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Antibacterial activity of shrimp chitosan against Escherichia coli, Salmonella, and Pseudomonas aeruginosa isolated from seafoods
(Society of Fisheries Technologists(India), Cochin, 2006) Kumar, R.; Surendran, P.K.; Thankappan, T.K.
Antibacterial activity of shrimp shell based chitosan was determined against three food borne pathogens viz., Escherichia coli, Salmonella, and Pseudomonas aeruginosa isolated from seafoods. Tests were carried out in phosphate buffered saline (PBS) at pH 6.0. Four different concentrations, 0.01%, 0.03%, 0.05% and 0.1% (w/v) of chitosan were used against E.coli and Salmonella, and 0.01%, 0.05%, 0.5% and 1% were against P.aeruginosa. Chitosan showed stronger bactericidal activity against Salmonella compared to E.coli and P.aeruginosa as evidenced by 3 log cfu.ml super(-1) reduction in Salmonella count with 0.01% chitosan concentration. There was complete elimination of 5 log cfu.ml super(-1) cell count of Salmonella within 24 h at 0.1% chitosan level. There was 3 log reduction in E.coli count at 0.01% chitosan, but not completely inhibited even at the maximum used concentration (0.1%). The inhibition concentration against P.aeruginosa was higher level compared to Enterobacteriaceae and it was found that reduction in 5 log cfu.ml super(-1) was achieved only at 72 h of chitosan exposure. Different chitosan concentrations 0.05%, 0.1% and 1% had almost same inhibition pattern for P.aeruginosa at 6h. 18h 36h and 72h of exposure. The effectiveness of chitosan against tested pathogens showed Salmonella was most susceptible towards chitosan followed by E.coli and P.aeruginosa.
Antibiotic Resistance Pattern of Staphylococcus aureus Isolated from Seafood
(Society of Fisheries Technologists(India), 2016) Murugadas, V.; Joseph, T.C.; Lalitha, K.V.
Staphylococcus aureus is a pathogen known to cause hospital, community acquired and foodborne illness with varying susceptibility to antibiotics. The selective pressure due to the use of antibiotics along with horizontal and vertical gene transfer has resulted in the evolution of multidrug resistant S. aureus. The purpose of the study was to determine the antibiotic resistance pattern in S. aureus isolated from seafood of Kerala, India. One hundred and thirty three S. aureus isolates obtained during 2012 to 2015 were included in this study. Antibiotic susceptibility testing was performed with a set of 20 antibiotics representing eleven classes of antibiotics by standard disk diffusion assay. The study revealed that 90.9% of the S. aureus isolates were resistant to at least one class of antibiotics. Resistance was found among 33.8, 27.8, 17.3, 6.8, 3.8, 0.7 and 0.7% isolates to one, two, three, four, five, six and eight classes of antibiotics respectively. Multidrug resistance was found in 29.3% of the S. aureus isolates with resistance to antibiotics ranging from 3 to 8 classes of antibiotics. The study reveals that S. aureus isolates were sensitive to aminoglycosides and phenicols.
Assessment of microbial quality of fish processing industrial effluent in bar-mouth at Bhidia landing site, Veraval, Gujarat, India
(Triveni Enterprises, 2016) Sivaraman, G.K.; Visnuvinayagam, S.; Jha, A.K.; Renuka, V.; Remya, S.; Deesha, V.
The present study was carried out to assess the microbial quality of fish processing industries effluent at Bhidia bar- mouth, Veraval, Gujarat during April, 2012 to March 2013. The total viable bacterial count (TVBC), total Enterobacteriaceae count, E.coli count (EC), Staphylococcus aureus and Fecal Streptococcal count in effluent ranged from 3.0 x 10 to 6.8 x 10 , 9.0 x 10 to 2.9 x 10 , 0 to 0. 5 x 10 , 0 to 0. 4 x 10 and 0.3 x 10 to 0. 1 x 10 cfu.ml , respectively. Significantly higher load of TEC ,E.coli, S.aureus , Fecal Streptococci, Total coliforms and Fecal coliforms were higher during summer whereas, TVBC was higher in the month of Sept.-Oct. Furthermore, the total coliform and fecal coliform counts were found to be higher with 1400+ /100ml MPN value throughout the year of the study, except in the month of August. Overall occurrence of pathogenic strains of E.coli, S.aureus and Fecal streptococci were 41.67%, 25.00% and 66.67% respectively during this period. The antibiogram of the isolated E.coli isolates show that almost 50% were resistant to Cefazidime/ Clavulanic acid (CAC), Amoxyclav (AMC), Ciprofloxacin (CIF) and Ampicillin (AMP). The present study indicated that the effluent of fish processing industry was heavily contaminated with E.coli, S. aureus and Fecal Streptococci which confirmed improper treatment of fish processing effluent. Moreover, the precedence of antibiotic resistant E.coli may pose threat to public health safety.
