Vibrio parahaemolyticus is one of the most reported food-borne pathogen along the south-west coas... more Vibrio parahaemolyticus is one of the most reported food-borne pathogen along the south-west coast of India, commonly occurring in marine fish. For epidemiological purposes, this pathogen is confirmed by various molecular typing methods, such as pulsed-field gel electrophoresis (PFGE) or ribotyping.These methods are labor intensive and time consuming. In the present study, rapid typing methods with specific sequences viz ., conserved ribosomal gene spacer sequence (RS), repetitive extragenic palindromic sequence (REP), and enterobacterial repetitive intergenic consensus sequence (ERIC) and RAPD (random amplified polymorphic DNA) were used. Marine fish / shellfish were collected from major landing centers located along the south-west coast of India and screened for V. parahaemolyticus . Following phenotypic characterisation, fingerprinting of bacterial strains was carried out by various typing methods viz., RAPD, ERIC, REP, and RS PCR. Cluster analyses revealed the conglomeration of ...
An open reading frame encoding the ectoine biosynthesis genes was cloned from the Bacillus halodu... more An open reading frame encoding the ectoine biosynthesis genes was cloned from the Bacillus halodurans genome. An expression plasmid containing the operon was introduced into Escherichia coli cells, and the recombinant ectoine was quantified. The secondary structure of ectoine biosynthesis proteins were predicted and was quite similar to that of reported proteins from eubacteria.
ABSTRACT Mass mortality was reported from cultured postlarvae of P. monodon from a farm in Ernaku... more ABSTRACT Mass mortality was reported from cultured postlarvae of P. monodon from a farm in Ernakulam District, Kerala, India. Yellow colonies from TCBS plates were randomly picked and were confirmed as V. cholerae by conventional biochemical tests and 16SrRNA sequencing.. The isolates were confirmed as V. cholerae O139 serogroup, generally considered as the causative agent of cholerae to human, using O139 serogroup-specific antiserum and by a PCR based assay targeting rfb-O139 gene. The isolates werefound to carry cholera toxin producing gene; ctx and genes coding for virulence determinants; zot and tcpA as revealed by PCR. Experimentally exposed shrimp larvae with V. cholerae exhibited significant mortalities that increased with increasing doses of bacteria. The LD50 value of one of the isolates was determined in postlarvae of P. monodon, Fenneropenaeus indicus and Litopenaeus vannamei and ranged from 4.6x104 for L. vannamei to 7.1x106 for P. monodon. V. cholerae was re-isolated from the larvae of experimentally infected moribund shrimps. Histopathological examination revealed rupture of basal laminae of hepatopancreatic tubules and severe necrosis. To our knowledge, this is the first report of V. cholerae O139 strain causing high mortalities in shrimp.
Foodborne outbreaks attributed to the contamination of foods with enterohemorrhagic Escherichia c... more Foodborne outbreaks attributed to the contamination of foods with enterohemorrhagic Escherichia coli (EHEC) O157:H7 are a growing global concern. Fish and shrimp samples obtained from different retail fish markets in Cochin, India, were screened by direct PCR assays targeting three important virulence markers of EHEC, the intimin protein (eaeA gene), enterohemolysin (hlyA gene), and Shiga toxin (stx gene). One shrimp (Fenneropenaeus indicus) sample was positive for all these virulence markers, and seven typical E. coli O157:H7 isolates were recovered from the marker-positive shrimp sample. This is the first report of recovery of typical E. coli O157:H7 from fish or shellfish in India. All the typical EHEC isolates had a characteristic reaction in eosin methylene blue agar and belonged to IMViC (indole, methyl red, Voges Proskauer, Simmons citrate reactions) biotype I. These isolates also were negative for sorbitol and methylumbelliferyl-beta-glucuronide and exhibited beta-hemolytic activity. One isolate showed self-agglutination for E. coli O157 antisera and produced a false-positive reaction with CHROMagar O157. These typical EHEC isolates belonged to a restricted biotype group and had a very low multiple antibiotic resistance index. Isolation of E. coli O157:H7 in fish and shellfish indicates that strict adherence to hygienic handling methods and proper cooking or processing is needed before consumption of these products.
