effect of different storage conditions on protein and ...joics.org/gallery/ics-1225.pdf · effect...

Effect of different storage conditions on protein and lipid

content of a common major carp Cirrhinus mrigala(Ham.)

in summer and winter season.

Vivek Kumar Anand1, Madhu Shree2, Vineet Kumar3, Dr. Ashok Kumar 1,#

1- Department of Zoology and Industrial Fish and Fisheries ,Ganga Singh

College, Chapra

2- Department of Zoology, Lok Mahavidyalaya, Hafizpur, Baniapur, Saran

3- Department of Zoology, Z.A. Islamia College, Siwan

#: Address of correspondence:

Dr. Ashok Kumar

Department of Zoology and Industrial Fish and Fisheries

Ganga Singh College, Chapra

Email- [email protected]

Abstract:

India is among top five nations in fish production & processing nations throughout the world.

She has about 2500 marine fishing villages spread near the sea-shore from Bengal (through

Tamilnadu, Kerala, Maharastra) to Gujrat states & about 1500 landing centres along the

coasts Fish is one of the most perishable human food which start spoiling the moment they

are caught and taken out from the water leading to their death. So it becomes essential to find

out some cost effective methods for preservation and transportation of fish from one place to

other. In the present study we observed changes in total protein content (mg/100g) of the

muscle of cirrhinus mrigala (Ham.). During selected days (18 days) of different conditions

(chilling, super chilling, sorbate treated and eviscerated condition) during summer and

winter months. The total protein ranges from 18.38±0.10 (Zero days) to 17.50±0.15 (on 18th

day) under chilling condition. The eviscerated super chilling condition caused comparatively

less decrease in total protein content as compared to other methods. The total fat ranges from

0.98±0.010 (Zero days) to 0.65±0.015 (on 18th day) under chilling condition. The eviscerated

super chilling condition caused comparatively more decrease in total fat content as

compared to other methods.

Keywords: Cirrhinus Mrigala, protein deficiency, Total protein content, Total lipid content

Journal of Information and Computational Science

Volume 9 Issue 8 - 2019

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mailto:[email protected]

Introduction

Fisheries play a very important role in Indian economy mainly because of its employment

potential and for earning valuable foreign exchange [1]. The Global Trade in fish & fishery

products has steadily increased in recent years and now India is among the seven largest fish

producing country in the world and also a major exporter of fish & fishery products to the

world market [2]. The sea food industry in India is in a transitional stage from the traditional

block freezing to the production of individually quick frozen (IQF) and value added frozen

items for export to major overseas markets. Indian products could not make a brand image as

we are exporting our products in block frozen form, which are reprocessed and repacked in

overseas markets under their brand names except a few exporters [3]. However, there is no

organised industry for exclusively fresh water fish preservations as some of these sea food

industries occasionally preserve major freshwater carps & prawns for export purposes in very

limited quantities, whereas, vast scope in this line also exists. Out of the total production from

fresh & brackish water aquaculture, about 75-78% is consumed in rural areas & remaining in

urban areas. About 2% to 3% production moves more than 100 km distance from production

centre, 25% to 28% moves up to 100 km while remaining is consumed locally in rural/urban

areas [4]. From reservoir fishery, only 7% to 10% of the production is consumed at urban

centres located away from the production centre. In these cases, ice is commonly used to

preserve the freshness of the fish and rarely the facility of cold storage is used [4]. Bamboo

baskets are commonly used followed by thermocol boxes in some cases. Due to poor &

unscientific preservation & handling, 40% to 50% of the fish brought into fresh fish markets

are in various stages of spoilage [5].

The chemical composition of fish varies from species to species & individual to individual

depending on age, size, sex, season, environment & health. However, the major chemical

constituents are: water/moisture (68-80%), Ash (0.4-1.5%), Carbohydrate (0.3-1.5%), Protein

(6-28%), Fat/Lipids (0.1-6.7%) and Vitamins & minerals (0.01-1%) [5]. The amount of

vitamins & minerals in fish muscle is species specific and varies with the season. Fish muscle

is also a good source of vitamin D and fatty species contains good amount of vitamin A & D

[6].

The intensity of spoilage may be categorised as biological changes, physical changes,

autolytic changes, chemical changes, and sensory change. Considering all the above

mentioned facts, the present investigation carried out to ascertain the shelf-life period of the

fish during chilled & super chilled conditions and in the same storage conditions after the

treatment of potassium sorbate and eviscerated fish during summer & winter seasons with

special reference to some biochemical & organoleptic changes in the fish muscle. The fish

under study is a very common major carp & in good demand, hence the findings may be

useful for the fish farmers to preserve their fish in fresh condition for longer periods &

transport to earn good profits.



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Material and Methods

Chemicals

All chemicals used were of highest purity grade purchased from SRL Pvt. Ltd, Mumbai.

Fish

Sufficient number of healthy & living specimen of the fish under study, Cirrhinus mrigala

(Ham.) belonging to family, Cyprinidae, Division, Cyprini of the order, Cypriniformes

(Teleost) in the weight group between 400-450 gms were procured from local fish farms

(Ponds), which had no connection with any contaminated substances or sewage, during

summer & winter season. These fishes were produced on every alternate day for different

storage conditions.

Chilling conditions: Fresh living fish were kept in ice in layers (one layer of ice followed by

a layer of fish & so on) in the tea-chest boxes & brought to the laboratory. During the storage,

care was taken to replace fresh ice at every 24 hours. As it is known that icing of fish

immediately after catch, keeps them fresh/acceptable condition for few days or weeks on

their inherent characteristics and the shelf-life of the fish may shortened due to delayed,

insufficient and/or improper icing. Therefore, immediate cooling of the catch becomes more

important as high environmental (atmospheric) temperature may result rapid spoilage.

Super chilling: The storage of fish at temperature between 0°C to -4°C is called

superchilling. In a very simple manner, superchilled temperature can be achieved with a

mixture of ice & salt (NaCl). The process taking place in it is melting of ice with

simultaneous dissolution of salt. The thawing temperature of ice depends on the amount of

salt contained in it. However, addition of salt to ice insures a maximum reduction of

temperature only to a certain limit. According to Bobkov’s approximate formula: tm= -0.7.

X, where in “tm” is the temperature of the mixture of ice & “X” is the amount of salt as % of

weight of ice. Considering this formula in present study, 1.5% of sodium chloride (Tata

Refined Salt) i.e. 375 gm salt for 25 kg of ice and 12-13 kg of fish were taken as this mixture

maintained the temperature at about -1°C during experiment. Higher salt concentration was

not used as they may cause freezing of the fish. Accordingly, 2-3 boxes were used for this

experiment. This mixture of ice and salt is also changed at every 24 hour during the

experiment.

