Comparative Clinical Pathology (2018) 27:1335–1342

https://doi.org/10.1007/s00580-018-2744-z

ORIGINAL ARTICLE

Effect of sex on haemocytobiochemical profiling of silver tiger fish

(Datnioides polota Hamilton, 1822)
Gayatri Acharya 1 & Prafulla Kumar Mohanty 2

Received: 3 April 2018 / Accepted: 16 May 2018 / Published online: 26 May 2018
# Springer-Verlag London Ltd., part of Springer Nature 2018

Abstract
This investigation presents an overview of the effect of sex on haematobiochemical and haemocytomorphometrical aspects
of Datnioides polota. For the present study, 15 males and 15 females of Datnioides polota were collected from Balugaon,
the central sector of Chilika lagoon of Odisha. Two millilitres of blood samples was collected from each sex through the
caudal vein. One millilitre of collected blood was transferred from the syringe into ethylenediaminetetraacetic acid
(EDTA) containing vials, for haematological studies and other 1 ml was taken in Eppendorf tube without anticoagulant
for analyses of biochemical parameters. Haematological parameters like red blood cell count (RBC), white blood cells
count (WBC), haemoglobin, haematocrit, mean cell volume (MCV), mean cell haemoglobin (MCH), mean cell
haemoglobin concentration (MCHC), biochemical parameters such as glucose, protein, albumin, globulin, cholesterol,
and morphometrical parameters namely length, breadth, area, and N/C ratio of erythrocytes, lymphocytes, monocytes,
eosinophils, and neutrophils are analysed in relation to the sex of the fish. Statistical analysis reveals the sex wise
significant difference in some haematological parameters like Hb which varies at P < 0.01 level while WBC deviate at
P < 0.05 level. All the biochemical parameters show higher value in females in comparison to males except albumin but
sex wise significant variation recorded in protein and globulin at P < 0.01 level. In males, the concentration of Hb is
positively correlated with RBC and PCV while in females negatively correlated. RBC is positively correlated with PCV in
both the sexes at R2 = 0.008 and R2 = 0.9 in male and female, respectively. The morphometry of blood cells such as
monocytes and lymphocytes shows significant sexual dimorphism. The cellular length and area of monocytes deviate at
the level of P < 0.01 and breadth at P < 0.05 level. The cellular and nuclear breadth of lymphocyte vary significantly at the
level of P < 0.05 and P < 0.001, respectively. The results of this study may serve as a reference value for haemocytological
and biochemical parameters of this species which in turn helps in diagnosing diseases and increasing fish production.

Keywords Sex . Haematological parameters . Blood cells . Morphometry . Chilika lagoon . Datnioides polota