The production of a lipopeptide surfactant from the sponge-associated eubacteria Bacillus licheni... more 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°C 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 g L(-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.
World Journal of Microbiology & Biotechnology, 2008
The work was aimed to study the microbial quality of the seafood sold in the domestic markets and... more The work was aimed to study the microbial quality of the seafood sold in the domestic markets and incidence of Vibrio parahaemolyticus. Samples comprising of shellfish, finfish, and cephalopods were collected from various fish markets in and around Cochin. Presumed V. parahaemolyticus were identified by standard biochemical tests, and further confirmed by polymerase chain reaction targeting species-specific tl gene (450 bp). About 81% of the samples were found to exceed the limits specified for total plate count while total presumptive V. parahaemolyticus count was above the limit in 71% of the samples ranging from 5.5 × 105 to 9.7 × 107 and 0.31 × 102 to 7.8 × 106 cfu/g, respectively. Pathogenicity of the identified isolates was confirmed by Kanagawa phenomenon and urease activity. A total of 10% of the isolates exhibited weak haemolysis on Wagatsuma agar, and 1% of the isolates showed urease activity using Christensen’s urea agar. Random amplified polymorphic DNA analysis revealed two major clusters based on the species rather than seasonality. The gel pattern revealed 8–10 bands ranging from 0.45 to 3.0 kb. Antibiogram results revealed 85% of the strains sensitive to chloramphenicol and nitrofurantoin. Multiple antibiotic resistance index was found to be 0.4 thus suggesting the risk potential involved in consuming seafoods. The present study has clearly demonstrated the need to adopt seafood safety measures for the products meant for human consumption.
Ectoine, a cyclic tetrahydropyrimidine (2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid),... more 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 confirmed 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.
World Journal of Microbiology & Biotechnology, 2010
An open reading frame encoding the ectoine biosynthesis genes was cloned from the Bacillus halodu... more An open reading frame encoding the ectoine biosynthesis genes was cloned from the Bacillus halodurans genome. An expression plasmid containing the operon was introduced into Escherichia coli cells, and the recombinant ectoine was quantified. The secondary structure of ectoine biosynthesis proteins were predicted and was quite similar to that of reported proteins from eubacteria.
Suppression Subtractive Hybridization was employed in order to identify the differentially expres... more Suppression Subtractive Hybridization was employed in order to identify the differentially expressed genes in the hepatopancreas of white spot syndrome virus infected Fenneropenaeus indicus. A forward subtracted cDNA library generated 356 clones following a white spot syndrome virus infection. A total of 345 clones with more than 100 nucleotides were selected for further analysis using bioinformatics tools after vector screening. Twenty-three contigs and 111 singletons were generated from a total of 134 consensuses. The consensuses, on a sequence homology search using BLASTX (NCBI), revealed that 74 (55%) of them had no significant match to reported sequences in the database, suggesting that they were found for the first time and are probably associated with shrimp immune function. Out of the remaining 60 (45%) consensuses, 43 had significant homology to known protein sequences in the database while 17 consensuses are homologous to unknown proteins in the database which are considered novel. The most abundant genes in the subtracted library were antimicrobial peptides accounting for 56 clones; among which one is a member of SNF2 family of proteins and another belonged to PfP1 family of proteins on analysis using Antimicrobial peptide predictor software. The other predicted genes in the subtracted library include signal transduction molecules (GTPase, Serine threonine kinase, Armadillo repeats etc), antioxidant enzymes (Cytochrome oxidase, Monomeric sarcosine oxidase and Catalase), active transporters (Nuclear Localization Signal [NLS], calcium ATPase, sodium glutamate symporter, Store-Operated Calcium Entry [SOCE] and ribonucleoprotein [RNP]) contributing to 19, 14 and 5 clones respectively. Three clones are homologous to reverse transcriptase; a first time report in shrimp and one each belong to cell adhesion molecule and Proteinase. InterProScan at EMBL, when used for an integrated search at PROSITE predicted; signal sequences and transmembrane regions for 13 clones. This is the first report on the differential gene expression in WSSV-infected F. indicus. The high expression of immune related genes in response to virus infection in shrimp will provide a new insight into the crustacean innate immunity. Further work on the functionality of the unknown genes in shrimps will give an overview on the role of the differentially expressed genes during viral infection and increase our understanding for developing antiviral measures by making use of the shrimp defense mechanism.