Sorbate treated chilling & superchilling : In these two storage conditions, the process are

same as described above save & except that in these two conditions, the fish were dipped in

5% Potassium sorbate soln (Aqueous) for 10-12 minutes prior to packing the fish in

respective boxes. In this study potassium sorbate has been choosen because it is one of the

safest preservative (Antimicrobial food additive), which is white fluffy powder & very much

soluble in water (139.2 gm/100ml).

Eviscerated chilled & Superchilled : For these experiments, the gill & viscera of each fish

were removed by a longitudinal cut on posterior side of abdomen, followed by proper



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washing with normal fresh water before placing in ice and ice+salt mixture (as described

above for chilled & superchilled processes in Tea-chests).

Total Protein (%) measurement

100 mg muscle was homogenized in 2.0 ml of double distilled water & repeated with residue.

1.0 ml of homogenate was taken & mixed up with 8.75 ml of 30 % NaOH soln. left for 20

minutes. To this, 0.25 ml of 20% Aq. CuSO4 soln. was added & mixed, Violet colour

developed due to formation of Cu-Na biurate compound. The OD/T of this solution was read

at 530 filter in colorimeter using 1.0ml of double distilled water in place of homogenate as

blank & proceeded as above like sample.

Total protein = OD of 1 ml sample x k-1 protein x4

= mg/100 mg(i.e.%)

Where in, k-1 protein = 48.661 mg/ml (constant)

“4” is the number of total homogenate of 100mg tissue

Fat/Total Lipid content (%) measurement

Some amount of moisture free muscle sample was transferred in an already weighed

extraction thimble & weighed. The thimble was then placed in the extraction tube of soxhlet

apparatus, which was connected to an already weighed empty flask on the lower end and a

condenser on the upper end. The extraction flask was filled with sufficient amount of

petroleum ether, until siphoning began. Once the lower flask got filled with ether, the heating

mantle was switched on to 35°C. The ether collected in the extraction flask was siphoned 5-6

times/hour. The ether was then allowed to collect in the extraction tube until the thimble was

immersed in the solvent & left overnight so as to extract the maximum possible fat. Next day,

the heating mantle was again switched on & the solvent siphoned for another 5-6 times. The

apparatus was allowed to cool & thimble removed. The heating system was set again & the

solvent was collected in the extraction flask below the siphoning level, the apparatus was

then cooled & ether was collected out. This process continued until traces of solvent

remained in the bottom flask. The flask was then removed & dried in the oven for 15-20

minutes at 45°C. The flask was then cooled in the desiccator & weighed.

Calculation: 100-% moisture = % dry mater (X)

Wt. of thimble with sample-wt. of empty thimble= Wt. of dry

sample (Y)

Wt. of wet sample = Y x 100/X =Z

Wt. of Flask with Fat – Wt. of Empty flask = Wt. of Fat(A)

% Fat = 100 x A/Z

Non protein Nitrogen (NPN) (mg/100gm) : 1.0 gm muscle sample was taken in a long

necked digestion flask. To this, 20.0 ml of 7%TCA & 2.0 ml of digestion mixture (Digestion

mixture contained 25.0mg Hgo : 50.0gm K2So4+300.0ml conc. H2So4 (Sp.gr.1.84) mixed &

made up to 1 liter with distilled water ) was added & flask was placed in an inclined position

& boiled for 2-3 hrs. till the soln. became clear. This clear soln. was allowed to boil further



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for 30 minutes. The flask was cooled & soln. was transferred to a 100 ml volumetric flask.

The inner side of the digestion flask was washed with distilled water & the volume was made

up to 100 ml mark & mixed.

4.0 ml of digested mixture was taken in Kjeldahl’s flask & treated with excess of 10 (N) Sod.

Hydroxide (NaOH) soln. A little distilled water was used to wash down the soln. in the flask.

A conical flask containing 10.0ml of 2% boric acid with 1 drop of mixed indicator (2.0 gm

boric acid dissolved in 50 ml hot water & cooled and 0.2 ml of 0.1 % bromocresol green in

alcohol was mixed & diluted to 100ml with distilled water) was placed on the receiving end

of the condenser. The tip of the condenser was dipped inside the soln. & about 20.0ml of

distillate was collected. Contact between the condenser tip & distillate was disconnected &

distillation continued for 5 minutes to steamout the condenser. The distillate was titrated with

N/70 Hcl to the point of getting light purple colour.

Calculation: 5 ml N/70 Hcl is equivalent to 1 mg nitrogen.

NPN (mg/100gm) = TV x 100x 100x 1

4x5x wt. of muscle

α- Amino Nitrogen (AAN) (mg/100gm) (Pope & Sevens, 1939) : 1.0 gm of muscle sample

was taken in 20 ml of ice cold 10% TCA. Homogenized, centrifuged & the residues were

rehomogenized & centrifuge. Now 15 ml of TCA extract was taken in a 50ml measuring

cylinder. Sufficient amount of 10N NaoH was added & mixed till the solution developed a

bluish tinge. Now 30ml of CuSo4 soln. (6 ml copper chloride+ 12 ml Sod. Phosphate + 12 ml

borate buffer) was added and the volume was made up to 50ml, mixed & filtered

immediately. Now 20 ml of filtrate was taken in a conical flask and to this 0.5ml of glacial

acetic acid and 200gm pot. Iodide was added. The flask was immediately covered &

placed in complete dark for 5 minutes to allow reaction to complete. Then L drop of

starch soln. was added & titrated to colourless using 0.01N Sod. Thiosulphate soln. (0.01N

Sod. Thiosulphate is equivalent to 280mg α-amino nitrogen).

Calculation:

α-Aminonitrogen (mg/100gm) = TVx 0.280 x50x100x100

20 x15x wt. of sample (muscle)

Total volatile base nitrogen (TVBN) (mg/100gm): 10.0 gm muscle sample (the

moisture/water content has already calculated as per moisture free sample as prepared above)

was blended with 30ml of 5% TCA. Filtered to obtain a clear extract. Now 5 ml of clear

extract was pipetted & placed in distillation apparatus. 5.0 ml of 2N NaoH was added &

distilled in to 15ml 0.01N Hcl containing 0.1 ml Rosolic indicator. After distillation, excess

acid in the receiving flask was titrated with 0.01N NaoH soln. up to pale to pink end point. A

blank was also used for the determination.

Calculation: TVBN (mg/100gm) = N X (A-B) (14)(30-w)/5

Where in, N= Normality of NaoH standard soln.