Background unique, spectacular, and peculiar because of varieties of

Fishes are the first successful aquatic vertebrates of Devonian
period of geological time scale that are characterised by the
presence of paired fins with fin rays and branchial respiration.
Among all the faunal diversity on the earth, the fish group is
* Gayatri Acharya
gayatri.acharya65@gmail.com
1
PG Department of Zoology, Research Scholar, Utkal University, Vani
Vihar, Bhubaneswar, India
2
PG Department of Zoology, Utkal University, Vani Vihar,
Bhubaneswar, India
morphological features. Among the various groups of verte-
brates, fishes are very sensitive to pollutants and regarded as
the most suitable laboratory test organism. Many clinical tools
which are used to evaluate mammalian health are not utilised
for the use of fishes. As the aquaculture industry expands,
there is a requirement of improved diagnostic methods.
Few tools are available to diagnose and monitor diseases
in fishes. One such tool is the analysis of blood, which
helps in detecting acute and chronic pathophysiological
changes in the body due to nutrition, water quality, toxi-
cants, and disease. The haematological characteristics can
be used as an effective and efficient tool to monitor phys-
iological and pathological changes in fishes. Normal
ranges for various blood parameters in fish have been
1336
established by different investigators in fish physiology and
pathology (Darvish et al. 2009; Satheeshkumar et al. 2011).
Haematological and biochemical parameters are being used as
an indicator in the assessment of health condition, toxicological
symptoms of organisms (Rao 2006) and it also indicates the
abnormal environmental condition (Elahee and Bhagwant
2007). Information about the existence, status, and degree of
possible sickness in organisms can be instantly obtained by the
use of haematological and biochemical parameters. One of the
difficulties in assessing the state of health of the natural fish
population is due to the lack of reliable references of
haematobiochemical parameters in the normal condition
(Kori-Siakpere et al. 2005; Satheeshkumar et al. 2011).
Although fish haematology acts as a valuable tool, but normal
range for blood parameters are lacking and literature in this area
is often incomplete (Satheeshkumar et al. 2011). Only a few
normal values for a limited number of haematological parame-
ters have been established for some teleosts, but these values
vary widely due to the lack of standardised collection and eval-
uation techniques. Number of factors cause normal and abnor-
mal variation in haematological data (Clauss et al. 2008) such as
species and strain (Langston et al. 2002), temperature (Langston
et al. 2002; Magill and Sayer 2004), age (Svetina et al. 2002),
season (Hofer et al. 2000), stress (Cnaani et al. 2004), photope-
riod (Leonardi and Klempau 2003), nutritional state (Svetina et
al. 2002; Lim and Klesius 2003), the cycle of sexual maturity,
and health condition (Rey Vazquez and Guerrero 2007).
Aim
Blood is a fluid connective tissue as well as a pathophysiolog-
ical reflector of the whole body. Therefore, blood parameters
are important in diagnosing the functional status of the animal.
Since the influence of sex on haemocytological and biochem-
ical parameters along with the morphometrical characteristics
of blood cells of Datinoides polota is inadequate and incon-
sistent, an attempt has been made to study the influence of sex
on silver tiger fish Datinoides polota, which is a common
edible and delicious fish of brackish water lagoon, Chilika.
Methods
In the present study, healthy, adult, and live specimens of both
the sexes having 15 from each sex of Datnioides polota were
collected from the Balugaon, the central sector brackish water
lagoon Chilika (Fig. 1), Odisha from the month of June to
August, 2016. All procedures used for analysis followed ap-
proved guidelines for the ethical treatment of animals. Before
the collection of blood, specimens are weighed and their total
length is measured. The blood samples are taken from the
caudal vein (Campbell 2006) using 22-gauge needle in the
morning hour to avoid diurnal variation. Care is taken not to
Comp Clin Pathol (2018) 27:1335–1342
puncture the caudal lymph vessels during the collection of
blood. Blood smears (three slides per individual fish) are
prepared immediately after the collection of blood to avoid
morphological changes. The smeared slides are air dried,
fixed with methanol, stained with Giemsa’s stain, and kept
for further morphological analyses of blood cells. The rest
of the collected blood is immediately separated into two
tubes out of which one contains ethylenediaminetetraacetic
acid (EDTA) for the determination of haematological
parameters and other without EDTA for the analysis of serum
biochemical parameters.
Total red blood cells (RBC) and total white blood cells
(WBC) are manually counted using a Neubauer ’s
haemocytometer. The haemoglobin concentration is estimated
by Sahli’s haemometer (Sahli 1909). Packed cell volume
(PCV) is determined using the microhaematocrit method
(Mcinroy 1953). Erythrocyte indices such as mean corpuscu-
lar volume (MCV), mean corpuscular haemoglobin (MCH),
and mean corpuscular haemoglobin concentration (MCHC)
are calculated as per following formulae.
MCV ¼ PCV=Erythrocytes count  10
MCH ¼ Haemoglobin=Erythrocytes count  10
MCHC ¼ Haemoglobin=PCV  100
Blood smears are used for differential WBC count (three
slides per fish) and classified as lymphocytes, neutrophils,
monocytes, and eosinophils. In order to obtain the size of
different blood cell types, 30 cells of each cell type for each
individual fish are photomicrographed with the help of micro-
scope eyepiece digital camera (CatCam130–1.3 MegaPixel
(MP), Code No. CC130, Catalyst Biotech, Maharashtra,
India,attached to Hund Wetzlar Microscope GmbH, Wetzlar-
Nauborn, Germany) and computer.
For biochemical analyses, the Eppendorf tubes with the
blood samples are centrifuged for 5 min at 5000 rpm. The
serum so obtained is used to determine biochemical parame-
ters such as glucose, cholesterol, protein, albumin, and glob-
ulin which are measured using standard commercial kits
(Crest Biosystem, India).
Statistical analyses
Haematological and biochemical parameters were expressed
as mean ± SE in both male and female fish and compared
according to sex using t test. All these statistical analyses were
performed using the Microsoft Office Excel 2007.
Results and discussion
This investigation focuses on the effect of sex on
haematobiochemocal parameters of Datnioides polota
Comp Clin Pathol (2018) 27:1335–1342 1337

