estimation of total protein content (mg/g) in body muscles of...

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Estimation of Total Protein content (mg/g) in body muscles

of selected fishes WALLAGO ATTU and SPERATA (MYSTUS)

SEENGHALA collected from selected habitats in different

seasons (reservoirs) by Lowry’s method (1951).

4(A) - INTRODUCTION

Fish are the important component in our diet and rich source of protein.

Fishes contains good quality, balanced and digestible protein. Protein content in fish

meat varied between 16 and 21%. In fish’s two types of muscles are present i.e. dark

muscles (red) and white muscles. Dark muscles contain low level of moisture and

protein than the white muscle. Proteins are classified on the basis of shape, solubility

and chemical structure. On the basis of solubility in salt solution, Proteins are of

three groups’ Sarcoplasmic protein, Myofibrillar protein and Stroma. Sarcoplasmic

protein (Albumin and Globulin) constitutes 25-30% of protein. Myofibrillar protein

(myosin, actin, tropomyosin, troponin) are structural protein constitute 65-70 % of

total protein. Stroma proteins are also known as connective tissue protein. It

constitutes 3% of the total protein.

Proteins are complex organic compounds and made up of Prolong chain of

amino acid bounded together by peptide bonds. Fresh fish meat provides good

source of protein for human diet, about 90-95% of fish protein is assimilated by

human. In this experiment, we need to know how much protein is present in our

sample or to measure the concentration or amount of protein in fish muscle samples.

Several investigators like Mohanty et. al., (2012), Memon et. al., (2011),

Marykutty Joseph et. al., (2011), Muzumder et. al., (2008), Siddiqui et. al., (2010),

Deka et. al., (2012), Emmanuel et. al., (2011), Debnath et. al., (2014) analyzed

protein in catfish Sperata seenghala, Wallago attu, Arius arius, Amblypharyngodon

mol, Gadusia chapra, Puntius sp., Chanda nama, Pseudeutropinus atherinoides and

in Ailia coila, Mystus tengara, Mystus cavasius, Mystus gulio, Labeo gonius, Clarias

gariepinus, Labeo bata in different regions. Protein content variation is affected by

breeding capability, planktonic diet and climatic changes in year. The highest and

lowest protein content associated to breeding, spawning and biological regression,

resting stages, respectively (Lee, 1995).

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Protein content was also estimated in fish species Thunnus albacares,

Parupeneus bifasciatus, Hyporhamphus dussumieri,Oncorhynchus myskiss,

Lepidocephalus guntea, Schizothorax niger, Schizothorax esocinus, Salmostoma

sardinella, Cirrhinus reba, Garra mullya, Rasbora daniconius, Puntitus conchonius,

Macrones leucophasis, Mystus cavasius, Clarius batrachus, Heteropneustes fossilis,

Anabas testudineus, Notopterus notopterus, Nandas nandas, Channa punctatus,

Mystus vittatus, Glossogobius giuris, Scomberoides commersonianus, Scomberoides

lysan, Catla catla, Labeo rohita, Cirrhinus mrigala, Cyprinus carpio,

Pangasianodon hypopthalmus, Chrysichthys nigrodigitatus, Labeo niloticus,

Mormyrops anguilloides, Marcusenius cyprinoides, Mormyrus niloticus, Clarias

lazera and Protopterus annectens, Thunnus alalunga, Evynnisjaponica,

Caulerpalentillifera, Orcynopsis unicolor and Euthynnusaffinis, Otelithes ruber,

Eleutheronema thetradactylium,Harpodon nehereus, Aristichthys nobilis, Polyodon

spathula, Lutjanus johni, Coregonus lavaretus, Lethrinus lentjan, Hemisynodontis

membranacea, Ailiichthys punctatus, Clupisoma psendeutropius atherinoides,

Puntius sarana, Gadusia chapra, Corica soborna, Tenualosa ilisha, Alestes nurse,

Oreochromis gallilaeus ,Cirrhinus mrigala, Lates niloticus, Tetraodon lineatus,

Heterotis niloticus, Mormyrus niloticus, Clarias lazera, Protopterus annectens.

4(B) - MATERIAL AND METHOD.

To estimate the total protein content from the selected fish species Wallago

attu and Sperata seenghala Lowry’s Method (1951) was used.

Protein was estimated by Lowry’s and his co-worker in 1951. This method is

sensitive enough to give a moderate constant volume. The blue colour developed by

the reduction of the Phosphomolybdic-Phosphotungustic components in the Folin-

Ciocalteau reagent by amino acids tyrosine and tryptophan present in the protein

plus the colour developed by biuret reaction of protein with alkaline cupric tartrate

are measured in the Lowry’s method.