W= Water content in the sample



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A= ml NaoH used for blank titration

B= ml NaoH used for sample titration

Fat/Total Lipid content (%) : Some amount of moisture free muscle sample was transferred

in an already weighed extraction thimble & weighed. The thimble was then placed in the

extraction tube of soxhlet apparatus, which was connected to an already weighed empty flask

on the lower end and a condenser on the upper end. The extraction flask was filled with

sufficient amount of petroleum ether, until siphoning began. Once the lower flask got filled

with ether, the heating mantle was switched on to 35°C. The ether collected in the extraction

flask was siphoned 5-6 times/hour. The ether was then allowed to collect in the extraction

tube until the thimble was immersed in the solvent & left overnight so as to extract the

maximum possible fat. Next day, the heating mantle was again switched on & the solvent

siphoned for another 5-6 times. The apparatus was allowed to cool & thimble removed. The

heating system was set again & the solvent was collected in the extraction flask below the

siphoning level, the apparatus was then cooled & ether was collected out. This process

continued until traces of solvent remained in the bottom flask. The flask was then removed &

dried in the oven for 15-20 minutes at 45°C. The flask was then cooled in the dessicator &

weighed.

Calculation: 100-% moisture = % dry maller (X)

Wt. of thimble with sample-wt. of empty thimble= Wt. of dry

sample (Y)

Wt. of wet sample = Y x 100/X =Z

Wt. of Flask with Fat – Wt. of Empty flask = Wt. of Fat (A)

% Fat = 100 x A/Z

Free Fatty Acid (FFA) (mg/gm) : The residue obtained from fat determination (above

mentioned expt.) was dissolved in 2.0 ml of 95% alcohol (Ethyle alcohol). One drop of 0.1%

alcoholic phenolphalin was added & titrated with N/50 KOH soln (Aqu.) until a pink colour

developed. This titration was done by using microburet with 0.01 ml titration capacity. For

blank, 2.0 ml of ethonol was titrated by same manner. The titration value of blank was then

substracted from the residue processed titration to get the titration value of the residue.

Net titration value x 32 = m Eq. free fatty acid/100 gm sample

and assuming an average mol. Wt. of 277 for fatty acid

m Eq/lit x 27.7 = mg free fatty acid (FFA)/gm sample.

Total plate count (TPC) in cfu x 10/gm:-

(i) Buffer solution : 34.0 gm KH2PO4 + 100ml distilled water

Adjusted to pH 7.0

(ii) Media preparation : Pepton – 3.0gm

Trypton – 5.0gm

Dextrose – 1.0gm



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Agar – 15.0gm

Distl. Water -100ml

pH adjusted to 7.0 ±0.1

Above substance were soaked for 15 minutes in 1000 ml distilled

water by gentle heating, then it was sterilized with container in Autoclave at 15 lbs

pressure (121°C) for 15 minutes. Some petridishes were also sterilized. Now, 15.0ml

of medium was taken in a sterilized petridish in an incubator at 42°C, keeping the

plate inverted for 20 minutes. Well dried plates were used for further experiment.

10.0gm muscle with skin was taken (accuratly weighed) in a pre-weighed

sterilized petridish & blended for 2 minutes at 1000 r.p.m. in a sterilized homogenizer

with working sterilized buffer (1.25 ml of stock buffer made up to 100ml with

distilled water) in the ratio of 1:9 (buffer).

A series of dilution of sample suspension i.e. 10-2,10-3,10-4 were prepared with

buffer soln. & 1.0 ml of each selected dilution were poured in separate petridishes

containing 9.0ml buffer soln. media (in duplicate). The media was spread by rotating

& tilting the petridish. Then the petridishes were incubated at 37±1°C in an incubator

in inverted position for 48 hours. Sometimes incubations were also done at room

temperature, covered by a big glass jar.

Coloney counting was done manually, plates were showing the counts

between 30-300 were enumarated. The total plate count was expressed in colony

forming units i.e. cfu/gm of fish muscle (10,000 cfu/gm).

Calculation : TPC (cfu/gm) = No. of colonies x Dilution

Weight of the sample

Organoleptic/Sensory evaluation : Though, many methods have been tested for

evaluating fish quality, but sensory methods are still most satisfactory way of

knowing quality deteriroration. In the present study, changes in appearance of body,

odour, eye & gills have been taken in to account for raw fish/muscle & cooked

muscles on different days in different storage conditions during both summer &

winter months, which are summarized in Table. 12.

Furhter, the sensory evaluation of raw & cooked meat (muscle) of the fish in

different storage conditions on different selected days was carried out by five different

persons on 10-0 point basis & average points have been noted & summarized in Table

– 13 & 14. The fish were cooked in 3% common salt solution after dressing the fish

muscle & cooled to room temperature before their tastes. The score for each of the

sample was given on the scale of 0 to 10 points, where 10 is for excellent, 9 – very

good; 8- good; 7- slightly good, 6- above borderline, 5- border line, 4- slightly poor

and 3 & below, unacceptable. Accordingly, the shelf-life of the fish in different

storage conditions were recorded up to 5 to 4.5 point score.

Results

Total Protein: The total protein content in the muscle of the fish in fresh condition (“0” day)

has been recorded to be 18.38±0.10 & 18.95±0.12 mg/100mg (i.e.%) in summer & winter

seasons respectively. The total protein content has been recorded significant increased



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(P<0.05) during winter than the summer season. A gradual decrease has been observed in

both season depended on the days of storage in selected storage conditions (Figure- 1).

During summer, the decline was found statistically significant (P<0.01) on 18th day (17.50±

0.16%) in chilled; on 20th & 22nd day (P<0.05, P<0.01) in superchilled (17.83±0.18 &

17.54±0.13%) & sorbate treated chilled condition (17.81±0.19 & 17.52±0.15%) while in

sorbate treated superchilled condition decline was found significant (P<0.05) on 22nd day and

P<0.01 on 24 to 26 days (17.54±0.15 & 17.45±0.17%), whereas, in Eviscerated chilled

condition P<0.05 & P<0.01 has been recorded on 26th (17.78±0.18%) & 28th day

(17.48±0.15%) and superchilled condition the same has been recorded on 28th (17.61±0.17%)

& 30 &31st day (17.53±0.15 & 17.45±0.11%) respectively.

Similarly, during winter season, the total protein content in the muscle of the fish under

different storage conditions showed a decreasing trend depended on the day of storage. The

fish stored in chilled, super chilled and sorbate treated chilled conditions showed a gradual

decline which were found statistically significant (P<0.05 & P<0.01) on 18th & 20th day

(18.25±0.14 & 18.00±0.16%) in chilled; and on 22nd & 24th day (18.26±0.15 & 18.12±0.13%

and 18.20±0.17 & 18.06±0.14%) in superchilled & sorbate treated chilled conditions

respectively. In sorbate treated superchilled significant decline (P<0.05 & P<0.01) were

observed on 26th & 28th day (18.26±0.19 & 18.06%). In eviscerated chilled condition

significant decrease (P<0.05) was observed on 28th day with maximum decline (P<0.01) on

31st day of storage, whereas, in eviscerated superchilled condition the same decline was

reported on 31st day (18.35±0.17%) with maximum on 34th day (18.06±0.13%).