Fig. 1 Chilika lagoon, the site of collection of fish

(Table 1). This study records that the males have more
concentration of haemoglobin than that of females and
varies significantly at P < 0.001 level which may possibly
be due to more metabolic activity of males in comparison
to females or probably due to the hormonal effect (Eisler
1965). RBCs play a vital role in respiration (Wells et al.
1980) and it is a part of the complete blood count (CBC).
This test helps in diagnosis of anaemia, and other conditions
affecting red blood cells (Bunn 2011). The current study re-
flects higher RBC count in males but the significant sexual
difference is not observed. The higher RBC count in males is
possibly because of their hyperactiveness compared to the
females. RBC count has proven to be a highly variable blood
parameter among fishes and shows a remarkable difference
with respect to sex (Svobodova et al. 2008). The number of
RBC is also related to the metabolic activity of fishes as well
as to the concentration of oxygen in the surrounding water
(Starmach 1970). In this study, the correlation of Hb with
RBC is statistically established. It is noted that the concentra-
tion of Hb in males is positively correlated with RBC at R2 =
3E-05 (Fig. 2a) while negatively correlated at negligible level
in females at R2 = 0.04 (Fig. 2b). It is found that the number of
RBC is positively correlate with Hb in male and negatively
correlated with the concentration of haemoglobin in female
means the concentration of haemoglobin increases but the
number of RBC decreases which may be due to inverse rela-
tionship between total erythrocyte count and size of erythro-
cytes in slow moving fishes. So, though number of RBC

Table 1 Haematobiochemical
parameters of Datnioides polota SL NO Haematobiochemical parameters Male Female P value
n = 15 n = 15

1 RBC(106mm3) 1.72 ± 0.06 1.58 ± 0.06 0.06

2 Hb (g/dl) 6.9 ± 0.13** 6.43 ± 0.13** 0.01
3 PCV (%) 21.86 ± 0.32 21.53 ± 0.66 0.32
4 MCV (fl) 129.24 ± 5.28 139.04 ± 6.11 0.11
5 MCH (pg) 40.81 ± 1.83 41.90 ± 2.20 0.35
6 MCHC (%) 31.55 ± 0.37 30.26 ± 1.09 0.13
7 WBC(103 mm3) 12.22 ± 0.78* 14.74 ± 0.89* 0.02
8 Lymphocyte (%) 66.18 ± 1.18 67.13 ± 1.03 0.23
9 Monocyte (%) 5.46 ± 0.37 5.33 ± 0.25 0.38
10 Eosinophil (%) 4.06 ± 0.28 4.33 ± 0.39 0.29
11 Neutrophil (%) 24.46 ± 1.09 23.2 ± 0.97 0.19
12 Protein (g/dl) 9.19 ± 0.31* 10.20 ± 0.50* 0.05
13 Albumin (g/dl) 2.81 ± 0.12 2.57 ± 0.08 0.48
14 Globulin (g/dl) 6.38 ± 0.35* 7.62 ± 0.51* 0.02
15 Glucose (mg/dl) 96.86 ± 2.10 97.08 ± 3.79 0.06
16 Cholesterol (mg/l) 280.15 ± 12.02 311.52 ± 16.68 0.069