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i) Reagents and Chemicals.

1) Reagent (A) -: 2% Sodium carbonate (Na2Co3.5H2O) in 0.1 N sodium

hydroxide.

2) Reagent (B) -: 0.5% Copper sulphate CuSo4.5H2O in % Potassium

sodium tartrate.

3) Reagent (C) -: Alkaline copper Solution - Mixed 50 ml of Reagent

(A) and 1ml of Reagent (B) prior to use.

4) Reagent (D) -: Folin - Ciocalteau Reagent.

5) Protein Solution (Stock Solution) -: 50 mg of BSA was accurately

weighed and dissolved in distilled water and made the volume up to

50 ml in a standard flask.

6) Working standard -: 100 ml of the stock solution was diluted into 50

ml of distilled water in a standard flask. 1ml of this solution contains

200 µg proteins.

ii) FLOW CHART SHOWING THE EXTRACTION OF PROTEIN

FROM FISH BODY MUSCLES.

Taken 500 mg of dry tissue (body muscle)

Added 6 ml of saline

Centrifuged for 15 - 20 min for 4000 rpm

Taken 1 ml of supernatant and added 5 ml of Reagent (C)

Incubated for 10 min.

Added 0.5 ml of Reagent (D)

Incubated for 30 min.

Measured the O. D. at 660 nm

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The selected fishes Wallago attu and Sperata seenghala were purchased in

fresh condition from fish market of Parbhani and selected reservoirs (Study area).

Washed the fishes in running water to remove the mucus from the outer surface of

the bodies of fishes. Then, weights of the fishes were recorded with the help of

weighing balance, noted down the reading of the fishes. Body muscles were

separated from fish body. 100 gm of muscles were weighed and dried in microwave

oven. After drying the dry muscle weight was weighed. Dried tissues were taken in

pestle and mortal and made into fine powder. 500 mg of dry muscle powder was

weighed in test tube and added 5 - 6 ml of saline. Centrifuged for 15 - 20 minutes for

4000 rpm. After some time, carefully 1ml of supernatant was taken and added 5ml

of reagent C in another test tube. This way the sample is ready for further chemical

analysis. In case of sample we have to make 10 time dilution, therefore pipette out

0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9 ml. sample solution and 0.9, 0.8, 0.7, 0.6, 0.5,

0.4, 0.3, 0.2, 0.1 ml of distilled water to make the volume up to 1ml. The samples

were incubated at room temperature for ten min. and shaked well 0.5 ml of Reagent

D in same test tube was added. Again incubated at room temperature for 30 min.

After 30 minute O. D. at 660 nm was recorded using UV- Spectrophotometer

(Shimadzu, UV - 1800).

iii) Statistical Analysis

The current results for two freshwater fishes Wallago attu and Sperata

seenghala are mean ± standard deviation acquired from the analysis of fish species.

The mean values of total protein content are submitted to two - way analysis of

variance using software Microsoft office excel 2007 statistically significant

difference (p<0.05) are recorded. If the entire F-test was significant (p<0.05), then a

Fishers T- test was performed to discern difference among the reservoirs and

seasons.

4(C) - RESULT

As shown in below table - 4.1, 4.2, 4.3 and 4.4 seasonal variation of total

protein (mg/g) in body muscles of selected fishes Wallago attu and Sperata

seenghala collected from selected habitats (reservoirs) during Feb. 2010 to Jan.

2011and Feb. 2011 to Jan. 2012.

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The total protein (mg/g) content in the body muscles of Wallago attu ranged

from 0.69±0.22 to 7.89±0.63 mg/gm collected from all selected habitats (reservoirs)

in three seasons during Feb. 2010 - Jan. 2011. Highest total protein content was

observed 7.89±0.63 mg/g in the body muscles of Wallago attu collected from

Masoli Reservoir during summer season. While, lowest total protein content in the

body muscles of Wallago attu was found 0.69±0.22 mg/g collected from

Siddheshwar Reservoir during monsoon season as compared to all selected

reservoirs from different seasons are presented in table - 4.1 and Fig. - 4.1.

The total protein content in the body muscles of W. attu was obtained

1.75±0.35 to 7.02±0.97 mg/g collected from all selected habitats in different seasons

during Feb. 2011 - Jan. 2012. Highest total protein content in the body muscles of

W. attu was found 7.02±0.97 mg/g collected from Masoli Reservoir during summer

season. While, lowest total protein content in the body muscles of W. attu was found

1.75±0.35 mg/g collected from Siddheshwar Reservoir during winter season as

compared to all selected reservoirs in all seasons are as shown in table - 4.2 and Fig.

