Occurrence of canine parvovirus type 2c in diarrhoeic faeces of dogs in Kolkata, India.

Canine parvovirus-2(CPV-2) causes a highly contagious disease of dogs characterised by acute hemorrhagic gastroenteritis, lethargy, vomiting, fever and usually bloody or mucoid diarrhoea. In the present study, 41 faecal samples collected from dogs exhibiting the signs of fever, vomition, bloody or mucoid diarrhoea in Kolkata, India were screened by haemagglutination test and PCR for detection of capsid protein coding VP2 gene. The viral genotype was detected by multiplex PCR and analysis of partial VP2 gene nucleotide sequences of selected PCR products with bioinformatics tool. Thirteen (31.71%) samples were found positive with HA titre ≥ 32 whereas 28 (68.29%) samples were positive by PCR of VP2 gene indicating higher sensitivity of PCR. Highest occurrence of CPV-2 was observed in the age group of 1-6 months (80.65%) and non-descript breeds with no history of vaccination (85%). Three samples were antigenic type CPV-2a, rest were CPV-2b/CPV 2c. Six CPV sequences were found to be highly similar to published CPV 2c sequences in BLAST analysis revealing a maximum identity of 99-100% with other CPV-2c strains and clustered together with CPV-2c strains of India and other countries in phylogenetic analysis. The present study highlights the need for continuous monitoring of samples to detect gradual changes in circulating CPV-2 genotypes in India.

Antibacterial effect of kitchen herbs against pathogenic multidrug-resistant E. coli isolates from calf diarrhoea.

Calf diarrhoea remains the biggest challenge both in the small and large farms. Infectious diarrhoea is associated with many pathogens, Escherichia coli being one, but majority are systematically treated with antibiotics. Since antimicrobial resistance (AMR) is a growing menace, the need to find alternative prophylactic solutions using popular kitchen herbs such as Trachyspermum ammi (carom seeds), Curcuma longa (turmeric) and cinnamon (Cinnamomum sp.) extracts is been investigated against virulent form of E. coli isolated from calf diarrhoea. The virulence factors identified in these isolates were ST (32.5%), LT (20%), eaeA (15%), stx1 (2.5%) and stx2 (5%) with the occurrence of the most common serogroups as O18 (15%) followed by O111 (12.5%). Highest resistance was seen with beta lactam + beta lactamase inhibitor (amoxicillin/clavulanic acid) followed by beta lactams (ampicillin, cefuroxime and cefepime). The zone of inhibition due to cinnamon (methanol) and carom seed (ethanol) extracts (500 to 250 µg/mL concentration) on E. coli bacteria was >19 mm, respectively. Turmeric, cinnamon and carom had the potency of inhibiting the pathogenic E. coli which maybe suggestive of its use in calf diets as prophylaxis against diarrhoea.

Comparative occurrence of ESBL/AmpC beta-lactamase-producing Escherichia coli and Salmonella in contract farm and backyard broilers.

The present study was conducted to detect the occurrence of beta-lactamase and biofilm-producing Escherchia coli and Salmonella in apparently healthy broiler birds reared in household and contract poultry farms. In total, 150 cloacal swabs were collected from apparently healthy broiler birds of various age groups reared in backyard (n = 100) and contract farms (n = 50) in West Bengal (India). The isolation rate of ESBL producers was significantly more (P < 0·05) reared in contract poultry farms than those reared in backyard. Majority of the E. coli isolates possessed blaCTX-M followed by blaSHV and blaTEM . Majority of the Salmonella strains possessed blaTEM followed by blaCTX-M , and no blaSHV was detected. The selected sequences of the PCR products were found cognate with blaCTX-M-1 , blaCTX-M-2 , blaCTX-M-9 , blaCTX-M-14 , blaCTX-M-15 , blaSHV-12 and blaTEM-52 . The study detected 46·8% of E. coli and 42·5% of Salmonella strains as biofilm producers. Beta-lactamase-producing Enterobacteriaceae strains showed resistance against tetracycline, chloramphenicol, doxycycline, co-trimoxazole and ampicillin and sensitivity to imipenem-EDTA, colistin and gentamicin. The study revealed the partial clonal relationship of ESBL sequences possessed by the poultry isolates of the present study and local clinical isolates available in the database. The study made consumer awareness about careful handling of live birds or poultry meat to avoid the zoonotic transmission of antimicrobial-resistant Enterobacteriaceae.