Bacterial Flora of EDTA Treated Oil Sardine (Sardinella longiceps), Indian Mackerel (Rastrelliger kanagurta) and Prawn (Mefapenaeus dobsoni) in Ice Storage
(Society of Fisheries Technologists(India), Cochin, 1982) Surendran, P.K.; Gopakumar, K.
The native flora of fresh oil sardine (Sardinella longiceps ) and mackerel (Rastrelliger kanagurta ) consisted mainly of Pseudomonas spp; Moraxella spp.; Acinetobacter spp. and Vibrio spp. During spoilage in ice, nearly 75% of their bacterial flora belonged to Pseudomonas spp. alone. But Na sub(2) EDTA treatment reduced the proportion of Pseudomonas) spp. considerably and the major bacterial groups at the time of spoilage were Moraxella spp. and Acinetobacter spp. In the case of fresh prawn (Metapenaeus dobsoni ) the native flora were constituted by Pseudomonas spp.; Moraxella spp.; Acenetobacter spp. and Vibrio spp. At the time of spoilage of prawn in ice, Moraxella spp. and Acinetobacter spp. predominated, together constituting 74% of the total population. Na sub(2) EDTA treatment did not alter significantly the spoilage flora of prawns. Morazella) spp. and Acinetobacter spp. accounted for 86% of the spoilage flora in ice storage of Na sub(2) EDTA treated prawns.
Bacteriological investigations of prawn canneries - II. Incidence of Clostridium perfringens
(Society of Fisheries Technologis(India), 1973) Narayanan Nambiar, V.; MahadevaIyer, K.
Prawn processing factories of the three major fish processing centres of the West Coast of India, viz., Cochin, Mangalore and Calicut were surveyed to determine the occurrence of Clostridium perfringens in processing areas, and in processed products. Direct plating on Sulphite -polymyxin- sulphadiazine Agar and enrichment techniques were used. Samples of prawn, prawn guts, frozen prawns, canned prawns, water, ice, swab from utensils and soil from the factory premises were examined. Among a total of 461 samples examined, only 32 (6.9%) gave positive results. The incidence of C. pe1jringens was more in prawn guts (80%), followed by soil (50%), prawn (38%), ice (33.3%), frozen prawns (11%), swab (5.0%) and water (1.1%). No C. perfringens was isolated from canned prawns.
Bacteriological quality and heavy metals in edible meat portion of Japanese threadfin bream (Nemipterus japonicus)
(2007) Prasad, M.M.; Imam Khasim, D.; Bandyopadhyay, J.K.; Gupta, S.S.
One hundred and fitly samples of Japanese threadfin bream (Nemipterus japonicus) were screened for bacteriological quality and for content of heavy metals. Out of 150 samples, 40.7% had the count above the stipulated limits for TVB (5.7 log cfulg-) The samples harboured 28,7% of faecal conforms which were present in the range of 2-3 log cfu/g, while 32 and 6% of the samples contained 3-4 and 5 lug cfu/g of the sample. Fecal Streptococci was present in 4N-. at 2-3-log cfu/g and 29,3% at 3-4 log cfu/g. The samples contained coagulase positive Staphylococci, E, coil, Salmonella and V cholera (Non 01) at 14.7, 44, 10 and 14%, respectively. The samples showed a significant decreasein TVB (3-4 log cfu/g of the sample), faecal coliforms and faecal Streptococci during 2 months storage at -18±1°C. Pathogenic bacteria were also not present alter the 2 months storage. The samples were free from hazardous levels of heavy metals., like Cd, Cu, Hg and Pb
Bacteriological quality of fresh and ice-stored farmed macrobrachium rosenbergii from central kerala
(Society of Fisheries Technologists(India),Cochin, 2007) Lalitha, K.V.; Surendran, P.K.
