksezgin, F., Altay, O., Uluturhan, E. and Kontas, A. (2001) Trace metal and
organochlorine residue levels in red mullet (Mullus barbatus) from the Eastern
Aegean, Turkey. Water. Res. 35: 2327–2332.
Kucuksezgin, F.A., Kontas, O., Altay, E. and Uluturhan, D. E. (2006) Assessment of
marine pollution in Izmir Bay; Nutrient heavy metal and total hydrocarbon
concentrations: Environ. Int. 32: 41-51.
Kumar, B., Rita, S. and Mukherjee, D. (2011) Geochemical distribution of heavy metals
in sediments from sewage fed fish ponds from Kolkata wetlands. India. Chem.
Speciation Avail. 23: 24–32.
Kurup, B.M. and Venu, S. (2006) Length-weight relationship of Priacanthus hamrur
(Forsskal) inhabiting the continental slopes beyond 300m depth along the west
coast of India. Fish. Tech. 43: 41-46.
Kurup, B.M., Harikrishnan, M. and Venu, S. (2006) Depth wise variations in the
maintenance of dimensional equality of the deep-sea fishes off southwest coast of
India. J. Mar. Biol. Ass. India. 48: 35-40.

Kurup, B.M., Jiji, T. and Venu, S. (2005) Distribution and biology of Chlorophthalmus
bicornis Norman, beyond 250m depth off the south west coast in the Indian EEZ.
J. Mar. Biol. Ass. India. 47: 57-62.
Kusumo, W.A. (1997) Profile fatty acid some pelagic fish and demersal from Pelabuhan
Ratu, West Java and Muara Angke, Jakarta . Thesis. Bogor. Faculty of Fisheries
and Marine Science. Bogor Agricultural University.
Kwon, Y.T. and Lee, C.W. (1999) Application of multiple ecological risk indices for the
evaluation of heavy metal contamination in a coastal dredging area. Sci. Total.
Environ. 214: 203–210.
Labonne, M., Basin, S., Othman, D. and Luck, J. (2001) Lead Isotopes in Muscels as
Tracers of Metal Sources and Water Movements in a Lagoon (Thau Basin, S.
France). Chem. Geol. 181: 181-191.
Lahnsteiner, F. (2000) Morphological, physiological and biochemical parameters
characterizing the over-ripening of rainbow trout. Eggs. Fish. Physiol. Biochem.
23: 107-118.
Lakshmanan, P.T. (1988) Levels of cadmium in seafood products. Fish. Tech. 25: 142-
146.
Landmark, K. and Alm, C.S. (2006) Alpha- linoleic acid, cardiovascular disease and
sudden death (in Norwegian). Tidsskr. Nor. laegeforen. 126: 2792-2794.
Latha, U., Sally, S., Meenakumari, B. and Devadasan, K. (2003) Trace metal
concentration in four species of edible fishes from the south coast of India. In: Sea
food safety, Proceedings of the symposium on sea food safety – status and
strategies. Cochin, p. 259-270.
Lead and health, in working party on lead in the environment (1980) Department of
Health and Social Security DHSS. London.
Lederer, W.H and Fensterheim, W.H, (1982) Arsenic. Van Nostrand Reinhold, New
York.
Leonard, A.E., Pereira, S.L., Sprecher, H. and Huang, H.S. (2004) Elogation of long-
chain fatty acids. Prog. Lipid. Res. 43:36-54.
Levy, L.S. and Venitt, S. (1986) Carcinogenicity and mutagenicity of chromium
compounds: the association between bronchial metaplasia and neoplasia.
Carcinogenesis. 7: 835-837.
Licata, P., Trombetta, D., Cristani, M., Naccari, C., Martino, D., Calo, M. and Naccari, F.
(2005) Heavy metals in liver and muscle of bluefin tuna (Thunnus thynnus)
caught in the straits of Messina (Sicily, Italy). Environ. Monit. Assess. 107: 239-
248.
Lidsky, I.T. and Schneider, S.J. (2003) Lead neurotoxicity in children: basic mechanisms
and clinical correlates. Brain. 100: 284-293.
Llobet, J.M., Granero, S., Torres, A., Schuhmacher, M. and Domingo, J.L. (1998)
Biological monitoring of environmental pollution and human exposure to metals
in Tarragona, Spain. Trace. Elem. Electrol.15: 76-80.

Love, R.M. (1957) The Biochemical composition of fish “In the physiology of fishes”
(Ed.) Brown, M.E. Academic Press. London and N.Y. 1: 401-418.
Love, R.M. (1970) The Chemical Biology of Fishes. Vol I. Acad. Press, London. 255-
262.
Love, R.M. (1980) The chemical biology of fishes. Vol II. Academic press, London.
2333-2342
Lucia, L.A., Sampaipo, G.R., Castellucci, C.M.N. and Torres, E.A.F.S. (2003) The
Influence of season on the lipid profiles of five commercially important species of
Brazilian Fish. Food. Chem. 83: 93-97.
Luoma, S.N. and Presser, T.S. (2000) Forecasting selenium discharges to the San
Francisco Bay-Delta Estuary: ecological effects of a proposed San Luis Drain
extension.Open-File Report 00-416, US Geological Survey, Menlo Park, CA.
Lytle, J.S. and Lytle, T.F. (1988) Fatty acid nutritional profiles in Gulf of Mexico fishes.
http://sst.ifas.ufl.edu.
Madhusudan, S., Fatima, L. and Nadim, C. (2003) Bioaccumulation of zinc and cadmium
in freshwater fishes. Indian. J. Fish. 50: 53-65.
Mai, H.X., Zhang, Y.W., Si, R., Yan, Z.G., Sun, L.D., You, L.P. and Yan, C.H. (2006)
High-Quality Sodium Rare-Earth Fluoride Nanocrystals: Controlled Synthesis and
Optical Properties. J. Am. Chem. Soc. 128: 6426-6436.
Manju, S., Chakraborty, K., Bineesh, K.K., Pillai, N.G.K., Abdussamad, E.M. and
Vijayan, K.K. (2011) Proximate composition and fatty acid profile of myctophid
(Diaphus watasei Jordan & Starks, 1904) from the Arabian Sea. Indian. J. Fish.
58: 103-107.
Mansour, S.A. and Sidky, M.M. (2002) Ecotoxicological studies. Heavy metals
containing water and fish from Fayoum Governate, Egypt. Food. Chem. 78: 15-
22.
Mansouri, B., Ebrahimpour, M. and Babaei, H. (2012b) Bioaccumulation and elimination
of nickel in the organs of black fish (Capoeta fusca). Toxicol. Ind. Health. 28:
361-368.
Marcelo, G.G., Racotta, I.S. and Humberto, V. (2003) Variation in lipid, protein, and
carbohydrate content during the embryonic development of the crayfish Cheraxq
uadricarinatus (decapoda: parastacidae) J. Crustacean. Biol. 23: 1-6.
Martin, J.H. and Fitzwater, S.E., (1988) Iron-deficiency limits phytoplankton growth in
the northeast Pacific subarctic. Nature. 331: 341-343.
Martin, K. and Scanes, P. (1996) Organochlorine compound and trace metal
contaminants in fish near Sydneys ocean outialls. Mar. Poilut. Bull. 33: 213-225.
McCay, C.M. and Tunison, A.V. (1936) Cortland Hatchery Report No.5 New York state
Cons. Dept. U.S. Bur. of fish and Cornell Univ.
Medale, F., Lefevre, F. and Corraze, G. (2003) Qualite nutritionnelle at dietetique des
poissons, constituants de la chair et facteurs de variations. Cahiers de Nutrition at
Dietetique 38: 37-44.

