Aims:  (i) To study the occurrence of Escherichia coli serotype O157 in cattle stool in West Bengal, India, and (ii) the virulence properties and antimicrobial resistance of the E. coli isolates. Methods and Results:  Following enrichment in modified EC broth and plating onto HiCrome MS.O157 agar, a total of 14 strains of E. coli serotype O157 was isolated from faecal samples from two (2.04%) slaughtered cattle and six (7.59%) diarrhoeic calves. By multiplex PCR, Shiga toxin genes were detected in all the isolates. The enterohaemolysin phenotype was found in all, but one strain. Among 14 strains, ten were resistant to at least one of the antimicrobial agents tested. Multiple antibiotic resistance was frequent. Conclusions:  The study showed that occurrence of Shiga toxin‐producing and multiple antibiotic‐resistant E. coli O157 among cattle population in this region of India is significant. Significance and Impact of the Study:  Considering routine human contacts with cattle, a large human population in this region may be at risk for exposure to Shiga toxin‐producing E. coli O157.

Background:Milk is considered as complete food and an important part of human diet throughout the world including India. Bacterial contamination of milk such as Escherichia coli due to unhygienic condition and poor udder health can cause infections, especially in infants and elders or in immunocompromised persons. Possession of antimicrobial resistance genes by commensal bacteria present in milk makes the issue more serious.Aim:The study was aimed to isolate and characterize extended-spectrum beta-lactamase (ESBL)-producing E. coli from milk samples collected from different parts of West Bengal, India, to assess the potential risk associated with the food.Materials and Methods:Around 182 milk samples were collected from apparently healthy cows reared by organized dairy farms in West Bengal. E. coli was isolated from collected samples as per standard methods followed by serotyping. The detection of ESBL-producing E. coli was done both phenotypically and genotypically by detecting the presence of blaCTX-M gene. Antibiogram of the ESBL-positive isolates was done using common 12 antibiotics by disc diffusion method.Results:A total of 22 (12.1%) samples were found to be positive for E. coli in this study. Different serotypes such as O11, O20, O22, O34, O35, O128, O149, and UT were isolated from the collected samples. 12 (54.5%) E. coli strains showed the capability of producing ESBL, both phenotypically and genotypically with the presence of blaCTX-M gene. Antibiogram of these ESBL-positive isolates revealed the drugs such as colistin (100%), levofloxacin (83.33%), and imipenem (66.67%) to be highly sensitive against this pathogen but drugs such as cefotaxime (100%), ceftazidime (91.67%), amoxicillin/clavulanic acid (83.33%), tetracycline (75.00%), and gentamicin (58.33%) to be very much resistant.Conclusion:More than 50% of the E. coli strains prevalent in the bovine milk samples were positive for ESBL production and are resistant to most of the common antimicrobials which may be alarming for human health.

Aim:Subclinical mastitis in bovines is mainly responsible for the huge economic loss of the dairy farmers, of which Pseudomonas aeruginosa is one of the causative agents. The study was aimed at a screening of suspected milk samples from different cattle farms of West Bengal for detection and confirmation of P. aeruginosa strains followed by their characterization.Materials and Methods:Around 422 milk samples were screened from different dairy farms primarily by on-spot bromothymol blue (BTB) test and then in the lab by somatic cell counts (SCC) to finally consider 352 samples for detection of P. aeruginosa. Selective isolation and confirmation of the isolates were done using selective media, viz., cetrimide and Pseudomonas agar followed by confirmation by fluorescent technique. Molecular characterization of the strains was done by polymerase chain reaction for the presence of toxA (enterotoxin A, 352 bp) and exoS (exoenzyme S, 504 bp) genes.Results:Approximately, 371 (87.9%) samples were positive in on-spot BTB test among which 352 (94.8%) samples revealed high SCC values (more than 3 lakh cells/ml) showing infection when screened. Among these, 23 (6.5%) samples yielded typical Pseudomonas sp. isolates out of which only 19 (5.4%) isolates were confirmed to be P. aeruginosa which showed characteristic blue-green fluorescence due to the presence of pigment pyoverdin under ultraviolet light. Out of these 19 isolates, 11 isolates were positive for toxA, 6 isolates for exoS, and 2 for both these pathogenic genes.Conclusion:Approximately, 5.4% cases of bovine subclinical mastitis infections in South Bengal were associated with P. aeruginosa which possess pathogenic genes such as toxA (63.2%) and exoS (36.8%).