Farmed freshwater Scampi (Macrobrachium rosenbergii de Man) collected from four different farms in central Kerala, India were analysed for bacteriological quality immediately after harvest and after icing for two weeks. Aerobic mesophilic bacteria (TPC), faecal streptococci, faecal coliforms, Escherichia coli and Staphylococcus aureus were enumerated. Total aerobic mesophilic bacterial counts (TPC) on fresh prawn (whole) were of the order of 10 super(6)to 10 super(7) cfu g super(-1). The TPC of fresh and ice stored prawn (whole and headless) were within the acceptable limit. The levels of faecal streptococci and faecal coliforms in fresh M. rosenbergii were above the acceptable limits. However, faecal coliform and E. coli numbers were within the limit ( <100 cfu super(-1)) in deheaded prawn during iced storage. Presence of excessively high TPC and faecal streptococci numbers has been a major microbiological problem in freshwater prawn. This study revealed that A. hydrophila, A. veronii biovar sobria and Enterococci grew well at refrigerated temperatures. A. hydrophila, A. veronii biovar sobria, S. putrefaciens and Pseudomonas were identified as potent spoilers of farmed M. rosenbergii.
The bacteriology of oil sardine (sardinella longiceps) and mackerel (rastrelliger kanagurta) caught from tropical waters off Cochin - I - quantitative aspects
(Society of Fisheries Technologis(India), 1982) Surendran, P.K.; Gopakumar, K.
The total aerobic viable plate counts (TPCs) of skin, gills and intestine of newly caught oil sardine (Sardinella longiceps) and Indian mackerel ( Rastrelliger kanagurta) at four different temperatures, namely 36 ± 1°C , 28 ±2°C (RT), 8 ± 1°C and 1 ±1°C , are reported. The total plate count at RT of the skin of oil sardine and Indian mackerel were in the range of lOS to 107 and 104 to 106 per cm2, that of gills in the range of 105 to lOB and 104 to lOB per g and that intestine in the range of 10s to 109 and 105 to 108 per g respectively. The TPCs were markedly affected by the incubation temperature. Incubation at 28±2°C gave the highest count; at 36± 1°C and 8 ± 1°C , the counts decreased by nearly 1-2 log. cycles from that at RT. Incubation at 1 ± 1°C registered the lowest count. The peak values for bacterial counts of these fishes occurred at different periods of the year.
Behaviour of marine microflora towards antibiotics - I. Sensitivity towards chlorotetracycline (CTC)
(Society of Fisheries Technologis(India), 1971) Surendran, P. K.; MahadevaIyer, K.
The sensitivity of 61 cultures of bacteria isolated from !fish towards chlortetracycline (CTC) at 5 ppm and 20 ppm levels lhas been determined on two solid media, viz; sea water agar (SW A) and a distilled water based medium consisting of peptone, beef extract, glucose and NaCI (PBGA). The cultures employed consisted of (i) gram-negative rods of marine origin viz; Achromobacler, Pseudomonas, Vibrio and Flavobacterium and (ii) gram positive orga• nisms viz; Micrococci and Corynebacterium. Depending on the inhibition zone diameter, the order of CTC sensitivity was found to be Pseudomonas < Vibrio < Achromobacter as determined on PBGA at CTC levels of 5 and 20 ppm. The SWA enhanced the development of resistance in majority of the cultures as compared to PBGA. PBG broth containing CTC at less than 10 ppm exerted a bacteriostatic effect only while that containing 10 ppm or more exerted a bactericidal effect on Achromobacter, Pseudomonas and Micrococci cultures. The significance of these results in fish preservation by CTC is discussed.