Multi-drug resistant, biofilm-producing high-risk clonal lineage of Klebsiella in companion and household animals.

Antimicrobial resistance is a global emergency which needs one health approach to address. The present study was conducted to detect the prevalence of beta-lactamase and biofilm-producing Klebsiella strains in rectal swabs (n = 624) collected from healthy dogs, cats, sheep and goats reared as companion or household animals in India. The dogs and cats were frequently exposed to third- or fourth-generation cephalosporins for therapy. The sheep and goats were occasionally exposed to antibiotics and had environmental exposure. Phenotypical ESBL (n = 93) and ACBL (n = 88)-producing Klebsiella were isolated significantly more (P < 0·05) from companion animals than household animals. Majority of the Klebsiella possessed blaCTX-M-15 . The sequences blaCTX-M-15.2 , blaCTX-M-197 and blaCTX-M-225 are reported first time from the companion animals. All ACBL-producing isolates possessed blaAmpC . The present study detected 65·8% of Klebsiella strains as biofilm producers possessing the studied biofilm associated genes. The isolates showed phenotypical resistance against chloramphenicol, tetracycline, doxycycline, co-trimoxazole, ampicillin, cefotaxime/clavulanic acid. The present study showed that companion and household animals (dogs, cats, sheep, goats) may act as a carrier of ESBL/biofilm-producing, multi-drug resistant, high-risk clonal lineage of Klebsiella.

Higher prevalence of multidrug-resistant extended-spectrum ß-lactamases producing Escherichia coli in unorganized pig farms compared to organized pig farms in Mizoram, India.

AIM: The present study was conducted to record the prevalence of multidrug-resistant (MDR), extended-spectrum ß-lactamases (ESBLs) producing Escherichia coli from pig population of organized and unorganized farms of Mizoram and to record the presence of ESBLs, non-ESBLs, and integrons. MATERIALS AND METHODS: Fecal samples were collected from pigs under organized (n=40) and unorganized (n=58) farms of Mizoram. Samples were processed for isolation and identification of E. coli by conventional techniques, BD Phoenix™ automated bacterial system, and polymerase chain reaction (PCR) based confirmatory test. All the isolates were subjected to antimicrobial sensitivity test by disk diffusion assay and ESBLs production by double-disk synergy test (DDST). The ESBLs producing isolates were subjected to PCR for determination of ESBLs genes and all the isolates were screened for non-ESBLs genes and integrons by PCR. RESULTS: A total of 258 E. coli was isolated and identified from organized (n=120) and unorganized farms (n=138). Majority of the E. coli isolates exhibited high level of resistance against amoxicillin (Ax) (81.78%), cefalexin (85.42%), co-trimoxazole (50.78%), sulfafurazole (69.38%), tetracycline (65.89%), and trimethoprim (TR) (51.94%). Statistically highly significant (p<0.01) variations in resistance among the isolates from organized and unorganized farms were recorded in case of Ax, ampicillin, cephalexin, ciprofloxacin, co-trimoxazole, gentamicin, piperacillin, and TR. By DDST, 65.89% isolates were recorded as ESBLs producer, of which 82/120 (68.33%) and 88/138 (63.77%) were from organized and unorganized farms, respectively. A total of 29/258 (11.24%) isolates were positive for at least one ESBLs gene. blaTEM was most frequently (9.69%) gene, followed by blaCTX -M (5.04%) and blaCMY (0.78%). Altogether, 6 (5.00%), 4 (3.33%), and 2 (1.67%) isolates from the organized farms were positive for blaCTX-M , blaTEM , and blaCMY genes, respectively. Similarly, 21 (15.22%) and 7 (5.07%) isolates from the unorganized farms were positive for blaTEM and blaCTX-M genes, respectively. None of them were positive for blaSHV genes. Altogether 57 (22.09%), 9 (3.49%), 66 (25.58%), 78 (30.23%), 21 (8.14%), and 18 (6.98%) isolates were positive for tetA, tetB, sul1, sul2, aadA, and dfrla genes, respectively. The prevalence of non-ESBLs genes was higher in the E. coli isolates from the unorganized farms than organized farms. CONCLUSION: MDR and ESBLs producing E. coli are circulating among the pigs and their environment in Mizoram. Pigs under unorganized farms exhibited higher level of resistance against majority of the antimicrobials, including third-generation cephalosporins, which might be an indication of overuse or misuse of antibiotics under the unorganized piggery sectors in Mizoram.

