08 Chapter1

Chapter 1: Nutritional value of acridids
Evaluation of nutrient composition

of acridids
Introduction:
ivestock industries such as poultry, fisheries and swine are the major
sources of protein for human consumption. With the rise in human
population, the need for these protein sources has increased. As a
result, these industries are growing day by day. But they demand fast-growing
animals and good quality of feed with high levels of energy, protein, vitamins
and essential minerals to support maximum growth of these animals before
they can be sold. At present the major conventional protein source that is used
to formulate various livestock feed is fish meal which is being over-exploited
and consequently has a very high price. As a result the cost of conventional
feed incurs about 5065% of the total cost of livestock production (Banerjee
1992; Sharma and Kishan 2006). Hence, various investigations are being
carried out to find alternative non-conventional supplementary diets for these
industries so that the over-exploitation of fish meal could be checked. Such
alternatives may include insects.
According to Prez et al., (1989) and Ramos-Elorduy et al., (1988),
these insects can be used as protein sources as well as natural pigments,
vitamins or minerals. Ledger (1987) also suggested the use of the brown locust
Locustana pardalina (Walker) as human and animal food. Ramos- Elorduy et
al., (1997) found that Sphenarium histrio Gerst. contains about 77% protein
and Melanoplus mexicanus (Saussure) about 71%. Wang et al., (2007)
estimated that the Chinese grasshopper Acrida cinerea (Thunberg) had more
than 65% crude protein and established it as a potential alternative food source
for poultry. Ramos- Elorduy et al., (1988) reported that poultry birds gave
better performance when reared on insect diets.
In this context Anand et al., (2008b) did a preliminary study on the
proximate composition and mineral content of four acridid species commonly
found in India. However, further studies are needed to look for the most
suitable species throughout the world. Moreover, for a complete picture on the
27

suitability of any material as food there is an obvious need to further explore
the contents of fatty acids, amino acids, vitamins and anti-nutritional factors.
Keeping this in mind the present study aimed to explore the nutritional value
of the three selected multivoltine acridids i.e., Oedaleus abruptus (Thunberg),
Spathosternum prasiniferum prasiniferum (Walker), and Oxya fuscovittata
(Marschall) in terms of proximate composition, minerals, vitamins, fatty acids,
amino acids, energy and anti-nutritional factors.
Materials and methods:

Collection and preparation of sample species:
Adult individuals of the selected acridid species were collected from
the nearby grasslands and croplands of Santiniketan (23 39 N, 87 42 E),
Birbhum, West Bengal, India by sweeping technique using standard insect
nets. They were freeze killed prior to the estimation of wet body weight. Then
they were oven dried at 60C until the dry body weight became constant.
Moisture was estimated using the formula:
% Moisture (M) = 100 [(dry weight 100)/wet weight]
Legs and wings were removed from the dried matter to get rid of
excess chitin, and after that they were crushed to powder form before they
were subjected to further analyses.
Estimation of proximate composition of the acridids:

Nutrient composition was estimated by standard procedures according
to AOAC (2000) on dry matter basis. The methods followed are discussed
below:
(a) Crude protein:
Kjeldahl method was followed for the determination of total nitrogen
in the samples. Accurately 0.5 g sample was weighed and transferred into the
digestion flask. The sample was then digested with 20ml conc. H2SO4 in the
presence of 5.0g of K2SO4, 1.0 g of Na2S2O3 and 0.1 g of Selenium till it
became colorless. The aliquot was then transferred into 800ml capacity
kjeldahl distillation flask along with its washings. Two to five drops of
phenolphthalein was added to it followed by 12N sodium hydroxide slowly till
28

the pink color persisted. Then distillation was carried out for about 30-40
minutes and the distillate was collected in a conical flask containing 25ml
N/10 H2SO4. After the distillation was completed the trap of the condenser
was washed repeatedly with distilled water into the conical flask. The distillate
was then titrated using methyl red as indicator with N/10 NaOH solution to
colorless/slightly yellow end point. The nitrogen content of the sample was
calculated as per following formula:
ml of N/10 H2SO4 taken (ml of N/10 NaOH
consumed Factor of NaOH) 0.0014
% Nitrogen content =
100
Sample taken in grams
% of crude protein = % of Nitrogen 6.25
Since NaOH is a secondary standard its strength was standardized against