Meek, R.P. and Childress, J.J. (1973) Respiration and the effect of pressure in the
mesopelagic fish Anoplogaster cornuta (Beryciformes). Deep. Sea. Res. 20:
1111–1118.
Meinke, W.W. and Mattil, K.F. (1973) Autolysis is a factor in the production of protein
isolates from whole fish. J. Food. Sci. 38: 864-866.
Mendil, D., Faruk, U.O., Tuzen, M. and Soylak, M. (2010) Determination of trace metals
in different fish species and sediments from the river Yesilırmak in Tokat,
Turkey. Food. Chem. Toxicol. 48: 1383-1392.
Menon, N.G. (1990) Preliminary investigation on the fish biomass in the deepsea
scattering layers of the EEZ of India. In: Proc. First Workshop Scient. Resul.
FORV Sagar Sampada, 1989, 5-7 June 1989, 273-280.
Menon, N.G., Pillai, N.G.K., Lazarus, S. and Nammalwar, P. (1996) Finfish resources in
the north eastern region in the Indian EEZ. In: Proc. Second Workshop Scient.
Resul. FORV Sagar Sampada, 1989, 295-304.
Merrett, N. and Haedrich, R. (1997) Deep-Sea Demersal Fish and Fisheries. Chapman
and Hall, London.
Mertz, W. (1969) Chromium occurrence and function in biological system. Physiol. Rev.
49: 163-239.
Meynier, L., Morel, P., Chilvers, L., Mackenzie, D., MacGibbon, A. and Duignan, P.
(2008) Temporal and sex differences in the blubber fatty acid profiles of the New
Zealand Sea lion Phocarctos hookeri. Mar. Ecol. Prog. Ser. 366: 271-279.
Mnari, A., Bouhlel, I., Chraief, I., Hammami, M., Romdhane, M.S., El Cafsi, M. and
Chaouch, A. (2007) Fatty acids in muscles and liver of Tunisian wild and farmed
gilthead sea bream, Sparus aurata. Food. Chem. 100: 1393-1397
Moeller, A., Mac Neil, S.D., Ambrose, R.F. and Hee, S.S.Q. (2003) Elements in fish of
Malibu Creek and Malibu Lagoon near Los Angeles, California. Mar. Pollut. Bull.
46: 424-429.
Mohammad, H.N.A. and Hossam, H.H.A. (2007) Accumulation and distribution of
copper and zinc in both water and some vital tissues of two fish species (Tilapia
zilli and Mugilcephalus) of lake Qarun, Fayoum Province, Egypt, Pak. J. Biol.
Sci. 10: 2106-2122.
Momtaz, M. (2002) Geochemical studies of Heavy Metals in the Seawater along Karachi
Makran Coast, in Chemistry. universituy of Karachi. p 414.
Mona, S., Zaki, N., Elbattrawy O.M., Fawzi, I.A. and Nagwa, S.A. (2011) Effect of
Mercuric Oxide Toxicity on some Biochemical Parameters on African Cat Fish
Clarias gariepinus Present in the River Nile. Life. Sci. 8: 363-368.
Monsen, E.R. (1985) NIH lauching major reaserch program on fish oil and health. Food
Chemical News. 6: 34-39.
Moore, J.W. (1991) Inorganic Contaminants of Surface Water. 2 ed. New York: Springer
Verlag.
Moore, J.W. and Ramamoorthy, S.N. (1984) Heavy metals in natural waters: Applied
Monitoring and impact Assessment. Springer, Berlin. p. 182-204.

Moore, J.W. and Ramamoorthy, S.N. (1984) Organic chemicals in Natural Waters,
Applied Monitoring and Impact Assessment, (Robert, D.S. Ed.), p 67-81, Springer
Verlang, New York.
More, T.G., Rajput, R.A. and Bandela, N.N. (2003) Impact of heavy metals on DNA
content in the whole body of freshwater bivalve, Lamelleiden marginalis.
Environ. Sci. Pollut. Res. 22: 605-616.
Morel, F.M.M., Reinfelder, J.R., Roberts, S.B., Chamberlain, C.P., Lee, J.G. and Yee. D.
(1994) Zinc and carbon colimitation of marine phytoplankton. Nature. 369:
740-742.
Mormede, S. and Davies, I.M. (2001) Heavy metal concentrations in commercial deep-
sea fish from the Rockall Trough. Cont. Shelf. Res. 21: 899-916.
Mozaffarian, D. and Rimm, E.B. (2006) Fish intake, contaminants and human health:
evaluating the risk and the benefits. JAMA. 296: 1885-1899.
Murray, H.M., Hew, C.L. and Fletcher, G.L. (2002) Skin-type antifreeze protein
expression in integumental cells of larval winter flounder. J. Fish Biol. 60: 1391-
1406.
Muthiah, C. (1994) Studies on the fishery and biology of lizardfish from Karnataka coast.
Ph.D. Thesis, Karnataka University. Karnataka, India.
Nair, B.N. and Thampy, D.M. (1980) A textbook of Marine Ecology. Mc Millan Co.,
India, 352 p.
Nair, K.V.S. and Reghu, R. (1990) Studies on the threadfin bream and the lizard fish
resources in the exclusive economic zone of India based on the demersal trawling
operations of FORV Sagar Sampada. In: Proc. First Workshop Scient. Result
FORV Sagar Sampada, 5-7 June 1989. 239-255.
Navaneethan, R., Vimaladevi, S., Ajeesh Kumar, K.K., Anandan, R., Niladri, S.C., Asha
K.K. and Suseela Mathew. (2016) Profiling of Omega-3 Polyunsaturated Fatty
Acids of Myctophid Fish Species Available in Arabian Sea. Fish. Technol. 53: 55-
58.
Navarro, P., Raposo, J.C., Arana, G. and Etxebarria, N. (2006) Optimasation of
microwave assisted digestion of sediments and determination of Sn and Hg. Anal.
Chim. Acta. 566: 37-44.
Nettleton, J. (1995) Omega-3 fatty acids and Health. Chapman and Hall, New York. p.
359.
Nielson, B.S. and Anderson, S.W. (1970) Copper ions as poison in the sea and in the
fresh water. Mar. Pollut. Bull. 6: 188-190.
Niimi, A.J. (1972) Changes in the proximate body composition of large mouth
(Micropterus salmoides) under starvation. J. Zool. 50: 815-819.
Nordgarden, U.B.E., Torstensen, Froyland, Hansen, T. and Hemre, G.I. (2003)
Seasonally changing metabolism rate in Atlantic salmon Salmo salar(L.). 2B-
oxidation capacity and fattyacid composition in muscle tissues and plasma
lipoproteins. Aquacult. Nutr. 9: 295-303.

Norwood, W.P., Borgmann, U. and Dixon, D.G. (2007) Chronic toxicity of arsenic,
cobalt, chromium and manganese to Hyalella azteca in relation to exposure and
bioaccumulation. Environ. Pollut. 147: 262-272.
Nriagu, J.O. (1988) A silent epidemic of environmental metal poisoning. Environ. Pollut.
50: 139-161.
Nriagu, J.O. (2002) Arsenic poisoning through the ages. In: Frankenberger WT Jr (ed)
Environmental Chemistry of Arsenic. Dekker, New York, p 1-26.
Nwaedozie, J.M. (1998) The determination of heavy metal pollution in some fish samples
from River Kaduna. J. Chem. Soc. Nig. 23:21-23.
Nybakken, J.W. (1992) Marine Biology, an Ecological Approach. Eidman M, interpreter.
Jakarta: Penerbit Gramedia. p. 222-290.
Oceanlink. (2006) Deep Sea Biology. http://www.oceanlink.island.net/ask/depsea.html
(2-06- 2006).
Odoemelam, S.A.. Iwuazor, C.C. and Ozuo, J.U. (1999) Tropical Environmental Forum
(TEF) held at Calabar, May 18th.
Oguize, F.A. (1999) Contribution of industrial effluents to the heavy metals load of the
lower Ikpoba river in Benin City, Nigeria. Afrian Journal of Applied Zoology. 2:
60- 63.
Oguzie, F.A. and Okosodo, C.I. (2008) Contribution of heavy metals in waste dumpsites
from selected markets to the heavy metals load of Ikpoba river in Benin City,
Nigeria. J. Aquat. Field. Stud. 4: 51-56.

