Antimicrobial Resistance Profiles of Campylobacter spp. Recovered from Chicken Farms in Two Districts of Bangladesh.

The rapid emergence of antimicrobial resistance (AMR) in Campylobacter has reinforced its status as a foodborne pathogen of significant public health concern. Resistant Campylobacter is typically transferred to humans via the consumption of contaminated animal products, particularly poultry. The genes associated with antimicrobial resistance in Campylobacter spp. are poorly understood. To address this knowledge gap, we conducted a prevalence survey of AMR Campylobacter across 84 chicken farms in two districts of Bangladesh. Pooled cloacal swabs were collected from chickens and underwent bacteriological testing for Campylobacter spp. with PCR confirmation. Antimicrobial susceptibility was tested against 14 antibiotics by disk diffusion method, and 12 resistance genes were screened in Campylobacter-positive isolates using multiplex PCR. A total of 34 (40.5%) farms were Campylobacter-positive of which 73.5% of isolates were resistant to at least 10 antibiotics. The antimicrobial susceptibility results indicate a high level of resistance against streptomycin (97.1%), clindamycin (97.1%), ampicillin (94.1%), tetracycline (94.1%), erythromycin (91.2%), ciprofloxacin (88.2%), nalidixic acid (85.3%), and imipenem (82.4%), and comparatively a low frequency of resistance to chloramphenicol (47.1%), ceftazidime (44.1%), and colistin (35.3%). Multidrug-resistant (MDR) and extensively drug-resistant Campylobacter were identified in 97.1%, and 50% of isolates, respectively. Ten resistance genes were identified including blaTEM (in 97.1% of isolates), strA-strB (85.9%), tetA (70.6%), tetB (32.4%), qnrS (23.5%), blaCTX-M-1 (20.6%), qnrB (20.6%), blaSHV (8.8%), aadB (5.9%), and qnrA (2.9%). Our findings demonstrate that resistance to ampicillin, tetracycline, and ceftazidime in Campylobacter isolates was significantly (p ≤ 0.05) associated with the presence of blaTEM, tetA, and blaSHV genes, respectively. The high rates of AMR in Campylobacter isolates from our study are not surprising given the liberal use of antimicrobials and incomplete biosecurity provisions on farms. Of particular concern are resistance rates to those classes of antibiotics that should be reserved for human use (azithromycin, ciprofloxacin, and colistin). AMR was more prevalent in chicken farms that used multiple antibiotics, engaged in prophylactic treatment of the birds, and improperly disposed of antibiotic packages. The high prevalence of MDR in chicken-derived Campylobacter isolates from the different regions of our study reinforces the need for more prudent use of antimicrobial compounds in Bangladeshi chicken farms.

Effective drying processes for Taikor (Garcinia pedunculata Roxb.) fruit by ultrasound-assisted osmotic pretreatment: Analysis of quality and kinetic models.

The present study aimed to analyze and establish an effective combination of ultrasound and immersion pretreatment processes for drying Taikor (Garcinia pendunculata Roxb.) fruits. Taikorslices were first immersed in 10 % sucrose, fructose, and glucose solution. Then, the immersed slices were treated in an ultrasonic bath at 30 °C for 10, 20, and 30 min. Drying operations were carried out at 50, 60, and 70 °C, with a fixed relative humidity of 30 %. The Page, Newton, Henderson and Pabis, and Weibull distribution models were fitted to the obtained drying data to determine the best kinetic model that effectively describes the drying properties ofTaikor. After drying operations, changes in quality parameters, e.g., ß-carotene, vitamin C, B vitamins, color, antioxidant activities, and microbial loads, were measured to obtain the best drying temperature and the most effective pretreatment combination with minimum loss of nutrients of the sample. Among different kinetic models, both Page and Weibull distribution models showed the best R2 values of 0.9867 and 0.9366, respectively. The chemical properties were preserved to the greatest extent possible by drying at 50 °C with glucose pretreatment. The color parameters were better preserved by fructose pretreatment. Sonication time also had profound effect on the quality parameters of dried Taikor slices. However, higher temperature drying required a shorter time for drying and exhibited better performance in microbial load reduction. This study's findings will help to establish an effective drying condition forGarcinia pedunculatafruits.

Sociodemographic determinants of use and misuse of antibiotics in commercial poultry farms in Bangladesh.