*Significant at p < 0.05

**Significant at p < 0.01
1338
Fig. 2 Correlation of Hb with
2.5
RBC in Datnioides polota. a 2.2
Male. b Female 1.9
RBC
1.6
1.3
1 1
5 6
decreases but size increases that fill with higher concentration
of haemoglobin.
Packed cell volume (PCV) is an essential blood parameter
to determine the presence of anaemia or polycythaemia in
fishes (Archer and Jeffcott 1977; Lowe 2004), and other ver-
tebrates. Impact of sex on PCV is also found in this study,
which shows higher value in males in comparison to females.
This study interprets that when concentration of haemoglobin
increases, the per cent of PCV also increases. So, a positive
correlation between Hb and PCV is observed at R2 = 0.59
(Fig. 3a). The relationship between PCV and haemoglobin
concentration is positively related probably due to the oxygen
carrying capacity of haemoglobin than the PCV itself (Rashidi
et al. 2012; Diyaware et al. 2013). However, in females, PCV
is negatively correlated with haemoglobin (Fig. 3b). The cor-
relation of PCV with RBC for both the sexes of fishes is also
interpreted. A strong positive relationship of RBC to PCV is
marked both in the male (Fig. 4a) and female ((Fig. 4b) fishes
at R2 = 0.008 and R2 = 0.9, respectively.
Erythrocyte indices like mean cell volume (MCV), mean
cell haemoglobin (MCH), and mean cell haemoglobin con-
centration (MCHC) define the size of red blood cells, and
haemoglobin content in the peripheral blood. Erythrocyte in-
dices are indicators of different types of anaemia. Depending
on erythrocyte indices, anaemia may be characterised as mi-
crocytic, normocytic, or macrocytic. According to the average
amount of haemoglobin of the red blood cells, the anaemia
may be further subdivided, as hypochromia, hyperchromia or
normochromia (Thika 1992). In the present study, except
MCHC, the value of other erythrocyte indices like MCV and
MCH is found to be more in the females in comparison to
males but does not vary significantly.
In fish, WBC is frequently used as an indicator of health status
because WBC is the key component of the innate immune sys-
tem (Duthie and Tort 1985; Gallardo et al. 2003; Ballarin et al.
2004) as in mammals. The Datnioides polota reflects higher
WBC count in females in comparison to males and deviates
Fig. 3 Correlation of Hb with
PCV in Datnioides polota. a 25
24
Male. b Female
PCV
23
22
21
20
5 6
Comp Clin Pathol (2018) 27:1335–1342
y = 0.0025x + 1.7087 y = -0.0992x + 2.2179
2.5
R² = 3E-05 2.2 R² = 0.0409
1.9
RBC
1.6
1.3
7 8 9 5 6 7 8 9
Hb a Hb b
significantly at P < 0.05 level. The differential blood cell count
is used as a tool in the diagnosis of haematological diseases.
Differential leucocyte count (DLC) of fishes is recorded in per-
centage. The presence of leucocytes is related to the health status
of fish and in many cases, these are also helpful in the evaluation
of the immune system. Therefore, variations in the proportion of
these defence cells in the blood are usually expected. In this
investigation, all the white blood cells show higher percentage
in males in comparison to females but do not deviate significant-
ly except for the per cent of lymphocytes, which is believed due
to the variation in sex. In this investigation, among all the
leucocytes, lymphocytes are the most abundant type of
leucocytes, which are present for more than 50% in the blood
of fish. This view corroborates with Campbell (2015).
Lymphocytes have highest percentage in blood as they play
a very important role in the innate immune system followed
by neutrophils, monocytes, and eosinophils. Basophils are
not detected in this study and it is believed to be absent
(Arnold 2005).
The biochemical parameters show more value in the
females in comparison to males except for albumin, but
statically significant sexual dimorphism is noted only in
protein and globulin at P < 0.05 level. This is similar to
the findings in Indian shad, Tenualosa ilisha (Jawad et al.
2004) but contradicts with the findings in Notopterus
notopterus by Kulkarni (2017) who reports that the bio-
chemical parameters have more value in males in compar-
ison to females. The serum biochemical parameters deviate
significantly with respect to sex and species in the current
finding which is due to variation in sex.
Study of vertebrate blood cells including those of fishes,
particularly teleosts has attracted the attention of ichthyolo-
gists. Gulliver (1875) undertakes a detailed study of the red
blood cells in scorpionfishes. Cytomorphometry of blood cells
include the measurement of length, breadth, and area of both
cell and nucleus as well as the estimation of N/C ratio. Due to
the presence of the lobed nucleus, it is impossible to measure
y = 1.8889x + 8.8333
25 y = -0.1186x + 22.297
R² = 0.591 24 R² = 0.0006
PCV
23
22
21
20
7 8 9 5 6 7 8 9
Hb a Hb b
Comp Clin Pathol (2018) 27:1335–1342
Fig. 4 Correlation of RBC with
PCV in Datnioides polota. a 25
24
Male. b Female
PCV
23
22
21
20
1 1.2 1.4 1.6 1.8
the nuclear length, breadth, and area of monocytes, neutro-
phils, and eosinophils. The morphology of blood cells has
great significance in the field of aquaculture and the
morphometrical observations of blood cells show significant
sexual variation. This study focuses on the morphometrical
parameters of Datnioides polota (Fig. 5a) (Table 2).
Fish erythrocytes are elongated and elliptical cells with an
oval, centrally located, nucleus and have the lifespan of 13–
150 days (Witeska 2013). It is observed that erythrocytes
(Fig. 5b) of Datnioides polota are also elliptical in their shape.
This corroborates with the findings of Srivastava (1968) and
Pandey et al. (1976) in Heteropneustes fossils. The
morphometrical parameters like cell length, breadth, and area
and N/C ratio of erythrocytes have more value in males in
comparison to the females while the nuclear metrical param-
eters are higher in the females. The sex wise significant
morphometrical deviation is not observed in erythrocytes.
Among the leucocytes, lymphocytes (Fig. 5c) are usually the
most common in the blood of fish, accounting as much as 85%
of the total leucocyte population (Groff and Zinkl 1999).
Some authors reveal the morphological diversity in leucocytes
a b
d
Fig. 5 a Adult Datnioides polota. b Erythrocyte. c Lymphocyte. d Monocyte. e Eosinophil. f Neutrophil, (bar = 40 × 10 mm)
1339
y = 0.4477x + 21.094
25 y = 3.0919x + 16.648
R² = 0.0081
24 R² = 0.0963
23
PCV
22
21
2 20
a 1 1.2 1.4 1.6 1.8 2b
RBC RBC
(Srivastava 1968; Blaxhall and Daisley 1973; Ezzat et al.
1973; Ellis 1975; Ferguson 1976; Imagawa et al. 1989;
Ueda et al. 2001). Radhakrishnan et al. (1976) reports three
types of lymphocytes in Diodon depending on their size as
large, medium and small. It is usually round in shape with
round nucleus. The morphometrical parameters of lympho-
cyte show more value in the females in comparison to males,
but significant variation is marked only in the breadth of cells
and nucleus at P < 0.05 and P < 0.0001 level, respectively.
Eosinophils (Fig. 5d) are round cells with bilobed nucleus.
The eosinophils are observed to be very rare (Kelenyi and
Nemeth 1969) and some authors are in doubt regarding their
presence in the blood (Ellis 1975; Rowley et al. 1988) and
others are unable to find eosinophils in channel catfish
(Tavares-Dias et al. 2007). However, eosinophils have been
reported in some fishes (Catton 1951). The eosinophils show
significant variation in its size with respect to sex. These cells
also participate in phagocytic activity along with monocytes
and neutrophils, but it is actively involved in metazoan para-
sitic infection and immune response to antigenic infections
(Stoskopf 1993). The morphometry of eosinophils is recorded
c
e f
1340 Comp Clin Pathol (2018) 27:1335–1342