Ojewola, G.S. and Annah, S.I. (2006) Nutritive and economic value of Danish fish meal
cray fish dust meal and shrimp waste meal inclusion in broiler diet. Int. J. Poult.
Sci. 5:390-394.
Okland, H.M.W., Stoknes, I.S., Remne, J.F., Kjerstad, M. and Synnes, M. (2005)
Proximate composition, fatty acid and lipid class composition of the muscle from
deep-sea teleosts and elasmobranches. Comp. Biochem. Physiol. B. 140: 437-443.
Oksuz, A., Ozyilmaz, A. and Turan, C. (2009) Fatty Acid Profiles of Anchovy of Black
Sea and Mediterranean Sea. 21: 3081-3086.
Oksuz, A., Ozyilmaz, A., Aktas, M., Gercek, G. and Motte, J. (2009) A comparative
study on proximate, mineral and fatty acid compositions of deep sea water Rose
Shrimp(Parapenaeus longirostris, Lucas 1846) and golden Shrimp (Plesionika
martia, A. Milne-Edwards, 1883). J. Anim. Vet. Adv. 8:183-189.
Olajire, A.A. and Imeokparia, F.E. (2000) A study of the water quality of the osun river:
Metal monitoring and geochemistry. Bull. Chem. Soc. Ethiopia. 14: 1-8.
Olsen, R.E. and Henderson, R.J. (1997) Muscle fatty acid composition and oxidative
stress indices of Arctic Charr, Salvelinus alpines (L.), in relation to delivery
polyunsaturated fatty acid levels and temperature. Aquacult. Nutr., 3: 227-238.
Omolara, O.O. and Omotayo, O.D. (2008) Preliminary studies on the effect of processing
methods on the quality of three commonly consumed marine fishes in Nigeria.
Biochem. 21:1-7.
Omotosho, O.E., Oboh, G. and Jweala, E.E.J. (2011) Comparative effects of local
coagulants on the nutritive value, invitro multi enzyme protein digestibility and
sensory properties of Wara cheese. Int. J. Dairy. Sci. 6: 58-65.
Orban, E., Nevigato, T., Di Lena, G., Masci, M., Casini, I., Gambelli, L. and Caproni, R.
(2008) New trends in the seafood market. Sutchi catfish (Pangasius
hypophthalmus) fillets from Vietnam: Nutritional quality and safety aspects.
Food. Chem. 110: 383-389.
Oremland, R.S. and Stolz, J.F. (2003) The ecology of arsenic. Science. 300: 939–944.
Osman, A., Wuertz, S., Mekkawy, I., Exner, H. and Kirschbaum, F. (2007) Lead Induced
Malformations in Embryos of the African Catfish Clarias Gariepinus (Burchell,
1822). Environ. Toxicol. 22: 375-389.
Osman, H., Suriah, A.R. and Law, E.C. (2001) Fatty acid composition and cholesterol
content of selected marine fish in Malaysian waters. Food. Chem. 73: 55-60.
Otitoloju, A.A. (2002) Evaluation of the joint action toxicity of binary mixtures of heavy
metals against the mangrove periwinkle Tympanotonus fuscatus var radula (L).
Ecotoxicol. Environ. Saf. 53: 404-415.
Owaga, E.E., Onyango, C.A. and Njoroge C.K. (2010) Influence of selected washing
treatments and drying Temperatures on proximate composition of Dagaa
(Rastrineobola argentea), a small pelagic fish species. Afr. J. Food Agric. Nutr.
Dev. 10:1-14.
Owen, S.F., Mc Carthy, I.D., Watt, P.W., Ladero,V. and Sanchez, J.A. (1999) Mosquito
fish as an environmental sentinel: parasite and histological In vivo rates of protein
synthesis in Atlantic Salmon smolts determined using a stable isotope flooding
dose technique. Fish. Physiol. Biochem. 2: 87
Ozagul, Y. and Ozagul, H. (2007) Fatty acid profiles of commercially important fish
species from the Mediterranean, Aegean and Black Seas. Food. Chem. 100: 1634-
1638.
Ozagul, Y., Ozagul, F. and Alagoz, S. (2007) Fatty acid profiles of commercially
important Seawater and Freshwater fish species of Turkey: A comparative study.
Food. Chem. 1003: 217-223.
Pal, M., Mukhapadhayay, T. and Ghosh, S. (2011) Proximate, fatty acid, and amino acid
composition of fish muscle and egg tissue of Hilsa (Tenualosa ilisha). J. Aquat.
Food. Prod. Technol. 20: 160-171.
Panicker, P.A., Boopendranath, M.R. and Abbas, M.S. (1993) Observations on deep sea
demersal resources in the Exclusive Economic Zone off Southwest coast of India.
Fish. Tech. 30: 102-108.
Parin, N.V. and Kotlyar, A.N. (1984) A new species of the family Aulopidae
(Myctophiformes, Osteichthyes) from the East Pacific. Zool. Zhur. 63: 1435-
1437.
Park, J. and Presley B.J. (1997) Trace metal contamination of sediments and organisms
from the swan lake area of Galveston bay. Environ. Pollut. 98: 209-221.
Paul, V.I. and Banerjee, T.K. (1997) Histopathological changes induced by ambient
ammonia (ammonium sulfate) on the opercular linings of the live fish
Heteropneustes fossilis (Bloch). Dis. Aquat. Organ. 28: 15-161.
Perez-Benito, J.F. (2006) Effects of Cr (VI) and V (V) on the lifespan of fish. J. Trace.
Elem. Med. Biol. 20:161-170.
Philip, K.P. (1994) Studies on the biology and fishery of the fishes of the family
Priacanthidae (Pisces: Perciformes) of Indian waters. Ph.D. Thesis, Dept. of
Industrial Fisheries, Cochin University of Science & Technology, Kerala, India.
169 p.
Phillips, D.J.H. (1990) Arsenic in aquatic organisms: a review, emphasiz, chemical
speciation. Aquatic. Toxicol. 16: 151-186.
Phillips, D.J.H. and Rainbow, P.S. (1994) Biomonitoring of trace aquatic contaminants.
Environmental Management Series, Chapman and Hall, London, p 371.
Phillips, D.J.H., Thompson, G.B., Gabuji, K.M. and Ho, C.T. (1982) Trace metals of
toxicological significance to man in Hong Kong seafood. Environ. Pollut. Ser. B.
3: 27-45.
Pillai, P.P. and Gopakumar, G. (2003) Tunas. Status of Marine Fisheries Resources of
India; CMFRI: 51-60.
Pinentel, C.R. and Joyeux, J.C. (2010) Diet and food partitioning between juveniles of
muton Lutjanus jocu and Lane Lutjanus synagris Snappers (Perciformes:

Lujanidae) in a mangrove-fringed estuarine environment. J. Fish. Biol. 76: 2299-
2317.
Pinot, F., Kreps, S.E., Bachelet, M., Hainaut, P., Bakonyi, M., Polla, B.S. (2000)
Cadmium in the environment: sources, mechanisms of biotoxicity, and
biomarkers. Rev. Environ. Hlth. 15: 299-323.
Pirestani, S., Sahari, A. and Barzegar, M. (2010) Fatty Acids Changes during Frozen
Storage in Several Fish Species from South Caspian Sea. J. Agr. Sci.Tech. 12:
321-329.
Piros, B., Glogowski, J., Kolman, R., Rzemienicecki, A., Domagala, J., Horvath, A.,
Urbanyi, B. and Ciereszko, A. (2002) Biochemical characterization of Siberian
sturgeon Acipenser ruthenus plasma and spermatozoa. Fish. Physiol. Biochem.
26: 289-295.
Plana. S., Sinquin, G., Maes, P., Paillard, C. and Le Pennec, M. (1996) Variations in
biochemical composition of juvenile Ruditapes philippinarum infected by a
Vibrio species. Dis. Aquat. Organ. 24: 205-213.
Pranoto, T. (2006) Unsaturated fatty acid as decrease of diseases coroner heart risk.
University of Indonesia.
Prasad, R.R. and Nair, P.V.R. (1973) India and Indian Ocean Fisheries, J. Mar. Biol. Ass.
India. 15:1-19.
Presley, B.J. (1997) A Review of Arctic Trace metal data with implications for biological
effects. Mar. Pollut. Bull. 35: 226-234.