- 4.2.

The total protein content in the body muscles of S. seenghala was observed

1.14±0.53 to 6.48±0.84 mg/g collected from all selected habitats during Feb. 2010 -

Jan. 2011. Highest total protein content in the body muscles of S. seenghala was

obtained 6.48±0.84 mg/g collected from Masoli Reservoir during summer season.

While, lowest total protein content in the body muscles of S. seenghala was found

1.14±0.53 mg/g collected from Siddheshwar Reservoir during winter season as

compared to all selected reservoirs in three seasons are as shown in table - 4.3 and

Fig. - 4.3.

The total protein content in the body muscles of S. seenghala was obtained

1.79±0.45 to 6.73±0.91 mg/g in all selected habitats during Feb. 2011- Jan. 2012.

Highest total protein content in the body muscles of S. seenghala was found

6.73±0.91 mg/g collected from Masoli Reservoir during summer season. While,

lowest total protein content in the body muscles of S. seenghala was observed

1.79±0.45 mg/g collected from Karpara Reservoir during monsoon season as

compared to all selected reservoirs in all three seasons are presented in table 4.4 and

Fig. 4.4.

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By using Two way ANOVA between the result obtained from different

selected reservoirs and seasons it was observed that there is significant difference

(p<0.05) in total protein content (mg/g) in body muscle of Wallago attu during Feb.

2010 to Jan. 2011 and Feb. 2011 to Jan. 2012 are given as in table - 4.5 (a) and (b);

4.6 (a) and (b) and S. seenghala during Feb. 2010 to Jan. 2011 as shown in table -

4.7 (a) and (b).

There is significant difference (p<0.05) in total protein content (mg/g) in

body muscle of S. seenghala in different seasons but, no significant difference

(p0.05) in total protein content (mg/g) in body muscle of S. seenghala collected

from selected different reservoirs during Feb. 2011 to Jan. 2012 as in table - 4.8 (a)

and (b).

Table 4.1 Seasonal variation of Total Protein content (mg/g) in

body muscles of Wallago attu collected from different

selected habitats during Feb. 2010 - Jan. 2011.

The data is shown as mean±standard deviation (n=10)




The data is shown as mean±standard deviation (n=10).

Seasons

Habitats

Karpara

Reservoir

Masoli

Reservoir

Yeldari

Reservoir

Siddheshwar

Reservoir

Summer 6.52±0.67 7.89±0.63 6.47±0.97 5.44±0.77

Monsoon 2.03±0.89 4.12±0.75 2.19±0.45 0.69±0.22

Winter 1.79±0.25 2.14±0.43 2.36±0.99 1.61±0.68

Seasons

Habitats

Karpara

Reservoir

Masoli

Reservoir

Yeldari

Reservoir

Siddheshwar

Reservoir

Summer 6.30±0.57 7.02±0.97 6.57±0.81 4.97±0.62

Monsoon 4.54±0.97 4.04±0.83 4.31±0.97 3.09±0.71

Winter 2.28±0.66 4.20±0.76 3.49±0.96 1.75±0.35

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body muscles of Sperata seenghala collected from

different selected habitats during Feb. 2010 - Jan. 2011.

The data is shown as mean±standard deviation (n=10)



different selected habitats during Feb. 2011 - Jan. 2012.

The data is shown as mean±standard deviation (n=10).

Seasons

Habitats

Karpara

Reservoir

Masoli

Reservoir

Yeldari

Reservoir

Siddheshwar

Reservoir

Summer 4.92±0.67 6.48±0.84 5.60±0.70 4.70±0.72

Monsoon 3.27±0.66 4.15±0.90 3.22±0.84 2.16±0.42

Winter 2.49±0.36 3.68±0.53 3.06±0.88 1.14±0.53

Seasons

Habitats

Karpara

Reservoir

Masoli

Reservoir

Yeldari

Reservoir

Siddheshwar

Reservoir

Summer 6.14±0.87 6.73±0.91 6.37±0.62 5.51±0.28

Monsoon 1.79±0.45 3.91±0.41 3.88±0.57 2.49±0.67

Winter 2.15±0.29 2.46±0.82 3.03±0.68 2.07±0.85

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0

1

2

3

4

5

6

7

8

9

Karpara

Reservoir

Masoli

Reservoir

Yeldari

Reservoir

Siddheshwar

Reservoir

To

tal

Pro

tein

(m

g/g

)

Habitats

Fig. 4.1 - Seasonal variation of Total Protein content (mg/g) in

body muscles of Wallago attu collected from different selected

habitats during Feb. 2010 - Jan. 2011 .

