Biochemical, Textural and Microbiological Quality of Squid Stored Under Conventional and Slurry Ice During Onboard Fishing

  • L. Narasimha Murthy
  • Girija G. Phadke
  • A. Jeyakumari
  • U. Parvathy
  • S. Visnuvinayagam
Research Article

Abstract

Careful handling of fish and shellfish onboard at the vessel is of prime importance in order to maintain the freshness. Use of slurry ice for onboard fish storage allows rapid heat transfer as well as reduces damage to the product by conventional ice. In the present investigation, the effect of onboard chilling with conventional ice and slurry ice on the quality of squid was evaluated. Samples stored at different time intervals (7th and 11th day) were collected and analyzed for their biochemical, textural and microbiological quality characteristics. Use of slurry ice, immediately after catch of squid reduced the formation of nitrogenous substances as well as the rate of pH increase during storage. Total volatile base nitrogen, tri methyl amine nitrogen and thio-barbituric acid values were within the acceptable limit in both the samples. Texture analysis showed higher hardness, springiness, gumminess and chewiness values for squid samples stored under slurry ice than the samples stored under conventional ice. Further, squid samples stored under slurry ice exhibited higher values of salt content. Microbiological analysis showed significant difference (p < 0.05) in psychrophilic counts of squid stored under slurry ice than the conventional iced ones.

Keywords

Squid Chilling Slurry ice Biochemical quality Microbiological quality Texture 

Introduction

Fish is highly perishable in nature and the qualities of fish deteriorate rapidly after the death [1]. The quality changes are dependent on various intrinsic and extrinsic parameters such as bacteriological, chemical and physical processes leading to spoilage. So, careful handling of fish and shellfish onboard at the vessel is of prime importance in order to maintain the freshness. Fish quality maintenance begins from catching and keeping the catch onboard the vessel. One of the major factors to keep the fish in fresh condition is to bring its temperature down to 0 °C during onboard storage conditions to get the prime quality and high commercial value for fish. Commercially, ice is used as chilling medium which allows degradation mechanism at slower rates [2]. The type of ice used in any particular fishery will generally depend on availability in particular locality. In general in shore side sub-cooled conventional ice or block ice are available for chilling.

Introduction of slurry ice for onboard storage of fish ensures the rapid and even heat transfer thereby preserving the freshness of fish. The two major characteristics of slurry ice are, a) its faster chilling rate, which is a consequence of its higher heat-exchange capacity, and b) the reduced physical damage caused to food products by its microscopic spherical particles as compared with the damage elicited by conventional ice. When technical advantages are considered, it is possible to pump the slurry ice mixture thus making fish handling more hygienic [3]. The application of slurry ice systems—consisting of ice water suspensions prepared from marine water and chilled at subzero temperatures to the storage of aquatic food products is receiving increasing attention [4]. Better quality of finfish during onboard storage under slurry ice conditions when compared with other traditional icing methods was reported [5]. Moreover, a huge quantity of normal ice has to be carried onboard i.e., double the amount of the fish catch is needed due to melting of the ice. But, the slurry ice can be prepared whenever the requirement/catch are available. In addition, better mileage of vessel is possible due to less weight of ice than conventional vessel.

Cephalopods are considered as seafood delicacy in foreign market and are important in international trade. Cuttlefish and squid have a major contribution in the marine products and are exported in frozen form from India. Squid belonging to the genus, Loligo is generally regarded as more valuable seafood item [6]. Hence, the maintenance of onboard quality is important in order to get higher export values. In the present investigation, squids stored onboard at the fishing vessel under conventional ice and slurry ice at different time intervals (7th and 11th day) were analyzed for physical, chemical and microbiological quality parameters.

Material and Methods

Refrigeration Systems

Slurry ice was prepared using ICEFLOW (Chirag Ice Factory Pvt. Ltd., Navi Mumbai, India) machine which was installed onboard at fishing vessel. The slurry ice machine comprised of BOCK F4 compressor with 1800 RPM wherein R-22 was used as refrigerant. The capacity of the machine is 5–6 tons per day and its efficiency is 2500 L/h. The slurry ice was prepared from filtered seawater (salinity: 3.5%). The temperature of the slurry ice mixture was −2 °C. In addition to the slurry ice machine, block ice was carried onboard and crushed using ice crusher for icing the squid.

