Peer Reviewed Journal Articles (Inter.) (M,F&B)

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A medium for enumeration of bacteria in foods containing swarming Bacillus spp.
(Blackwell Publishing., 1993) Thampuran, Nirmala; Surendran, P.K
Swarming of Bacillus spp. on agar plates may interfere with enumeration or isolation of bacteria in pickled, canned or fried fish products. A new medium containing sodium tripolyphosphate, EDTA disodium salt, dihydrate, sodium desoxycholate, supplemented with minerals and nutrient sources was developed, which was effective in inhibiting swarming of Bacillus spp. This medium polyphosphate-EDTA-desoxycholate agar (PEDA) was compared with tryptone glucose extract agar (Oxoid) for determining total plate count of various semi-conserved fishery products. While promoting discrete colony formation of Bacillus spp., PEDA provided good bacterial recovery. PEDA is recommended for isolation or enumeration of bacteria in [sea foods] dominated by swarming Bacillus spp.
Occurrence and distribution of vibio vulnificus in tropical fish and shellfish from Cochin (India)
(1998) Nirmala Thampuran; Surendran, P.K.
The incidence and distribution of Vibrio vulnificus in marine and brackish-water fish and shellfish from coastal areas of Cochin on the west coast of India were studied. For marine fish collected in very fresh condition from vessels, a level of incidence of 16.6% was noted. Frequency of isolation was greater from the intestine than from the muscle. The greater Most Probable Number (MPN) count determined by the three tube method ranged from 15 to 910 g-1 in the positive samples.
Distribution and ecology of Clostridium botulinum in fish and aquatic environments of atropical region
(Academic Publications, 2000) Lalitha, K.V.; Gopakumar, K.
Growth and toxin production by Clostridium botulinum in fish (Mugil cephalus and shrimp (Penaeus indicus) tissue homogenates stored under vacuum
(2001) Lalitha, K.V.; Gopakumar, K.
Toxin production by Clostridium botulinum types A-E in mullet and shrimp tissue homogenates stored between 4 and 30 C under vacuum for 6 weeks was evaluated. Mullet and shrimp tissue homogenates were inoculated with C. botulinum spores, then packaged under vacuum and held at 4, 10,15 and 30 C. At 30 and 15C storage, growth and toxin production by types A-E resulted in both mullet and shrimp tissue homogenates. Only type E toxin was observed in the homogenates held at 4 and 10 C. The storage temperature and the growth substrate markedly influenced the toxin titres.
Reduction of halophilic bacterial load in solar salt by sun drying
(2007) Prasad, M.M.; Seenayya, G.
Preservation of fish by salt curing is the most popular method. However, the spoilage of cured fish due to red discoloration by halophilic bacterial contamination limits the shelf life. The source of contamination is mainly the salt itself. The present study was undertaken to evaluate a method for reducing the halophilic bacterial load from solar salt. Sun drying of solar salt for 16 days caused a reduction in counts of red halophiles, slime producing halophilic bacteria and Halomonas sp. at the rate of 97.2, 97.2 and 93.5% on ordinary and 97.8, 98.7 and 89.4% on black cemented platforms, respectively
Prevalence of enterotoxigenic Staphylococcus aureus in fishery products and fish processing factory workers
(Elsevier Science, 2007) Simon, S.S.; Sanjeev, S.
Of the 168 fishery products and 87 samples from fish processing factory workers analyzed, 21 (17%) and 54 (62%) were found positive for enterotoxigenic Staphylococcus aureus, respectively. Among the products, incidence was high in frozen peeled un-deveined (PUD) prawns and frozen fish cutlets (33% each) compared to frozen peeled and deveined (PD) prawns and frozen squids (25% and 20%, respectively). 21 out of 51 strains (41%) isolated from products and in 15 out of 54 strains from processing workers (28%) were found to be enterotoxigenic. The enterotoxin type Staphylococcal enterotoxin C (SEC) (57%) was predominated followed by Staphylococcal enterotoxin A (SEA) (43%) in products and Staphylococcal enterotoxin B (SEB) (40%), Staphylococcal enterotoxin C (SEC) (40%) and Staphylococcal enterotoxin A (SEA) (20%) in workers.
Effects of sodium acetate dip treatment and vacuum-packaging on chemical, microbiological, textural and sensory changes of Pearlspot (Etroplus suratensis) during chill storage
(Elsevier, 2007) Manju, S.; Jose, L.; Gopal, T.K.S.; Ravishankar, C.N.; Lalitha, K.V.