Ecology of bats and their role in emerging zoonotic diseases: a review.

The order Chiroptera is the second largest order of mammals and shows great physiological and ecological diversity. These animals play significant ecological roles as prey and predator as well as facilitating pollination, seed dispersal, arthropod reduction and nutrient distribution and reutilisation in nature. Bats act as hosts to a range of viral, bacterial, fungal and parasitic zoonoses. Human activities increase the likelihood of exposure to bats, thereby increasing the opportunity for infections to spill over. Continuing ecological processes, emergence and spillover of novel pathogens in naïve hosts, including humans, along with other complex natural phenomena require proper understanding that may help in predicting the next spillover. This review will discuss the ecology of bats and their role in the emergence of different zoonoses, particularly those of viral origin, in an organised manner to increase understanding of the factors that may play significant roles in spillover of these pathogens from bats to other animals, including humans.

L'ordre des Chiroptères, le deuxième des mammifères en nombre d'espèces, présente des caractéristiques physiologiques et écologiques très diverses. Ces animaux jouent un rôle écologique majeur à divers titres, en tant que proies et prédateurs, mais aussi parce qu'ils interviennent dans la pollinisation, la dispersion des semences, la réduction des populations d'arthropodes et la répartition et réutilisation des nutriments dans la nature. Les chauves-souris sont les hôtes d'un large éventail de virus, bactéries, champignons et parasites zoonotiques. Les activités humaines entraînent une probabilité accrue d'exposition aux chauves-souris, ce qui à son tour multiplie les situations à la faveur desquelles les maladies infectieuses peuvent se transmettre d'une espèce réservoir à des hôtes accidentels. Les processus écologiques continus, l'émergence d'agents pathogènes nouveaux et l'atteinte accidentelle d'une espèce réceptive mais habituellement non touchée, y compris l'homme, ainsi que d'autres phénomènes naturels complexes, doivent être correctement appréhendés afin de pouvoir prédire la prochaine occurrence de ce type de transmission inter-espèces. Les auteurs examinent l'écologie des chauves-souris et leur rôle dans l'émergence de plusieurs zoonoses, notamment d'origine virale, en les présentant sous forme raisonnée afin de mieux faire comprendre les facteurs susceptibles de jouer un rôle significatif dans la transmission de ces agents pathogènes des chauves-souris à d'autres espèces, y compris l'homme.

El orden de los quirópteros, que es el segundo más grande de los órdenes de mamíferos, presenta una gran diversidad fisiológica y ecológica. Estos animales cumplen importantes funciones ecológicas como presas y predadores, además de facilitar la polinización, la dispersión de semillas, la reducción de las poblaciones de artrópodos y la distribución y reutilización de nutrientes en el medio natural. Los murciélagos son hospedadores de una serie de zoonosis víricas, bacterianas, fúngicas y parasitarias. Las actividades humanas aumentan la probabilidad de exposición a murciélagos, acrecentando con ello las posibilidades de extensión secundaria de una infección. Los continuos procesos ecológicos, la aparición de nuevos patógenos y su extensión a hospedadores no expuestos previamente, incluidas las personas, junto con otros fenómenos naturales complejos, obligan a conocer cabalmente estos procesos para ayudar a predecir dónde puede darse la siguiente extensión secundaria. Los autores examinan la ecología de los murciélagos y la función que estos cumplen en el surgimiento de diferentes zoonosis, en especial las de origen vírico, de manera organizada para conocer mejor los factores que pueden influir sustancialmente en el paso de esos patógenos del murciélago a otros animales, en particular el ser humano.

Extended spectrum beta-lactamase producing Shiga-toxin producing-Escherichia coli in piglets, humans and water sources in North East region of India.