Materials

The fishing trip was for 18 days and squids (Loligo species) were caught on 7th and 11th day of fishing trip and the caught squids were stored onboard at the fishing vessel in conventional ice and slurry ice for a period of 11 and 7 days, respectively. The squid samples were collected from the vessel on landing and carried to laboratory for further analysis. Squid had an average weight of 42.36 g and length of 9.37 cm. All the chemicals and glasswares used were of analytical grade.

Chemical Analysis

Proximate composition of the squid stored under conventional ice as well as slurry ice conditions was determined by AOAC method [7]. The pH of homogenate was determined by using a glass electrode digital pH meter. Tri methyl amine nitrogen (TMA-N) and total volatile base nitrogen (TVB-N) content was estimated by the Conway micro diffusion method [8]. Alpha amino nitrogen (AAN) content of the sample was determined from the TCA extract of the samples according to the standard method of Pope and Stevens [9]. The non-protein nitrogen (NPN) content of meat was analyzed according to the described method of Velankar and Govindan [10] and expressed as mg N/100 g meat. Salt soluble nitrogen (SSN) was estimated as per the method of Ironside and Love [11].

Textural Analysis

Squid samples were subjected to instrumental texture profile analysis using TVT6700 (Perten Instruments of Australia Pty Limited, NSW, Australia). A cylindrical probe with 20 mm diameter equipped with 50 N load cell was used to assess the texture of squid samples in uniform shape. Two cycle compression test was performed with initial speed of 1 mm/s, test speed of 1 mm/s and post-test speed of 10 mm/s using 50% compression. The values were generated using TexCalc software attached to the instrument as a result of generated curve for deformation of sample due to the load. Hardness (N), cohesiveness, springiness (mm), chewiness (kgf.mm) and gumminess (kgf) of the squid samples were calculated as defined in the texture analyzer user manual.

Microbiological Analysis

Pour plate technique was followed to enumerate the APC using the plate count agar (HiMedia, # M091). Plates were incubated at 35 °C and the colonies were enumerated after 48 h [12]. Streptomycin Thallous Acetate (S.T.A.) Agar (Hi Media: M1299) is supplemented with S.T.A. Selective supplement (Hi Media: FD127) was incubated at 22 °C for 2 days. White or semi-transparent convex colonies appear as masses of woven threads with oxidase negative were enumerated as a Brochothrix thermosphacta. Pseudomonas Agar Base (Hi Media: M085) supplemented with CFC (Hi Media: FD036) was incubated at 20 °C for 5 days. Oxidase positive colonies were enumerated as a Pseudomonas species. H2S producing bacteria Shewanella putrefaciens is considered for the enumeration. Peptone Iron age (PIA) (Hi Media: M440) incubated at 20 °C for 5 days, then the black colour colonies were considered as H2S producing bacteria [13]. Lactobacillus MRS agar (Hi Media: M440) is prepared and the required amount of diluted sample is spread over the surface, after the drying again the same medium to overlaid on the plate surface and incubated at 37 °C for three days in anaerobic Jar with 5% CO2. Pure white colonies were enumerated as a Lactobacillus species. Tributyrin Agar Base (Hi Media: M157) added with Tributyrin (FD081) was incubated at 37 °C for 2 days. Colonies with clear halo zone are enumerated as lipolytic bacteria. H2S producing bacteria S. putrefaciens is considered for the enumeration. Peptone Iron agar (PIA) (Hi Media: M440) was supplemented with 1.5 ml of 2% ferrous ammonium sulphate (FAS) and 4 ml of 1% l-cystine hydrochloride. All plates were incubated at 20 °C for 5 days. Black colour colonies are considered as H2S producing bacteria [13].

Statistical Analysis

All the analyses were done in triplicate (n = 3). The data was subjected to ANOVA by statistical software, SPSS version 16.0 (SPSS, Inc., Chicago, IL, USA). Students t test was used to compare the means.