Pringle, B.H.D.E., l-lissong, Katz E.L. and Muslawka, S.T. (1968) Trace metal
accumulation by estuarine molluscs. J. Sanit. Engg. Div. Am. Soc. Civ. Eng. 94:
455-475.
Prudente, M., Kim, E.Y., Tanabe, S. and Tatsukawa, R. (1997) Metal levels in some
commercial fish species from Manila Bay, Phulippines. Mar. Pollut. Bull. 34:
671-674.
Ptashynski, M.D., Pedlar, R.M., Evans, R.E., Baron, C.L. and Klaverkamp, J.F. (2002)
Toxicology of dietary nickel in lake whitefish (Coregonus clupeaformis). Aquat.
Toxicol. 58: 229-247.
Qyvind, L., Einar, L., Amund, M., Leif, R.N. and Kjartan, S. (1994) Nutrient content in
fish and shellfish. Fisk. Div. Skr. Ernaering. 6: 1-101.
Radhakrishnan, A.G. (1993) Studies on the trace metal content of fish and shell fish
including bivalves, Devadasan, K. et al. (Eds) Nutrients and bioactive substances
in aquatic organisms Society Of Fisheries Technologists (India) Cochin. 266-270.
Radhika, R. (2004) Systematics, fishery, resource characteristics and bionomics of deep
sea prawns off Kerala. Ph. D Thesis, Cochin University of Science and
Technology, Cochin, 385 p.
Radhika, R. and Madhusoodana K.B. (2005) Some observations on the carotenoid
distribution and proximate composition in deep sea prawns off Kerala (South
India) Paper presented in the International Symposium Sustainfish. Cochin, India.

Rahman, M.A., Gheyasuddin, S., Rashid, M.H. and Chowdhury, M.F.Z. (1982)
Proximate composition and nutritive quality of freshwater zeal fishes of
Bngladesh. Bangladesh J. Fish. 2-5: 37-43.
Rahman, M.A., Hasegawa, H. and Lim, R.P. (2012) Bioaccumulation, biotransformation
and trophic transfer of arsenic in the aquatic food chain. Environ. Res. 116: 118-
135.
Rai, D., Sass, B.M. and Moore, D.A. (1987) Chromium(III) hydrolysis constants and
solubility of chromium (III) hydroxide. Inorg. Chem. 26: 345-349.
Rainuzzo, J.R., Reitan, K.I. and Olsen, Y. (1997) The significance of lipids at early
stages of marine fish: a review. Aquacult. 155: 103-116.
Raja, P., Veerasingam, S., Suresh, G., Marichamy, G. and Venkatachalapathy, R., (2009)
Heavy metals concentration in four commercially valuable marine edible fish
species from Parangipettai Coast, South East Coast of India. Int. J. Ani. Veter.
Adv. 1: 10-14.
Rajamoorthy, K., Pradeep, K., Anandan, R., Libin, B., Sankar, T.V. and Lakshmanan,
P.T. (2013) Biochemical composition of myctophid species Diaphus watasei and
Myctophum obtusirostre caught from Arabian sea. Fish. Technol. 50: 41-44.
Rajan, K.N., Nandakumar. G.K. and Chellappan, K. (2001) Innovative exploitation of
deep-sea crustaceans along the Kerala coast. Mar. Fish. Infor. Serv. T and E Ser.
168: 1-5.
Ramesh, R. Chakraborty, S.K. and Venkateshwarlu, G. (2015) Seasonal Changes in
Total Lipid Content and Fatty acid Profiles of Ribbon Fish, Lepturacanthus
savala (Cuvier,1829). Fish. Technol. 52: 95-100.
Rao, K.R., Viji, P., Sreeramulu, K. and Sreedhar, U. (2016) Proximate Composition and
Heavy Metal Accumulation in Deep-Sea Crustaceans from Selected Stations in
the Indian Exclusive Economic Zone (EEZ). Fish. Technol. 53: 155-161.
Rao, N.K.V. (1965) On a record of Epinnula orientalis Gilchrist and Von Bonde, a
bathypelagic fish, from the Konkan coast. J. Mar. Biol. Ass. India. 7: 217-218.
Rao, P.S. and Rao, L.M. (2002) Variations in bio-chemical composition of Glossogobius
giuris (Hamilton) from Gosthan with observation on distribution (Caranx
sexfasciatus host India). Curr. Sci. (India). 35: 101-102.
Rasmussen, B., Vessby, B., Uusitupa, M., Berglund, L., Pedersen, E., Riccardi, G.,
Rivellese, A.A., Tapsell, L. and Hermansen, K. (2006) Effects of dietary
saturated, monounsaturated and n-3 fatty acids, on blood pressure in healthy
subjects. Am. J. Clin. Nutr. 83: 221-226.
Ravichandran, S., Kumaravel, K. and Pamela Florence, E. (2011) Nutritive Composition
of Some Edible Fin Fishes. Inter. J. Zool. Res. 7: 241-251.
Ravichandran, S., Kumaravel, K., Rameshkumar, G. and Ajithkumar, T.T. (2010)
Antimicrobial peptides from the marine fishes. Res. J. Immunol. 3: 146-156.
Regunathan, A., Glesne, D.A., Wilson, A.K., Song, J., Nicolae, D., Flores, T.,
Bhattacharyya, M.H. (2003) Microarray analysis of changes in bone cell gene

expression early after cadmium gavage in mice. Toxicol. Appl. Pharmacol. 191:
272–293. doi:10.1016/S0041-008X(03)00163-7
Reiffel, J.A. and McDonald, A. (2006) Antiarrhythmic effects of omega-3 fatty acids.
Am. J. Cardiol. 98: 50-60.
Richardson, E.W., Stobbe, E.D. and Bernstein, S. (1980) Ion exchange traps chromates
tor reuse. Environ. Sci. Technol. 2: 10-16.
Rina Sharlinda Zabri. (2009) Accumulation of Selected Heavy Metals (Zn, Pb, As and
Hg) In Some Marine Fishes from Straits of Malacca, Malaysia. Master Thesis.
International Islamic University Malaysia.
Rist, L., Muellerv, A., Barthel, C., Snijders, B., Jansen, M., Simoeswust, A.P., Huber, M.,
Kummeling, I., Von Mandach, U., Steinhart, H. and Thijs, C. (2007) Influence of
organic diet on the amount of conjugated linoleic acids in breast milk. British. J.
Nutr. 97: 735-743.
Romeo, M., Siau, Z., Sidoumou, Y. and Gnassia, M. B. (1999) Heavy Metal Distribution
in Different Fish Species from the Mauritania Coast. Sci. Total. Environ. 232:
169-175.
Rossano, R., Caggiano, M.A., Mastrangelo, L., Di Lauro, R., Ungaro, N., Ettorre, M. and
Riccio, P. (2005) Proteins, Fatty Acids, and Nutritional Value in the Muscle of the
Fish Species Mora moro (Risso, 1810). Mol. Nutr. Food. Res. 49: 926-931.
Roynette, C.E., Calder, P.C. and Dupertuis, Y.M. (2004) N-3 polyunsaturated fatty acids
and colon prevention. Clin. Nutr. 23: 139-151.