Objectives: This study was conducted to assess poultry farmers' knowledge and practices regarding antibiotics, antimicrobial usage (AMU), and antimicrobial resistance (AMR), and to identify the sociodemographic factors of inappropriate use of antibiotics in commercial poultry farms in Bangladesh. Methods: A qualitative survey of 140 farmers in Bangladesh was conducted from March to May 2019. A logistic regression model was used to identify factors associated with the inappropriate use of antibiotics. Results: 47.1% of farmers were unable to explain antibiotics, 42.9% used antibiotics for preventive purposes, 4.3% used them as growth promoters, 25.7% used them as suggested by veterinarians, 42.9% used leftover antibiotics, 50% did not maintain antibiotics residual withdrawal period, and 98.6% did not know about AMR. In bivariable regression analysis, sex and primary occupation of poultry farmers, their knowledge about withdrawal periods for antibiotics, and no contact with veterinary surgeons (VS) were found to be significantly associated with the inappropriate use of antibiotics, while only 'no contact with VS' was identified in multivariable regression analysis. Conclusions: The findings suggest an urgent need to improve understanding of antibiotics and AMR. Adequate supervision by veterinarians would ensure adherence to appropriate AMU patterns, and would limit the misuse of antibiotics and associated AMR development in farms.

Sink survey to investigate multidrug resistance pattern of common foodborne bacteria from wholesale chicken markets in Dhaka city of Bangladesh.

Antimicrobial resistance (AMR) among foodborne bacteria is a well-known public health problem. A sink survey was conducted to determine the AMR pattern of common foodborne bacteria in cloacal swab of broiler chickens and sewage samples from five wholesale chicken markets of Dhaka city in Bangladesh. Bacteria were identified by culture-based and molecular methods, and subjected to antimicrobial susceptibility testing. Resistance genes were identified by multiplex PCR and sequencing. Multidrug resistance (MDR) was observed in 93.2% of E. coli, 100% of Salmonella spp., and 97.2% of S. aureus from cloacal swab samples. For sewage samples, 80% of E. coli, and 100% of Salmonella and S. aureus showed MDR. Noteworthy, 8.3% of S. aureus from cloacal swab samples showed possible extensively drug resistance. Antimicrobial resistance genes (beta-lactamase-blaTEM, blaSHV; quinolone resistance gene-qnrS) were detected in a number of E. coli and Salmonella isolates from cloacal swab and sewage samples. The methicillin resistance gene (mecA) was detected in 47.2% and 25% S. aureus from cloacal swab and sewage samples, respectively. The findings envisage the potential public health risk and environmental health hazard through spillover of common foodborne MDR bacteria.

Epidemiology and antimicrobial resistance of Escherichia coli in broiler chickens, farmworkers, and farm sewage in Bangladesh.

BACKGROUND: Antimicrobial resistance (AMR) has become an emerging threat worldwide, and developing countries such as Bangladesh are considered to be at greater risk of disseminating the resistant bacteria between human-animal interfaces. OBJECTIVES: The present study was carried out to determine the prevalence and AMR profile of Escherichia coli isolated from broiler chickens, the environment, and farmworkers. This study also aimed to identify the risk factors associated with multidrug-resistant (MDR) E. coli infection in broiler chickens. In addition, the presence of carbapenem resistance gene (NDM-1) was assessed. METHODS: A total of 114 E. coli isolates, recovered from 150 samples (cloacal swabs = 50, farm sewage = 50, and hand washed water of farmworkers = 50) collected from 50 broiler farms, were identified by biochemical examination and polymerase chain reaction (PCR) assay. Antimicrobial susceptibility test was performed for 10 antibiotics by disk diffusion test. Carbapenem resistance gene (NDM-1) was detected by PCR. Risk factors were identified through multivariable logistic regression. RESULTS: The highest prevalence of E. coli was recorded in broiler chickens (86%) and the lowest in farmworkers (66%). For MDR E. coli infection, 'winter season', 'absence of specific shoes for staff', and 'use of antibiotics without veterinarian's prescription' were the significant risk factors. High resistance of the E. coli isolates was observed against levofloxacin (81.6%), doxycycline (78.1%), cefotaxime (78.1%), and ciprofloxacin (70.2%). About 76% of the isolates demonstrated MDR. None of the isolates were positive for the NDM-1 gene. CONCLUSIONS: The high level and similar pattern of antibiotic resistance in E. coli isolates from broiler chickens, farmworkers, and sewage in poultry farms indicates a good possibility of spreading the antibiotic-resistant E. coli in such settings.

Epidemiology and antimicrobial resistance profiles of Salmonella in chickens, sewage, and workers of broiler farms in selected areas of Bangladesh.