N/10 succinic acid and N/10 H2SO4 before titration and the factor of NaOH
was calculated as per the following formula:
Exact strength of NaOH in relation to succinic acid

Factor of NaOH =
Exact strength of H2SO4
(b) Crude fat:

Crude fat content was determined using Soxhlet apparatus. Two grams
of oven dried sample was wrapped with a Whatman filter paper (No. 1) and
placed in a thimble connected with Soxhlet apparatus. The initial weight of the
Soxhlet flask was recorded and filled up with 200 ml petroleum ether (boiling
point 60 - 80 C). The total apparatus was then placed over a mantle and the
petroleum ether was allowed to boil for 6 - 8 hr and circulate through the
thimble by siphon process. After boiling, the flask was taken out and the
petroleum ether was allowed to evaporate. The crude fat was determined by
the following formula:
29
Final weight of the flask Initial weight of the flask

% Crude fat =
100
Sample taken in grams
(c) Crude fiber:

The residual sample after fat extraction was digested for exactly half
an hour with 200 ml of 1.25% H2SO4 and washed free from acid by filtering
through a linen cloth of standard mesh (18 threads per cm). The sample was
then digested with 200 ml of 1.25% NaOH for the same time and washed free
from alkali. The residue was transferred to a vitreosil crucible, dried overnight
at 100C and weighed. The dried sample was then put into a muffle furnace
and ignited at 550C till the sample burnt to ashes. The percentage of crude
fiber was calibrated using the following formula:
Weight of the crucible with oven dried digested

residue Weight of the crucible with ash
% Crude fiber =
100
Fat free sample taken in grams
(d) Total ash:

The ash content was estimated by igniting the weighed amount of
sample in a muffle furnace at 550 50C for 6 hours. The ash content was
determined by the following formula:
Weight of ash
% Ash =
100
Weight of sample
(e) Total carbohydrate:

This constituent was not determined analytically, but was calculated
mathematically by the difference method. The sum of the percentages of crude
protein, crude fat and total ash on dry matter basis was subtracted from 100 to
30

get the percentage of carbohydrate. The calculation could be expressed by the
following formula:
% carbohydrate = 100 (%CP + % EE + %TA)
Where, CP = crude protein
EE = crude fat (ether extract)
TA = total ash
(f) Nitrogen free extract (NFE):
Like that of total carbohydrate, nitrogen free extract was also
calculated mathematically by the difference method. In this case the sum of
the percentages of crude protein, crude fat, crude fiber and total ash on dry
matter basis was subtracted from 100 to get the percentage of NFE. The
calculation could be expressed by the following formula:
% NFE = 100 (%CP + % EE + % CF + %TA)
Where, CP = crude protein
EE = crude fat (ether extract)
CF = crude fiber
TA = total ash
Estimation of minerals, amino acids, fatty acids and vitamins:

To determine the mineral content such as Ca, Fe, Zn, Mg, Cu and Mn
in the tissue of acridids, the specimens were freeze killed and dried in a hot air
oven at 50 C for 48 hours. The dried samples were finely ground to powder
and digested in aqua regia at 140 C for 3 hours. After cooling, the clear
samples were diluted and metal concentration was determined by Atomic
absorption Spectrophotometry (Varian Tectron AA575 series).
The extraction of fatty acids from the acridid samples and their methyl
ester preparation were performed according to the method of Bettelheim and
Landesberg (1997). Methyl esters of fatty acid mixture were further purified
by thin layer chromatography (TLC). The purified methyl esters of fatty acids
were subjected to gas chromatographic (GC) analysis. Percent compositions
of the samples were computed from the GC peak areas.
Amino acids were analyzed according to Ghosh et al., (1995) and
Wang et al., (2007). Samples were first hydrolyzed with 6N HCl containing
31

1% phenol for 22 hours at 105C. Then the amino acid contents were
determined by high performance liquid chromatography (HPLC) at 38C.
Quantitative estimation of tryptophan could not be done by afore mentioned
method, so it was determined by colorimetric method, following the strategies
proposed by Fischl (1960). Vitamins like retinol, ascorbic acid, niacin,
riboflavin and thiamin were estimated by colorimetric method according to
AOAC (2000).
Estimation of anti-nutritional factors:

Anti-nutritional factor of phenolic polymers like tannin was
determined chemically with vanillin-HCl reagent and catechin solution
according to Gupta et al., (1988). Content of oxalate was determined by
simple titration using methyl red as indicator following the procedures
proposed by Gupta et al., (1988). Titration was again used to measure the
amount of phytin phosphorus using ferric chloride (FeCl3) as indicator
according to Agbede and Aletor (2004). Phytin content was calculated with a
multiplication of the value of phytin phosphous by 3.55 (Agbede and Aletor,
2004).
Statistical analysis:
The experiments were carried out in three replicates. For completely
randomized designs all the data were statistically analyzed by one-way
analysis of variance (ANOVA) using S Plus (version 4.0) software. Results
were subjected to Duncans multiple range test (DMRT) to understand the
significant difference between the data within a sample group. Data are
presented as means SD.
32
Results:
Proximate composition of the selected acridid species on dry matter basis:
Proximate composition of the selected acridid species has been
tabulated in Table 1.1. The results revealed about 67-70% moisture content in
the selected acridids with the highest value of 70.46% in case of O. abruptus
(P<0.001, one-way ANOVA, DMRT). Crude protein values ranged between
59% and 66% where the highest value (65.26%) was obtained in S.
prasiniferum prasiniferum (P<0.001, one-way ANOVA, DMRT). The highest
percentage of crude fat (7.78%) was also observed in the latter species. On the
other hand rest of the other parameters showed lowest values in S.
prasiniferum
prasiniferum
(P<0.001,
one-way
ANOVA,
DMRT).
Carbohyhrate, crude fiber and nitrogen free extract (NFE) were highest in O.
abruptus (28.51%, 8.29% and 20.22% respectively), whereas the percentage
of total ash was highest (5.27%) in O. fuscovittata (P<0.001, one-way
ANOVA, DMRT).
Content of minerals in mg/kg of dried acridid body tissues:

Total six minerals were estimated. Among them calcium (Ca) and
magnesium (Mg) was found to be present in 5-8mg/kg, whereas manganese
(Mn) was present in the least amounts (in between 0.25-0.60 mg/kg) in all of
the acridids (Figs 1.1, 1.2, 1.3). When the data were compared within minerals
between acridid species it was observed that calcium and iron (Fe) was present
in the highest amount in O. fuscovittata (about 8.0 and 3.9 mg/kg) (Figs 1.1,
1.4), whereas magnesium (Mg), zinc (Zn), and copper (Cu) (Figs 1.2, 1.5, 1.6)
were present in the highest amount in S. prasiniferum prasiniferum (near about
33

3.75, 7.6 and 2.2 mg/kg respectively) (P<0.001, one-way ANOVA, DMRT).
On the other hand in case of manganese (Mn) the results of O. fuscovittata and
S. prasiniferum prasiniferum did not vary significantly (DMRT), but
significantly lowest (P<0.001, one-way ANOVA, DMRT) value of nearly 0.28
mg/kg was obtained in O. abruptus.
Content of fatty acids in g/100g (%) of fat in dried acridid body tissues:
Total eight fatty acids were detected in O. abruptus (Table 1.2).
Among them myristic acid, palmitic acid and stearic acid were saturated and
rest of the five fatty acids were unsaturated. However, total six fatty acids
were detected in S. prasiniferum prasiniferum and O. fuscovittata where
myristic acid and arachidonic acid were absent in S. prasiniferum
prasiniferum and stearic acid and arachidonic acid were absent in O.
fuscovittata. It is noteworthy that all the three acridid species contained two
very important polyunsaturated fatty acids (PUFAs) i.e. linoleic acid and
linolenic acid. It is also notable that both of these PUFAs were present in quite
high amounts in all the three acridids. The content of linoleic acid was highest
in S. prasiniferum prasiniferum whereas linolenic acid was highest in O.
abruptus (P<0.001, one-way ANOVA). On the other hand the contents of both
of these fatty acids were almost similar in case of O. fuscovittata (27.25%
linoleic acid and 27.49% linolenic acid respectively).
34
Table 1.1. Proximate composition of the selected acridid species on dry matter basis
Acridid species
% moisture %CP ( SD ) %EE ( SD ) %C ( SD ) %CF ( SD ) %TA ( SD ) %NFE ( SD )
O. fuscovittata
69.682.09a
64.070.23b
6.480.31a
24.180.47b
7.470.09b
5.270.09b
16.710.51b
S. prasiniferum prasiniferum 67.840.42a
65.260.14c
7.780.06c
21.970.28a
7.010.11a
4.990.03a
14.960.28a
59.320.11a
7.150.19b
28.510.31c
8.290.13c
5.020.06a
20.220.31c
O. abruptus
70.46.86a
Note: CP = crude protein, EE = ether extract (crude fat), C = carbohydrate, CF = crude fiber, TA = total ash, NFE = nitrogen free extract.
Values with different letters within a column are significantly different (P<0.001) using DMRT.
35