Table 2 Cytomorphometry of blood cells of Datnioides polota

SL NO Cell type Cytoplasm/nucleus Sex P value

Male (n = 30) Female (n = 30)

1 Erythrocyte Cytoplasm Mean length in μm 9.19 ± 0.21 9.61 ± 0.20 0.08

Mean breadth in μm 7.86 ± 0.16 8.17 ± 0.23 0.14
Mean area in μm2 57.44 ± 2.20 62.95 ± 3.02 0.07
Nucleus Mean length in μm 7.06 ± 0.72 6.49 ± 0.28 0.23
Mean breadth in μm 5.25 ± 0.21 4.79 ± 0.24 0.08
Mean area in μm2 25.82 ± 2.33 24.38 ± 1.49 0.30
N/C ratio 0.44 ± 0.03 0.40 ± 0.02 0.22
2 Lymphocyte Cytoplasm Mean length in μm 9.40 ± 0.15 9.28 ± 0.16 0.29
Mean breadth in μm 8.02 ± 0.16* 7.45 ± 0.24* 0.03
Mean area in μm2 14.76 ± 0.24 14.57 ± 0.25 0.29
Nucleus Mean length in μm 5.90 ± 0.23 5.84 ± 0.16 0.41
Mean breadth in μm 4.76 ± 0.18*** 3.87 ± 0.14*** 0.0001
Mean area in μm2 9.27 ± 0.37 9.17 ± 0.26 0.41
N/C ratio 0.62 ± 0.02 0.63 ± 0.015 0.48
3 Eosinophil Cytoplasm Mean length in μm 9.89 ± 0.19 9.88 ± 0.29 0.49
Mean breadth in μm 8.38 ± 0.21 8.32 ± 0.29 0.44
Mean area in μm2 15.53 ± 0.30 15.52 ± 0.46 0.49
4 Monocyte Cytoplasm Mean length in μm 11.86 ± 0.35** 10.75 ± 0.37** 0.01
Mean breadth in μm 10.01 ± 0.35* 9.17 ± 0.35* 0.04
Mean area in μm2 18.63 ± 0.54** 16.88 ± 0.59** 0.01
5 Neutrophil Cytoplasm Mean length in μm 10.39 ± 0.39 10.63 ± 0.28 0.30
Mean breadth in μm 7.62 ± 0.17 7.68 ± 0.19 0.41
Mean area in μm2 16.31 ± 0.61 16.69 ± 0.44 0.30