Extended spectrum ß-lactamases (ESBL) producing Shiga toxin producing E. coli (STEC) are posing a constant threat to public health throughout the world leading to serious infections and raising key therapeutic issues. A total of 219 fecal samples were collected from piglets with diarrheoa, pig farmers and water sources in North East India; and were processed for isolation of Escherichia coli. The isolates were screened for antimicrobial resistance and suspected isolates for ESBLs production by double-disk synergy test (DDST). Escherichia coli isolates positive for DDST were subjected for detection of selected ESBL/beta-lactamase genes and virulence associated genes by PCR. By DDST, 337 (67·94%) E. coli isolates were detected as ESBLs producer, of which 211 (66·98%), 117 (70·91%) and 9 (56·25%) isolates were from piglets, humans and water sources respectively. A total of 64 (12·90%) isolates were recorded as STEC, of which 48 (9·68%), 6 (1·21%) and 10 (2·02%) were from human, piglets and water respectively. Majority of the STEC isolates (64·06%) possessed multiple virulence genes, of which 59·38% also harboured ESBL/beta-lactamase genes with 32·81% STEC isolates being positive for multiple ESBL/beta-lactamase genes. SIGNIFICANCE AND IMPACT OF THE STUDY: Multidrug resistant (MDR) enteric bacteria are global concern. Association of MDR traits in STEC isolates are another rising issue in human and animal health perspective. The interaction of such organisms among the human, domestic animals and adjoining water sources require to be analysed systematically. The present study exhibited the possible transmission of MDR-STEC among the human, domestic animals and water sources in the North eastern states of India. To the best of our knowledge, this is the first report of such kind in India.

Characterization of beta-lactamase and biofilm producing Enterobacteriaceae isolated from organized and backyard farm ducks.

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.

Detection and characterization of pathogenic Pseudomonas aeruginosa from bovine subclinical mastitis in West Bengal, India.

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%).

Detection of dermatophytes in healthy companion dogs and cats in eastern India.

In recent times increasing occurrence of dermatophytosis, especially among the school children in eastern India was evidenced along with increased tendency of keeping companion animals such as dogs and cats. This study was undertaken to detect the occurrence of dermatophytes with antifungal susceptibility among the companion animals. A total of 1501 healthy companion animals comprising 1209 dogs and 292 cats belonged to individual owners in and around Kolkata (West Bengal, India) were examined for the evidence of dermatophytosis during 2011-2013. The collected samples were subjected to direct examination by standard KOH mount technique. The samples were inoculated into both Sabouraud dextrose agar (SDA) with 0.05% chloramphenicol and 0.5% cycloheximide and dermatophyte test medium (DTM). Each of the fungal isolate was identified based upon its colony characteristics and hyphal and conidial cells it produced. Antifungal susceptibility of the isolates was tested by broth micro dilution assay using fluconazole, ketoconazole, itraconazole, miconazole, griseofulvin and amphotericin-B antifungals. Among the 1209 samples from dogs and 292 samples from cats, 253 (20.93%) and 109 (37.33%) samples were positive for dermatophytes by direct examination. Three identified species of dermatophytes with predominant occurrence were Microsporum canis, Microsporum gypseum and Trichophyton mentagrophytes. Ketoconazole (0.06-0.5 µgm/ml), itraconazole (0.03-0.5 µgm/ml) and amphotericin-B (0.03-0.5 µgm/ml) showed lowest MIC values against M. canis, T. mentagrophytes and M. gypseum, respectively. This is the first systemic report of dermatophytes in healthy companion animals with large numbers of samples in India.

Molecular signature of extended spectrum ß-lactamase producing Klebsiella pneumoniae isolated from bovine milk in eastern and north-eastern India.

The present study reports on 23 extended spectrum ß-lactamase producing Klebsiella pneumoniae (KP), isolated from milk samples (n=340) of healthy cows (n=129) and cows with subclinical (n=159) and clinical (n=52) mastitis, from three different states of India viz. West Bengal, Jharkhand and Mizoram. Seven of them were AmpC type ß-lactamase producers, as well. The ESBL producing KP were significantly (P=0.006, χ2=10.04, df=2) and more frequently detected in milk samples of mastitic cows than healthy ones. The ß-lactamase genes - blaCTX-M, blaTEM and blaSHV were detected in 19, 8 and 3 isolates, respectively. In all but one CTX-M positive isolates, the genetic platform - ISEcp1-blaCTX-M-orf477 was detected. Ten of the isolates carried plasmid mediated quinolone resistance gene - qnrS and 1 isolate possessed qnrB. Again 11 of them were found to have sulfonamide resistance gene - sul1 and 12 possessed class I integron. Sequencing of the class 1 integron revealed the presence of dfrA12/dfrA17 and aadA2/aadA5 gene cassettes conferring resistance to trimethoprim and aminoglycosides, respectively. All the isolates, characterized by enterobacterial repetitive intergenic consensus (ERIC) PCR, yielded distinct fingerprinting profile. However, most of the isolates from Jharkhand were clustered along with two isolates each from West Bengal and Mizoram indicating their clonal relatedness even though isolated from geographically different areas. Isolation of ESBL producing KP from bovine milk samples implies its public health significance; as such pathogens may enter the human food chain causing severe health hazards.