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Figure 1: Graph showing protein content in the muscle of Cirrhinus mrigala at selected days

of different storage conditions during summer and winter seasons

Non Protein Nitrogen (NPN): Like total protein content, the non protein nitrogen content in

the muscle of the fish also showed a gradual decrease (Figure - 2). The declines were

statistically significant (P<0.05) on 18th day in chilled condition (250±7 mg/100gm) and on

22nd day (242±9 & 246±8mg/100gm) in superchilled & sorbate treated chilled conditions

respectively, when compared with the value of fresh fish (314±13mg/100gm) during summer

season. However in sorbate treated super chilled & eviscerated chilled fish & superchilled

fish, the declines were significant (P<0.05) on 24th, 26th & 28th day (235±12, 244±11 &

246±12 mg/100gm) with a maximum decline (P<0.01) on 26th,28th & 31st day (220±10,

213±9 & 210±11mg/100gm) respectively.

Similarly, the NPN during winter season were also found statistically significant (P<0.05) on

20th (212±11mg/100gm) 22nd (240±10mg/100gm), 24th (232±10mg/100gm), 26th

(225±10mg/100gm), 28th (232±14) & 31st (228±14mg/100gm) days in chilled, superchilled,

sorbate treated chilled & superchilled and eviscerated chilled & superchilled conditions with

maximum decline (P<0.01) on 24th day (221±8 mg/100gm) in superchilled, 28th day (210±8

mg/100gm) in sorbate treated superchilled and on 31st & 34th day (202±11 & 200±9

mg/100gm) in eviscerated chilled & superchilled conditions. Thus, the decrease in NPN

contents might be due to the consumption of NPN fraction by bacteria.



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Figure 2: Graph showing non protein nitrogen in the muscle of Cirrhinus mrigala at selected

days of different storage conditions during summer and winter seasons

α-Amino Nitrogen : The α-amino nitrogen in the muscle of the fresh fish during summer &

winter seasons have been recorded to be 46.48±1.47 & 55.90±1.56 mg/100gm respectively. It

was comparative more during winter than the summer season and the difference of 9.42

mg/100gm has been recorded significantly (P<0.05) high in winter (Figure- 3). An initial

increase from 5 to 18 days has been recorded in both seasons. During summer, maximum

increase has been recorded (P<0.05) on 5th day in chilled (53.94±1.39 mg/10gm), on 10th day

in superchilled & sorbate treated chilled (54.52±1.26 & 53.80±1.22mg./100gm) conditions

followed by a decrease. The declines were statistically significant on 18th day in chilled &

22nd day in superchilled & sorbitate treated chilled conditions (35.98±1.26, 39.65±1.42 &

38.97±1.39mg/100gm respectively). Similarly, significantly increased values (P<0.05 &

P<0.01) have been observed on 10 to 15th days in sorbate treated superchilled (53.25± 1.30 to

56.70± 0.28mg/100gm), eviscerated chilled (52.76± 1.25 to 55.94± 1.22mg/100gm) and on

15th to 18th day in eviscerated superchilled (53.10± 1.28 to 56.68± 1.37mg/100gm)

conditions. After reaching the maximum increase, it gradually decreased and the declines

were statistically significant during flag ends (P<0.01) than the normal value of the fresh fish.

The decreases are up to 35.98±1.26 mg/100gm. On 18th day in chilled, 39.65±1.42 &

38.97±1.39 mg/100gm in superchilled & sorbate treated chilled, 26th, 28th & 28th to 31st day



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in sorbate treated superchilled, eviscerated chilled & superchilled conditions (33.10±1.20,

36.80±1.41 and 38.54±1.22 to 31.36±1.24 mg/100gm) respectively.

Similarly, during winter season also a significant increase (P<0.05) has been observed on 5th

day in chilled (62.48±1.30mg/100gm), on 10th day in superchilled & sorbate treated chilled

(61.54±1.40 &63.10±1.26mg/100gm) and on 15th & 18th day in eviscerated chilled &

superchilled conditions (63.00±1.40 & 63.90±1.30mg/100gm) respectively. These are

followed by a gradual decrease and the declines were statistically significant (P<0.05 &

P<0.01) in all storage conditions depended on the day of storage as the maximum decline up

to 48.78±1.26, 38.50±1.22 & 42.06±1.29mg/100gm have been recorded on 20th day in

chilling and 24th day in superchilling & sorbate treated chilling conditions respectively,

whereas in sorbate treated, eviscerated chilling & superchilling conditions the maximum

decrease up to 38.54±1.32, 39.00±1.31 & 34.54±1.27mg/100gm respectively were recorded

on 28th, 31st & 34th day of storages.

Thus, the initial increase observed up to certain period of storage may be attributed to

maintain good flavour & texture of the fish but maximum decline thereafter during flag days

of the experiment may be related to the unacceptability or spoilage of the fish and/or linked

with bacterial proteolytic action as suggested by Shewan (1961).

Figure 3: Graph showing alpha amino nitrogen in the muscle of Cirrhinus mrigala at

selected days of different storage conditions during summer and winter seasons.

Total volatile base nitrogen : The total volatile base nitrogen content in the muscle of the

fish (in fresh condition) during summer & winter seasons has been recorded to be 10.02±0.44



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& 7.95±0.30 mg/100gm respectively. The difference of 2.07 mg/100gm indicates that the

TVBN content during summer is significantly (P<0.05) more than that of the winter season

(Figure- 4). An initial increase in TVBN content has been observed upto 10 to 15 days of the

storage followed by a gradual decrease in almost all the storage conditions during summer &

winter seasons. However such increases were not found statistically significant at any stage,

but a gradual decrease thereafter observed were found statistically significant

(P<0.05/P<0.01) on 18th day of storage in chilled (7.40±0.30mg/100gm), 20 to 22nd day in

superchilled & sorbate treated chilled (8.08±0.25 to 7.60±0.27 and 8.46±0.28 to 7.55±

0.30mg/100gm respectively), on 24 to 26 day 8.02±0.22 to 7.56±0.26mg/100gm in sorbate

treated superchilled and on 26th to 28th and 28th to 31st day of eviscerated chilled and

superchilled (8.20±0.29 to 7.64±0.22 and 8.47± 0.30 to 7.56±0.27mg/100gm respectively)

conditions during summer season.