The effects of sodium acetate dip treatment, followed by vacuum-packaging, on the shelf life of beheaded, scaled and gutted Pearlspot (Etroplus suratensis) during chill storage were examined. Sodium acetate (2%, w/v) solution was used for the dip treatment. pouches (size: 15 x 22 cm) made of 12p.-polyester laminated with 300 gauge low-density polyethylene were used for packing fish. After packing, all thepacks were iced with flake ice in the ratio (1:1) fish: ice in an insulated box and were kept in a cold room maintained at 0-2 °C. The control and the treated packs were analysed periodically for chemical (pH, TBA, TMA, TVB-N), microbiological (total viable count), textural and sensory characteristics. Changes in Staphylococcus aureus, Enterobacteriacea and Feacal streptococci were determined for fresh fish and for fish samples at the time of sensory rejection. Air packed samples were found to have a shelf life of about 8 days; vacuum-packed samples were found to be acceptable up to 10 days. whereas sodium acetate-treated vacuum packed samples were found to be acceptable up to 15 days. Thus, vacuum-packaging, in combination with sodium acetate, was found to delay the spoilage, thereby significantly extending the shelf life of Pearlspot at refrigeration temperatures
Characterization and phylogenitic analaysis of ecotine biosynthesis genes from bacillus halodurans
(Springer, 2008) Rajan, A.; Joseph, C.T.; Thampuran, N.; James, R.; Viswanathan, C.; Bansal, K.C.
Ectoine, a cyclic tetrahydropyrimidine (2- methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid), is a natural compound, which serves as a protective substance in many bacterial cells. In this study, the putative ectABC gene cluster from Bacillus halodurans was heterologously expressed in E. coli and the production of ectoine was con- Wrmed by HPLC analysis. The activity of the enzymes coded by the ectA, B and C genes were found to be higher in induced transgenic cells compared to the uninduced cells. Phylogenetic analysis revealed sequence identities ranging from 36–73% for ectA gene, 55–81% for ectB gene and 55–80% for ectC gene indicating that the enzymes are evolutionarily well conserved.
Antibiotic susceptibility of staphylococci isolated from labeo rohita sold in burla fish market, orissa
(Asian Fisheries Society, 2009) Sahoo, R.S.; Sahu, J.; Prasad, M.M.
Screening of commercial sold freshwater Labeo rohita revealed that the staphylococcal count decreased from one to two log cycles/g upon washing the fish. The fish samples harboured more number of staphylococci in edible meat portion than in guts. The microscopic, biochemical and oxidative fermentations tests of the isolates by conventional and rapid detection methods revealed that they are Staphylococcus haemolyticus, S. auricularius and S. caseolyticus. When the isolates were compared with 45 different antibiotics the results varied from susceptible to resistant in comparison with American Type Culture Collection cultures. In comparison to Clinical Laboratory Standards Institute isolates both the staphylococcal test cultures were resistant to penicillin (G) and vancomycin. With other antibiotics viz., Pristinamycin, Ticarcillin/Clavulanic acid, Gatifloxacin, Clindamycin, Clarithromycin, Levofloxacin, Linezolid, Cefeprime, Erythromycin, Streptomycin, Fosfomycin, and Piperacillin/Tazobactam the results varied from intermediate to sensitive.
Effect of modified atmosphere packaging on chemical, textural, microbiological and sensory quality of seer fish (Scomberomorus commerson) steaks packaged in thermoformed trays at 0-2C
(Wiley, 2009) Yesudhason, P.; Gopal, T.K.S.; Ravishankar, C.N.; Lalitha, K.V.; Ashok Kumar, K.
The effect of modified atmosphere on chemical, textural, microbiological and sensory quality of seer fish (Scomberomorus commerson) steaks stored at 0-2C for 23 days was studied. Of the chemical indicators of spoilage, significant differences (P < 0.05) in trimethylamine and thiobarbituric acid values were observed between air- and modified atmosphere (MA)-packaged samples. Seer fish under MA was acceptable on the basis of sensory and microbiological parameters even though the K value was more than 95%, indicating that K value was not a suitable quality indicator. Based on the inosine to hypoxanthine observed in this trial, seer fish was classified as hypoxanthine producers. Bacterial counts of air-packaged seer fish were always higher than MA-packaged samples. Based on sensory score and microbiological data, the shelf life of fish steaks packaged in MA 21 days whereas air-stored samples had a shelf life of 12 days only.