Summer

Monsoon

Winter

0

1

2

3

4

5

6

7

8

9

Karpara

Reservoir

Masoli

Reservoir

Yeldari

Reservoir

Siddheshwar

Reservoir

Tota

l P

rote

in (

mg/g

)

Habitats




Summer

Monsoon

Winter

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0

1

2

3

4

5

6

7

8

Karpara

Reservoir

Masoli

Reservoir

Yeldari

Reservoir

Siddheshwar

Reservoir

To

tal

Pro

tein

(m

g/g

)

Habitats

Fig. 4.3 - Seasonal varition of Total Protein content (mg/g) in

body muscles of Sperata seenghala collected from different


Summer

Monsoon

Winter

0

1

2

3

4

5

6

7

8

9

Karpara

Reservoir

Masoli

Reservoir

Yeldari

Reservoir

Siddheshwar

Reservoir

Tota

l P

rote

in (

mg/g

)

Habitats


body muscles of Sperata seenghala collected from different


Summer

Monsoon

Winter

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selected habitats during Feb. 2010 - Jan. 2011 as in table

4.5 (a) and (b) shown below.

Table 4.5 (a) - ANOVA: Two factors without replication for Total

Protein content (mg/g).

SUMMARY Count Sum Average Variance SD

Summer season 4 26.32 6.58 1.010467 1.00522

Monsoon season 4 9.03 2.2575 1.994092 1.412123

Winter season 4 7.9 1.975 0.1143 0.338083

Karpara Reservoir 3 10.34 3.446667 7.098433 2.664289

Masoli Reservoir 3 14.15 4.716667 8.532633 2.921067

Yeldari Reservoir 3 11.02 3.673333 5.873233 2.423475

Siddheshwar

Reservoir 3 7.74 2.58 6.3463 2.519186

Table 4.5 (b) - ANOVA (Analysis of variance) for Total Protein

content (mg/g)

Source of

Variation SS df MS F P-value F crit

Seasons 53.29312 2 26.64656 66.39278 8.08E-05 5.143253

Reservoirs 6.948492 3 2.316164 5.770973 0.033484 4.757063

Error 2.408083 6 0.401347

Total 62.64969 11

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selected habitats during Feb. 2011 - Jan. 2012 as in table

4.6 (a) and (b) shown below.




Summer season 4 24.86 6.215 0.7771 0.881533

Monsoon season 4 15.98 3.995 0.405767 0.636998

Winter season 4 11.72 2.93 1.247133 1.116751




Siddheshwar

Reservoir 3 9.81 3.27 2.6164 1.617529


content (mg/g).

Source of


Seasons 22.4718 2 11.2359 42.36942 0.000289 5.143253

Reservoirs 5.698867 3 1.899622 7.16328 0.020794 4.757063

Error 1.591133 6 0.265189

Total 29.7618 11

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different selected habitats during Feb. 2010 - Jan. 2011

as in table 4.7 (a) and (b) shown below.




Summer season 4 21.7 5.425 0.641433 0.800895

Monsoon season 4 12.8 3.2 0.663133 0.81433

Winter season 4 10.37 2.5925 1.173825 1.083432




Siddheshwar

Reservoir 3 8 2.666667 3.360933 1.833285


content (mg/g).

Source of


Seasons 17.79032 2 8.895158 96.34615 2.75E-05 5.143253

Reservoirs 6.881225 3 2.293742 24.84421 0.000879 4.757063

Error 0.55395 6 0.092325

Total 25.22549 11

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different selected habitats during Feb. 2011 - Jan. 2012

as in table 4.8 (a) and (b) shown below.




Summer season 4 24.75 6.1875 0.262958 0.512795

Monsoon season 4 12.07 3.0175 1.108492 1.052849

Winter season 4 9.71 2.4275 0.189625 0.43546




Siddheshwar

Reservoir 3 10.07 3.356667 3.521733 1.876628


content (mg/g).

Source of


Seasons 32.7128 2 16.3564 67.99113 7.55E-05 5.143253

Reservoirs 3.239825 3 1.079942 4.489158 0.056114 4.757063

Error 1.4434 6 0.240567

Total 37.39603 11

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4(D) - DISCUSSION

As per the results obtained on the total protein content in the selected

fish species W. attu and S. seenghala, there is significant difference (p<0.05) in total

protein content (mg/g) in body muscle of Wallago attu during Feb. 2010 to Jan.