Almost similar trend has also been observed during winter season. A gradual increase

followed by a decrease has been observed in all the storage conditions and the decreases were

recorded significant (P<0.05/P<0.01) on 20th day in chilled (6.35±0.21mg/100gm), 22nd to

24th days in superchilled & sorbate treated chilled (6.25±0.30 to 6.05±0.25 and 6.68±0.26 to

5.90±0.31mg/100gm respectively) conditions, on 26th & 28th day in sorbate treated

superchilled (6.79± 0.23 to 5.90±0.26mg/100gm) and on 30th & 31st day and 32nd to 34th day

of storages in eviscerated chilled (6.40±0.23 to 6.14±0.20mg/100gm) & eviscerated

superchilled (6.80±0.25 to 6.02±0.25mg/100gm) conditions respectively. The TVBN value is

expected to increase gradually with the storage period but uncontrolled leaching with ice-melt

water makes this parameter ineffective as an index of spoilage & thus this values should not

be considered as an index of spoilage.



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Figure 4: Graph showing total volatile base nitrogen in the muscle of Cirrhinus mrigala at

selected days of different storage conditions during summer and winter seasons.

Fat (Total Lipid) : The average changes in fat (total lipid) content in the muscle of the fish

have been recorded to be 0.93±0.02 & 1.09±0.05mg/100gm(%) during summer and winter

seasons. (Figure- 5) However, the difference in between the two seasons is not statistically

significant. The fat content has been recorded gradually decreased depended on day of

storage & storage conditions. During summer season, the decrease were found statistically

significant (P<0.05 & P<0.01) on 15th & 18th day (0.71±0.06 & 0.64±0.03%) in chilled, on

18th to 22nd day (0.74±0.04 to 0.69±0.04%) in superchilled, on 20th and 22nd day (0.76±0.05

& 0.69± 0.03%) in sorbate treated chilled, on 24 & 26th day (0.73±0.06 & 0.68±0.04%) in

sorbate treated superchilled, on 26th & 28th day in eviscerated chilled (0.71±0.05 &

0.63±0.03%) and on 28th to 31st day (0.73±0.05 to 0.60±0.04%) in eviscerated superchilled

conditions.

Similarly, the fat content during winter season also showed a gradual decrease which became

statistically significant (P<0.05 &P<0.01) on 18th & 20th day (0.79±0.07 & 0.70±0.05%) in

chilled, on 20 to 24 days (0.92±0.04 to 0.71±0.04%) in superchilled, on 22nd & 24 days

(0.80±0.04 & 0.74±0.05%) in sorbate treated chilled, on 24th to 28th day (0.81±0.05 to

0.69±0.03%) in sorbate treated superchilled, on 28th to 31st days (0.78±0.05 to 0.66±0.05%)

in eviscerated chilled and on 31st to 34 days (0.90±0.04 to 0.68± 0.05%) in eviscerated

superchilled conditions.



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Figure 5: Graph showing total fat in the muscle of Cirrhinus mrigala at selected days of

different storage conditions during summer and winter seasons.

Free Fatty Acid (FFA): Contrary to the fat content, free fatty acid content in the muscle of

the fish has been recorded comparatively more during summer than the winter seasons, but

the difference in between them are not found statistically significant. The free fatty acid in the

muscle has been recorded to be 3.10±0.33 & 2.72±0.29 mg/gm during summer & winter

seasons when the fish were in fresh (“0-day”) condition (Figure- 6). Further, in both summer

& winter seasons, a gradual increase followed by a decrease after considerable periods has

been noticed but even at the flag end, the decreases were always significantly more in content

than the amount present during fresh condition of the fish on”0” day of storages.

During summer season, a gradual increase in the FFA content has been noticed in almost all

storage conditions, which became statistically significant (P<0.05/P<0.01) on 15 day

(5.07±0.27mg/gm) followed by decrease but significantly more than fresh value (P<0.05) on



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18th day (4.45±0.30mg/gm) in chilled; on 15 to 18th day in superchilled & sorbate treated

chilled (4.94±0.32 to 5.42±0.36 and 4.76±0.37 to 5.38±0.29mg/gm) followed by significant

decrease (P<0.05) on 22nd day (4.56±0.29 and 4.25±0.33mg/gm) conditions respectively. In

sorbate treated superchilled, eviscerated chilled & superchilled conditions, significant

increase (P<0.05-P<0.01) have been observed on 20th day onward to 26th & 28th days

(4.88±0.04 to 5.62±0.37; 5.02±0.36 to 5.55±0.38 and 4.67±0.37 to 5.46±0.33mg/gm

respectively) of storage, followed by a significant decrease (P<0.05) but more than the

normal value on 26th, 28th & 31st day (4.42±0.28, 4.39±0.32 & 4.35±0.30mg/gm) in sorbate

treated super chilled, eviscerated chilled & eviscerated super chilled conditions respectively.

Similarly, during winter season, significant increase (P<0.05 & P<0.01) has been recorded on

15th to 18th day & 18th to 20th day (4.74±0.36 to 5.05±0.37 and 4.40±0.36 to

5.16±0.29mg/gm) in chilled & super chilled conditions, on 20th to 22 & 23rd day (4.70±0.34

to 5.15±0.33 and 4.34±0.32 to 5.19±0.34mg/gm) in sorbate treated chilled and super chilled

conditions and on 24th to 28th & 28th to 30th day (4.50±0.36 to 5.44±0.32 & 4.56 to

5.38±0.37mg/gm) in eviscerated chilled & super chilled conditions respectively, whereas a

gradual decrease thereafter have been noticed, but those were significantly (P<0.05) more

than that of the normal value as it was recorded on 20th day in chilled (4.68±0.42mg/gm), on

24th day in superchilled (4.04±0.30mg/gm) & sorbate treated chilled (4.77±0.36mg/gm), on

28th, 31st & 34th days (4.50±0.27, 4.45± 0.32 & 4.62±0.29mg/gm) in sorbate treated

superchilled, eviscerated chilled & superchilled conditions respectively. Thus the data

indicates that the decrease in fat content with an increase in free fatty acid have some

relationship in between them. The increase in free fatty acid content might be due to

hydrolysis of fat/lipids due to low temperature.



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Figure 6: Graph showing total free fatty acid in the muscle of Cirrhinus mrigala at selected

days of different storage conditions during summer and winter seasons.