Fig 1.1. Contents of calcium in mg/kg of dried acridid body tissues:
9
8
mg/kg
5
4
3
2
1
0
O. fuscovittata
O.abruptus
S. pr. pr
Note: Values are means SD. Bars with different letters are significantly
different (P<0.001) using DMRT.
Fig 1.2. Contents of magnesium in mg/kg of dried acridid body tissues:
*c
8
7
6
O. fuscovittata
O.abruptus
mg/kg
5
4
3
2
1
0
S. pr. pr
36

Fig 1.3. Contents of manganese in mg/kg of dried acridid body tissues:
0.7
0.6
b
b
mg/kg
0.5
0.4
0.3
0.2
0.1
0
O. fuscovittata
O.abruptus
S. pr. pr
Fig 1.4. Contents of iron in mg/kg of dried acridid body tissues:
4.5
4
3.5
mg/kg
2.5
2
1.5
1
0.5
0
O. fuscovittata
O.abruptus
S. pr. pr
37

Fig 1.5. Contents of zinc in mg/kg of dried acridid body tissues:
4.5
4
3.5
mg/kg
3
2.5
2
1.5
1
0.5
0
O. fuscovittata
O.abruptus
S. pr. pr
Fig 1.6. Contents of copper in mg/kg of dried acridid body tissues:
2.5
mg/kg
1.5
0.5
O. fuscovittata
O.abruptus
S. pr. pr
38

Table 1.2. Content of fatty acids in g/100g (%) of fat in dried acridid body
tissues:
Name of the
fatty acids
% content in
O. abruptus
% content in
S. pr. prasiniferum
% content in
O. fuscovittata
Myristic acid
0.37
----
1.12
Palmitic acid
4.55
3.50
0.97
Oleic acid
4.27
5.07
7.24
Stearic acid
1.12
1.05
----
Arachidonic acid
1.62
----
----
Eicosenoic acid
7.91
15.18
15.39
Linoleic acid
15.34
27.91
27.25
Linolenic acid
39.45
21.52
27.49
Content of amino acids in g/100g (%) of protein in dried acridid body

tissues:
Among the amino acids detected in this study, threonine (thr), proline
(Pro) and tyrosine (Tyr) were present in very high amount (more than 9%)
(Figs 1.7, 1.8, 1.9), whereas aspartic acid+asparagine (Asx) and cysteine (Cys)
were detected to be very low (less than 1%) (Figs 1.10, 1.11). When the results
were compared between the species comparatively higher values were
obtained in O. abruptus for Tyr, Asx, valine (Val), leucine (Leu), arginine
(Arg), isoleucine (Ile), phenylalanine (Phe), methionine (Met), glutamic acid +
glutamine (Glx), and serine (Ser) (P<0.001, one-way ANOVA, DMRT) (Figs
1.9, 1.10, 1.12-1.19). But S. prasiniferum prasiniferum showed highest values
for Pro, Cys, histidine (His), glycine (Gly) and alanine (Ala) (P<0.001, oneway ANOVA, DMRT) (Figs 1.8, 1.11, 1.20, 1.21, 1.22). On the other hand O.
fuscovittata only showed highest values for Thr, lysine (Lys), and tryptophan
(Trp) (P<0.001, one-way ANOVA, DMRT) (Figs 1.7, 1.23, 1.24).
39

Fig 1.7. Contents of threonine in g/100g (%) of protein in dried acridid
body tissues:
c
18.5
% Amino acid content
18
17.5
17
16.5
16
15.5
15
14.5
O.abruptus
O.fuscovittata
S.pr.pr.
Note: Bars with different letters are significantly different (P<0.001) using
DMRT.
Fig 1.8. Contents of proline in g/100g (%) of protein in dried acridid

body tissues:
16
15.9
15.8
15.7
15.6
15.5
15.4
15.3
15.2
15.1
15
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
40