Prevalence of molecular characterization of Vibrio cholerae 01, Non-01 and Non-0139 in tropical seafood, Cochin, India
(Mary Ann Libert Inc, 2009) Kumar, R.; Lalitha, K.V.
The objective of this study was to determine the prevalence of 01, 0139, and non-01 and non-0139 Vibrio cholerae, which were associated with fresh and raw seafood samples harvested from Cochin, India waters during 2009-2011. Results from V. cholerae—specific biochemical, molecular, and serological assays identified five El Tor V. cholerae 01 Ogawa strains and 377 non-01, non-0139 V. cholerae strains from 265 seafood samples. V. cholerae 0139 strains were not isolated. Polymerase chain reaction assays confirmed the presence of V. cholerae 01 El Tor biotype in seafood. Antibiotic susceptibility analysis revealed that the V. cholerae 01 strains were pansusceptible to 20 test antibiotics, whereas 26%, 40%, 62%, and 84% of the non-01, non-0139 V. cholerae strains were resistant to cefpodoxirne, ticarcillin, augmentin, and colistin, respectively. Detection of virulence and regulatory genes in V. cholerae associated with seafood revealed the presence of virulence and regulatory genes (i.e., ctx, zot, ace, toxR genes) in V. cholerae 01 strains, nevertheless, presence of ace and toxR genes were detected in non-01, non-0139 in 9.8 and 91% strains, respectively. In conclusion, the presence of pathogenic V. cholerae in seafood harvested from local Cochin waters warrants the introduction of a postharvest seafood monitoring program, which will lead to a greater understanding of the distribution, abundance, and virulence of diverse pathogenic Vibrio populations that inhabit these different coastal regions so that a risk management program can be established.
Virulence genes, serobiotypes and antibiotic resistance profile of Escherichia coli strains isolated from aquaculture and other sources
(Wiley, 2010) Surendraraj, A.; Thampuran, N.; Joseph, T.C.
In order to determine the prevalence of pathogenic Escherichia coli, a total number of 155 E. coli isolates from aquaculture, clinical and veterinary sources were screened for seven pathogenic virulence markers and a house-keeping gene by a polymerase chain reaction. The targeted virulence genes included eaeA of enteropathogenic E. coli, elt and est of enterotoxigenic E. coli (ETEC), ipaH of enteroinvasive E. coli, pCVD432 of enteroaggregative E. coli, stx, hlyA and eaeA of shigatoxigenic E. coli (STEC) and Enterohaemorrhagic E. coli. All the isolates were positive for phoA, the house-keeping gene for E. coli. Among the 155 isolates, seven numbers (4.5%) harboured the virulencemarkers belonging to the pathogenic group ETEC and STEC. The virulent genes detected in these groups were elt, est, hlyAand stx.The sources of these virulence genes were ¢sh (hlyA), shrimp (elt), feeder canal water (hlyA and elt) of aquaculture origin and from diarrhoea a¡ected cow (hlyA, est and stx). The isolates with pathogenic traits belonged to the serogroups O6 or O29 and the remaining could not be typed. They showed resistance to two to four antibiotics out of the 12 antibiotics tested. Biotyping revealed that three isolates belonged to a single biotype (7333) and the remaining isolates were of diverse types. In conclusion, a molecular tool such as PCR proves as more e¡ective tool for detection of this pathogen than the conventional methods. Detection of these emerging pathogens in aquaculture samples warrants for strict adherence to hygienic handling at retail outlets and proper cooking by the consumer before consumption
Isolation and characterization of Listeria monocytogenes from tropical seafood of Kerala, India
(springer, 2013) Das, S.; Lalitha, K.V.; Thampuran, N.; Surendran, P.K.
Listeria monocytogenes, which is an intracellular pathogen, causes various illnesses in human as well as in animals. The pathogenicity of this organism depends upon the presence of different virulence genes. A total of 324 tropical seafood and fishery environmental samples were screened for L. monocytogenes. The incidence of the human pathogenic species L. monocytogenes was 1.2 % of the samples. Listeria spp. was detected in 32.3, 27.1, and 5%of fresh, frozen, and dry fish samples, respectively. Listeria innocua was found to be the most prevalent species of Listeria in the tropical seafood and environmental samples of Kerala. Listeria monocytogenes and L. innocua isolates were confirmed by multiplex PCR. L. monocytogenes isolates from the four positive samples showed phosphatidylinositol-specific phospholipase C reaction on Chromocult® Listeria selective agar. Molecular characterization of L. monocytogenes isolates for virulence genes revealed the presence of β-hemolysin (hly), plcA, actA,metalloprotease (mpl), iap and prfA genes in all the isolates recovered from the positive samples.