Total Plate count (TPC): The total plate count in the muscle+skin (with skin) in fresh fish

during summer & winter seasons has been recorded to be 5.38x103 & 3.94x103cfu/gm

respectively. In both seasons, a gradual decrease in TPC has been recorded during initial days

of different storage conditions, which after a considerable period gradually increased with

maximum on the flag days of the storages. During summer season, maximum decrease has

been recorded on 5th day in chilled (5.12x103cfu/gm), superchilled (4.69x103cfu/gm), sorbate

treated chilled & superchilled (3.75x103 & 4.10x103cfu/gm) and in eviscerated chilled and

super chilled (3.70 x 103 & 3.35x 103 cfu/gm) conditions, whereas, maximum increase has

been recorded on 18th day in chilled (6.76x106cfu/gm), on 22nd day in superchilled & sorbate

treated chilled (2.32x107 & 1.23x107cfu/gm), on 26th,28th & 31st day in sorbate treated

superchilled, eviscerated chilled & superchilled (2.72x107, 2.20x107 & 1.96x107cfu)

conditions respectively. Similarly, during winter season also maximum decrease has been

observed on 5th day (3.18x103, 3.10x103, 3.70x103, 3.60x103, 3.54x103 and 3.30x103)

followed by a gradual increase up to 20,24,24,28,31 & 34 days in chilled, superchilled,

sorbate treated chilled and superchilled, and eviscerated chilled & superchilled (5.09x106,

7.84x 106, 2.08x107, 1.68x107, 2.33x107 and 1.46x107cfu/gm respectively) conditions. The

initial decrease observed in different storage conditions may be due to leaching & cold shock



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due to which the number of mesophilic bacteria decreased and during later stages, due to

increase in cold loving psychrophilic bacteria, the TPC increased and the maximum increase

on the boarder day/flag days might be related to the spoilage of fish (Figure- 7)..

Figure 7: Graph showing average changes in total plate count (cfu/gm) in log value in the

muscle of Cirrhinus mrigala at selected days of different storage conditions during summer

and winter seasons

Discussion

Proteins are basically complex nitrogenous organic substances of very high molecular weight

and are directly involved in muscular contractions during postmortem changes. It is an

important constituent of fish muscle and varies from 18 to 25% except in exceptional cases

such as Bombay duck, in which it is about 9% only. Power et al (1969) have noted a decrease

in extractable protein nitrogen & free fatty acid contents in the fillets from mechanically

superchilled whole cod. Rao (1998) studied changes in L. rohita actomycin during frozen

storage & opined protein coagulation. Kumar (2002) observed a gradual decrease in total



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protein content in the muscle of C. catla & C. mrigala in chilled & superchilled storage

conditions. In the present study, the total protein content in fresh fish has been recorded to be

18.38±0.10 & 18.95±0.12% during summer & winter months. Thus, the protein content has

been recorded significantly (P<0.05) higher during winter than the summer months. Furhter,

the gradual decrease in the total protein contents has been observed in the different storage

conditions during both seasons & the decreases were found statistically significant

(P<0.05/P<0.01) on 18th day (17.50%) & on 20-22 days (17.83% & 17.54%) in chilled &

superchilled, on 20th & 22nd day (17.81 & 17.52%) & on 22nd & 24-26th day (17.76 &

17.45%) in sorbate treated chilled & superchilled and on 26th & 28th day (17.78 & 17.48%) &

28th and 31st day (17.61 & 17.45%) in eviscerated chilled & superchilled conditions

respectively during summer months, whereas, significant decline during winter months has

been observed on 18th & 20th day (18.25 & 18.00%) in chilled, on 22nd & 24th day (18.26 &

18.12%) in superchilled as well as (18.20 & 18.06%) in sorbate treated chilled, on 26th & 28th

day (18.26 & 18.06%) in sorbate treated superchilled and on 28th & 30-31st day (18.18 &

17.84%) in eviscerated chilled and on 31st & 33-34th day (18.35 & 18.06%) in eviscerated

superchilled conditions respectively. The decrease observed may be due to the breakdown of

protein into aminoacids and taking active role in muscular contractions during rigor mortis

stage of the fish. In fish muscle, the non-protein nitrogen compounds normally comprises

ammonia, trimethylammonium bases, guanidine & imidazol derivatives apart urea, amino

acids, purines & pyrimidines. These influence the flavor & keeping qualities of the fish,

however in freshwater fishes, trimethyl amine oxide is not present. Lerke et al ( 1967), have

observed an initial decrease in NPN content in the muscle of different fish species in different

storage conditions, followed by a slight increase & thereafter again decrease and stated that

initial fall was probably due to consumption of NPN fractions by bacteria & afterwards due

to breakdown of protein. The slow rate of increase & further decrease may be due to constant

leaching effect of the ice-melt water, Perigreen et al (1987) in Channa striatus & Joseph et al

(1980) in L. rohita stored in ice, observed a decrease in NPN value depended on storage

period. In the present study the NPN (mg/100gm) content in fresh C. mrigala has been

recorded to be 314±13 & 282±11 mg/100gm during summer & winter months. In both

seasons, the NPN contents have been recorded comparatively decrease for few days of

storage, followed by an increase and thereafter a decline in later stages of storage dependent

on storage condition. Though, the decreases follow by an increases were not found

statistically significant when compared with their respective normal (fresh) conditions, but

the decrease became statistically significant at flag days of different storage conditions as

these were recorded to be 250±7 & 252±9 mg/100gm on 18th & 22nd day in chilled &

superchilled conditions, on 22nd & 24-26day (246±8 & 235-220±10 mg/100gm) in sorbate

treated chilled & superchilled and on 26-28th & 28-31 day (244& 213±9 & 246 & 210±11

mg/100gm) in eviscerated chilled & superchilled conditions during summer, while on 20th &

22-24th day (212±11 & 240-221±8 mg/100gm) in chilled & superchilled, on 24th & 26-28th

day (232±10 & 225-210±8 mg/100gm) in sorbate treated chilled & superchilled and on 28th -

30th day & 31st to 34th day (232-202±11&228-200±9 mg/100gm) in eviscerated chilled &

superchilled conditions respectively during winter months. Thus, the initial decreases may be

due to consumption of NPN fraction by bacteria and/or due to breakdown of protein followed



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by increase & again significant decrease may be due to constant leaching effect of ice melt

water.

Kannaiyan (2014) has stated that attractive flavours in Prawn & other crustacean is

due to the presence of comparatively higher amount of free α- amino acids in them as the

rapid falls occurring in these compounds in iced prawn are accompanied by parallel fall in

their characteristics flavor. The comparatively quicker rates of spoilage occurring in

invertebrates than that of teleosts may be attributed to the presence of larger quantity of α-

amino acids in their muscle which provides excellent substrate for rapid proliferation of

spoilage micro-organisms especially during early stages of spoilage. Bacterial breakdown of

these compounds in to lower volatile fatty acids & ammonia, generally serves as indices of

spoilage. Further, Lerke et al (1967) have stated that a gradual increase in α-amino nitrogen

showed the extent of protein degradation by enzymes & bacteria during storage period. The

increase may be attributed to stepwise breakdown of protein as Protein – Polypeptide – Free

amino acid. In the present study, the AAN content in fresh fish has been recorded

comparatively less during summer (46.48±1.47 mg/100gm) than the winter

(55.90±1.56mg/100gm) months. A significant initial increase in AAN content has been

recorded up to certain days, followed by a decline, which were recorded statistically

significant during flag days depended on storage condition both in summer & winter months.