Fig 1.9. Contents of tyrosine in g/100g (%) of protein in dried acridid
body tissues:
% amino acid content
c
10.4
10.2
10
9.8
9.6
9.4
9.2
9
8.8
8.6
8.4
8.2
b
a
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
Fig 1.10. Contents of aspertic acid+asperagine in g/100g (%) of protein

in dried acridid body tissues:
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
41

Fig 1.11. Contents of cysteine in g/100g (%) of protein in dried acridid
body tissues:
c
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
Fig 1.12. Contents of valine in g/100g (%) of protein in dried acridid

body tissues:
c
6.25
6.2
6.15
6.1
6.05
6
5.95
5.9
5.85
5.8
5.75
b
a
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
42

Fig 1.13. Contents of leucine in g/100g (%) of protein in dried acridid
body tissues:
5.3
5.2
5.1
5
4.9
4.8
4.7
4.6
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
Fig 1.14. Contents of arginine in g/100g (%) of protein in dried acridid

body tissues:
8.2
8
7.8
7.6
7.4
7.2
7
6.8
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
43

Fig 1.15. Contents of isoleucine in g/100g (%) of protein in dried acridid
body tissues:
c
1.45
1.4
1.35
1.3
1.25
1.2
1.15
1.1
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
Fig 1.16. Contents of phenylalanine in g/100g (%) of protein in dried

acridid body tissues:
4.2
4.15
4.1
4.05
4
3.95
3.9
3.85
3.8
3.75
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
44

Fig 1.17. Contents of methionine in g/100g (%) of protein in dried acridid
body tissues:
c
2
1.95
1.9
1.85
1.8
1.75
1.7
1.65
1.6
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
Fig 1.18. Contents of glutamic acid+glutamine in g/100g (%) of protein

in dried acridid body tissues:
4.5
3.5
3
2.5
2
1.5
1
0.5
0
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
45

Fig 1.19. Contents of serine in g/100g (%) of protein in dried acridid
body tissues:
c
5.1
4.9
4.8
4.7
4.6
4.5
4.4
4.3
4.2
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
Fig 1.20. Contents of histidine in g/100g (%) of protein in dried acridid

body tissues:
c
8.4
8.2
8
7.8
7.6
7.4
7.2
7
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
46

Fig 1.21. Contents of glycine in g/100g (%) of protein in dried acridid
body tissues:
c
8.4
8.3
8.2
8.1
8
7.9
7.8
7.7
7.6
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
Fig 1.22. Contents of alanine in g/100g (%) of protein in dried acridid

body tissues:
3.16
3.14
3.12
3.1
3.08
3.06
3.04
3.02
3
2.98
2.96
2.94
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
47

Fig 1.23. Contents of lysine in g/100g (%) of protein in dried acridid
body tissues:
c
3.5
2.5
2
1.5
1
0.5
0
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
Fig 1.24. Contents of tryptophan in g/100g (%) of protein in dried acridid

body tissues:
c
2.55
2.5
2.45
2.4
2.35
2.3
2.25
2.2
2.15
2.1
O.abruptus
O.fuscovittata
S.pr.pr.
DMRT.
48

Content of vitamins in mg/100g of dried acridid body tissues:
Total five vitamins, namely: retinol, ascorbic acid, thiamine, riboflavin
and niacin were detected in the body tissues of the selected acridids (Figs
1.25-1.29). Among the B vitamins thiamine was found to be present in very
low amount in all the three species (less than 0.50 mg/100g) (Fig 1.27).
Riboflavin was present in moderate amounts (just above 1mg/100g) (Fig
1.28), whereas retinol, ascorbic acid and niacin content were quite high (more
than 4 mg/100g) in all the acridids. Comparison of the values between species
revealed that retinol and riboflavin was highest in S. prasiniferum
prasiniferum (P<0.001, one-way ANOVA, DMRT) (Figs 1.25, 1.28), ascorbic
acid and niacin was highest in O. fuscovittata (Figs 1.26, 1.29) and only
thiamine was highest in O. abruptus (P<0.001, one-way ANOVA, DMRT)
(Fig 1.27).
Content of anti-nutritional factors in g/100g of dried acridid body tissues:

Four anti nutritional factors oxalate, tannin, phytin, and phytin
bound phosphorus (phytin P) were analyzed for each of the acridid of interest
(Figs 1.30-1.33). All of them were found to be present in very low amount.
Among the analyzed anti-nutritional factors, tannin was present in the highest
amount, which was in between 1g/100g and 2.45g/100g (Fig 1.31). On the
contrary phytin P was present in the least amount (less than 0.025%) (Fig
1.33). Statistical comparison between the acridid species within each antinutrient revealed that oxalate content was least in O. fuscovittata (P<0.001,
one-way ANOVA, DMRT) (Fig 1.30), tannin contents were similar in O.
fuscovittata and S. prasiniferum prasiniferum but significantly higher values
were obtained in O. abruptus (P<0.001, one-way ANOVA, DMRT) (Fig
1.31). Similarly, in case of phytin-P and phytin O. abruptus showed highest
values (Figs 1.32, 1.33).
49
Fig 1.25. Contents of retinol in mg/100g of dried acridid body tissues:
c
6
b
a
mg/100g
4
3
2
1
O. fuscovittata
O. abruptus
S.pr. pr
DMRT.
Fig 1.26. Contents of ascorbic acid in mg/100g of dried acridid body

tissues:
c
8
mg/100g
6
5
4
3
2
1
0
O. fuscovittata
O. abruptus
S.pr. pr
DMRT.
50

Fig 1.27. Contents of thiamin in mg/100g of dried acridid body tissues:
c
0.5
mg/100g
0.4
0.3
0.2
0.1
0
O. fuscovittata
O. abruptus
S.pr. pr
DMRT.
Fig 1.28. Contents of riboflavin in mg/100g of dried acridid body tissues:
mg/100g
1.5
1
0.5
O. fuscovittata O. abruptus
S.pr. pr
DMRT.
Fig 1.29. Contents of niacin in mg/100g of dried acridid body tissues:
mg/100g
6.5
5.5
5
4.5
O. fuscovittata
O. abruptus
S.pr. pr
DMRT.
51

Fig 1.30. Contents of oxalate in g/100g of dried acridid body tissues
0.8
O.abruptus
S. pr. pr
0.7
0.6
g/100g
0.5
0.4
0.3
0.2
0.1
0
O.fuscovittata
DMRT.
Fig 1.31. Contents of tannin in g/100g of dried acridid body tissues

3
2.5
g/100g
1.5
a
1
0.5
0
O.fuscovittata
O.abruptus
S. pr. pr
DMRT.
52
Fig 1.32. Contents of phytin in g/100g of dried acridid body tissues

0.1
0.09
0.08
g/100g
0.07
0.06
0.05
0.04
0.03
0.02
0.01
0
O. fuscovottata
O. abruptus
S.pr.pr
DMRT.
Fig 1.33. Contents of phytin-phosphorus in g/100g of dried acridid body

tissues
0.03
0.025
g/100g
0.02
0.015
0.01
0.005
0
O. fuscovottata
O. abruptus
S.pr.pr
DMRT.
53
Discussion:
Dietary protein is one of the most important constituents in feed of
vertebrates, and fish is of no exception. In general, higher protein levels and
higher protein to energy (P/E) ratios can produce significantly better growth
and feed utilization in fish (Ali and Jauncey, 2005). Our results revealed that
the acridid grasshoppers are high in protein contents which prove this group of
insects to be a good protein supplement.
Protein is composed of mainly twenty amino acids. Yang et al., (2010)
reported that the amino acid profile of dietary protein plays crucial role for the
increase
of
protein
retention
and
flesh
quality
for
fishes
like,
Ctenopharyngodon idella. Among these, total ten amino acids are important in
fish physiology namely: arginine, histidine, isoleucine, leucine, lysine,
methionine, phenylalanine, threonine, tryptophan and valine (Yang et al.,
2010). The presence of all these amino acids was observed along with the
others in the selected acridids of our interest. This finding was in concert with
the report of Chen et al., (2009) where more than hundred edible insects in
China showed that they contain all the necessary amino acids. De Silva and
Anderson (1998) reported the amount of optimal need of these amino acids in
different fishes. Our results were very encouraging because they revealed that
six of the ten essential amino acids (i.e. Val, Leu, Arg, Trp, His and Thr) were
present in the acridids in more amounts than the fishes need. However, despite
having a good amount of amino acid content, reports say that some of the
edible insects are deficient in some amino acids. As for example studies on
other edible insects like Mormon cricket (Anabrus simplex) and house cricket
(Acheta domesticus) showed that they were deficient in methionine (DeFoliart
et al., 1982; Finke et al., 1985; Nakagaki et al.,1987). Similarly, as reported by
Landry et al., (1986) the essential amino acids of six lepidopteran species were
also deficient in the same amino acid. Our result corresponds with the above
stated reports as the content of methionine, cysteine and lysine were found to
be quite low (below 3%). Therefore we support the conclusion of Wang et al.,
(2007) that insects might be unsatisfactory as the only source of dietary
protein due to limiting amino acids, but would be extremely beneficial as a
supplement.
54