*Significant at p < 0.05

**Significant at p < 0.01
***Significant at p < 0.001

to be higher in males, but does not vary significantly with
females.
Monocytes (Fig. 5e) are recorded to be the largest of the
formed blood elements (Barber et al. 1981). These are distin-
guished from granulocytes by their kidney-shaped nucleus.
The nomenclature used to describe monocytes in fishes is
variable. Monocytes have been termed haemoblasts and mac-
rophages (Barber et al. 1981), while other authors have been
unable to find monocytes (Blaxhall and Daisley 1973).
However, Campbell and Ellis (2007) report the presence of
monocytes in the blood of fish. The piscian monocytes are
phagocytic cells and involved in inflammatory response
(Rowley et al. 1988). The metrical parameters of monocytes
show higher value in males in comparison to females like cell
length and area which deviate significantly at P < 0.01 and
cellular breadth at the level of P < 0.05.
Neutrophils (Fig. 5f) are larger cells with bilobed and
sometimes with the multilobed nucleus. In teleosts, the
neutrophils are the most frequent granulocytes. These are
round cells with an eccentric nucleus and are common in
the circulating blood. Neutrophils show variation in size
and shape in different species, different habitat, and also
in respect to sex (Kumar 2016). The morphometrical mea-
surement of neutrophils is noted to be more in females, but
does not vary significantly in males.
The literatures reveal that basophils are not found in a ma-
jority of the fish. Saunders (1968) examines the formed blood
elements of 171 species of fishes and basophils are observed
only in five species. In this study, basophils are not noticed.
Similar results are also found in the blood of plaice (Ellis
1975) and rainbow trout (Blaxhall and Daisley 1973).
Conclusion
The reference haemocytological and biochemical parameters
obtained from Datnioides polota may enhance the current
understanding of the haematological values, which in turn
may assist ichthyologists and fish farmers with the ability to
Comp Clin Pathol (2018) 27:1335–1342
establish an accurate interpretation of laboratory data, critical
in diagnosis, prognosis, and treatment of diseases in fishes.
Funding This study was funded by the Department of Science and
Technology, New Delhi, India (grant number IF 130 457).
Compliance with ethical standards
This investigation followed all the guidelines for the care and use of
animals.
Conflict of interest The authors declare that they have no conflict of
interest.
References
Archer RK, Jeffcott LB (1977) Comparative clinical haematology.
Blackwell Scientific Publishers, London, p 737
Arnold JE (2005) Hematology of the sandbar shark, Carcharhinus
plumbeus: standardization of complete blood count techniques for
elasmobranchs. Vet Clin Pathol 34:115–123
Ballarin L, Dall’Oro M, Bertotto D, Libertini A, Francescon A, Barbaro
A (2004) Haematological parameters in Umbrina cirrosa (Teleostei,
Sciaenidae): a comparison between diploid and triploid specimens.
Comp Biochem Physiol 138A:45–51
Barber DL, Westermann JEM, White MG (1981) The blood cells of the
Antarctic icefish Chaenocephalus aceratus Lonnberg: light and
electron microscopic observations. J Fish Biol 19:11–28
Blaxhall PC, Daisley KW (1973) Routine haematological methods for
use with fish blood. J Fish Biol 5:771–781
Bunn HF (2011) Approach to the anaemias. In: Goldman L, Schaffer AI
(eds) Cecil medicine, 24th edn. Elsevier, Philadelphia, p 161
Campbell TW (2006) Clinical pathology of reptiles. In: Mader DR (ed)
Reptile medicine and surgery, 2nd edn. Saunders, Philadelphia, pp
453–470
Campbell TW (2015) Exotic animal haematology and cytology, 4th edn.
John Wiley and Sons, Inc, Oxford, pp 97–111
Campbell TW, Ellis CK (2007) Avian and exotic animal hematology and
cytology, 3rd edn. Blackwell Publishing, Ames, p 304
Catton (1951) Blood cell formation in certain teleost fishes. Blood 6(1):