Results and Discussion

Proximate Composition

The moisture content of squid samples stored under conventional and slurry ice conditions ranged from 82.75 to 84.55% respectively. Protein content ranged from 10.33 to 12.79%. Fat content didn’t show significant difference; whereas, ash content of squid samples stored under slurry ice condition was higher than the conventional ice stored squid samples on 11th day of storage. Expressible moisture content of squid samples stored for 11th day showed higher value than squid samples stored for 7th day in both chilled system (Table 1). Slurry ice samples exhibited higher ash content than conventional ice stored squid samples. The results were well comparable with the literature with slight variations in protein content and ash content [6, 14]. There was little variation in the constituents which may be attributed to fish-to-fish variations and not to storage conditions [15].
Table 1

Proximate composition of squid stored under conventional and slurry ice conditions

Sample/parameters

Conventional iced squid

Slurry iced squid

A

B

A

B

Moisture (%)

83.67 ± 0.03b

84.20 ± 0.03c

82.75 ± 0.12a

84.55 ± 0.00c

Crude protein (%)

12.79 ± 0.07c

12.67 ± 0.13c

11.93 ± 0.14b

10.33 ± 0.06a

Crude fat (%)

0.51 ± 0.02a

0.50 ± 0.00a

0.52 ± 0.02a

0.48 ± 0.13a

Ash (%)

0.82 ± 0.02b

0.40 ± 0.05a

3.01 ± 0.03c

2.94 ± 0.01c

Expressible moisture (%)

12.61 ± 0.45b

21.42 ± 3.04d

11.26 ± 0.92a

15.26 ± 0.52c

Results are expressed as mean ± standard deviation; N = 3, Different lower case letters indicate significant difference between the different samples for the same parameter (p < 0.05)

A, 7th day of storage; B, 11th day of storage

Changes in Biochemical Quality Indices

Changes in biochemical quality indices (pH, TVB-N, TMA-N and TBA) of squid samples are given in Table 2. The pH of squid samples stored under slurry ice was lower (6.77 and 6.85) when compared with conventional ice stored squid samples (7.13–7.18). Several researchers reported that the pH increase in conventional ice samples may be observed due to induction of ammoniacal compounds owing to a significant growth of alkalinizing bacteria [16, 17, 18]. Live fish muscle pH is close to the value of 7.0. However, seasonal variations may occur [19]. Moreover, TVB-N content of squid samples stored under slurry ice was lower as compared to squid samples stored under conventional ice indicating maintenance of freshness of slurry ice stored squid samples. Variations in TVB-N values often result from spoilage due to bacteria [20]. The level of 35–40 mg TVB-N/100 g of fish muscle is usually regarded as an acceptable limit for human consumption [21]. In the present investigation, TVB-N values ranged from 4.2 to 7.0 mg %. Results are in agreement with similar results reported for horse mackerel [22]. TMA values were found to be nil in squid sample stored for 7th day and it increased slightly in squid sample stored for 11th days in both chilled system. Recommended limit of TMA-N/100 g is 10–15 mg for human consumption [23]. In the present study, TMA-N and TVB-N values for all the squid samples were within the acceptable range. Oxidative rancidity is one of the major causes of flavor deterioration in fish and shellfish. The TBA index is a measure of secondary lipid oxidation in food products. TBA values of 1–2 mg of MDA/kg are usually regarded as the acceptable limit for human consumption of fish and fishery products [23]. It was observed that TBA values showed lower value and were within in acceptable limit in squids stored both in conventional ice and slurry ice conditions. Lower TBA values were observed in slurry ice stored frozen pelagic fish as compared to flake ice [24]. It can be observed that, slurry ice system onboard fishing vessel reduced the formation of thio- barbituric acid reactive substances in squid.
Table 2

Biochemical quality of squid stored under conventional and slurry ice conditions

Sample/parameters

Conventional iced squid

Slurry iced squid

A

B

A

B

pH

7.13 ± 0.00b

7.18 ± 0.00b

6.77 ± 0.00a

6.85 ± 0.00a

TVB-N (mg %)

6.30 ± 0.00c

7.00 ± 0.00d

5.60 ± 0.00b

4.20 ± 0.49a

TMA (mg %)

0.00 ± 0.00a

1.40 ± 0.00c

0.00 ± 0.00a

0.70 ± 0.00b

TBA (mg malonaldehyde/kg)

0.09 ± 0.01b

0.06 ± 0.01a

0.03 ± 0.03a

0.14 ± 0.04c

Results are expressed as mean ± standard deviation; N = 3, Different lower case letters indicate significant difference between the different samples for the same parameter (p < 0.05)

