Detection of Campylobacter spp. in farmed sheep in Mymensingh division of Bangladesh: Prevalence, risk factors, and antimicrobial susceptibility status.

Background and Aim: Campylobacter infections in sheep may be asymptomatic or cause enteritis, ileitis, infertility, and abortion. Thus, this study aimed to estimate the prevalence of Campylobacter spp. in farming sheep and to detect risk factors, molecular patterns, and antimicrobial susceptibility status of these pathogens. Materials and Methods: Four hundred and eight fecal samples were collected from 12 flocks in the Mymensingh and Sherpur districts. Samples were tested by both basic (culture and biochemical tests) and molecular (initially 16S rRNA and later hipO gene-based polymerase chain reaction). Furthermore, the antimicrobial susceptibility status of Campylobacter jejuni was confirmed using disk diffusion. Flock- and animal-level data were captured using semi-structured interviews with farm owners under bivariate and multivariate logistic regression analyses to confirm the risk factors for Campylobacter-positive status. Results: The prevalence of C. jejuni staining at the animal and flock levels was 8.82% (36/408) and 66.70% (8/12), respectively. The age of sheep was identified as an important risk factor. Up to 1 year of age, sheep were 3.78 times more likely to be infected with C. jejuni (95% confidence interval: 1.0736-13.3146, p = 0.038). Of the 36 isolates of C. jejuni, all were found to be fully susceptible (100%) to gentamicin and ciprofloxacin. In this study, three antimicrobial agents, oxytetracycline, azithromycin, and ceftriaxone, were fully resistant (100%). The majority of isolates were resistant to a combination of 4-6 antimicrobial agents. Conclusion: The present study highlights the predominant maintenance of zoonotic Campylobacter species in sheep, and their burden on human health is enormous. Therefore, environmental, animal, and human health needs to be focused under a One Health lens to mitigate the occurrence of Campylobacter in farm settings and to prevent further introduction to animals and humans.

Investigation of Clostridium perfringens in small-scale commercial broiler flocks in Mymensingh district of Bangladesh.

BACKGROUND AND AIM: Necrotic enteritis (NE) is one of the most prevalent diseases in broiler poultry caused by Clostridium perfringens connected with significant economic losses. A cross-sectional survey was conducted in Mymensingh district of Bangladesh to assess the prevalence of C. perfringens through toxinotyping molecular assay and confirm the risk factors for NE, including antimicrobial-resistant (AMR) status of the isolates. MATERIALS AND METHODS: We included 40 small-scale commercial broiler farms randomly selected from two subdistricts of Mymensingh district of Bangladesh. As an individual sample, 240 cloacal swabs, and as a pooled sample, 40 drinking water, 40 workers' hand washing, 40 litter swab, and 40 feed samples were collected and evaluated by culture, biochemical, and molecular assays. A pretested semi-structured interview questionnaire was employed to capture flock-level data on risk factors from the farm owners. The flock-level data on risk factors were assessed through univariable and multivariable logistic regression analyses with p<0.05 was considered statistically significant. RESULTS: Overall flock-level prevalence of C. perfringens was estimated to be 10.3% (95% confidence interval [CI] 7.5-13.6%). Litter swab (pooled) was found to be highly contaminated with C. perfringens (25.0%, 95% CI: 12.7-41.2%) followed by the cloacal swab (10.4%, 95% CI: 6.9-15.0%) and feed sample (5.0%, 95% CI: 0.6-16.9%). History of coccidia infection (Adjusted odds ratio =33.01, 95% CI: 2.14-507.59, p=0.01) was significantly associated with flock-level C. perfringens infection status. In this study, 78.1% isolates were found as multidrug-resistant as they demonstrated resistance to 3-5 antimicrobial agents. CONCLUSION: Evidence-based control options need to be taken through the uses of prebiotics and probiotics, biosecurity, and hygienic measurement, including control of coccidia infection, is needed to lessen the NE infection and AMR related to this pathogen in small-scale commercial broiler poultry.

Prevalence and Antimicrobial Resistance of Campylobacter Species in Diarrheal Patients in Mymensingh, Bangladesh.

Campylobacter enteritis is the leading cause of gastroenteritis in humans worldwide including Bangladesh. The objectives of this study were to estimate the prevalence and antimicrobial-resistance status of Campylobacter spp. in human diarrheal samples collected from Surya Kanta Hospital, Mymensingh, Bangladesh. In this study, we evaluated a total of 330 clinical samples for the presence Campylobacter spp. via cultural and biochemical tests and molecular assays. Furthermore, antimicrobial susceptibility testing for Campylobacter species was accomplished by the standard agar disc diffusion technique against eight commercially available antimicrobial agents. A pretested semistructured questionnaire was used to capture the data on socioanthropological factors from the diarrheal patients. Pearson's chi-square test was performed, and a p value of <0.05 was considered for the level of significance. Nearly one in three diarrheal patients admitted in this hospital were infected with Campylobacter spp. Overall prevalence of Campylobacter spp. was estimated to be 31.5% (104/330) that comprised the prevalence of C. jejuni, 21.8% (n = 72), and C. coli, 9.6% (n = 32). Among the positive cases, the prevalence of Campylobacter was higher in the age group 0-5 years (52%) followed by 6-18 years (42.7%), 19-40 years (34.0%), 41-60 years (25.4%), and >60 years (10.5%). Age, family level's personal hygiene, and involvement with animal husbandry were captured as potential determinants to be associated with the Campylobacter positive status. Among the isolates, 27.3% (n = 20) of C. jejuni and 31.2% (n = 10) of C. coli demonstrated as multidrug-resistant (MDR) to three or more antimicrobial agents. The present study shows that Campylobacter spp. is most prevalent among the hospital-admitted diarrheal patients, and proper measures should be taken to reduce the burden focusing on the potential determinants.