INTRODUCTION: A cross-sectional study was conducted to determine the prevalence and AMR pattern of Salmonella isolated from broiler chickens, farm sewage, and farm workers. This study also aimed at identifying the risk factors for Salmonella infection in chickens. METHODOLOGY: Cloacal swabs (n = 50) from broiler chickens, farm sewage (n = 50), and hand washed water of farm workers (n = 50) were collected along with data on farm management and antimicrobial usage from 50 broiler farms of Mymensingh and Gazipur districts. All samples were analysed for the presence of Salmonella using selective media and PCR assay. Antimicrobial susceptibility test was done for ten antimicrobials by disk diffusion test. Risk factor analyses were carried out by multivariable logistic regression using SPSS. RESULTS: The overall Salmonella prevalence was 66% (99/150). Salmonella prevalence were 82% and 72% in cloacal swabs and farm sewage samples, respectively. From hand washed water, 44% of the samples were positive for Salmonella. Salmonella infection in broiler chickens was significantly associated with farming experience (≤ 5 years) and age of birds (≥ 11 days). Similar pattern of antimicrobial resistance was observed in Salmonella isolated from three types of samples, and high resistances were observed to colistin, doxycycline, ciprofloxacin and ceftazidime. Moreover, isolates from all the three sources showed high percentage of multidrug-resistance (80.6% to 97.6%). CONCLUSIONS: The findings of this study reveal that antimicrobial-resistant Salmonella are prevailing at animal-human-environment interface, which needs an integrated approach to limit further spread of AMR.

Prevalence and Multidrug Resistance Pattern of Methicillin Resistant S. aureus Isolated from Frozen Chicken Meat in Bangladesh.

Infections by methicillin-resistant Staphylococcus aureus (MRSA) are continuously expanding within the community. Chicken meat is usually contaminated by MRSA, and this contaminated chicken meat is an important source of foodborne infections in humans. In this study, a cross-sectional supershop survey was conducted to determine the prevalence and antimicrobial resistance pattern of MRSA in 113 domestic frozen chicken meat samples purchased from nine branded supershops available in five divisional megacities of Bangladesh. The study also focused on the determination of methicillin resistance gene in MRSA isolates. S. aureus was identified by standard culture-based and molecular methods, and subjected to antimicrobial susceptibility testing. MRSA was screened by cefoxitin disk diffusion test. Methicillin resistance gene was identified by PCR. Of samples, 54.9% were positive for S. aureus, and, of these, 37.1% isolates were identified as MRSA. All the isolates were multidrug resistant (MDR): 52.2% were resistant to 6-8 antimicrobial classes, and 47.8% isolates to 9-12 classes. Three (3.2%) isolates of S. aureus were possible extensively drug resistant. The highest rates of resistance were observed against cefoxitin (100%), followed by nalidixic acid, ampicillin and oxacillin (97.7%), colistin (91.3%), amoxicillin-clavulanic acid and amoxicillin (87%), penicillin-G and cloxacillin (82.6%), oxytetracycline (78.3%), and cefixime (73.9%). Screening of methicillin resistance gene revealed that 43.5% isolates of MRSA were positive for mecA gene. The high prevalence of MDR MRSA in frozen chicken meat samples in this study emphasizes the need for better sanitary education of food handlers in hygienic practices focusing on their potential role as reservoirs and spreaders of MRSA.

Dietary supplementation of Neem (Azadirachta indica) leaf extracts improved growth performance and reduced production cost in broilers.

BACKGROUND AND AIM: Global trend to remove the antibiotic growth promoter (AGP) from animals contributes to the exploration of successful measures to sustain production and reduces the intestinal diseases in the post-AGP era. Plant extracts, therefore, have been used to improve performance and intestinal health. Here, we conducted a study to evaluate the effects of neem (Azadirachta indica) leaf extracts (NLE) as alternatives to AGPs in broiler chickens. MATERIALS AND METHODS: Sixty day-old broiler chicks were assigned to 12-floor pens, each stocked with five birds and divided into three treatment groups of four pens per treatment. The groups were: Negative control, basal diet without additives; positive control, basal diet with antibiotics and vitamins; and NLE treated group, basal diet supplemented with 0.1% aqua extract of neem leaves. RESULTS: Overall feed intake was significantly (p≤0.05) highest in the positive control. Higher body weight, higher dressing percentage, and lower feed conversion ratio were observed in birds treated with NLE compared to the negative control group (p≤0.05) but not the positive control group (p>0.05). There was no significant variation in hematology between different groups. Furthermore, the economic evaluation indicated that the NLE treatment was found cheaper than control and antibiotic treatment in cost-benefit analysis. CONCLUSION: We suggest NLE might be a cheaper alternative to antibiotics in broiler production as indicated by improved body weight and feed efficiency.

Antimicrobial Resistance Pattern of Escherichia coli Isolated from Frozen Chicken Meat in Bangladesh.