2011 and Feb. 2011 to Jan. 2012 are given as in table - 4.5 (a) and (b); 4.6 (a) and

(b) and S. seenghala during Feb. 2010 to Jan. 2011. There is significant difference

(p<0.05) in total protein content (mg/g) in body muscle of S. seenghala in different

seasons but, no significant difference (p0.05) in total protein content (mg/g) in

body muscle of S. seenghala collected from different selected reservoirs during Feb.

2011 to Jan. 2012 as in table - 4.8 (a) and (b).

The total protein content in various tissues in fishes were investigated

by various authors and correlated their results with different factors like seasons,

habitat differences, processing methods, sex differences, breeding seasons and non-

breeding seasons, size and age differences etc. which are summarized as below.

There are very few reports on the variations in the total protein content in the fish

tissues with difference in the habitat of fish.

Siddiqui et. al., (2010) examined the protein level of Mystus tengara,

Mystus cavasius, Mystus gulio at fresh condition were 16.26 %, 15.52 % and 14.80

%. After twenty days of freezing protein level decreased as 14.97 %, 13.91 %, 13.43

%. Amount of moisture for these fishes increased 76.12 % to 78.02 %, 75.35 % to

77.25 % and 76.03 % to 78.23 % respectively. In fresh condition the lipid content

level of Mystus tengara, Mystus cavasius and Mystus gulio at were 6.15%, 5.98%,

and 6.20% and at last lipid were 4.9%, 5.37% and 4.86%. The reduction in protein

level is connected with denaturation of fish protein that is associated with freezing.

Dhaneesh et. al., (2012) analyzed the fish species Thunnus albacares, Parupeneus

bifasciatus, Hyporhamphus dussumieri and maximum protein level was found in

Thunnus albacares (13.69%) and minimum protein level was in Hyporhamphus

dussumieri (10.51%).The lipid content was ranged from 2.96% in Thunnus

albacares to 6.97% in Hyporhamphus dussumieri.The lipid was negatively

correlated with protein (r = − 0.765, p<0.05).

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S. Kandemir et. al., (2007). Studied total lipid from muscles of

Oncorhynchus myskiss reached the lower level in May and increased from June to

September. Marykutty Joseph (2011) examined the highest protein 17.61% in May

(pre-monsoon) and lowest in 16.0% in September (post-monsoon) in catfish Arius

arius. The average protein value was 17.36% during pre-monsoon, 16.9% during

monsoon (June and July) and 16.3% during Post-monsoon months. The highest

amount of lipid content was 6.26% in April and lowest amount in August 3.7%. The

average lipid value was 6.1% in pre-monsoon, 3.9% in monsoon and in post-

monsoon 5.6%. Islam et. al., (2008) analyzed the lipid from the body muscle of

Lepidocephalus guntea (Ham.) in three size group size-I (below 8.5 cm), size-II

(8.5- 9.5 cm) and size-III (above 9.5 cm), among these groups, highest amount of

lipid was found in size-III i.e. 3.455% and lowest in size-I i.e. 2.515%.

Okonji et. al., (2013) Studied the mean value of fat content of Clarias

gariepinus (19.68%) was higher than Oreochromis niloticus (14.4%). Fat content

ranged from 19.50 to 19.86 in C. gariepinus and 12.58 to 16.22 in Oreochromis

niloticus. Significantly higher concentration of total fatty acids 16.54 gm/100 gm in

Clarias gariepinus and lower in 15.72 gm/100 gm in Oreochromis niloticus.

Various fatty acids were higher in Clarias gariepinus than Oreochromis niloticus

with values of 49.82% and 48.66%. Palmitic acid had higher concentration among

the fatty acid types found in both species ranging from 25.465 to 26.115 gm/100gm

in Clarias gariepinus and24.480 to 25.405 gm/100gm in Oreochromis niloticus.

Concentration of Oleic acid in both species ranging from 16.320 to 17.235 gm/100

gm in Clarias gariepinus and16.455 to 17.210 gm/100 gm in Oreochromis niloticus.

Deka et. al., (2012) reported that the higher level of total protein in

muscle of Labeo gonius was observed in pre- monsoon season 138.22 in Lotic

habitats and lowest level was found in retreating monsoon 42.8. Comparatively

lower protein content was observed in all the seasons of Lentic habitat whereas in

the present study the protein content in the muscles of Wallago attu has significant

difference between seasons and reservoirs selected the study. Ulfat Jan et. al., (2012)

reported monthly variation of the total protein content ranged from 0.0947 to 0.220

gm/gm tissue in Schizothorax niger. Highest total protein was found in month of

July and lowest was in January and December. Seasonal variation of total protein in

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the muscle tissue of Schizothorax niger was observed in summer season (19.25%)

and lowest in winter season, from Dal Lake of Kashmir.