The genes involved in the biosynthetic pathway of ectoine (2-methyl-1,4,5,6-tetrahydropyrimidine-... more The genes involved in the biosynthetic pathway of ectoine (2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid) from Bacillus halodurans were cloned as an operon and expressed in E. coli. Analysis of the deduced ectoine biosynthesis cluster amino acid sequence revealed that the ectoine operon contain 2,389 bp, encoded by three genes; ectA, ectB and ectC that encode proteins of 189, 427 and 129 amino acids with deduced molecular masses of 21,048, 47,120 and 14,797 Da respectively. Extracts of induced cells showed two bands at 41 kDa and 17 kDa, possibly corresponding to the products of the later two genes. However the expression of ectA gene could not be ascertained by SDS-PAGE. The activity of the ectA protein was confirmed by an acylation assay. The transgenic E. coli accumulated upto 4.6 mg ectoine/l culture. This is the first report of an engineered E. coli strain carrying the ectoine genes of the alkaliphilic bacterium, B. halodurans.
In order to determine the prevalence of pathogenic Escherichia coli, a total number of 155 E. col... more 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 virulence markers belonging to the pathogenic group ETEC and STEC. The virulent genes detected in these groups were elt, est, hlyA and stx. The sources of these virulence genes were fish (hlyA), shrimp (elt), feeder canal water (hlyA and elt) of aquaculture origin and from diarrhoea affected 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 effective 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.
Vibrio parahaemolyticus is one of the most reported food-borne pathogen along the south-west coas... more Vibrio parahaemolyticus is one of the most reported food-borne pathogen along the south-west coast of India, commonly occurring in marine fish. For epidemiological purposes, this pathogen is confirmed by various molecular typing methods, such as pulsed-field gel electrophoresis (PFGE) or ribotyping.These methods are labor intensive and time consuming. In the present study, rapid typing methods with specific sequences viz ., conserved ribosomal gene spacer sequence (RS), repetitive extragenic palindromic sequence (REP), and enterobacterial repetitive intergenic consensus sequence (ERIC) and RAPD (random amplified polymorphic DNA) were used. Marine fish / shellfish were collected from major landing centers located along the south-west coast of India and screened for V. parahaemolyticus . Following phenotypic characterisation, fingerprinting of bacterial strains was carried out by various typing methods viz., RAPD, ERIC, REP, and RS PCR. Cluster analyses revealed the conglomeration of ...
An open reading frame encoding the ectoine biosynthesis genes was cloned from the Bacillus halodu... more An open reading frame encoding the ectoine biosynthesis genes was cloned from the Bacillus halodurans genome. An expression plasmid containing the operon was introduced into Escherichia coli cells, and the recombinant ectoine was quantified. The secondary structure of ectoine biosynthesis proteins were predicted and was quite similar to that of reported proteins from eubacteria.