39–60
Clauss T, Dove ADM, Arnold J (2008) Haematological disorders of fish.
Vet Clin Exot Anim Pract 11(3):445–462
Cnaani A, Tinman S, Avidar Y, Ron M, Hulata G (2004) Comparative
study of biochemical parameters in response to stress in O. aureus,
O. mossambicus and two strains of O. niloticus. Aquac Res 35:
1434–1440
Darvish BK, Haji MA, Mohamadi ZA, Salehi SV, Shakiba MM (2009)
Measurement of some haematological characteristics of the wild
carp. Comp Clin Pathol 18(3):321–323
Diyaware MY, Haruna AB, Abubakar KA (2013) Some haematological
parameters of inter-generic hybrid of African catfish (Clarias
anguillaris × Heterobranchus bidorsalis) juveniles and their pure
lines in north eastern Nigeria. J Fish Aquat Sci 8:33–42
Duthie GG, Tort L (1985) Effects of dorsal aortic cannulation on the
respiration and haematology of Mediterranean living Scyliorhinus
canicula. Comp Biochem Physiol 81A:879–885
Eisler R (1965) Erythrocyte counts and haemoglobin content in nine
species of marine teleosts. Chesap Sci 6:119–120
Elahee KB, Bhagwant S (2007) Hematological and gill histopathological
parameters of three tropical fish species from a polluted lagoon on
the west coast of Mauritius. Ecotoxicol Environ Saf 68:361–371
1341
Ellis AE (1975) Leucocytes of fish: a review. J Fish Biol 11:453–492
Ezzat AA, Shabana MB, Farghaly AM (1973) Studies on the blood char-
acteristics of Tilapia zilli (Gervais). I. Blood cells. J Fish Biol 6:1–12
Ferguson HW (1976) The ultrastructure of the plaice (Pleuronecte
platessa) leucocyte. J Fish Biol 8:139–412
Gallardo MA, Sala-Rabanal M, Ibarz A, Padrós F, Blasco J, Fernández-
Borra J, Sánchez J (2003) Functional alterations associated with
Bwinter syndrome^ in gilthead sea bream (Sparus aurata).
Aquaculture 223:15–27
Groff JM, Zinkl JG (1999) Haematology and clinical chemistry of cypri-
nid fish, common carp and goldfish. Vet Clin North Am Exot Anim
Pract 2:741–776
Gulliver G (1875) Observations on the sizes and shapes of the red cor-
puscles of the blood of vertebrates, with drawings of them to a
uniform scale, and extended and revised tables of measurements.
Proc Zool Soc London:474–495
Hofer R, Stoll M, Romani N, Koch F, Sordyl H (2000) Seasonal changes
in blood cells of Artic char (Salvelinus alpinus L.) from a high
mountain lake. Aquat Sci 62:308–319
Imagawa T, Hashimoto Y, Kitagawa H, Kon Y, Kudo N, Sugimura N
(1989) Morphology of the cell in carp (Cyprinus carpio L.). Jpn J
Vet Sci 51:1163–1172
Jawad LA, Al-Mukhtar MA, Ahmed HK (2004) The relationship be-
tween haematocrit and some biological parameters of the Indian
shad, Tenualosa ilisha (family Clupidae). Anim Biodivers Conserv
27:478–483
Kelenyi G, Nemeth A (1969) Comparative histochemistry and electron
microscopy of the eosinophil leukocytes of vertebrates. I. A study of
avian, reptile, amphibian and fish leukocytes. Acta Biol Acad Sci
Hung 20:405–422
Kori-Siakpere O, Ake JEG, Idoge E (2005) Haematological characteris-
tics of the African snakehead, Parachanna obscura. Afr J
Biotechnol 4:527–530
Kulkarni RS (2017) Sex differences in the blood biochemical parameters
of the freshwater fish, Notopterus notopterus (Pallas, 1789). World
News of Natural Sciences 6(2017):44–51
Kumar MV (2016) Morphometric studies of blood cells in Cyprinus
carpio, Ctenopharyngodan idella and Hypophthalmichthys molitrix
cultured fish in west Godavari region of Andhra Pradesh.
International Journal of Fisheries and Aquatic Studies 4(5):489–493
Langston AL, Hoare R, Stefansson M, Fitzgerald R, Wergeland H,
Mulcahy M (2002) The effect of temperature on non-specific de-
fence parameters of three strains of juvenile Atlantic halibut
(Hippoglossus hippoglossus L.). Fish Shellfish Immunol 12:61–76
Leonardi MO, Klempau AE (2003) Artificial photoperiod influence on
the immune system of juvenile rainbow trout (Oncorhynchus
mykiss) in the southern hemisphere. Aquaculture 221:581–591