A, 7th day of storage; B, 11th day of storage

Changes in Other Biochemical Quality Parameters

Changes in nitrogen content, non-protein nitrogen content, salt soluble nitrogen and water soluble nitrogen contents of squid samples are given in Table 3. Salt content of squid samples stored under slurry ice conditions was higher than that of conventional ice stored samples. In slurry ice system, chilling medium comprises NaCl which led to increase in salt content of squid muscle which is higher but the increase in salt concentration is lower than that in refrigerated sea water system [25] or salted fish [26]. The increase in NaCl content in the slurry ice system stored samples of chilled fish has been previously reported [15, 27].
Table 3

Nitrogen, non-protein nitrogen, salt content, salt soluble nitrogen and alpha amino nitrogen content of squid stored under conventional and slurry ice conditions

Sample/parameters

Conventional iced squid

Slurry iced squid

A

B

A

B

Nitrogen content (%)

2.05 ± 0.07b

2.03 ± 0.13b

1.91 ± 0.14b

1.65 ± 0.06a

Non-protein nitrogen content (mg %) (NPN)

350.70 ± 2.47c

311.50 ± 8.91b

312.90 ± 1.48b

266.00 ± 0.00a

Salt (NaCl) content (%)

0.56 ± 0.02a

0.62 ± 0.02a

2.68 ± 0.03b

2.77 ± 0.02b

Salt soluble nitrogen content (g %) (SSN)

17.15 ± 0.64b

16.95 ± 0.35b

20.70 ± 0.85c

12.10 ± 0.14a

Alpha amino nitrogen content (mg %) (AAN)

106.40 ± 0.00b

126.00 ± 0.00c

137.20 ± 0.00d

95.20 ± 0.00a

Results are expressed as mean ± standard deviation; N = 3, Different lower case letters indicate significant difference between the different samples for the same parameter (p < 0.05)

A, 7th day of storage; B, 11th day of storage

Nitrogenous substances of non-protein nature ranged from 266 to 350 mg %. The AAN content was found to be in higher range of 95–137 mg % which may be due to the liberation of amino acids as a result of enzymatic proteolytic degradation and action of microbial enzymes on muscle proteins. Salt soluble nitrogen exhibited decreasing trend with the progression of storage period in both the samples. Significantly lower values of SSN were observed as a function of storage period (p < 0.05).

Changes in Textural Quality Attributes

Changes in textural quality attributes such as hardness, springiness, cohesiveness, gumminess and chewiness are given in Table 4. Hardness refers to peak force during compression [28]. Cohesiveness measures capability of samples to withstand deformation during the compression. Springiness determines recovered height of the food during the time between end of first compression and the start of second compression. Cohesiveness indicates degree of deformation before breaking. Gumminess is the product of hardness and cohesiveness [29]. Hardness, cohesiveness, gumminess and chewiness values of squid samples stored under slurry ice were significantly higher than conventional ice (p < 0.05) with comparable springiness values for both the batches. Hardness and gumminess of slurry iced squid samples increased significantly from day 7 to day 11 whereas, the same showed a decreasing trend in conventional iced squid. More softened muscles in conventional ice squids may have resulted from the biochemical and microbiological reactions in those samples.
Table 4

Texture profile of squid stored under conventional and slurry ice conditions

Sample/parameters

Conventional iced squid

Slurry iced squid

A

B

A

B

Hardness 1 (N)

71.98 ± 7.09b

59.44 ± 12.03a

97.44 ± 4.48c

131.94 ± 10.11d

Hardness 2 (N)

57.55 ± 8.45b

46.02 ± 6.42a

82.50 ± 5.02c

110.84 ± 8.66d

Springiness (mm)

0.79 ± 0.16b

0.71 ± 0.26a

0.79 ± 0.04b

0.83 ± 0.04b

Cohesiveness

0.38 ± 0.06a

0.37 ± 0.07a

0.40 ± 0.03a

0.42 ± 0.02a

Gumminess (kgf)

27.47 ± 4.70b

21.54 ± 1.18a

39.30 ± 3.58c

55.15 ± 6.40d

Chewiness (kgf mm)

21.34 ± 3.99b

15.20 ± 5.19a

31.02 ± 1.99c

45.59 ± 5.31d

Results are expressed as mean ± standard deviation; N = 3, Different lower case letters indicate significant difference between the different samples for the same parameter (p < 0.05)