Fat is one of the major nutrients for any living organism. Authors like
Feng et al., (2000a,b; 2001a,b,c), Chen and Feng (1999), He et al., (1999) etc.
reported that insects are higher in fat content at the larva and pupa stages,
while adults contained relatively lower amount of fat. Among grasshoppers
the fat content of adult Oxya chinensis was found to have only 2.2% (Chen et
al., 2009); likewise Melo et al., (2011) reported that the same of Sphenarium
purpuracens was also quite low (5.75%). The present study showed lower fat
content (nearly 5-7%) in the experimental acridids. Although the fat content of
most of the edible insects is lower than the optimal need of 10-20% for fish
(Cowey and Sargent, 1979), still they could play an important role as a
supplementary diet. Chen et al., (2009) reported that unlike other animal fat,
edible insects have a higher essential fatty acid content which is necessary for
animal nutrition, and hence edible insect fat has a high nutritive value. The
results of fatty acid content of our species when compared to that of the
variegated grasshopper Zonocerus variegatus showed that O. abruptus
contained a total of eight fatty acids, and O. fuscovittata and S. prasiniferum
prasiniferum contained six fatty acids each, compared to five of Z. variegatus
(Adeye, 2011). On the other hand Wang et al., (2007) also reported the
existence of eight fatty acids in A. cinerea where they found palmitotelic and
lauric acid that was not observed in our results; instead we got arachidonic and
eicosenoic acids. According to Chen et al., (2009), edible insects are rich in
protein and fat but not so rich in carbohydrate content; however the authors
also added that carbohydrates of edible insects differ from species to species
ranging from 1-10% in China. On the other hand larva of Cirina forda which
is relished as a human food in Nigeria contains near about 38% of
carbohydrate (Akinnawo and Ketiku, 2000). The three acridid species of our
interest also showed a moderate amount of carbohydrate content.
Edible insects have been found to contain good amount of minerals.
Among them sodium, potassium, calcium, zinc, iron and magnesium are
prevalent in literature (Akinnawo and Ketiku, 2000; Ramos-Elorduy et al.,
2002; Ojewola and Udom, 2005; Ekop et al., 2010). Ramos-Elorduy (2005)
reported that edible insects under the orders Orthoptera, Lepidoptera and
Hymenoptera have much lower variation in mineral contents. The author
55

continued stating that these insects are mostly low in Na, and sometimes high
in Ca, Zn, Fe, K and Mg. Our results support this view as Mg is present in the
highest amount (more than 6mg/kg) followed by Ca and Zn. Fe was observed
in a moderate amount. Though Na and K were not detected, a small trace of
Mn (less than 1%) was obtained. Reports are there that show the edible insects
to be rich in vitamin contents too. As for example Kodondi et al., (1987b)
reported rich vitamin content in some edible caterpillars of Attacidae family in
Zaire. Ramos-Elorduy et al., (1988) observed that edible insects are rich in
vitamin B group such as thiamine, riboflavin and niacin. The presence of these
three vitamins were also observed in case of the selected acridid species where
niacin was present in high amount but the rest of the two were a little lower. In
addition to this, two other vitamins i.e. ascorbic acid and retinol were also
present in quite high amount.
Anti-nutritional factors are usually present in plant materials. However,
many phytophagous insects have been identified to retain these materials in
quite a good amount (Berenbaum, 1993). Hence it is recommended to analyze
these anti-nutrients if a phytophagous insect is being considered as food,
though this kind of study is not much frequent in the literature. Among the
anti-nutritional factors tannin, oxalate and phytate were detected in the
grasshoppers of our interest. Adeduntan (2005) reported tannin percentage in
grasshoppers of Ondo state of Nigeria to be 1.05g/100g, which is almost
similar to the amounts observed in O fuscovittata and S. prasiniferum
prasiniferum, but a little higher amount (about 2.5g/100g) was observed in O.
abruptus. However, this amount is itself quite lower than that found in cereal
grains and other plant food materials (Ogunlade et al., 2011; Hassan et al.,
2011). On the other hand the percentage of phytate was reported to be quite
high in grasshoppers by Adeduntan (2005) which was about 1.1g/100g
compared to quite lower amount of nearly 0.017- 0.025g/100g of phytin-P and
nearly 0.06 - 0.09g/100g of phytin in the present study. We could not find any
literature describing the oxalate content of any grasshopper species but it has
been reported in some edible beetles such as adults of Oryctes monoceros and
Cirina forda (Ifie and Emerua, 2011; Omotoso, 2006). Both of these papers
revealed that those beetles contain less than 0.005% oxalate, much lower than
56