ABSTRACT Mass mortality was reported from cultured postlarvae of P. monodon from a farm in Ernaku... more ABSTRACT Mass mortality was reported from cultured postlarvae of P. monodon from a farm in Ernakulam District, Kerala, India. Yellow colonies from TCBS plates were randomly picked and were confirmed as V. cholerae by conventional biochemical tests and 16SrRNA sequencing.. The isolates were confirmed as V. cholerae O139 serogroup, generally considered as the causative agent of cholerae to human, using O139 serogroup-specific antiserum and by a PCR based assay targeting rfb-O139 gene. The isolates werefound to carry cholera toxin producing gene; ctx and genes coding for virulence determinants; zot and tcpA as revealed by PCR. Experimentally exposed shrimp larvae with V. cholerae exhibited significant mortalities that increased with increasing doses of bacteria. The LD50 value of one of the isolates was determined in postlarvae of P. monodon, Fenneropenaeus indicus and Litopenaeus vannamei and ranged from 4.6x104 for L. vannamei to 7.1x106 for P. monodon. V. cholerae was re-isolated from the larvae of experimentally infected moribund shrimps. Histopathological examination revealed rupture of basal laminae of hepatopancreatic tubules and severe necrosis. To our knowledge, this is the first report of V. cholerae O139 strain causing high mortalities in shrimp.
Foodborne outbreaks attributed to the contamination of foods with enterohemorrhagic Escherichia c... more Foodborne outbreaks attributed to the contamination of foods with enterohemorrhagic Escherichia coli (EHEC) O157:H7 are a growing global concern. Fish and shrimp samples obtained from different retail fish markets in Cochin, India, were screened by direct PCR assays targeting three important virulence markers of EHEC, the intimin protein (eaeA gene), enterohemolysin (hlyA gene), and Shiga toxin (stx gene). One shrimp (Fenneropenaeus indicus) sample was positive for all these virulence markers, and seven typical E. coli O157:H7 isolates were recovered from the marker-positive shrimp sample. This is the first report of recovery of typical E. coli O157:H7 from fish or shellfish in India. All the typical EHEC isolates had a characteristic reaction in eosin methylene blue agar and belonged to IMViC (indole, methyl red, Voges Proskauer, Simmons citrate reactions) biotype I. These isolates also were negative for sorbitol and methylumbelliferyl-beta-glucuronide and exhibited beta-hemolytic activity. One isolate showed self-agglutination for E. coli O157 antisera and produced a false-positive reaction with CHROMagar O157. These typical EHEC isolates belonged to a restricted biotype group and had a very low multiple antibiotic resistance index. Isolation of E. coli O157:H7 in fish and shellfish indicates that strict adherence to hygienic handling methods and proper cooking or processing is needed before consumption of these products.
The production of a lipopeptide surfactant from the sponge-associated eubacteria Bacillus licheni... more 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°C 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 g L(-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.
World Journal of Microbiology & Biotechnology, 2008
The work was aimed to study the microbial quality of the seafood sold in the domestic markets and... more The work was aimed to study the microbial quality of the seafood sold in the domestic markets and incidence of Vibrio parahaemolyticus. Samples comprising of shellfish, finfish, and cephalopods were collected from various fish markets in and around Cochin. Presumed V. parahaemolyticus were identified by standard biochemical tests, and further confirmed by polymerase chain reaction targeting species-specific tl gene (450 bp). About 81% of the samples were found to exceed the limits specified for total plate count while total presumptive V. parahaemolyticus count was above the limit in 71% of the samples ranging from 5.5 × 105 to 9.7 × 107 and 0.31 × 102 to 7.8 × 106 cfu/g, respectively. Pathogenicity of the identified isolates was confirmed by Kanagawa phenomenon and urease activity. A total of 10% of the isolates exhibited weak haemolysis on Wagatsuma agar, and 1% of the isolates showed urease activity using Christensen’s urea agar. Random amplified polymorphic DNA analysis revealed two major clusters based on the species rather than seasonality. The gel pattern revealed 8–10 bands ranging from 0.45 to 3.0 kb. Antibiogram results revealed 85% of the strains sensitive to chloramphenicol and nitrofurantoin. Multiple antibiotic resistance index was found to be 0.4 thus suggesting the risk potential involved in consuming seafoods. The present study has clearly demonstrated the need to adopt seafood safety measures for the products meant for human consumption.
Ectoine, a cyclic tetrahydropyrimidine (2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid),... more 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 confirmed 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.