Molecular characterization and antibiotic susceptibility pattern of caprine Shiga toxin producing-Escherichia coli (STEC) isolates from India.

The present study was conducted to detect the occurrence, serotype, genotype, phylogenetic relationship and antimicrobial resistance pattern of STEC from healthy goats of West Bengal, India. From the 125 faecal samples collected from healthy goats, 245 isolates were identified as Escherichia coli. The E. coli harbouring any gene for Shiga toxins (stx 1/stx 2) was detected in 36 (14.7%) of the 245 E. coli isolates. These STEC strains belonged to 22 different serogroups (O2, O5, O20, O21, O22, O25, O41, O44, O45, O60, O71, O76, O84, O85, O87, O91, O103, O112, O113, O120, O156, and O158) and three were untypeable. The stx 1 and stx 2 was detected in 26 (72.2%) and 21 (58.3%) of Shiga toxin producing-E. coli (STEC) isolates, respectively. Further, E. coli harbouring eaeA only (Enteropathogenic E. coli) and ehxA was detected in 22 (61.1%) and 28 (77.7%) isolates, respectively. Whereas the saa was present in 8 (22.2%) E. coli isolates. The subtyping of the 26 E. coli strains possessing stx 1 showed that 73.% (19/26) of these isolates were positive for stx 1C subtype. Of the 21 isolates with the stx 2 gene, 42.8% (9/21) were positive for stx 2C, and 38.1% (8/21) were positive for stx 2d gene. The phylogenetic analysis of STEC strains after RAPD reveals eight major clusters. However, no serogroup specific cluster was observed. Resistance was observed most frequently to erythromycin (80.5%), amikacin (52.7%), cephalothin (50%), kanamycin (41.6%), neomycin (36.1%) and gentamycin (36.1%) and less frequently to norfloxacin (2.7%), enrofloxacin (2.7%), and ciprofloxacin (2.7%). Multidrug resistance was observed in eleven STEC isolates.

Comparative possession of Shiga toxin, intimin, enterohaemolysin and major extended spectrum beta lactamase (ESBL) genes in Escherichia coli isolated from backyard and farmed poultry.

The present work was conducted to compare the occurrence of Escherichia coli possessing virulence and ESBL genes in backyard and farmed poultry. Three hundred and sixty samples from the poultry kept in backyard system and 120 samples from the farmed birds were collected from West Bengal, India. Among the E. coli isolates of backyard poultry (O2, O10, O25, O55, O60, O106, UT), none of them possessed any of the Shiga toxin genes and eight E. coli isolates (8/272; 2.9%) harboured eaeA gene alone. Whereas among the E. coli isolated from the farmed poultry (O17, O20, O22, O102, O114, O119, rough, UT), four isolates (4/78, 5.1%) harboured stx 1/stx 2 gene and 11 isolates (11/78, 14.1%) possessed eaeA gene. None of the E. coli isolates from the backyard poultry harboured any studied ESBL gene. Whereas 29.4% of E. coli isolates from the farmed poultry were found to possess the ESBL genes.

Evaluation of egg production after adoption of biosecurity strategies by backyard poultry farmers in West Bengal.