As a significant increase upto 53.94±1.39 & 54.52±1.26mg/100gm during summer and

62.48±1.30 62.54±1.40 mg/100gm during winter on 5th & 10th day in chilled & superchilled

storage conditions; 53.80±1.22 & 52.25mg/100gm on 5th & 10th day during summer and

60.75±1.35 & 60.75±1.35 & 62.67±1.28mg/100gm on 10th & 15th day during winter in

sorbate treated chilled & superchilled conditions and on 15th & 18 days (55.94±1.22 &

54.68±1.37mg/100gm) during summer & on 15th & 18th day (63.00±1.40 &

63.90±1.30mg/100gm) during winter months in eviscerated chilled & superchilled conditions

respectively, whereas significant decreased valves were obtained on 18th & 22nd day onwards

with maximum 35.98±1.26 & 39.65±1.42mg/100gm during summer & 18th & 24th day

(4078±1.26 & 41.50±1.22mg/100gm) during winter in chilled & superchilled conditions, on

20th & 26th day (38.97±1.39 33.10±1.20mg/100gm) in summer & 24th & 28th day

(42.06±1.27 & 38.24±1.32mg/100gm) in winter in sorbate treated chilled & superchilled

conditions and on 28th & 31st day (36.80±1.41 & 31.36±1.24mg/100gm) during summer & on

26 to 31st & 30th to 34th day (39.00±1.31 & 34.54±1.27mg/100gm) during winter in

eviscerated chilled & superchilled storage conditions respectively. Hence, the present finding

is contrary to the observation made by Bandopadhyaya et al (1986) but in aggrement with the

findings of Kumar (2005) in Labeo rohita.

It is well known that total volatile nitrogenous compounds impart undesirable flavour

to the product while generally free α- aminonitrogen impart desirable one. TVBN is normally

below 20mg% in fishes, molluscs & crustacean an is mainly constituted by ammonia in the

fresh muscle produced by deamination of muscle adenylic acid and by process leading to

denaturation of muscle protein. In spoilage fish, the TVBN are produced by putrefactive

processes and are determined as measure of the extent of spoilage. As these compouds are

highly soluble in water, the storage of fish in ice interferes with their application in quality



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control in indian environment, where melting rate of ice is high. Further, Mahanta &

Muzaddadi (2013) in Shidal (salt free semi-fermented traditional fish product using minor

carps) observed significant increase in NPN, AAN & TVBN compounds at the end of storage

period indicating hydrolysis & degradation of protein. In the present study, the TVBN

content in the muscle of fresh fish has been recorded to be 10.02±0.44 & 7.95±0.30

mg/100gm during summer & winter months respectively, indicating that the TVBN contents

in the fish muscle are comparatively more during summer than the winter months. A gradual

insignificant increase has been noticed during initial days, followed by a gradual decline

which were found statistically significant in flag days of different storage conditions during

both summer & winter months as these were recorded maximum increase upto 10th day in

iced & superchilled condition (11.10 & 10.96mg/100gm during winter & 8.48 &

8.56mg/100gm during winter months respectively, on 10th & 15th days in summer (11.05 &

10.45mg/100gm) and on 10th day in winter (8.54 & 8.42mg/100gm) in sorbate treated chilled

& superchilled condition and on 10th & 15th day in summer (11.04 & 11.12mg/100gm) & on

15th day in winter (8.38 & 8.45mg/100gm) in eviscerated chilled & superchilled condtions

respectively. Thereafter it decrease gradually and the declines were found statistically

significant with maximum on 18th day (7.40±0.30mg/100gm) in chilled, 22nd day

(7.60±0.27mg/100gm) in superchilled & 7.55±0.30mg/100gm in sorbate treated chilled, on

26th day (7.56±0.26mg/100gm) in sorbate treated superchilled and on 28th & 31st day

(7.64±0.22 & 7.56±0.27mg/100gm) in eviscerated chilled & superchilled conditions

respectively during summer months whereas, during winter months it was recorded

maximum significant decline on 20th & 24th day (6.35±0.21 & 6.05±0.25mg/100gm) in

chilled and super chilled, on 24th & 28th day (5.90±0.31 & 5.90±0.36 mg/100gm) in sorbate

treated iced & superchilled and on 31st & 34th day (6.14±0.20 & 6.02±0.25mg/100gm) in

eviscerated chilled & superchilled conditions respectively. Thus, the intial increase up to

certain days followed by decline during flag days in different storage conditions observed,

suggests that the spoilage pattern & bacterial flora associated with spoilage is entirely

different from that of marine fishes which generally showed an increase in TVBN values

during different storage condition. Hence, the present finding is almost similar as suggested

by Joseph et al (1988) & Kumar (2002).

Fish fats have high content of polyunsaturated long chain fatty acids with 4-6 double

bonds, which is not common in mammalian fats. As far fatty acid in seawater & freshwater

fishes are concerned, a noteworthy difference is that the former have abundant in C18 C20 &

C22 acids with varying degree of unsaturation while in latter, fatty acids carry predominantly

C16 & C18 atoms. A high level of unsaturation of the fish oil renders susceptibility to easy

oxidative rancidity. Bandopadhyaya et al (1985) in some fresh water fishes of Hirakund

reservoir observed a gradual increase in free fatty acid contents in the muscle during ice

storage conditions. In the present study, the fat (Total lipid) content in the muscle of the fish

has been recorded significantly (P<0.05) high during winter (1.09±0.05%) than the summer

months (0.93±0.02%). A gradual decrease has been observed in all different storage

conditions during both summer & winter months and the decline became statistically

significant on 15th day with maximum (0.64±0.03%) on 18th day; 18th day with maximum on

22nd day (0.65±0.04%) in chilled & superchilled; on 20th day with maximum (0.69±0.03) on



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22nd day & 24th day with maximum on 26th day (0.68±0.04%) in sorbate treated chilled &

superchilled and on 26th day with maximum (0.63±0.03%) on 28th day & on 28th day with

maximum (0.60±0.04%) on 31st day in eviscerated chilled & superchilled conditions during

summer months, whereas, during winter months maximum decreases were observed on 18-20

days (0.70±0.05) & on 20-24 day (0.71±0.04%) in chilled & superchilled; on 20-24th day

(0.74±0.05) & 24-28th day (0.69±0.03%) in sorbate treated chilled & superchilled and on 28-

31st day (0.66±0.05%) & on 31-34th day (0.68±0.05%) in eviscerated chilled & superchilled

conditions. The FFA (Free fatty acid) contents in the fresh fish have been recorded to be