Aim:The ringworms of pet dogs, cats, and stray animals (dogs, cats, and other animals) could be a potential source of zoonotic infections causing a serious public health problem in the busy city Kolkata. The pet owners are more susceptible to get this infection from their pets, because of the close contact with them as dermatophytosis is very much prevalent in those pets. So, this study was aimed to check the prevalence of dermatophytosis in dogs, cats, and in pet owners.Materials and Methods:A total of 362 clinically suspected cases of dermatophytosis from dogs (123 in number), cats (202 in number), and human beings (37 in number) were collected and studied from in and around Kolkata to detect the presence of significant dermatophytes. Direct microscopy and cultural examination of the isolates were performed following standard methodology. Identification and characterization of the isolates were done by different biochemical tests.Results:Samples (n=285) having significant dermatophytic fungal infections were found to be of highest number in cats (158, 55.5%) than in dogs (108, 37.8%) and humans (19, 6.7%), respectively. The incidence of Microsporum canis (60.0%) was the highest from affecting dogs, cats, and human beings in comparison to Microsporum gypseum (22.5%), Trichophyton mentagrophytes (15.8%) and Trichophyton rubrum (1.7%). Detection of T. rubrum was only from human cases in this study, whereas the presence of rest three were slightly higher in cats than that of the dogs and humans in this present study. The incidences were higher in young animals and in humans of the age group of 21-30 years, during the rainy season (from April to August) and also in in-contact human beings.Conclusion:M. canis was the most commonly pathogen among all causing dermatophytosis in animals and also in the pet owners. M. gypseum and T. mentagrophytes were other pathogens associated with these infections. These infections were more prevalent in the rainy seasons and in in-contact human patients or pet owners.

Aim:The aim was to characterize Salmonella enterica serovar Gallinarum isolated from backyard poultry by polymerase chain reaction (PCR) detection of virulence genes invasion (invA) and Salmonella plasmid virulence C (spvC).Materials and Methods:Two strains of Salmonella serovar Gallinarum isolates used in this study were obtained from an outbreak of fowl typhoid in backyard Vanaraja fowl. PCR technique was used for detection of invA and spvC genes using standard methodology. The invA PCR product from one representative isolate was sequenced and compared with other related Salmonella serovars in GenBank data.Results:Salmonella Gallinarum produced expected amplicons of invA and spvC gene products. Nucleotide sequence of 285 bp invA gene was deposited in GenBank with accession no. KX788214. Sequence analysis of invA gene was found conserved in Salmonella serovars and demonstrated 100% homology with closely related serovars of Salmonella.Conclusion:Invasion gene (invA) was found to be highly conserved in Salmonella Gallinarum and highly similar with closely related serovars. The isolates also contained plasmid-mediated spvC gene indicating possession of virulence plasmid.