Quality characteristics and shelflife of Sutchi cat fish (Pangasiandon Hypophthalmus) steaks during refrigerated storage
(Research India Publications, 2014) Viji, P.; Tanuja, S.; Zynudheen, A.A.; Lalitha, K.V.
Fish is more vulnerable to deterioration than chicken and red meat as it contains relatively larger amounts of free amino acids, a higher water activity and a higher final pH, limiting the shelf life of the product. Hygienic handling practices can control the deterioration of fish quality during storage and distribution. The quality changes and shelf life of iced (0°C) and chill (4°C) stored sutchi catfish (Pangasianodon hypophthalmus) steaks was assessed by evaluating the changes in biochemical, textural, microbiological and sensory attributes during storage. The study revealed that all the biochemical quality indices studied like pH, TVBN, TBA and PV were within the acceptable range for both the treatments throughout the storage period. However, the chill stored steaks showed higher degree of textural deterioration than the ice stored steaks. Total mesophilic and Enterobactereaceae counts were lower in ice stored steaks compared to chill stored steaks. Based on the sensory and microbiological analysis, the shelf life of chilled and ice stored sutchi catfish steaks is determined as 14 and 17 days, respectively, during storage.
Functional and molecular characterization of a lipopeptide surfactant from the marine sponge-associated eubacteria bacillus licheniformis NIOT-AMKV06 of Andaman and Nicobar islands, India
(Elsevier, 2014) Lawrance, A.; Balakrishnan, M.; Joseph, T.C.; Sukumaran, D.P.; Valsalan, V.N.; Gopal, D.; Ramalingam, K.
The production of a lipopeptide surfactant from the sponge-associated eubacteria Bacillus licheniformis NIOT-AMKV06 from the Andaman and Nicobar Islands was investigated. The highest production was attained with glucose and yeast extracts as the carbon and nitrogen sources (1.789 mg mL-1), respectively. The surfactant was highly stable over a pH range of 5.0–10 and a temperature range of 20–70 0C with high NaCl concentrations. Excellent emulsification activity was exhibited by the purified surfactant with crude oil, kerosene, and diesel. A two-fold increase in surfactant production (3.0 mg mL-1) was observed using the newly formulated medium in this study. The surfactant biosynthesis gene cluster (sfp, sfpO, and srfA) from B. licheniformis NIOT-AMKV06 was heterologously expressed in Escherichia coli, and the production was increased three-fold (11.78 gL-1) over the original strain. The results confirm the potential of the surfactant for use in bioremediation of hydrocarbons in a marine environment and for enhanced oil recovery. To our knowledge, this is the first report on the ability of a hydrocarbon degrading B. licheniformis from marine sponges for the biosynthesis of a potent lipopeptide surfactant possessing characteristics of maximum stability, outstanding surfactant activity, and exceptional emulsifying capability.
Draft genome sequence of the halophilic and highly halotolerant gammaproteobacteria strain MFB021
(American Society of Microbiology, 2014) CIFT
Retention of shelf life and microbial quality of seer fish stored in modified atmosphere packaging and sodium acetate pretreatment
(Elsevier, 2014) Yesudhason, P.; Lalitha, K.V.; Gopal, T.K.S.; Ravishankar, C.N.
Preliminary shelf life tests were performed to select the best gas composition and the best sodium acetate concentrations in the modified atmosphere. Bacterial quality assessment was carried out by the monitoring of total aerobic bacteria, H2S producing bacteria, lactic acid bacteria, pseudomonads, Enterobacteriaceae, Staphylococcus aureus, streptococci, sulphite reducing clostridia and botulinum toxin by mouse bioassay. Bacteria grew most quickly in seer fish stored in air, followed by those in MAP and the lowest counts were with MAP pre-treated with sodium acetate. The application of sodium acetate treatment to seer fish steaks resulted in a bacteriostatic effect, contributing to the improvement of the microbiological quality of seer fish steaks. There was less trimethylamine in the modified atmosphere packed samples pretreated with sodium acetate than in air packed samples by the factor of seven, at the end of the shelf life. Modified atmosphere packaging alone increased the shelf life from 8 (air pack) to 22 days; however, addition of sodium acetate further extended the shelf life to 28 days. The results showed that the combined effect of MAP (70 vol.% CO2:30 vol.% O2) and sodium acetate (1%, w/v) is a valuable tool to allow an effective extension of the shelf life of raw fish products.