3.10±0.33 & 2.72±0.29mg/100gm during summer & winter months, i.e. contrary to total fat

content, which was more during winter than the summer months. Similarly the FFA content

in the fish muscle in different storage conditions were recorded a gradual increase depended

on storage conditions as the increases were found statistically significant during flag days of

respective storage conditions. The maximum increases were observed on 18th day

(4.45mg/gm) & 22nd day (4.56±0.29mg/gm) in chilled & superchilled, on 22nd day

(4.25±0.33mg/gm) & 26th day (4.42±0.28mg/gm) in sorbate treated chilled & superchilled

and on 28th day (4.39±0.32mg/gm) & 31st day (4.35±0.30mg/gm) in eviscerated chilled &

superchilled conditions, during summer months, whereas, during winter months, maximum

increases were recorded on 20th & 24th day (4.68±0.42 & 4.04±0.30mg/gm) in chilled &

superchilled, on 24th & 28th day (4.77±0.36 & 4.50±0.27mg/gm) in sorbate treated chilled &

superchilled and on 31st & 34th day (4.45±0.32 & 4.62±0.29mg/gm) in eviscerated chilled &

superchilled conditions respectively. Almost all sample stored in superchilled conditions

showed increase percentage of lipid hydrolyzed to free fatty acids during storage period.

Lovern (1961) has reported that maximum rate of hydrolysis of lipids to free fatty acids takes

place between -2.4°C to -10°C. Castell et al(1966) have reported a relationship between the

production of free fatty acids in the muscle & the resistance to oxidative rancidy of the lipids.

Hence, the decrease in fat content with an increase in FFA indicates a relationship between

the two parameters & the increase in FFA may be due to hydrolysis of fat/total lipids due to

low temperature.

Bacteria, a unicellular microscopic organism invariably occurring in nature (in both

animals & plants, terrestrial & aquatic environment) are responsible for decay of dead

animals & spoilage of food substances apart causing diseases in animals & plants including

human beings. The bacteria are divided in three groups: spherical, rod-shaped & spiral

ranging from 1-3micron in rods & 0.5 micron in diameter in spherical one. They are either

Gram(+), retaining violet colouration of crystal violet stain or Gram(-), which gives away the

violet colouration after iodine & ethanol treatment. According to optimum temperature of

growth, they may be devided in psychrophilic or cold loving (0°-20° range), mesophilic (20°-

40°C range) and thermophilic or loving (45°-90° range).

Nair et al (1971) reported that fish stored in refrigerated sea water (RSW) treated with Co2,

inhibited the growth of bacteria & increased storage life of rockfish & shrimps by one weak.

Garg & Stephan (1982) in Katifish stored in ice, observed a fall in bacterial count during

initial stage, which might be due to washing out of surface slime and a gradual rise in

bacterial number after 5 days, might be due to proliferation of psychrophilic bacterial flora.



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An initial decrease in total plate count in early stages of ice storage has been reported by

several workers in different fish species like Ramchandran et al (1990). Joseph et al (1980) in

Labeo rohita observed an increase in total bacterial count with the day of storage at room

temperature. The maximum recommended bacterial counts for marginally acceptable quality

fish is 107g-1. Fish flesh containing 108 bacteria g-1 is considered unsuitable for human

consumption. About 80 to 90% of total bacterial population is constituted by mesophils, but

during ice storage, these mesophils gradually decreaed as it was only 1-2% by 13th day of ice

storage. Kokane et al. (2010) have studied the Total viable bacteria in some fishes from

Kolkata fish market and reported the counts were 5.00-6.00 log cfug-1 in Cirrhinus mrigala,

6.00-7.00 log cfug-1 in Labeo rohita & Catla catla. In the present study, the TPC in the

muscles of fresh fish during summer & winter months has been recorded to be

5.38x103&3.94x103cfu/gm respectively. The values have been found more during summer

than the winter months. The total plate count has been recorded less than the normal fresh

conditions in the fishes stored in different storage conditions for a certain period varying

between 5-10 days storage period and thereafter a gradual increases were observed in both

seasons depended on storage conditions. The incerases were up to 6.76x106 & 2.32x107

cfu/gm on 18th & 22nd day in chilled & superchilled;1.23x107 & 2.72x103 cfu/gm on 22nd &

26th day in sorbate treated chilled & superchilled, on 28th & 31st day (2.20x107 & 1.96x107

cfu/gm) in eviscerated chilled & superchilled conditions during summer and on 20th & 24th

day (5.09x106 & 7.84x106 cfu/gm) in chilled & superchilled, on 24th & 28th day (2.08x107 &

1.68x107 cfu/gm) in sorbate treated chilled & superchilled and on 31st &34th day (2.33x107 &

1.46x107 cfu/gm) in eviscerated chilled & superchilled conditions respectively during winter

months. The initial decrease in TPC observed, may be due to leaching & cold shocks due to

which, the number of mespohilic bacteria decreased but thereafter due to increase in

psychophilic bacteria, the TPC number increased & reached to maximum number which

caused spoilage. Comparatively, lower count of bacteria on different days in sorbate treated

chilled and super chilled conditions than the corresponding values in normal ice conditions,

may be due to antimicrobial action of sorbate compound. Huss et al. (1995) has reported that

the slower spoilage of some fish species has been attributed to the slower bacterial growth &

different spoilage rate might be partly related to the rate of increase of bacteria on them.

Similarly comparatively less number of bacterial count in eviscerated storage conditions

might be removal of gills & viscera of the fish prior to storing them as these organs contain

maximum bacteria.

Reference

Bandopadhyay, J.K., Chattopadhyay, A.K. & Bhattacharya, S.K. 1985 : “Harvest & Post

harvest technology of fish”. (Eds. Ravindran, K., Unnikrishnan, N.N.; Perigreen, P.A.;

Madhavan, P.; Gopal Krishna, P.A.G.; Panicker, P.A. & Thomas, M.), Soc. of Fish

Technologists, India, Cochin. 1986: On ice storage

charecteristics of Catla catla & Labeo fimbriatus. Fish Technol. 23 : 140-142.

Garg, D. K. & Stephen, J. 1982 : Ice storage studies of Kati fish (Pallonia spp). Fish

Technol., 19 : 45-47.



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Huss, H. H. 1995 : Quality & quality changes in fresh fish F.A.O. Tech. Paper No. 348.

Rome pp. 65.

Joseph, J.; Perigreen, P. A., George, C. & Govindan, T. K. 1980 : studies on ice

storage of cultured rohu (L. rohita). Fish Technol. 25(2) : 105-109.

Kannaiyan, S. K.; Annamalai, J.; Kannuchamy, N & Gudipati, V. 2014 : Self life

extension of Tuna fillets using using natural preservatives isolated from garlic. Fish.

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Kokane, M. R. 2000 : Studies on enhancement of shelf life of fish preserved in ice

containing permitted bactericidal chemicals. M. F. Sc. Dissertation , C. I. F. E, I. C. A.

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