Saha, K.F. and Guha, B.C. (1940) Nutritional investigations of Bengal fish. Indian J.
med. Res. 27: 873-876.
Saify, Z.S., Akhtar, S., Hassan, S., Arif, M., Ahmed, F. and Siddiquis. (2000) A study on
fatty acid composition of fish oil from two marine fish, Eusphyra blochii and
Carcharhinus bleekehi. Pak. J.Pharma. Sci. 13: 5-12.
Saiki, M., Okano, S. and Mori, T. (1955) The radioactive material in the radiologically-
contaminated fish caught in the Pacific Ocean in 1954. Nippon Suisan, Gakkaishi
(Bull. Japan. Soc. Sci. Fisheries). 20: 902-906.
Saito, H., Ishihara, K. and Murase, T. (1997) The fatty acid composition in tuna (bonito,
Euthynnus pelamis) caught at three different localities from tropics to temperate.
J. Sci. Food. Agric. 73: 53-59.
Saito, M.A. and Offett, J.W.M. (2001) Complexation of cobalt by natural organic
ligands in the Sargasso Sea as determined by a new high-sensitivity
electrochemical cobalt speciation methodsuitable for open ocean work. Mar.
Chem.75: 49-68.
Sankar, T.V., Zynudheen, A.A., Anandan, R. and Viswanathan Nair, P.G., (2006)
Distribution of organo-chlorine pesticides and heavy metal residues in fish and
shellfish from Calicut region, Kerala, India. Chemosphere. 65: 583-590.
Sargent, J.R. (1995) Origin and functions of eggs lipids: nutritional implications. In: N.R.
Bromage and R.J. Roberts (Eds.), Broodstock Management and Egg and Larval
Quality. Blackwell Science, London: 353–372.

Sargent, J.R., Gatten, R.R. and Mcintosh, (1973) The distribution of neutral lipids in
shark tissues. J. Mar. Biol. Ass. U.K. 53: 649-656.
Sastry, K.V. and Das Gupta, A. (1991) Effect of Nwvacron on the nutritive value of fresh
water teleost fish channa punctatus. J. Envi. Biol. 12: 243-248.
Satheeshkumar, P. and Kumar, D.S. (2011) Identification of heavy metals contamination
by multivariate statistical analysis methods in Pondicherry mangroves. India. J.
Environ. Earth. Sci. 1: 30-48.
Satheeshkumar, P., Senthilkumar, D., Ananthan, G., Soundarapandian, P. and Khan, A.B.
(2011) Measurement of hematological and biochemical studies on wild marine
carnivorous fishes from Vellar estuary, Southeast Coast of India. Comp. Clin.
Path. 20: 127-134.
Schoeters, G., Hond, E., Zuurbier, M., Naginiene, R., Hazael, P., Stilianakis, N.,
Ronchetti, R. and Koppe, J. (2006) Cadmium and children: exposure and health
effects. Acta. Paediatrica. 95: 50-54.
Sebastine, M., Chakraborty, K., Bineesh, K.K., Pillai, N.G.K., Abdusamad, E.M. and
Vijayan, K.K. (2011) Proximate composition and fatty acid profile of the
myctophid Diaphus watasei Jordan & Starks from the Arabian Sea. Ind. J. Fish.
58: 103-107.
Secrieru, D. and Secrieru, A. (2002) Heavy Metal Enrichment of Man-made Origin of
Superficial Sediment on the Continental Shelf of the Northwestern Black Sea.
Estuar. Coast. Shelf. Sci. 54(3): 513-526. doi:10.1006/ecss.2000.0671.

Seixas, T.G., Kehrig, H.A., Fillmann, G., Ana Paula, M., Di, B., Souza, C.M., Secchi,
E.R., Moreira, I. and Malm, O. (2007) Ecological and biological determinants of
trace elements accumulation in liver and kidney of Pontoporia blainvillei. Sci.
Total. Environ. 385: 208- 220
Shamsan, E.F. and Ansari, Z.A. (2010) Biochemical composition and caloric content in
sand whiting Sillago sihama (Forsskal), from Zuari Estuary, Goa. Indian J. Fish.
57: 61-64.
Shamsudin, S. and Salimon, J. (2006) Physicochemical Characteristics Of Aji-Aji Fish
Seriola nigrofasciata Lipids. Mal. J. Anal. Sci. 10: 55-58.
Shotton Roger (UNCDF). (2005) Local Development Programme Guidelines. Based on
accumulated experience of past and current LDPs across the globe. United
Nations Capital Development Fund.Two United Nations Plaza, 26th Floor. New
York, NY 10017.
Shraf, M., Tariq, J. and Jaffar, M. (1991) Contents of trace metals in fish, sediment and
water from three freshwater reservoirs on the Indus River, Pakistan. Fish. Res. 12:
355-364.
Silas, E G. and Regunathan, A. (1974) Studies on demersal fishes of the deep neritic
waters and the continental slope. 3. On the occurrence of the oil fish, Rivettus
pretiosus Cocco (Gempylidae: Pisces) on the upper continental slope along the
south west coast of India. J. Mar. Biol. Ass. India. 16: 291-294.
Simopoulos, A.P. (1991) Omega-3 fatty acids in health and disease and in growth and
development. Am. J. Clin. Nutr. 54: 438-443.
Singh, M. (2005) Essential fatty acids, DHA and the human brain. J. Pediatr. 72: 239-
242.
Sivakami, S. (1990) Observations on the demersal fishery resource of the coastal and
deep sea areas of the Exclusive Economic Zone of India. Proc. First Wiorkshop
Scient. Resul. FORV Sagar Sampada, 5-7 June, 1989. 215-231.
Sivakami, S., Marichamy, P., Livingston, P., Gopakumar, G., Thiagarajan, R.,
Vivekanandan, E., Vidyasagar, K., Selvaraj, G.S.D., Muthusamy, S., Pillai
N.G.K. and Khan, M.Z. (1996) Distribution of finfish resources along
southeastcoast of India in relation to certain environmental parameters. In: Proc.
Second Workshop Scient. Resul. FORV Sagar Sampada. 315-330.
Sivakami, S., Vivekanandan, E., Nammalwar, P., Khan, M. F., Zacharia, P. U., Mohanraj,
G., Mathew, G., Jayasankar, P. (1998) The nonconventional finfish resources of
the Indian EEZ. In: M.S. Hameed and B.M. Kurup (Eds.) Technological
Advancements in Fisheries. Publn. No. 1- School Indl. Fish., Cochin University of
Science and Technology, Cochin, 243-255.
Sivani, G. (1994) Studies on biology, nutritive value and fishery of some commercially
important fishes of Gosthani estuary near Visakhapatnam. Ph.D. thesis, Andhra
University.
Sivaperumal, P., Sankar, T.V. and Viswanathan Nair, P.G. (2007) Heavy metal
concentrations in fish, shellfish and fish products from internal markets of India
vis-a-vis international standards. Food. chem. 102: 612-620.
Sivaprakasam, T.E. (1986) Potential trawl fishery for mackerel off Orissa and West
Bengal coasts. 0cc. Pap. Fish. Surv. India, 2: 7 p.
Skorupa, P. (1998) Environmental impact statement: selenium and fish. Mar. Poll. Bull.
46: 424-429.
Smith, F.E. and Arsenault, E.A. (1996) Microwave assisted sample preparation in
analytical chemistry. Talanta, 43:1207-1268.
Smith, P.K., Mahdavi, J., Carvalho, M., Fisher, S., Russell, S. and Tippett, N. (2008)
Cyberbullying: Its nature and impact in secondary school pupils. J. Child.
Psychol. Psychiatry. 49: 376-385.
Sobha, K., Porinima, A., Harini, P. and Veeraiah, K. (2007) A study on biochemical
changes in the freshwater fish, Catla catla (Hanailton) exposed to the heavy metal
toxidant Cadmium Chloride. Kathmandu University. J. Sci. Eng. Technol. 1: 112-
117.
SOFIA. (2016) The State of World Fisheries and Aquaculture 2016. Rome. p 10 & 20.
(http://www.fao.org/3/a-i5798e.pdf).
Soltan, S.S.A.M. and Gibson, R.A. (2008) Levels of omega-3 fatty acids in Australian
seafood. Asia. Pac. J. Clin. Nutr. 17: 385-390.