Jain et. al., (2002) showed the correlation between the absorbance

obtained at 700 nm and the concentration of curve is linear from 2.5 to 200µg/ml.

The microwave micro assay tolerates the presence of higher concentrations of

various known substances to interfere with Lowry’s method. Ulfat Jan et. al., (2012)

examined the protein content in Schizothorax esocinus was higher in the month of

July 0.358 gm/gm and lowest in the month of December 0.089 gm/gm. Seasonal

variation in the protein content was observed in summer season (28%) and lowest in

winter season (10%) from Dal Lake of Hazratbal Shrinagar, Kashmir. Pawar et. al.,

(2013) was examined the protein content in Salmostoma sardinella 25.64%,

Cirrhinus reba 24.19%, Garra mullya 28.69%, Rasbora daniconius 32.79% and

Puntitus conchonius 27.10%. The highest protein content was found in Rasbora

daniconius 32.79%. Malviya et. al., (2011) reported that the amount of protein

percentage in the muscles of Macrones leucophasis fluctuated between 22.09 to

26.47 in female and in male from 22.07 to 25.63.

Niamke et. al., (2005) reported the Lowry method to be unreliable in the

acidic and increasing vitamin C rich medium. It was suggested to evaporate organic

solvents (compounds) when they are used to extract protein prior to protein

measurement the Lowry method. The influence of phenolic compounds on the

Lowry’s method was found to be governed by the structure of their molecules while

ammonium sulphate was a major agent used in the enzyme purification process.

Ashashree et. al., (2013) reported the protein levels of muscle in male catfish Mystus

cavasius varied from 0.42 to 5.27, highest in December and lowest in July.

Deka et. al., (2012) analyzed the higher amount of Total lipid (301mg/g) in

lotic habitat in retreating monsoon in the muscle tissue of Labeo gonius. Higher

mean values in lotic and lentic habitat were found in premonsoon and monsoon

season (97.51, 146.3mg/g) and showed a significance impact of habitat and seasonal

variation on the nutritional quality of lipid. Kamal et. al., (2007) revealed that mean

value of protein 14.87% and Fat 7.90% in Clarius batrachus, protein 17.34% and fat

3.45% in Heteropneustes fossilis, 19.63% protein and 7.79% fat in Anabas

testudineus, 18.30% protein and 4.98% fat in Notopterus notopterus, 16.09% protein

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and 7.34% fat in Nandas nandas, 19.13% protein and 4.55% fat in Channa

punctatus, 15.62% protein and fat 7.53% fat in Mystus vittatus from river Mouri,

Khulna, Bangladesh.

Islam (2005) studied protein content in Glossogobius giuris ranged from

14.09% (February) to 16.03% (August) in male and in female 13.88% (February) to

15.56% in (November). The average protein content of female 14.61% Glossogobius

giuris was lower than the male 15.23%. Lipid percentage in female Glossogobius

giuris increased during pre-spawning season and reach to minimum (1.06%) in June,

in its Spawning peak. In male, lipid content maximum 1.52% in March and

decreased in August 0.95% during post breeding period. Sutharshiny et. al., (2011)

studied the protein value was recorded higher in Scomberoides commersonianus

21.68% and lower in Scomberoides lysan 19.47%. The amount of lipid was higher in

Scomberoides commersonianus 1.003% and lower in Scomberoides tol 0.594% from

Sri Lankan water.

Saranya et. al., (2014) studied the biochemical composition of cultural and

natural fishes Catla catla, Labeo rohita and Cirrhinus mrigala. The concentration

muscle protein was maximum in March 19.920 gm/gm, 20.021 gm/gm, 19.560

gm/gm in natural fishes. But in Cultural fishes, maximum protein concentration in

muscle was 18.516 gm/gm in Catla catla, 19.210 in Labeo rohita and 18.327 gm/gm

in Cirrhinus mrigala. While, lipid concentration in muscles of Catla catla 3.961

gm/gm, Labeo rohita 4.231 gm/gm, Cirrhinus mrigala 3.731 gm/gm, in natural

fishes. In cultured fishes, it was 2.967 gm/gm in Catla catla, 3.160 gm/gm in Labeo

rohita, and 2.532 gm/gm in Cirrhinus mrigala.