A, 7th day of storage; B, 11th day of storage

Changes in Microbiological Quality

Changes in microbial quality of squid are given in Table 5. It was observed that except aerobic plate count, there was no significant changes (p > 0.05) in other microbiological parameters for sample stored for 7 days. On seventh day, the conventional ice stored fish’s APC was significantly lesser than the slurry iced ones. In case of 11-day old samples there is a significant difference (p < 0.05) in psychrophilic count (2.3 times higher count) in samples stored under conventional ice conditions when compared with slurry ice stored squid samples. Moreover, results indicated that in all the samples aerobic plate counts (APC) were within the acceptable limit. The results are in agreement with similar studies reported for oil sardine stored under conventional ice and slurry ice and lobster [16, 30]. The present study indicated that during initial days, the conventional ice gave better results in terms of APC; but on further storage, slurry ice proved to be better in controlling total bacterial count. Further, there was no significant difference in H2S producing bacteria, Pseudomonas, B. thermosphacta and Lactobacillus counts of squid samples stored under conventional and slurry ice for 7 days. However, samples stored for 11 days exhibited considerable Lactobacillus count when compared with 7 days stored samples.
Table 5

Microbiological quality of squid stored under conventional and slurry ice conditions on seventh and eleventh day

Sample/parameters

7 days stored squid

11 days stored squid

Conventional iced squid (log10)

Slurry iced squid (log10)

Conventional iced squid (log10)

Slurry iced squid (log10)

Aerobic plate count (APC)

2.30 ± 2.15a

3.27 ± 3.08b

3.59 ± 2.15a

3.08 ± 2.15a

Psychrophilic count

4.15 ± 3.93a

3.99 ± 2.45a

5.33 ± 5.18a

3.62 ± 2.45b

Enterobacteriaceae

0.00 ± 0.00

0.00 ± 0.00

0.00 ± 0.00

0.00 ± 0.00

H2S forming bacteria

3.54 ± 2.90a

3.20 ± 0.00a

4.02 ± 0.00a

2.56 ± 1.75a

Pseudomonas sp.

4.53 ± 0.00a

4.67 ± 0.54a

4.20 ± 0.90a

3.02 ± 2.35a

Lactobacillus sp.

0.00 ± 0.00

2.08 ± 0.00a

4.02 ± 3.90a

1.60 ± 0.00a

Brochothrix thermosphacta

2.56 ± 0.00a

1.90 ± 0.00a

1.60 ± 0.00a

2.08 ± 0.00a

Different lower case letters indicate significant difference between the different samples for the same parameter on seventh and eleventh day of storage under conventional and slurry ice (p < 0.05)

Results are expressed as mean ± standard deviation; N = 3

Conclusion

Due to perishable nature of fish, it is necessary to maintain the freshness of the fish by means of preservation of fish using ice onboard at the fishing vessel. Use of conventional block type of ice is practiced onboard at the fishing vessel, in general. Slurry ice system lowers the temperature rapidly as a result of quick chilling compared to conventional ice. Application of slurry ice may be beneficial when food quality and safety are concerned which will allow automation and hygienic handling of seafood onboard. Quality of squid in terms of aerobic plate count, H2S forming bacteria, and psychrophilic count, textural attributes, pH and TVB-N were slightly beneficial compared to conventional ice. However, the use of slurry ice storage system resulted in higher salt uptake compared to conventional ice stored squid samples. From the study, it can be concluded that usage of slurry ice onboard will reduce the rate of microbiological and biochemical degradation mechanisms compared to conventional ice system and maintain the quality of squid as preferred by consumer.

Notes

Acknowledgements

The authors express sincere thanks to Dr. C. N. Ravishankar, Director, ICAR-CIFT, Cochin for the encouragement and granting permission to publish this research article.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© The National Academy of Sciences, India 2017

Authors and Affiliations

  • L. Narasimha Murthy
    • 1
  • Girija G. Phadke
    • 1
  • A. Jeyakumari
    • 1
  • U. Parvathy
    • 1
  • S. Visnuvinayagam
    • 1
  1. 1.Mumbai Research Center of Central Institute of Fisheries TechnologyCIDCONavi MumbaiIndia

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