Sreedhar, U., Sudhakar, G.V.S. and Meenakumari, B. (2007) Deep-sea fish catch from 16
stations off southeast coast of India. J. Mar. Biol. Ass. India. 49: 183-187.
Sreenivasa Rao, C. and Neeraja, P. (1989) Transmission pattern and protein levels in
selected tissues of fish Tilapia mossambica on chromic Ammonia stress. Environ.
Ecol. 7: 915-917.
Srivastava, V.K. (1959) Limnological studies of freshwater bottom II. Qualitative
composition and variation of the available food supply for fishes. Proc. Nat. Sci.
India. 29: 207-210.
Stanby, M.E. and Olcott H.S. (1963) Composition of Fish in Industrial Fishery
Technology. J.A. Dassow, Ed. Chapman & Hall. London.
Stange, K., Maage, A. and Klungsoyr, J. (1995) Contamination in fish and sediment in
the North Atlantic Ocean. TemaNord 1996:522. Nordic Council of Ministers,
Copenhagen.
Stansby, M. (1985) Fish or fish oil in the diet and heart attack. Mar. Fish. Rev. 46: 60-63.
Stansby, M.E. (1981) Reliability of fatty acids purporting to represent composition of oil
from different species of fish. J. Am. Oil. Chem. Soc. 58: 13-16.
Stark, A.K., Sriskantharajah, S., Hessel, E.M. and Okkenhaug, K. (2015) PI3K inhibitors
in infl ammation, autoimmunity and cancer. Curr. Opin. Pharmacol. 23: 82-91.
Stephan, G., Dreanno, C., Guillaume, J. and Arzel, J. (1996) Incidence of different
amounts of proteins, lipids and carbohydrates in diets on the muscle lipid
composition in the turbot (Scophthalmus maximus). Ichtyophysiol. 19: 11-30.

Stokens, I.S., Okland, H.M.W., Falch, E. and Marianne, S. (2004) Fatty acid and lipid
class composition in eyes and brain from teleosts and elasmobranches. Biochem.
Molecular. Biol. 138: 183-191.
Stone, N.J. (1996) Fish consumption, fish oil, lipids and coronary heart disease.
Circulation. 94: 2337-2340.
Stouthart, X.X., Hans, J.M., Lock, R.C. and Wendelaar, S.E. (1996) Effects of water pH
on copper toxicity to early life stages of the common carp (Cyprinus carpio).
Environ. Toxicol. Chem. 15: 376-383.
Subramanian, K.S. (1988) Lead. in Quantitative Trace Analysis of Biological Materials.
Mckenzie, H.A. and Smythe, A. (Eds.), Amsterdam: Elsevier.
Sugeng, H., Suseno, R., Permata, S., Hayati, R. and Nugraha, S. (2013) Fatty Acid
Composition and Steroid Content of Selected Deep Sea Fishes (Escolar
(Lepidocibium plavobrunneum) and Deep Sea Lobster (linuparus somniosus))
from Southern Java Ocean–Indonesia. 2nd International Conference on
Ecological, Environmental and Bio- Sciences (ICEEBS'2013) Dec. 20-21, 2013
Bali (Indonesia).
Sulochanan, D. and John, M.E. (1988) Offshore, deep sea and oceanic fishery resources
of Kerala coast. Bull. Fish. Surv. India. 16: 27-48.
Suman, A., Wudianto. and Bintoro, G. (2006) Species composition, distribution, and
potential yield of deep sea shrimp resources in the Western Sumatera of the Indian
Ocean EEZ of Indonesian waters. J. Ind. Fish. Res. 12: 159-167.

Sunda, W.G. (2002) Bioavailability and bioaccumulation of iron in the sea, p. 41–84. In
D. R. Turner and K. A. Hunter [eds.], The biogeochemistry of iron in seawater.
Wiley.
Sunda, W.G. and and Huntsman, S.A. (1995) Cobalt and zinc interreplacement in marine
phytoplankton: Biological and geochemical implications. Limnol. Oceanogr. 40:
1404-1417.
Suseno, S.H., Suman, A., Wudiyanto. and Damayanti, A. (2007) The nutrient and steroid
content of some deep sea fish species from west Sumatera ocean. Prosiding
Konferensi Sains Perikanan dan Kelautan Indonesia I Kampus FPIK IPB
Dramaga 18-17 July. p 24-36.
Suseno, S.H., Tajul, A.Y., Nadiah, W.A., Hamidah, A. and Ali, S. (2010) Proximate,
fatty acid and mineral composition of selected deep sea fish species from
Southern Java Ocean and Western Sumatra Ocean, Indonesia. Int. Food. Res. J.
17: 905-914.
Tang, H., Chen, L., Xiao, C. and Wu, T. (2009) Fatty acid profiles of muscle from large
yellow croaker (Pseudosciaena crocea R.) of different age. J. Zhejiang. Univ. Sci.,
10: 154-158.
Tariq, J. Jaffar, M. and Ashraf, M. (1994) Trace metal concentration, distribution and
correlation in water, sediment and fish from the Ravi River, Pakistan. Fish. Res.
19: 131-139.
Tawari-Fufeyin, P. and Ekaye, S.A. (2007) Fish species diversity as indicator of pollution
in Ikpoba river, Benin City, Nigeria. Rev. Fish Biol. Fisheries. 17: 21-30.
Teal, J. (1971) Pressure effects on the respiration of vertically migrating decapod
crustacea. Amer. Zool. 11: 571-576.
Telisman, S., Cvitkovic, P., Jurasovic, J., Pizent, A., Gavella, M. and Rocic, B. (2000)
Semen quality and reproductive endocrine function in relation to biomarkers of
lead, cadmium, zinc, and copper in men. Environ. Hlth. Perspect. 108: 45-53.
Thirumilu, P. and Rajan, S. (2003) Commercial exploitation of deep-sea fishes and
crustaceans along Tamilnadu and Pondycherry coast. Mar. Fish. Infor. Serv. T &
E ser. 178: 6-8.
Thomas, J., Venu, S. and Kurup, B.M. (2003) Length-weight relationship of some deep-
sea fish inhabiting the continental slope beyond 250m depth along the West Coast
of India. NAGA, ICLARM. 26: 17-21.
Thompson, B., Adelsbach, T., Brown, C., Hunt, J., Kuwabara, J., Neale, J., Ohlendorf,
H., Schwarzbach, S., Spies, R. and Taberski, K. (2007) Biological effects of
anthropogenic contaminants in the San Francisco Estuary. Environ. Res. 105:
156-174.
Timmermans, K.R., Gerringa, L.J.A., de Baar, H.J.W., van der Wagt, B., Veldhuis,
M.J.W., de Jong, J.T.M., Croot, P.L. and Boye, M. (2001) Growth rates of large
and small Southern Ocean diatoms in relation to availability of iron in natural
seawater. Limnol. Oceanogr. 46: 260-266.

Topcuoglu, S., Kırbasoglu, C. and Gungor, N. (2002) Heavy metals in organisms and
sediments from Turkish Coast of the Black Sea, 1997–1998. Environ. Int. 27:
521-526.
Tornaritis, M., Peraki, E., Georgulli, M., Kafatos, A., Charalambakis, G., Divanack, P.,
Kentouri, M., Yiannopoulos, S., Frenaritou, H. and Argyrides, R. (1994) Fatty
acid composition and total fat content of eight species of Mediterranean fish. Int.
J. Food. Sci. Nutr. 45: 135-139.
Torres, J.J. and Somero, G.N. (1988) Metabolism, enzymatic activities and cold
adaptation in Antarctic mesopelagic fishes. Mar. Bio. 98: 169-180.
Torres, J.J., Belman, B.W. and Childress, J.J. (1979) Oxygen consumption rates of
midwater fishes as a function of depth of occurrence. Deep. Sea. Res. 26: 185-
197.
Tracey, D.M. and Horn, P.L. (1999) Background and review of ageing orange roughly
(Hoplostethus atlanticus Trachichthyidae) from New Zealand and elsewere. N.Z.J.
Mar. Freshwater. Res. 33: 67-86.
Turan, H., Sönmez, G. and Kaya, Y. (2007) Fatty acid profile and proximate composition
of the thornback ray (Raja clavata L. 1758) from the Sinop coast in the Black Sea.
J. Fish. Sci. 1: 97-103.
Turkmen, A., Turkmen, M., Tepe, Y. and Akyurt, I. (2005) Heavy metal in tree
commercially valuable fish species from Iskenderun Bay, Northern East
Mediterranean Sea, Turkey. Food. Chem. 91: 167-172.