Aims:  To (i) study the serogroup distribution and virulence characteristics of non‐sorbitol‐fermenting Escherichia coli isolates from foods of animal origin and cattle faeces and (ii) re‐examine the true sorbitol and β‐d‐glucuronidase (GUD) reactions of sorbitol‐negative (Sor−) strains from MacConkey sorbitol agar (SMAC) to assess their phenotypic similarity with E. coli O157. Methods and Results:  One hundred and thirty Sor−E. coli were isolated from 556 food samples and 177 cattle stool samples using cefixime tellurite–supplemented SMAC (CT‐SMAC) and chromogenic HiCrome MS.O157 agar respectively. Based on typing of somatic antigen, the isolates were classified into 38 serogroups. PCR results identified about 40% strains, belonging to O5, O8, O20, O28, O48, O60, O78, O82, O84, O101, O110, O123, O132, O156, O157, O‐rough and OUT as Shiga toxigenic. Majority of O5, O84, O101, O105, O123, O157, O‐rough and OUT strains were enterohaemolytic. Further, 39·2% and 63·1% of Sor− isolates from CT‐SMAC fermented sorbitol in phenol red broth and hydrolysed 4‐methylumbelliferyl‐β‐d‐glucuronide (MUG) respectively. Members of serogroups O5, O28, O32, O81, O82, O84, O101, O‐rough lacked both the sorbitol fermentation (broth test) and GUD activity and might create confusion in phenotypic identification of E. coli O157. Conclusions:  Sor−E. coli isolates from raw meat, milk, shrimp and cattle stool belonged to 38 serogroups, with E. coli O157 constituting only 14·6% of the isolates. Many of these nonclinical Sor− strains were potentially pathogenic. Nearly 39% of these Sor−E. coli from CT‐SMAC fermented sorbitol in broth, indicating the need for confirmation of sorbitol reaction in broth. Significance and Impacts of the Study:  Classical sorbitol utilization and GUD tests are not likely definitive tests for E. coli O157. Further improvement of differential media based on these phenotypic properties is necessary for detection of pathogenic serotypes from foods and environmental samples.

Significance and Impact of the Study: Characterization of antimicrobial resistance is an utmost necessity in animals having direct contact with human as a part of the one health approach. This is the first large-scale monitoring of ESBL/biofilm-producing Klebsiella in companion/household animals in India sharing human habitat. Majority of the Klebsiella possessed bla CTX-M-15 which is associated with infection in both humans and animals throughout the world. Few nucleotide sequences of bla CTX-M in the present study are reported for the first time from the companion animals. The emergence of resistance determinants in Klebsiella isolated from companion and household animals was correlated with therapeutic antibiotic exposure and contaminated environment, respectively.

This study was undertaken to detect the occurrence of beta‐lactamase and biofilm producing Enterobacteriaceae in healthy ducks. A total 202 cloacal swabs were collected from ducks kept in organized (n = 92) and backyard (n = 110) farms in West Bengal (India). The ducks had no history of antibiotic intake. Among the 87 phenotypically beta‐lactamase producing Escherichia coli, 19 (17·43%), 6 (5·05%) and 15 (13·76%) isolates possessed blaTEM, blaSHV and blaCTX‐M respectively. Whereas, 5 (38·46%) Salmonella isolates were found to harbour blaCTX‐M. In K. pneumoniae 10 (33·33%), 3 (13·33%), 4 (13·33%) isolates possessed blaTEM, blaSHV and blaCTX‐M respectively. The sequences of selected PCR products were found 98% cognate with blaCTX‐M‐9, blaSHV‐12 and blaTEM‐1. Beta‐lactamase producing E. coli isolates belonged to 14 different serogroups such as O1, O2, O3, O5, O7, O8, O35, O83, O84, O88, O119, O128, O145 and O157. Moreover, 87 E. coli (79·82%), six Samonella (46·15%) and 13 K. pneumoniae (43·33%) isolates were detected as AmpC producers possessing blaAmpC. Majority of E. coli (46·79%), Salmonella (46·15%) and K. pneumoniae (70%) isolates were detected as biofilm producers and possessed the associated genes (csgA, sdiA, rcsA, rpoS). Significantly higher occurrence of beta‐lactamase and biofilm producing Enterobacteriaceae isolates was detected in backyard ducks than organized farms. Significance and Impact of the Study Consumption of antibiotic through feed or during therapy is considered as potential reason for generation of antimicrobial resistant bacteria in birds. This study provides valuable evidence that exposure to contaminated environment may be an additional source for generation of antimicrobial resistant bacteria in backyard ducks. The backyard ducks are reared by marginal farmers in India who cannot offer antibiotics to them either through feed or during therapy due to high cost. The study also reveals a significant correlation between biofilm formation and possession of antimicrobial resistance genes in the bacterial isolates from the ducks.