Yeganeh et al. (2012) reported that protein content of Cyprinus carpio

decreased from summer to spring season 17.6- 15.9% in the farmed carp samples

and 18.2-17.9% in the wild carp samples and lipid content also decreased from

summer to spring season 5.1-1.5% in farmed carp and 3.8-2.8% in wild carp.

Muzumder et. al., (2008) analyzed protein in Amblypharyngodon mola (18.46%),

Gundusia chapra (15.23%), Puntius chola (14.08%), Chanda nama (18.26%),

Pseudeutropinus atherinoides (15.84%) and in Ailia coila (16.99%). Whereas, Fat

content was recorded highest in Gundusia chapra (5.41%) and lower in

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Amblypharyngodon mola (4.10%) and 3.05% in Puntius chola, 1.53% in Chanda

nama, 2.24% in Pseudeutropinus atherinoides, 3.53% in Ailia coila.

Shabani et. al., (2014) analyzed the total fat content in fatty tissue of male

fish Pangasianodon hypopthalmus was 73.23% and in female fish 77.64%. The fatty

acids in catfish Pangasianodon hypopthalmus was Palmitic acid (C16:0), Oleic acid

(C18:1n-9) and Linoleic acid (C18:2n-6). Sabry S. El-Serafy et. al., (2005) showed

the fat content in fish muscle of Orechromis niloticus was high in winter and early

spring in adult specimens, it was decreased after breeding season. The protein

content concentration varied from 73.1 to 85.6%, 73.0 to 87.5%, 73.5 to 88.0 % and

73.8 to 85.2% from four stations i.e. El- Kanates, Benha, Zefta and Talka. Lowest

protein content (73.0%) was found in female during autumn from Behna and highest

(88.0%) was from Zefta during spring season.

Olopade et. al., (2015) reported the protein content varied from 19.47 -

20.14 for Chrysichthys walkeri and 18.62 - 20.54 for Chrysichthys nigrodigitatus.

The amount of fat content was varied from 1.24% in February to 1.63% in April for

Chrysichthys walker, 9.00 %- 9.53% for Chrysichthys nigrodigitatus. Elagha (2013)

studied mean percentage of protein and lipid Labeo niloticus, Mormyrops

anguilloides, Marcusenius cyprinoides, Mormyrus niloticus, Clarias lazera and

Protopterus annectens. The protein content in muscle tissues ranged from 66.00-

84.00% and higher mean percentage of protein was found in muscle tissues of Labeo

niloticus 66.00% and Protopterus annectens 84.00%. While lipid percentage ranged

from 2-26% in muscle tissues of fishes and higher amount of mean percentage of

lipid was found in Marcusenius cyprinoides and Mormyrus niloticus.

Ali Aberoumand (2014) analyzed the protein content in Thunnus

alalunga 22%, Evynnisjaponica 13.02%, Caulerpalentillifera 26.9%, Orcynopsis

unicolor 22% and Euthynnusaffinis 24%. The amount of protein was higher in

Caulerpa lentillifera 26.9% and lower in Evynnis japonica 13.02%.While, Fat

content were recorded as 23.3%, 0.24%, 15%, 16% and 14%. The highest fat content

was recorded in Thunnus alalunga (23.3%). Sahar Jalili et. al., (2014) recorded the

protein and lipid content levels in Tiger Tooth Croaker (Otelithes ruber) were

21.524% and 2.916%. For four Finger threadfin (Eleutheronema thetradactylium)

were 20.732% and 4.935%. Kumar V. V. et. al., (2012) reported protein content

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level in immature and mature group of Harpodon nehereus were 7.10% and 7.43%.

Whereas lipid level in those were 1.73% and 2.0%.

Shingadia (2013) reported the total protein in muscle tissues of Harpodon

nehereus ranged from 15g % in September 37g % in March. While, The highest

lipid content in the month of April 0.26 g % and lower 0.14% in September.

Debnath et. al., (2014) was reported the highest protein content in Labeo bata

(75.43%) and lowest in Catla catla (39.37%). Shi et. al., (2013) reported the protein

contents of bighead carp (Aristichthys nobilis) muscle were slightly higher than

those of Paddlefish (Polyodon spathula) but no significance difference were

observed (p0.05). However, the amount of fat content in paddlefish was

significantly higher than that of bighead carp (p<0.01).