AIM: On the basis of identified source of major bacterial infections at four agro-climatic zones in West Bengal the cost-effective biosecurity strategy was formulated for backyard poultry farmers. The aim of the present study was to assess the adoption. So, the study was aimed to detect the adoption level of the formulated biosecurity strategy to mitigate the Salmonella and Escherichia coliweek post-hatch period chicks were contamination level in the sources and its correlation with egg production in West Bengal. MATERIALS AND METHODS: A questionnaire was prepared querying regarding the biosecurity measures presently followed by the farmers, if any and egg production of their birds. Subsequent to the interview the formulated biosecurity strategy was conveyed. After 3 months, the interview with the same questionnaire was conducted to the same farmers to detect their adoption level. RESULTS: The change in practices were noted in certain parameters which differs significantly (p<0.01 or p<0.05). As a consequence, the average egg production/flock was increased in 3 months after adoption of the strategy (618.2±37.77/flock) in comparison to last 3 months average before adoption of the strategy (495.3±30.00/flock) which also differs significantly (p<0.01). CONCLUSION: The present study detected the implementation of the biosecurity strategy in backyard poultry farming in West Bengal can substantially benefit the farmers in terms of increased egg production.

Isolation, molecular characterization and antibiotic resistance of Shiga Toxin-Producing Escherichia coli (STEC) from buffalo in India.

In total, 363 Escherichia coli were isolated from 165 faecal samples of healthy buffaloes in West Bengal, India. Twenty-four of these isolates (6·61%) were found to carry at least one gene characteristic for Shiga toxin-producing Escherichia coli (STEC). These STEC strains belonged to 13 different O-serogroups. The stx1 gene was present in 23 (95·8%) of total STEC isolates, whereas 20 (83·3%) STEC isolates carried the gene stx2. Twelve strains of E. coli (50% of total STEC isolates) possessed enterohaemolysin (ehxA) gene in combination with others. Fourteen (58·33%) isolates found to possess saa gene. However, no E. coli was detected harbouring gene for intimin protein (eaeA). Of 23 stx1 -positive isolates, seven (30·43%) were positive for genes of the stx1C subtype. Of the 20 isolates with the stx2 gene, 25% (5/20) possessed stx2C and 10% (2/20) possessed stx2d gene. The phylogenetic analysis after RAPD of STEC strains revealed six major clusters. The isolated STEC strains were resistant most frequently to erythromycin (95·83%), cephalothin (62·5%), amikacin (54·17%), kanamycin (45·83%) and gentamicin (41·67%) group of antibiotics. No ESBL-producing (blaCTXM , blaTEM , blaSHV ) or quinolone resistance gene (qnrA) was detected in the STEC isolates.

Characterization of shiga toxin producing (STEC) and enteropathogenic Escherichia coli (EPEC) in raw yak (Poephagus grunniens) milk and milk products.

Thirty-one shiga toxin-producing (STEC) and 6 enteropathogenic Escherichia coli (EPEC) were isolated from 87 raw yak milk and 63 'churpi' samples. Of 18 stx(1) positive isolates (48.6%), 14 carried stx(1c) (77.7%). Subtyping of 28 stx(2) positive isolates (75.7%) revealed the presence of stx(2c) (9, 32.1%), stx(2d) (3, 10.7%), stx(2e) (1, 3.57%) and stx(2f) (3, 10.7%) variants. Furthermore, intimin (eaeA), enterohaemolysin (ehxA), autoagglutinating adhesin (saa), iha (adherence conferring protein), efa1 (EHEC factor for adherence), bundle forming pilli (bfpA) and toxB (type III secreted protein encoded on LEE Island, similar to toxin B of Clostridium difficile) genes were detected in 14, 16, 12, 4, 3, 2 and 2 isolates, respectively. Univariate and multivariate analysis depicted that both stx(1) and stx(2) or their variants were more likely to occur in isolates from Arunachal Pradesh (p<0.04) rather than Sikkim. Dendogram constructed on the basis of RAPD and ERIC PCR profile distributed the STEC and EPEC isolates in separate clusters irrespective of their sources and serotypes. The STEC and EPEC isolates exhibited resistance against erythromycin, amikacin, azithromycin, amoxicillin, ampicillin+cloxacillin, cephalothin, furazolidone, gentamicin, kanamycin, streptomycin and tetracycline. This is the first ever report on occurrence and characterization of STEC and EPEC isolated from yak milk and milk products.

Assessment of yeast cell wall as replacements for antibiotic growth promoters in broiler diets: effects on performance, intestinal histo-morphology and humoral immune responses.