Chemical and microbiological quality of dry fish from Kakinada
(Society of Fisheries Technologists (India), 1994) Prasad, M.M.; Rao, C.C.; Gupta, S.S
Chemical and microbiological qualities of 75 samples of dry fish from local markets were studied. In most of the samples, moisture content was found to be higher and salt content lower as compared with ISI specifications for cured dried fish. Potential pathogens, Escherichia coli and coagulase positive staphylococci, were isolated from some of the samples.
Comparative analysis of RAPD, RFLP and PFGE for the characterization of pathogenic vibrio parahaemolyticus isolated from seafood and coastal environment
(Society of Fisheries Technologists (India), Cochin, 2019) Minimol, V.A.; Kishore, P.; Nadella, R.K.; Panda, S.K.; Nayak, B.B.; Prasad, M.M.; Kumar, H.S.
Vibrio parahaemolyticus is ubiquitous in coastal marine environment. Certain strains of V. parahaemolyticus are pathogenic to humans and harbors either or both thermostable direct hemolysin (tdh) gene and tdh related hemolysin (trh) gene. Pathogenic V. parahaemolyticus isolates from the aquatic environment from Mumbai, India were characterized with respect to their virulence genotypes by employing random amplified polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP) and pulsed field gel electrophoresis (PFGE). Out of 140 samples, twenty five samples harboured virulent V. parahaemolyticus. Pandemic group specific PCR assays viz., GSVP, GSOLD, GSOK, ORF8 and PGS PCR showed differential performances which revealed difficulties in the confirmation of pandemic clone from environmental samples. Results from the molecular characterization employing RAPD and RFLP indicated that the PGS PCR positive isolates (2) showed identical banding pattern in RFLP analysis, whereas GS:OK positive isolates (4) showed similar RAPD banding pattern with 80% similarity among 11 pathogenic strains studied. However, in whole genome comparison by PFGE using notI restriction enzyme, these isolates were scattered in different groups at 40% similarity level. This study suggests that the genetically diverse pathogenic strains of V. parahaemolyticus with characteristic of pandemic clones are present in the coastal environment.
Detection of ctx gene positive non-O1/non-O139 V. CCholerae in shrimp aquaculture environments
(Springer, 2013) Rao, B.M.; Surendran, P.K.
Water and post-larvae samples from black tiger (Penaeus monodon) shrimp hatcheries; pond water, pond sediment and shrimp from aquaculture farms were screened for the presence of V cholerae. A V cholerae-duplex PCR method was developed by utilizing V cholerae species specific sodB primers and ctxAB genes specific primers. Incidence of V cholerae was not observed in shrimp hatchery samples but was noticed in aquaculture samples. The incidence of V cholerae was higher in pond water (7.6%) than in pond sediment (5.2%). Shrimp head (3.6%) portion had relatively higher incidence than shrimp muscle (1.6%). All the V cholerae isolates (n=42) belonged to non- 01/non-0139 serogroup, of which 7% of the V cholerae isolates were potentially cholera-toxigenic (ctx positive). All the ctx positive V cholerae (n=3) were isolated from the pond water. Since, cholera toxin (CT) is the major contributing factor for cholera gravis, it is proposed that the mere presence of non-01/non-0139 V cholerae need not be the biohazard criterion in cultured black tiger shrimp but only the presence of ctx carrying non-01/non-0139 V cholerae may be considered as potential public health risk.
Detection of multi drug resistant bacteria in retail fish market water samples of Vashi, Navi Mumbai
(2018) Visnuvinayagam, S.; Murthy, L.N.; Parvathy, U.; Jeyakumari, A.; Adiga, T.G.; Saravanan, G.K.
Occurrence of faecal indicator bacteria (FIB) is being reported regularly by various researchers. But there is no information about the microbiological quality of the water used in the retail fish markets. Hence to understand the hygienic status of the water used in retail outlets, about 51 water samples were collected from the retail outlets of Navi Mumbai region and microbiological parameters such as aerobic plate count (APC), faecal indicator bacteria viz., Escherichia coli, faecal streptococci (FS) and sulphite reducing clostridia (SRC) were enumerated. Antibiogram was also carried out for 57 isolates against 20 antibiotics. Results indicated that all water samples harboured higher levels of APC, E. coli, FS and SRC with an average value of 558, 41, 51 cfu/mL and 2.42/20 mL respectively. Ratio between the FS and E. coli i.e., 1:1.25 indicates the multiplication of E. coli in the water used in the retail market. Higher level of resistance was observed for Augmentin and Colistin. Four multi drug resistant (MDR) E. coli were observed in the water samples. Checking of water in the retail fish market is the most neglected area where high level of contamination and MDR bacteria have been detected. So, it was inferred that the repeated use of same water without replacement is a major cause of higher levels of FIB and MRD bacteria in the retail fish market water samples that further leads to their transfer from water to fish. Hence, running water facility should be used to clean the fish.