Tuzen, M. (2003) Determination of heavy metals in fish samples of the middle Black Sea
(Turkey) by graphite furnace atomic absorption spectrometry. Food. Chem. 80:
119-123.
Unlu, E., Akba, O., Sevim, S. and Gumgum, B. (1996) Heavy metal levels in mullet, Liza
abu (Heckel, 1843) (mugilidae) from the Tigris river, Turkey. Fresen. Environ.
Bull. 5: 107-112 .
Usero, J., Izquierdo, C., Morillo, J. and Gracia, I. (2003) Heavy metals in fish (Solea
vulgaris, Anguilla anguilla and Liza aurata) from salt marshes on the southern
Atlantic coast of Spain. Environ Int. 29: 949-956.
USFDA (United States Food and Drug Administration). (1993) Guidance Document for
Chromium in Shellfish. DHHS/PHS/FDA/CFSAN/Office of Seafood,
Washington, DC.
Vaccaro, A.M., Buffa, G., Messina, C.M., Santulli, A. and Mazzola, A. (2005) Fatty acid
composition of a cultured sturgeon hybrid (Acipenser naccarii · A. baerii). Food.
Chem. 93: 627-631.
Valeria, S., Smith, C.M.M. and Donovan, A. (2003) Microwave digestion of sediment,
soils and urban particulate matter for trace metal analysis. Talanta, 60: 715-723.
DOI: 10.1016/S0039-9140(03)00131-0
Vas, P., Gordon, J.D.M., Fielden, P.R. and Overnell, J. (1993) The trace metal ecology of
ichthyofauna in the Rockall Trough, Northeastern Atlantic. Marine. Poll. Bull. 26:
607-612.
Vaulyeva, O.B., Sidorov, V.S. and Lizenko, K.I. (2004) Lipid composition of
Lipoproteins in female rainbow trout Salmo gairelnerii in annual cycle. Institute
of Biology, Karelian Research Centry Russian Academy of Sciences, U1.
Pushkinskaya. Russ. J. Ichthyol., 44: 4-9.
Velusamy, A., Satheesh Kumar, P., Ram, A. and Chinnadurai, S. (2014) Bioaccumulation
of heavy metals in commercially important marine fishes from Mumbai Harbor,
India. Mar. Pollut. Bull. 81: 218-224.
Venkiteswaran, K. (2007) India second largest producer of farmed fish.
http://www.hindu.com/2007/02/04 stories/2007020401111500.htm.
Venu, S. and Kurup B.M. (2006a) Life history traits of silver roughy Hoplostethus
mediterraneus (Cuvier) (Family: Trachichthydae) from the continental slope of
south west coast of India. Fish. Tech. 43: 204-211.
Venu, S. and Kurup, B.M. (2002a) Distribution and abundance of Deep Sea fishes along
the west coast of India. Fish. Tech. 39: 20-26.
Venu, S. and Kurup, B.M. (2002b) Distribution and biology of the deep sea fish
Psenopsis cyanea (Alcock) inhabiting continental slope of the west coast of India.
J. Mar. Biol. Ass. India. 44: 176-186.
Venu, S. and Kurup, B.M. (2006b) Distribution and biology of deep-sea fishes
Neoepinnula orientalis Gilchrist and von Bonde 1924 and Psenes squamiceps
(Lloyd, 1909 from west coast of Indian EEZ. J. Mar. Biol. Ass. India. 48: 24-28.
Verghese, B. and Jayasankar, P. (1999) Electrophoretic patterns of sarcoplasmic proteins
in Carangid fishes. Indian. J. Fish. 46: 383-389.
Vijayakumar, P., Lavanya, R., Veerappan, N. and Balasubramanian, T. (2011) Heavy
Metal Concentrations in Three Commercial Fish Species in Cuddalore Coast,
Tamil Nadu, India. J. Exp. Sci. 2: 20-23.
Vijayakumaran, K. and Naik, S.K. (1988) A study of the stock of Priacanthus hamrur
(Forsskal) during March and September between Lat. 11-16 N along the west
coast of India. Fish. Surv. India, Spl. Pub. No. 2: 106-119.
Vijayakumaran. K., and Philip, K.P. (1988) Deep sea demersal fishery resources off the
north Kerala ,Karnataka and Konakan coast. Symp. Tropical Marine Living
Resources, J. Mar. Biol. Assoc. India. 51: 12-16.
Vijayan Divya, K., Jayarani, R., Dilip Kumar, S., Chatterjee, N.S., Suseela M., Mohanty,
B.P., Sankar, T.V. and Anandan, R. (2016) Comparative studies on nutrient
profiling of two deep sea fish (Neoepinnula orientalis and Chlorophthalmus
corniger) and brackish water fish (Scatophagus argus). J. Basic. Appl. Zool. 77:
41-48.
Vivekanandan, E. (2001) Predatory diversity of two demersal fish species in the trawling
grounds off Veraval. Indan. J. Fish. 48: 133-143.
Vivekanandan, E. (2011) Climate Change and Indian Marine Fisheries. CMFRI Special
Publication, 105: p. 97.

Vivekanandan, E., Srinath, M., Pillai, V.N., Immanuel, S. and Kurup, K.N. (2003)
Trophic model of the coastal fisheries ecosystem of the southwest coast of India.
In: Assessment, Management and Future Directions for Coastal Fisheries in Asian
Countries (Silvestre, C., Garces, L., Stobutzki, I., Ahmed, M., Valmonte-Santos,
R.A., Luna, C., Lachica-Alino, L., Munro, P., Christensen, V. and Pauly, D. Eds.)
World Fish Center Conference Proceedings, Penang. 67: 281-298.
Vlieg, P., Lee, J. and Grace, N.D. (1991) Elemental concentration of marine and
freshwater finfish and shellfish from New Zealand waters. J. Food. Compos.
Anal. 4: 136-147.
Wang, S., Xu, L., Zhao, Z., Wang, S., Jia, Y., Wang, H. and Wang, X. (2012) Arsenic
retention and remobilization in muddy sediments with high iron and sulfur
contents from a heavily contaminated estuary in China. Chem. Geol. 314: 57-65.
Waqar, A. (2006) Levels of selected heavy metals in tuna fish. Arab J Sci Eng. 31: 89-92.
Watanabe, Y., Oozeki, Y. and Kitagawa, D. (1997) Larval parameters determining
preschooling juvenile production of Pacific saury (Cololabis saira) in the
northwestern Pacific. Can. J. Fish. Aquat. Sci. 54: 1067-1076.
Watling, H.R. (1983) Accumulation of seven metals by Crassostrea gigas, C.
margaritacea, Perna perna and Chromytillus meridionalis. Bull. Environ. Contam.
Toxicol. 30: 213-320.
Whelan, J. (2008) Polyunsaturated fatty acids (n-6 and n-3) and the aging brain: Food for
thought. J. Nutr. 138: 2521-2522.

WHO (World Health Organisation). (1989) Environment Health Criteria. Heavy metals-
environmental aspects, Geneva, Switzerland.
WHO (World Health Organisation). (2003) Diet, nutrition and the prevention of chronic
diseases, WHO Technical Series Report 916 (Geneva). Accessed January 2015 at:
http://whqlibdoc.who.int/trs/who_trs_916.pdf.
WHO (World Health Organisation). (2006) Health aspects of plumbing. Geneva, World
Health Organization and World Plumbing Council. Chemical fact sheet. Available
at
http://www.who.int/water_sanitation_health/publications/plumbinghealthasp/en/
Winchester, R.V. (1988) Trace metal levels in fish from the Manukau Harbour,
Auckland, New Zealand, related to water pollution incident. New. Zeal. J. Mar.
Fresh. Res. 22: 621-624.
Windom, H., Stein, D., Sheldon, R. and Smith, R. (1987) Comparison of trace metal
concentrations in muscle tissue of a benthopelagic fish (Coryphenoides armatus)
from the Atlantic and Pacific Oceans. Deep. Sea. Res. 34: 213-220.
Wittmann, F. (1974) Toxic Metals. ln: Forstner, U. and G.T.W. Wittmann (Eds) Metal
Pollution in the aquatic environment - Springer-verlag, Berlin. p 3-68.
Yamada, M. and Hayashi, K. (1975) The lipids of marine animals from various habitat
depths. I. Fatty acid composition of lipids from 22 species of fish and molluscs.
Bull. Jpn. Soc. Sci. Fish. 41: 1143-1152.