Daniel (2015) examined the mean values for protein and fat were 14.2%

and 10.61% in Chrysichthys nigrodigitatus, 18.56% and 2.57% in Cynoglossus

senegalensis, 17.83% and 3.39% in Polydactylus quadrifilis. Mohanty et. al., (2012)

reported crude protein 20.06% and crude fat 1.40 % in edible muscle tissue of

Sperata seenghala (Skyes). Memon N. N. et. al., (2011) examined protein and lipid

content in Wallago attu 21.06 % and 2.21%and Aorichthys aor 19.05% and lipid

1.78%. Pilla et. al., (2014). Reported that the highest level of protein and lipid in

muscles of Lutjanus johni was observed 85.56 mg/g, 8.24 mg/g in monsoon season

and lowest in post monsoon season as 75.01 mg/g, 5.01 mg/g during 2010-2011.

During 2011 - 2012, highest 84.92 mg/g, 8.30 mg/g in monsoon and lowest as 73.99

mg/g, 5.04% in post monsoon from Vishakhapatnam coast.

M. Dorucu (2000) examined the protein and lipid mean percentage of muscle

as 86.00% and 5.03% in Powan (Coregonus lavaretus) from Loch Lomond,

Scotland. Mathana et. al,. (2012) reported the protein and lipid content in muscle of

Lethrinus lentjan ranged from 18.75 to 23.84 and 7.28 to 9.88 mg/gm. Olagunju et.

al., (2012) studied maximum level of protein 20.26% and lipid content was low as

3.13% in catfish Hemisynodontis membranacea of the other three fishes. Ayeloja et.

al., (2013) studied the crude protein and lipid in Clarias gariepinus as 16.375%,

2.033% and 14.328%, 1.298% in Oreochromis niloticus. Adewumi A. A., (2015)

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examined the crude protein 65.94%, 67.77% and lipid 0.61%, 0.79% in wild and

cultured Clarias gariepinus.

Begum et. al., (2012) reported that Protein content as 16.78%, 17.31%,

18.17%, 16.69%, 16.73% and Lipid as 4.55%, 3.99%, 4.5%, 1.87%, 9.00% in

Ailiichthys punctatus, Clupisoma psendeutropius atherinoides, Puntius sarana,

Gundusia chapra, Corica soborna. S. M. Moniruzzaman et. al., (2014) et. al., (2014)

reported protein content and fat content as17.20%, 19.60%, 16.68%, 17.09% and

16.07%, 16.94%, 15.24%, 14.43% in young or spent Hilsa (Tenualosa ilisha) flesh.

In gravid or ripe Hilsa (Tenualosa ilisha) flesh protein and fat value with 15.74%,

16.29%, 18.07%, 19.44% and 14.38%, 13.84% 14.67%, 14.33%. Mba et al., (2011)

reported the crude protein level in freshwater fish varied from 20.26% in Alestes

nurse to 21.52% in Oreochromis gallilaeus. Shakir et. al., (2013) reported the crude

protein was19.30%, 17.37%, 16.86%and fat level as 2.71%, 1.75%, 2.16% in Labeo

rohita, Cirrhinus mrigala, and Catla catla.

Elagba (2013) studied Lates niloticus (protein 74.2%, lipid 2.00%),

Tetraodon lineatus (protein 71.5%, lipid 4.9%), Heterotis niloticus (protein 78.8%,

lipid 12.5%), Mormyrus niloticus protein 73.4%, lipid 3.8%), Clarias lazera (protein

59.9%, lipid 16.6%), Protopterus annectens (protein 68.00%, lipid 4.5%).

Emmanuel et. al. (2011) reported the highest level of protein 30.31% for young adult

tail Clarias gariepinus and least value 23.06% Juvenile tail Clarias gariepinus and

the highest level of lipid 0.06% for young adult tail Clarias gariepinus and least

value 0.31% Juvenile tail Clarias gariepinus.

4(E) - CONCLUSION

Total protein content in body muscles of Wallago attu from Masoli

Reservoir was higher in summer season and lower in winter season

from Siddheshwar Reservoir.

Total protein content in body muscles of Sperata seenghala from

Masoli Reservoir was higher in summer season and fluctuations in

the results were found. Lower total protein content in body muscles

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of the fish from Siddheshwar Reservoir in monsoon season and from

Karpara Reservoir in winter season.

There is significant difference (p<0.05) in total protein content of

body muscles of Wallago attu during Feb. 2010 to Jan. 2011 and Feb.

2011 to Jan. 2012 collected from different selected reservoirs in the

selected study area in different seasons.

There is significant difference (p<0.05) in total protein content of

body muscles of Sperata seenghala in all seasons during Feb. 2010 to

Jan. 2011 and Feb. 2011 to Jan. 2012 but there is no significant

difference (p0.05) in total protein content of body muscles of

Sperata seenghala collected from different selected reservoirs during

Feb. 2011 - Jan. 2012.

estimation of total protein content (mg/g) in body muscles of...

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