Distribution of pathotypes of Escherichia coli in seafood from retail markets of Kerala, India
(Indian Council of Agricultural Research, 2016) Murugadas, V.; Toms C. Joseph; Lalitha, K.V.
Escherichia coli is an indicator species of bacteria to determine faecal/sewage contamination in food and water. Some strains of E. coli have the potential to be pathogenic to humans. The food and its environment should regularly be monitored for the prevalence of pathogenic E. coli. A study was conducted to determine the prevalence of pathogenic E. coli from different fish markets and landing centres in three districts of Kerala, India. Isolated E. coli were pathotyped as enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC) and Shiga toxin producing E. coli (STEC) by PCR, targeting the genes viz., bfpA, eaeA, stx1 stx2, Lt1, Lt2, ST, phoA and rfbeO157. Pathogenic E. coli was found in 23.4% of the 123 seafood samples screened and ETEC, EPEC, and STEC were prevalent in 18.6, 4.0 and 0.8% of the samples respectively.
Distribution of potentially pathogenic Vibrio parahaemolyticus in seafood and the aquatic environment in Mumabai, India
(Society of Fisheries Technologists (India), Cochin, 2018) Minimol, V.A.; Balange, A.K.; Nayak, B.B.; Kumar, S.
The occurrence of total and pathogenic Vibrio parahaemolyticus in fresh seafood and the coastal environment of Mumbai, India was examined in this study. Samples comprising of fish, shellfish, coastal sediment and coastal waters were analyzed for V. parahaemolyticus by selective enrichment and isolation. Biochemically identified isolates were tested for the presence of tlh (thermolabile hemolysin), tdh (thermostable direct hemolysin) and trh (tdh-related hemolysin) by polymerase chain reaction (PCR). While Chromogenic Vibrio (CV) agar yielded V. parhaemolyticus from all samples (100%), TCBS agar yielded V. parahaemolyticus from 57.1% of the samples. The incidence of trh+ V. parahaemolyticus was high, being isolated from 16.4% of the samples analyzed, while the tdh+ V. parahaemolyticus were isolated from 1.4% of samples. Two tdh-, trh+ isolates from the coastal water were positive by a in pandemic group-specific (GS) PCR. The study suggests that seafood and coastal environment may harbor pathogenic V. parahaemolyticus with characteristics of pandemic clones, although their incidence is very low. Further, sodium taurocholate (ST) broth and chromogenic Vibrio (CV) agar combination is highly suitable for the isolation of total and pathogenic V. parahaemolyticus from seafood and environmental samples.
Effect of different organic acids on survival of larvae and control of water microflora in milkfish (Chanos chanos) hatchery system
(Society of Fisheries Technologists (India), Cochin, 2018) Kumar, N.R.; Kumar, P.P.; Siddaiah, G.M.; Murugadas, V.; Basha, K.A.; Sivaraman, G.K.; Prasad, M.M.