Lim C, Klesius PH (2003) Influence of feed deprivation on haematology,
macrophage chemotaxis, and resistance to Edwardsiella ictaluri
challenge of channel catfish. J Aquat Anim Health 15:13–20
Lowe CJ (2004) The effect of acute and chronic elevation of temperature
on aspects of the physiology of Antarctic Nototheniid fishes. Ph.D.
Thesis, University of Canterbury, New Zealand
Magill AH, Sayer MDJ (2004) The eject of reduced temperature and
salinity on the blood physiology of juvenile Atlantic cod. J Fish
Biol 64:1193–1205
Mcinroy RA (1953) A micro-haematocrit for determining the packed cell
volume and haemoglobin concentration on capillary blood. J Clin
Pathol 7:32–36
Pandey BN, Pajjdey PK, Choubey BJ, Dattamunshl JS (1976) Studies on
blood components of an air breathing siluroid fish Heteropneustes
fossilis in relation to body size. Folia Haemato 103:101–116
Radhakrishnan S, Stephen M, Balkrishnan N (1976) A study on the blood
cells of marine teleost fish Diodon hystrix, together as a suggestion
as to the origin of lymphocytes. Proc Indian Natl Sci Acad 42(4):
212–226
1342
Rao JV (2006) Biochemical alterations in euryhaline fish, Oreochromis
mossambicus exposed to sub-lethal concentrations of an organo-
phosphorus insecticide, monocrotophos. Chemosphere 65(10):14–
20
Rashidi Z, Khara H, Mousavi-Sabet H (2012) Haematological profile of
the mature Rutilus frisii kutum (Cyprinidae) migrated to the Tajan
River in the southern Caspian Sea. World Journal of Fish and Marine
Sciences 4:665–671
Rey Vazquez G, Guerrero GA (2007) Characterization of blood cells and
hematological parameters in Cichlasoma dimerus. Tissue Cell 39:
151–160
Rowley AF, Hunt TC, Page M, Mainwaring G (1988) Fish. In: Rowley
AF, Ratcliffe NA (eds) Vertebrate blood cells. Cambridge University
Press, Cambridge, pp 19–127
Sahli H (1909) Lehrbuch d.klin. untersuchungen methode, 5th edn.
Leipsic, pp 846
Satheeshkumar P, Ananthan G, Senthil KD, Jagadeesan L (2011)
Haematology and biochemical parameters of different feeding be-
haviour of teleost fishes from Vellar estuary, India. Comp Clin
Pathol 21:1187–1191
Saunders DC (1968) Differential blood cell counts of 50 species of fishes
from the Red Sea. Copeia 1968:491–498
Srivastava AK (1968) Studies on the hematology of certain freshwater
teleosts 1. Erythrocytes. Anat Anz 123:233–249
Starmach J (1970) The number of erythrocytes in blood of Cottus
poecilopus Heckel and Cottis gobio L. Acta Biol Cracon Ser Zool
13:243–249
Comp Clin Pathol (2018) 27:1335–1342
Stoskopf MK (1993) Clinical pathology. In: Stoskopf MK (ed) Fish
Medicine. Saunders, Philadelphia, PA, USA. pp 113–131
Svetina A, Matasin Z, Tofant A, Vucemilo M, Fijan N (2002)
Haematology and some blood chemical parameters of young carp
till the age of three years. Acta Vet Hung 50:459–467
Svobodova H, Kroupova H, Modra H, Flajshans M, Randak T, Savina
LV, Gela D (2008) Haematological profile of common carp
spawners of various breeds. J Appl Ichthyol 24:55–59
Tavares-Dias M, Barcellos JFM, Marcon JL, Menezes GC, Ono EA,
Affonso EG (2007) Haematological and biochemical parameters
for the Pirarucu Arapaima gigasschinz, (Osteoglossiformes,
Arapaimatidae) in net cage culture. Electron J Ichthyol 2:61–68
Thika R (1992) Anemia among children and adolescents in the Keewatin
region of the Northwest territories. MSc thesis, Department of
Community Hea1t.h Sciences Faculty of Medicine University of
Manitoba .Winnipeg, Manitoba
Ueda IK, Egami MI, Sasso WS, Matushima ER (2001) Cytochemical
aspects of the peripheral blood cells of Oreochromis (Tilapia)
niloticus.(Linnaeus, 1758) (Cichlidae, Teleostei)—part II. Braz J
Vet Res Anim Sci 38(6):273–277
Wells RM, Ashby MD, Duncan SJ, McDonald JA (1980) Comparative
study of the erythrocytes and haemoglobin in Nototheniid fishes
from Antarctica. J Fish Biol 17:517–5279
Witeska M (2013) Erythrocytes in teleost fishes: a review. Zool Ecol
23(4):275–281