The study compared the effects of an antibiotic growth promoter (AGP), yeast (Saccharomyces cerevisiae) and yeast cell wall (YCW) on performance, microbiology and histo-morphology of the small intestine and humoral immune responses in Ross 308 broilers. The treatments (eight replicates/treatment, n = 12/replicate) were negative control (NC, without AGP), positive control (PC, supplemented with bacitracin methylene disalicylate, 400 mg/kg), Y and YCW (supplemented with yeast and YCW, respectively, 1000 mg/kg). Live weight at 42 days improved (p = 0.086) in the PC, Y and YCW groups. Feed conversion ratio was better (p = 0.039) in the YCW group compared with the other groups. Antibiotic growth promoter in the PC group shortened the villi in duodenum (p = 0.044). Mucosal Escherichia coli number was higher in the PC group (p < 0.001), whereas in the digesta E. coli number was lower (p = 0.001) in the PC, Y and YCW groups in relation to the NC. Mucosal Salmonella populations increased (p = 0.0001) in the PC group, whereas in the digesta, all treatments reduced the Salmonella (p = 0.0001). Following oral challenge with Salmonella pullorum, YCW increased E. coli numbers on the mucosa (p < 0.001) whereas in the digesta the Y group had lower (p < 0.0001) number of E. coli. In the digesta, Salmonella count was lower in the YCW group compared with the other treatments (p < 0.01). Yeast cell wall -treated birds exhibited better (p < 0.05) humoral immune response against Newcastle disease which was far more persistent over time than in any other treatments. It was concluded that the yeast and the yeast cell wall may have effects identical to BMD on performance of broilers and thus may constitute an effective replacement strategy in the dietary regimens for broiler chickens.

Virulence repertoire of Shiga toxin-producing Escherichia coli (STEC) and enterotoxigenic Escherichia coli (ETEC) from diarrhoeic lambs of Arunachal Pradesh, India.

A total of 107 faecal samples were collected from diarrhoeic lambs of high altitude terrains (2,000 to 5,000 m above the mean sea level) of Tawang and West Kameng districts of Arunachal Pradesh, India. Total 234 Escherichia coli were isolated and further subjected to PCR for the study of virulence repertoire characteristics of Shiga toxin-producing E. coli (STEC) and enterotoxigenic E. coli (ETEC). Out of the 234 isolated E. coli, 32% were found positive for STEC, and 9% were carrying virulence gene for ETEC. The isolated STEC serogroups were O159, O127, O120, O113, O60, O30, O25, O8 and O2. Of all the 74 STEC strains, PCR showed that 18% isolates carried stx ( 1 ), 26% possessed stx ( 2 ) and 47% produced positive amplicon for both. Other virulent attributes like intimin (eaeA), enterohaemolysin (ehxA) and STEC auto-agglutinating adhesin (saa) were present in 18%, 43% and 44% of the isolates, respectively. The isolated ETEC serogroups were O172, O170, O159, O146, O127, O120, O113, O86, O75, O60, O30, O25, O8, O2, OR and OUT. Of the 22 ETEC-positive isolates, 23%, 18% and 4.5% possessed the gene only for LT, STa and STb, respectively, whereas 54% carried genes for both LT and STb. Some serogroups of E. coli like O159, O127, O120, O113, O60, O30, O25, O8 and O2 possessed genes for both Shiga toxin and enterotoxin. This study is the first report of ETEC isolation from diarrhoeic lambs in India. The moderately high proportion of STEC and ETEC in the diarrhoeic lambs implicated that these animals are important reservoir of STEC and ETEC. This is really a grave concern for the 'brokpas' and nomads (shepherds) who share a close relationship with this animals for their livelihood. This study also indicates that ETEC may be a major cause for frequent diarrhoeal episodes in lambs of this region.

Serological evidence of antibodies against Chlamydophila abortus in free-ranging yak (Poephagus grunniens) in Arunachal Pradesh, India.

Serum samples were randomly collected from 172 free-ranging yak (Poephagus grunniens, presently Bos grunniens) from six different yak tracts of Arunachal Pradesh, India, and subjected to enzyme-linked immunosorbent assay to detect the presence of specific antibodies against Chlamydophila abortus. The overall prevalence of this disease in yak was 35%. The prevalence of Cp. abortus-specific antibodies was significantly higher in yak cows (41%) than among bulls (25%). The highest prevalence (39%: 95% confidence interval [Cl] = 27, 55) was found in yak between one and three years of age, while the lowest prevalence (20%: 95% CI = 10, 41) was reported in yak below one year of age.