This study reports the use of organic acids (acetic, malic, formic and citric) in fish larval rearing systems for survival and controlling the microflora of rearing water. The milk fish (Chanos chanos) larvae was treated for 10, 15, 30 and 60 min with seven concentrations (%) viz., 1, 0.2, 0.1, 0.075, 0.05, 0.025 and 0.02 of organic acids respectively. The percentage survival was zero (1, 0.2, and 0.1% conc) for all the organic acid at various time periods. The effective concentrations for treating fish larvae are 0.05, 0.05, 0.025 and 0.075% concentrations of acetic, malic, formic and citric acid, respectively where 100% survival was observed. Similarly, when the larval rearing water was treated with different concentrations of same organic acids and time period, complete inhibition of microbial flora was observed (1, 0.2 and 0.1% conc) for all organic acids at various time periods. However, at concentrations less than 0.1%, total bacterial count (TBC) and presumptive Vibrio count (PVC) were found in the range of 3.19 to 5.75 and 2.22 to 3.58 log 10 cfu ml-1. In contrast, TBC and PVC in control group were found in the range of 6.37 to 6.90 and 4.35 to 4.74 log 10 cfu ml-1. The acid treatment with concentrations of 0.075, 0.05 and 0.025 except formic acid at 0.025, 0.020 significantly (p<0.05) improved the survival rates of fish larvae and reduced both TBC and PVC in larval rearing water. The present findings strongly recommend the use of organic acids for treating fish larvae for improved survival and also for reducing pathogenic bacterial load from larval rearing waters.
Effect of Monsoon on Sulphite Reducing Clostridia Levels in Fish from Retail Outlets in Maharashtra
(Society of Fisheries Technologists (India) Cochin, 2015) Visnuvinayagam, S.; Viji, P.; Binsi, P.K.; Chakrabarti, R.; Triveni, A.; Joshy, C.G.; Lalitha, K.V.
Enteropathogenic E. coli and other coliforms in marine fish
(Society of Fisheries Technologis(India), 1978) Rao, C.C.P.
One hundred and twenty six samples of marine fish (96 from landing centre and 30 from retail market) and swabs from deck surfaces of 34 fishing boats were examined for coliforms including enteropathogenic E. Coli. Forty out of 96 fish samples from landing centre, 24 out of 30 from retail market and 11 out of 34 fishing boats revealed coliforms. On further tests, 5, 7 and 4 coliform isolates from the three groups respectively were found to be E. coli. Two of the E. coli. isolates, one from sciaenids and one from cat fish, were found to be enteropathogenic serotypes 055 and 0111. Enteropathogenic serotypes of E. coli are reported from sciaenids and cat fish for the first time in this country.
Expression Study of Pluripotency Marker Genes in Gold Fish, Carassius auratrus
(International Journal of Current Microbiology and Applied Sciences (IJCMAS), 2020) Kumar, Abhay; Tripathi, Gayatri; Vimal, Bhartendu; Bedekar, Megha K.; Kumar, A. Pavan
Aquarium fishes, like medaka, zebrafish and goldfish offer an excellent model to conduct studies on molecular mechanisms of embryonic developmental pathways, patterns of genes involved in pluripotency and cell fate decision. To determine the possible candidate gene as an ideal pluripotency marker in goldfish, the comparative study on quantitative expression of a set of genes were conducted in the mid-blastula derived inner cell mass (ICM) and adult somatic cells. Present work explored the expression pattern of four standard pluripotency genes i.e. Nanog, Oct4, Sox and Pou2 in blastocytes (ICM) as well as somatic tissues of kidney, liver, heart, brain, muscle and mature gonads of goldfish using RT-qPCR. Absolute copy number of mRNA transcripts was calculated for each gene in every tissue. Significantly high (P<0.05) mRNA transcript number of all the genes was recorded in ICM, compared to other somatic tissues. Maternally inherited genes like Nanog, Pou2 and Oct4 were found to be highly expressive in ICM as well as in oocytes. The expression profile showed that Nanog andOct4 could belong to similar group where expression is limited to ICM and gonads whereas Pou2expressing in almost all the tissues, including ICM and adult gonads. Because of the low expression level in ICM than in the ovary, Sox2 exhibited expression in several somatic tissues apart from the gonad. Therefore these genes could be characterized as stem cell marker but not a pure pluripotency marker. The results showed that although all the genes compared in this study can be considered as stem cell markers but Nanog gene could be employed as a pure pluripotency marker for ES cell lines from goldfish.
Identification of Salmonella enterica subsp. enterica serovar Paratyphi B dT+ strains from seafood of Cochin retail markets
(Society of Fisheries Technologists (India), 2021) S. S, Greeshma; Pillai, Devika; Sebastian, Mathew; U, Prinetha; Toms. C. Joseph

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