Development and characterization of an LDPE/chitosan compositeantimicrobial film for chilled fish storage
(Elsevier, 2015) Reesha, K.V.; Panda, S.K.; Bindu, J.; Varghese, T.O.
tAn antimicrobial packaging material was developed by uniformly embedding 1, 3 and 5% chitosan (w/w)in low density polyethylene matrix using maleic anhydride grafted LDPE as a compatible agent. Thematerials were mixed by compounding and blown into monolayer films via blown film extrusion. Thedeveloped films showed good barrier properties against oxygen. Characterization of the composite filmswith Fourier transform infrared spectroscopy revealed that chitosan and LDPE interacted well with eachother. Overall migration showed better release of chitosan adduct from the LDPE matrix which enhancedthe antibacterial properties of the films. The interaction between the LDPE/CS and maleic anhydridegrafted LDPE had a decreasing effect on the tensile strength and heat sealing properties. Investigationon antimicrobial properties of LDPE/CS films showed 85–100% inhibition of Escherichia coli. Efficacy ofLDPE/CS films was evaluated by using them as packaging material for chilled storage of Tilapia (Ore-ochromis mossambicus). Analysis of storage quality indices (peroxide value, free fatty acid, total volatilebase nitrogen and aerobic plate count) revealed good antibacterial property and extension of shelf life ofTilapia in the chitosan incorporated novel composite films compared to virgin LDPE film.
Bacteriophages for aquaculture: Are they beneficial or inimical
(Elsevier, 2015)
World fisheries production is projected at 164 million tonnes in 2020, and a major increase in the quantity of fish produced is expected to originate from aquaculture. Diseases represent a severe threat to aquaculture productivity and natural/organic strategies to combat infectious diseases are gaining new ground to address the issues of antibiotic resistance of bacteria and issues related to antibiotic residues in aquatic food animals. Phage therapy is currently considered as a viable alternative to antibiotics for treatment of bacterial infections in aquaculture systems. A cocktail of lyric phages and a synergistic combination of phages and other antimicrobials are viable options to control bacterial infections and at the same time evade phage resistance. However, lysogenic phages have the ability to transform non-virulent bacterial strains in to virulent strains which may cripple aquatic food production and also threaten food safety. The review focuses on the roles of bacteriophages and their implications for aquatic food production and food safety.
Synthesis of polyaniline hybrid composite: A new and efficient sensor CrossMark for the detection of total volatile basic nitrogen molecules
(Elsevier, 2015) Ashraf, P.M.; Lalitha, K.V.; Edwin, L.
The detection of fish spoilage and adulteration is a challenging problem faced by consumers and traders. A new polyaniline-curcumin-copper-cobalt hybrid composite was synthesized and evaluated for its ability to detect the total volatile basic nitrogen (TVBN) content. The molecule was characterized by spectroscopic, chemical, Fourier transform infrared (FTIR) and atomic force microscope (AFM) methods. The molecule exhibited excellent sensor responses to ammonia and mono-, di-, and trimethyl amines when assessed through visual, spectroscopic and electrochemical methods. The cyclic voltammetric and electrochemical impedance response of the composite to TVBN showed an excellent linear relationship of concentration vs. current and concentration vs. real/imaginary impedance. Dimethyl amine exhibited an unusual cyclic voltammetric response behavior with the current decreasing with increasing concentration. The individual TVBN content can be distinguished by using the electrochemical impedance spectroscopy technique based on the intensity of impedance response. The composite showed distinct color variation with increasing concentration of TVBN from 1 to 200 p.g g-.1 and the finding was supported by the spectrophotometric response. The spatial spa Tr electron cloud from the organic part and partially filled d orbital electron from the metals played an important role in a more rapid sensing response to the polyaniline hybrid composite. The molecule can be a potential candidate for developing strip sensors, electronic devices, quantitative analytical reagent and TVBN odour absorbers.

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