Escherichia coli is known as one of the most important foodborne pathogens in humans, and contaminated chicken meat is an important source of foodborne infection with this bacterium. The occurrence of extended-spectrum ß-lactamase (ESBL)-producing E. coli (ESBL-Ec), in particular, in chicken meat is considered a global health problem. This study aimed to determine the magnitude of E. coli, with special emphasis on ESBL-Ec, along with their phenotypic antimicrobial resistance pattern in frozen chicken meat. The study also focused on the determination of ESBL-encoding genes in E. coli. A total of 113 frozen chicken meat samples were purchased from 40 outlets of nine branded supershops in five megacities in Bangladesh. Isolation and identification of E. coli were done based on cultural and biochemical properties, as well as PCR assay. The resistance pattern was determined by the disc diffusion method. ESBL-encoding genes were determined by multiplex PCR. The results showed that 76.1% of samples were positive for E. coli, of which 86% were ESBL producers. All the isolates were multidrug-resistant (MDR). Resistance to 9-11 and 12-13 antimicrobial classes was observed in 38.4% and 17.4% isolates, respectively, while only 11.6% were resistant to 3-5 classes. Possible extensive drug resistance (pXDR) was found in 2.3% of isolates. High single resistance was observed for oxytetracycline (93%) and amoxicillin (91.9%), followed by ampicillin (89.5%), trimethoprim-sulfamethoxazole, and pefloxacin (88.4%), and tetracycline (84.9%). Most importantly, 89.6% of isolates were resistant to carbapenems. All the isolates were positive for the blaTEM gene. However, the blaSHV and blaCTX-M-2 genes were identified in two ESBL-non producer isolates. None of the isolates carried the blaCTX-M-1 gene. This study provided evidence of the existence of MDR and pXDR ESBL-Ec in frozen chicken meat in Bangladesh, which may pose a risk to human health if the meat is not properly cooked or pickled raw only. This emphasizes the importance of the implementation of good slaughtering and processing practices by the processors.

Prevalence and risk factors of Campylobacter infection in broiler and cockerel flocks in Mymensingh and Gazipur districts of Bangladesh.

Campylobacter spp. is one of the most frequent causes of foodborne gastroenteritis. This study aimed to estimate the prevalence and to identify the risk factors of farm-level Campylobacter infection in meat-type chicken flocks. A cross-sectional study was conducted in two selected districts of Bangladesh over the period of January to July 2019. A total of 84 pooled cloacal swab samples were collected from 84 broiler and cockerel farms. Data on farm management, biosecurity, and hygiene practices were collected using a structured questionnaire through a face-to-face interview during sampling. Thereafter, Campylobacter spp. were isolated through bacteriological culture and identified by Gram staining and biochemical tests. Furthermore, the isolates were confirmed using the polymerase chain reaction by targeting the 16S rRNA gene. The risk factors were analyzed at the farm level using multivariable logistic regression with the significant levels of P-value ≤ 0.05. Among the 84 farms, 34 were positive to Campylobacter spp.; thus, the prevalence was estimated to be 40.5% (95% CI: 30.1%-51.8%). In risk factor analysis, the following factors were found to be significantly associated with Campylobacter infection: shed older than five years, birds older than 30 days, flock size with more than 1500 birds, downtime less than seven days, no disinfection of shed surroundings during rearing, rice husk as litter materials, and less than 10 years of farming experience. The study identified the factors that could lead to the setting of effective interventions in controlling Campylobacter infection in chickens to reduce campylobacteriosis in humans through meat consumption.

Antibacterial efficacy of ethanolic extract of Camellia sinensis and Azadirachta indica leaves on methicillin-resistant Staphylococcus aureus and shiga-toxigenic Escherichia coli.

OBJECTIVE: This study aims at investigating the antibacterial potential of ethanolic extract of Camellia sinensis (common name: Green tea) and Azadirachta indica (common name: Neem) leaves on methicillin-resistant Staphylococcus aureus (MRSA) and shiga-toxigenic Escherichia coli (STEC). MATERIALS AND METHODS: Fresh leaves were processed and extracted by 99% ethanol and reconstituted with 50% ethanol before testing. Disk diffusion and broth microdilution methods were used to determine zone diameter of inhibition (ZDI) and minimum inhibitory concentration (MIC), respectively. Nutrient agar plate was used to estimate the minimum bactericidal concentration (MBC). RESULTS: Maximum ZDI value was observed for green tea against MRSA (7.5 mm) and minimum for neem (4.9 mm). Moreover, the highest ZDI against STEC was also for green tea and the combination of green tea and neem (4.5 mm). The MIC values of green tea extract were 15.625 and 31.25 mg/ml against MRSA and STEC, respectively, whereas the MIC of neem was 31.25 and 125 mg/ml, respectively. The combination had similar MIC (46.87 mg/ml) against both organisms. Green tea showed the lowest MBC values, 31.25 and 62.5 mg/ml, against MRSA and STEC, respectively. However, MBC of neem and the combination against MRSA and STEC were found >250 mg/ml, >500 mg/ml and 93.75 mg/ml, >375 mg/ml, respectively. CONCLUSION: Green tea and neem leaves showed good antimicrobial effects and can be used to explore novel antimicrobial compounds against MRSA and STEC.