the insects of our interest that ranged between 0.54 - 0.69g/100g. However this
amount is also under the tolerance limit because much higher amounts have
been observed in various plant food materials (Kalita et al., 2007).
Conclusion:
Proximate composition of the present study reveals that the selected
acridid grasshoppers are nutritious with high protein contents. Amino acids,
fatty acids, vitamins and minerals were present in high amount. Though antinutritional factors were present but were found to be in very low amount and
within the tolerance limit. Thus it can be safely stated that acridids could be a
good alternative source of not only protein, but also important vitamins and
minerals. This proves that an effort should be made so that these grasshoppers
could be mass reared in controlled conditions in farms as a mini-livestock so
that it could supply more and more amount of nutritious alternative food for
the formulation of protein rich diet for the livestock industry; thus lowering
down the over-exploitation of fish meal.
57

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Chapter 1: Nutritional value of acridids

Evaluation of nutrient composition

Chapter 1: Nutritional value of acridids

Materials and methods:

Estimation of proximate composition of the acridids:

Chapter 1: Nutritional value of acridids

% of crude protein = % of Nitrogen 6.25

Since NaOH is a secondary standard its strength was standardized against

Exact strength of NaOH in relation to succinic acid

(b) Crude fat:

Chapter 1: Nutritional value of acridids

Final weight of the flask Initial weight of the flask

(c) Crude fiber:

Weight of the crucible with oven dried digested

(d) Total ash:

(e) Total carbohydrate:

Chapter 1: Nutritional value of acridids

Estimation of minerals, amino acids, fatty acids and vitamins:

Chapter 1: Nutritional value of acridids

Estimation of anti-nutritional factors:

Chapter 1: Nutritional value of acridids

Content of minerals in mg/kg of dried acridid body tissues:

Chapter 1: Nutritional value of acridids

Chapter 1: Nutritional value of acridids

% moisture %CP ( SD ) %EE ( SD ) %C ( SD ) %CF ( SD ) %TA ( SD ) %NFE ( SD )

S. prasiniferum prasiniferum 67.840.42a

Chapter 1: Nutritional value of acridids

Fig 1.2. Contents of magnesium in mg/kg of dried acridid body tissues:

Chapter 1: Nutritional value of acridids

Fig 1.4. Contents of iron in mg/kg of dried acridid body tissues:

Chapter 1: Nutritional value of acridids

Fig 1.6. Contents of copper in mg/kg of dried acridid body tissues:

Chapter 1: Nutritional value of acridids

Content of amino acids in g/100g (%) of protein in dried acridid body

Chapter 1: Nutritional value of acridids

% Amino acid content

Fig 1.8. Contents of proline in g/100g (%) of protein in dried acridid

Chapter 1: Nutritional value of acridids

% amino acid content

Fig 1.10. Contents of aspertic acid+asperagine in g/100g (%) of protein

% Amino acid content

Chapter 1: Nutritional value of acridids

% Amino acid content

Fig 1.12. Contents of valine in g/100g (%) of protein in dried acridid

% Amino acid content

Chapter 1: Nutritional value of acridids

% Amino acid content

Fig 1.14. Contents of arginine in g/100g (%) of protein in dried acridid

% amino acid content

Chapter 1: Nutritional value of acridids

% Amino acid content

Fig 1.16. Contents of phenylalanine in g/100g (%) of protein in dried

% Amino acid content

Chapter 1: Nutritional value of acridids

Fig 1.18. Contents of glutamic acid+glutamine in g/100g (%) of protein

% Amino acid content

Chapter 1: Nutritional value of acridids

% amino acid content

Fig 1.20. Contents of histidine in g/100g (%) of protein in dried acridid

Chapter 1: Nutritional value of acridids

% Amino acid content

Fig 1.22. Contents of alanine in g/100g (%) of protein in dried acridid