Prevalence, Risk Factors, and Molecular Detection of Campylobacter in Farmed Cattle of Selected Districts in Bangladesh.

A cross-sectional survey was conducted in selected districts of Bangladesh to estimate prevalence, risk factors, and molecular detection of Campylobacter isolates from 540 farmed cattle of 90 herds. As an individual sample, 540 feces, and as a pooled sample, 180 milk samples, 90 feed samples, 90 water samples, 90 manure samples, and 90 animal attendants' hand-rinse water were collected and tested via culture, biochemical, and molecular assays. A pretested semi-structured questionnaire was used to collect herd-level data on risk factors with the herd owners. The herd-level data on risk factors were analyzed through univariate and multivariate analyses, and a p-value <0.05 was considered statistically significant for all analyses. Overall, farm-level prevalence of bovine Campylobacter was enumerated to be 53.3% (95% confidence interval [CI]: 42.5-63.9%). The feces sample was found to be a high level of contamination of 30.9% (95% CI: 27-35%) followed by the manure swab (pooled) at 15.6% (95% CI: 8.8-24.7%). Campylobacter jejuni was documented as an abundant species (12.6%), followed by Campylobacter coli (5.1%), and Campylobacter fetus (0.3%). Older farms (>5 years of age), no/minimum cleaning and disinfection practices, along with animal roaming outside of the farm, were documented as significant risk factors for farm-level Campylobacter occurrence. Evidence-based control measures need to be taken through stringent biosecurity and hygienic measurement to lessen the load of the Campylobacter pathogen in the farm environment and prevent further transmission to animals and humans.

Salmonella Gallinarum in Small-Scale Commercial Layer Flocks: Occurrence, Molecular Diversity and Antibiogram.

Salmonella Gallinarum is one of the most important bacterial pathogens associated with diminished egg production in poultry. The aim of this study was to understand the occurrence, molecular traits and antimicrobial resistance patterns of Salmonella Gallinarum strains isolated from small-scale commercial layer flocks with low level biosecurity standards in Bangladesh. A total of 765 samples, including cloacal swabs (535), visceral organs (50), and droppings (180), were collected from chickens of 12 layer flocks in 11 districts. Salmonella Gallinarum was isolated and characterized through culture-based method, followed by biochemical tests, sero-grouping, PCR assays, sequencing, and antibiogram. The identity of biochemically detected isolates of Salmonella Gallinarum was confirmed via genus-specific 16S rRNA gene based PCR, followed by invA and spvC genes based PCR assays. Occurrence of Salmonella Gallinarum was detected in overall 25.75% (197/765) samples, with a significantly (p < 0.05) higher incidence in visceral organs (42%) in comparison to cloacal swab (24%) and droppings (26%). Sequencing and subsequent phylogenetic analysis of invA and spvC genes in representative strains of Salmonella Gallinarum revealed a close genetic lineage, with a sequence similarity of 98.05-99.21% and 97.51-99.45%, respectively, to previously published sequences of the corresponding genes from the same serogroup strains. Remarkably, 66.5% (131/197) of the isolated strains of Salmonella Gallinarum were found to be resistant to 3 to 6 antimicrobial agents, and interpreted as multidrug resistant (MDR). The findings of this study underscore an inherent need of appropriate control measures to curb the widespread incidence of MDR Salmonella Gallinarum in small-scale commercial layer flocks, thereby, facilitating enhanced egg production and further support to the food security and safety in low resource settings.

Occurrence of Campylobacter spp. in Selected Small Scale Commercial Broiler Farms of Bangladesh Related to Good Farm Practices.

Poultry origin Campylobacter is considered as one of the leading causal agents of human foodborne illness. This study was conducted to estimate the occurrence, molecular identification, and antimicrobial resistance (AMR) of Campylobacter species from the broiler farms in Bangladesh. Samples (352) were collected from 32 farms and comprised of 128 cloacal swab, 64 feed, 64 drinking water, 64 attendants' hand rinsed water, and 32 whole carcasses. All samples were tested for the presence of Campylobacter via cultural, biochemical, and PCR. The AMR was determined via the disc diffusion method. An overall occurrence of Campylobacter spp. was estimated as 26.4%. The level of Campylobacter contamination was found to be higher in conventional farms (36.4%) than the good practice farms (16.5%) including all sample categories (p = 0.000). Of 93 isolates, 67.74% and 32.26% were confirmed as C. jejuni and C. coli respectively, of which 34.92% C. jejuni, and 30% C. coli were shown to be multidrug-resistant. A significant occurrence of Campylobacter contamination in broiler farms with multidrug resistant patterns might be cogitated as serious food safety and public health concern linking to poultry food chain. A risk reduction approach through good farming practices targeting the prudent use of antimicrobials for broiler production is thus necessitated.