Yancey, P.H., Gerringer, M.E., Drazen, J.C., Rowden, A.A. and Jamieson, A. (2014)
Marine fish may be biochemically constrained from inhabiting the deepest ocean
depths, Proceedings of the National Academy of Sciences. 111: 4461-4465.
Yehezkel, G., Chayoth, R., Abdu, U., Khalaila, I. and Sagi, A. (2000) High-density
lipoprotein associated with secondary vitellogenesis in the hemolymph of the
crayfish Cherax quadricarinatus. Comp. Biochem. Physiol. 127: 411-121.
Yılmaz, A.B. (2003) Levels of heavy metals (Fe, Cu, Ni, Cr, Pb, and Zn) in tissue of
Mugil cephalus and Trachurus mediterraneus from Iskenderun Bay, Turkey.
Environ. Res. 92: 277-281.
Yoneyama, S., Miura, K., Sasaki, S., Yoshita, K., Morikawa, Y., Ishizaki, M., Kido, T.,
Naruse, Y. and Nakagawa, H. (2007) Dietary intake of fatty acids and serum C-
reactive protein in Japanese. J. Epidemiol. 17: 86-92.
Yongmanitchai, W. and Ward, O.P. (1989) Omega-3 fatty acids: alternative sources of
production. Process. Biochem. 24: 117-125.
Zaboukas, N., Miliou, H., Megalofunou, P. and Maraitou Apostolopoulou, M. (2006)
Biochemical composition of the Atlantic bonito, Sarda sarda from the Aegean Sea
in different stages of sexual maturity. J. Fish. Biol. 69: 347-362.
Zezina, O.N. (1985) Some rare species of the recent brachiopoda and the problem of
"living fossils" [in Russian]. In: Ecology of the ocean coastal zone benthic fauna
and flora, P. P. Shirshov Inst. Oceanol., Akad. Sci., Moscow. 51: 9-14.
Zhang, Z., He, L., Li, J. and Wu, Z.B. (2007) Analysis of heavy metals of muscle and
intestine tissue in fish in Banan section of Chongqing from three Gorges reservoir,
China.. Polish J. Environ. Stud. 16: 949-958.
Zhou, J.L., Salvador, S.M. and Liu, Y.P. (2001) Sequeria M. Heavy metals in the tissues
of common dolphins (Delphinus delphis) stranded on the Portuguese coast. Sci.
Total. Environ. 273: 61-76.

Rushinadha Rao Kakara

Загружено:

Сведения о документе

Оригинальное название

Авторское право

Доступные форматы

Поделиться этим документом

Поделиться или встроить документ

Параметры публикации

Этот документ был вам полезен?

Это неприемлемый материал?

Авторское право:

Доступные форматы

Rushinadha Rao Kakara

Загружено:

Авторское право:

Доступные форматы

BIOCHEMICAL, HEAVY METAL AND FATTY ACID PROFILING STUDIES

ON SOME SELECTED DEEP-SEA FISHES ALONG THE CONTINENTAL

RUSHINADHA RAO KAKARA

Under the Supervision of

Dr. U. SREEDHAR Prof. K. SREERAMULU

THESIS SUBMITTED TO ANDHRA UNIVERSITY, VISAKHAPATNAM IN

The work incorporated in this thesis is a part of the research project

“Exploration and Assessment of demersal fishery resources along the continental

laboratory of ICAR - Central Institute of Fisheries Technology (CIFT),

Visakhapatnam. I hereby declare that this is the novel work entitled

BIOCHEMICAL, HEAVY METAL AND FATTY ACID PROFILING

STUDIES ON SOME SELECTED DEEP-SEA FISHES ALONG THE

CONTINENTAL SLOPE (200 TO 1200 M DEPTH) OF INDIAN EXCLUSIVE

DR. U. SREEDHAR (Internal Research Director), ICAR-Central Institute of

Fisheries Technology, Visakhapatnam and Prof. K. SREE RAMULU (External

Research Director), Department of Zoology, Andhra University, Visakhapatnam and

Institution for the award of any Degree

Date: (RUSHINADHA RAO KAKARA)

out by Shri. RUSHINADHA RAO KAKARA under my guidance.

out by Shri. RUSHINADHA RAO KAKARA under my guidance as an extramural

(K. SREE RAMULU)

It is an immense pleasure to express my deep sense of gratitude to my Internal Research

I am sincerely thankful to Dr. N. RAVI SANKAR, Director, ICAR-Central Institute of

I am much indebted to Dr. G. RAJESWARI, Principal Scientist and Scientist-In-

My sincere thanks to Dr. R. RAGHU PRAKASH, Principal Scientist, ICAR-Central

My profound thanks to Dr. B. MADHUSUDHANA RAO, Principal Scientist, ICAR-

I humbly express my sincere gratitude to Dr. M.M. PRASAD, Dr. L. N.

I sincerely acknowledge Ministry of Earth Sciences, New Delhi for providing

I am very grateful to express my special gratitude to Shri. R. UMA MAHESWARA

I also express my sincere gratitude to Shri. P. RADHA KRISHNA, PRASANNA

It is my pleasure to acknowledge my tenants R.S. NAIDU & R. SATYAVATHI at

Rushinadha Rao Kakara

pelagic-oceanic The pelagic-oceanic fisheries with which we are concerned here

mechanized fishing has encouraged many entrepreneurs to do the same, resulting in a

47,000 (unpublished). This has resulted in intensive non-selective fishing in order to

regulations for responsible fishing (Gopakumar, 1997).

improvement in efficient fishing technology, that is becoming available to even smaller

gear and time to reproduce. Biological overexploitation of fishery resources leads to

attracted to the structures.

Oceanographic Research Vessel) Sagar Sampada, Ministry of Earth Sciences, India.

Data collected during deep-sea fishery expeditions of FORV (Fisheries

Oceanographic Research Vessel) Sagar Sampada is depicted. The information provided

in this investigation is new and would be of a great interest and curiosity. By

more amount of protein accumulation was found in Chelidoperca investigatoris Cubiceps

in cruise 322 is Fe>Zn>Cu>Cr>Co>As>Cd>Pb>Se, whereas in cruise 332 is

Fe>Zn>Cu>Co>Cr>Cd>Pb>As>Se and the order of metal content in cruise 338 is

Fe>Zn>Cu>Co>Cr>Cd>As>Pb>Se. The obtained results indicate that environmental

and scanty literature has been found.

definitely also compete when it comes to taste.

development and creation of employment as it stimulates enlargement of a number of

subsidiary industries. It is an imperative source of low-priced and nutritious food,

moreover it provides as an instrument of livelihood for a large section of economically

divisions, namely pelagic-neritic fisheries and pelagic-oceanic fisheries. The previous

decade (1.5% versus 2.5%) (SOFIA, 2016).

Trichiurus auriga, Chlorophthalmus agassizi, Neoepinnula orientalis and Cubiceps sp.

against dreadful human pathogens (Ravichandran et al., 2010).

(Mona et al., 2011).

specific and species-specific changes in composition. The composition of the fish

analysis of the same is referred to as proximate composition (Love, 1970).

supplement to prevent a number of coronary heart diseases (Edirisinghe et al., 1998).

complimentary to each other. As lipids increase, the water content decreases

surface-living and mesopelagic (midwater) species, fewer studies have considered

biochemistry and metabolism of deep-sea species. In fact, proximate composition, energy