World Journal of Microbiology & Biotechnology, 2010
An open reading frame encoding the ectoine biosynthesis genes was cloned from the Bacillus halodu... more An open reading frame encoding the ectoine biosynthesis genes was cloned from the Bacillus halodurans genome. An expression plasmid containing the operon was introduced into Escherichia coli cells, and the recombinant ectoine was quantified. The secondary structure of ectoine biosynthesis proteins were predicted and was quite similar to that of reported proteins from eubacteria.
Suppression Subtractive Hybridization was employed in order to identify the differentially expres... more Suppression Subtractive Hybridization was employed in order to identify the differentially expressed genes in the hepatopancreas of white spot syndrome virus infected Fenneropenaeus indicus. A forward subtracted cDNA library generated 356 clones following a white spot syndrome virus infection. A total of 345 clones with more than 100 nucleotides were selected for further analysis using bioinformatics tools after vector screening. Twenty-three contigs and 111 singletons were generated from a total of 134 consensuses. The consensuses, on a sequence homology search using BLASTX (NCBI), revealed that 74 (55%) of them had no significant match to reported sequences in the database, suggesting that they were found for the first time and are probably associated with shrimp immune function. Out of the remaining 60 (45%) consensuses, 43 had significant homology to known protein sequences in the database while 17 consensuses are homologous to unknown proteins in the database which are considered novel. The most abundant genes in the subtracted library were antimicrobial peptides accounting for 56 clones; among which one is a member of SNF2 family of proteins and another belonged to PfP1 family of proteins on analysis using Antimicrobial peptide predictor software. The other predicted genes in the subtracted library include signal transduction molecules (GTPase, Serine threonine kinase, Armadillo repeats etc), antioxidant enzymes (Cytochrome oxidase, Monomeric sarcosine oxidase and Catalase), active transporters (Nuclear Localization Signal [NLS], calcium ATPase, sodium glutamate symporter, Store-Operated Calcium Entry [SOCE] and ribonucleoprotein [RNP]) contributing to 19, 14 and 5 clones respectively. Three clones are homologous to reverse transcriptase; a first time report in shrimp and one each belong to cell adhesion molecule and Proteinase. InterProScan at EMBL, when used for an integrated search at PROSITE predicted; signal sequences and transmembrane regions for 13 clones. This is the first report on the differential gene expression in WSSV-infected F. indicus. The high expression of immune related genes in response to virus infection in shrimp will provide a new insight into the crustacean innate immunity. Further work on the functionality of the unknown genes in shrimps will give an overview on the role of the differentially expressed genes during viral infection and increase our understanding for developing antiviral measures by making use of the shrimp defense mechanism.
The genes involved in the biosynthetic pathway of ectoine (2-methyl-1,4,5,6-tetrahydropyrimidine-... more The genes involved in the biosynthetic pathway of ectoine (2-methyl-1,4,5,6-tetrahydropyrimidine-4-carboxylic acid) from Bacillus halodurans were cloned as an operon and expressed in E. coli. Analysis of the deduced ectoine biosynthesis cluster amino acid sequence revealed that the ectoine operon contain 2,389 bp, encoded by three genes; ectA, ectB and ectC that encode proteins of 189, 427 and 129 amino acids with deduced molecular masses of 21,048, 47,120 and 14,797 Da respectively. Extracts of induced cells showed two bands at 41 kDa and 17 kDa, possibly corresponding to the products of the later two genes. However the expression of ectA gene could not be ascertained by SDS-PAGE. The activity of the ectA protein was confirmed by an acylation assay. The transgenic E. coli accumulated upto 4.6 mg ectoine/l culture. This is the first report of an engineered E. coli strain carrying the ectoine genes of the alkaliphilic bacterium, B. halodurans.
In order to determine the prevalence of pathogenic Escherichia coli, a total number of 155 E. col... more 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 virulence markers belonging to the pathogenic group ETEC and STEC. The virulent genes detected in these groups were elt, est, hlyA and stx. The sources of these virulence genes were fish (hlyA), shrimp (elt), feeder canal water (hlyA and elt) of aquaculture origin and from diarrhoea affected 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 effective 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.
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