Biosecurity, health and disease management practices among the dairy farms in five districts of Bangladesh.

Herd health management is a planned program to optimize health, welfare, and production of dairy cows. A cross-sectional study focused on herd health management of dairy farms, was carried out to assess the biosecurity, health, and disease management practices with associated socio-demographic factors. A structured questionnaire with closed- and open-ended questions was employed for farmers' interviews to acquire information on dairy farms. Data were collected from randomly selected 205 dairy farms in five districts (Dhaka-Savar, Mymensingh, Gazipur, Rangpur, and Sirajganj) of Bangladesh. Overall, 1.5% farms had "good", 17% had "moderate" and 81.5% had "poor" levels of biosecurity, health and disease management practices (BHDMP). Analysis of variance revealed that mean BHDMP scores were significantly (p < 0.05) higher in farms of Rangpur (27.6 ± 0.8) and Gazipur (26.7 ± 0.6) than those of the other three districts. Educational status had a significant effect on BHDMP scores. The farms owned by graduated farmers had the highest mean BHDMP score (30.4 ± 1.1) while the farms operated by illiterate farmers had the lowest BHDMP score (20.4 ± 0.4). Multiple linear regression analysis also revealed that educational status (secondary to post-graduation), training experience, area of study (Rangpur district), and herd size were significantly associated with BHDMP score of the farms. The findings lay a basis for improving dairy farms' biosecurity, health and disease management practices, which further envisage the adoption and implementation of herd health management programs.

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.

Detection of antimicrobial resistance genes in Lactobacillus spp. from poultry probiotic products and their horizontal transfer among Escherichia coli.

The study was conducted to identify the antimicrobial resistance genes (ARGs) in Lactobacillus spp. from poultry probiotic products and their potential to spread among Escherichia coli. Lactobacillus spp. were isolated and identified from 35 poultry probiotic samples based on the cultural, biochemical, and molecular findings. All the isolates (n = 35) were screened for the presence of some ARGs such as ß-lactamases encoding genes (blaTEM, blaCTXM-1, and blaCTXM-2), plasmid-mediated quinolone resistance gene (qnrA, qnrB, and qnrS), and tetracycline resistance genes (tetA and tetB). Five Lactobacillus spp. isolates from three brands were positive for one or more ARGs. The qnrS was detected in four isolates. The blaTEM and tetB were detected in two isolates. One isolate contained blaCTX-M-1, blaCTX-M-2, and tetA genes. Brand-wise analysis revealed that one isolate from Brand 4 contained blaTEM, blaCTX-M-1, blaCTX-M-2, qnrS, and tetA genes, one isolate from Brand 2 contained blaTEM gene, and three isolates from Brand 7 harbored qnrS gene. The co-culture of Lactobacillus spp. and E. coli resulted in the transmission of qnrS, CTX-M-1, and tetA from Lactobacillus spp. to E. coli. Results of antimicrobial susceptibility test revealed that the highest resistance was observed to cefepime and cefotaxime followed by penicillin G, oxacillin, cefuroxime, and ofloxacin. The findings of the present study indicate the potential risk of horizontal spread of antimicrobial resistance through probiotic bacteria among the poultry population. Therefore, it is very necessary to check for ARGs along with other attributes of probiotic bacteria to avoid the inclusion of resistant strains in probiotics.

Epidemiology and economic impact of lumpy skin disease of cattle in Mymensingh and Gaibandha districts of Bangladesh.

Lumpy skin disease (LSD) is an emerging viral disease of cattle that negatively impacts livestock by reducing animals' production and increasing management costs. Like other countries, Bangladesh has also faced the ominous effects of LSD. Because of this pressing concern, the present study was undertaken to determine the attack rate and risk factors along with economic losses of the recent LSD outbreaks from October 2019 to June 2020 in Bangladesh's Mymensingh and Gaibandha districts. A total of 1187 cattle from 150 randomly selected cattle farms were surveyed. The diagnosis was made both clinically and molecularly. The overall attack rate, mortality and case fatality were 34%, 1.10% and 3.23%, respectively, but in calves up to 1 month of age, the attack rate, mortality and case-fatality were 41.6%, 12.5% and 30%, respectively. In risk factor analysis using multiple logistic regression, the following factors were significantly (p < .05) associated with LSD occurrence: no administration of goat pox vaccine, frequency of goat pox vaccination - single dose, introduction of new cattle, land type (low land and medium-high land), use of common syringe and needle, and use of common utensils. The average economic loss per case was 9384.41 BDT (110.40 US $). The higher economic loss per case was accounted in Mymensingh district (10248.91 BDT ≈ 120.58 US $) than Gaibandha district (8211.52 BDT ≈ 96.61 US $). The loss was higher in crossbred cattle (9709.58 BDT ≈ 114.23 US $) compared to indigenous cattle (7595.94 BDT ≈ 89.36 US $). The total estimated annual loss due to LSD in Mymensingh and Gaibandha districts was 7763.25 million BDT (91.33 million US $). In Gaibandha district, the estimated annual loss was 2666.20 million BDT (31.37 million US $) while in Mymensingh district it was 5097.05 million BDT (59.97 million US $). Findings of the study could provide useful epidemiological data with losses incurred due to LSD to the livestock owners, field veterinarians and government-level livestock regulators, which will help to formulate prevention and control strategies and minimize the negative impact of LSD's probable return or spread.

Prevalence of Methicillin and ß-Lactamase Resistant Pathogens Associated with Oral and Periodontal Disease of Children in Mymensingh, Bangladesh.

Oral and periodontal diseases (OPD) is considered one of the main problems of dentistry worldwide. This study aimed to estimate the prevalence of oral and periodontal pathogenic bacteria along with their antimicrobial resistance pattern in 131 children patients aged between 4-10 years who attended in Mymensingh Medical College Hospital during October 2019 to March 2020. OPD pathogens were identified through isolation, cultural and biochemical properties, and nucleic acid detection. The isolates were subjected to antimicrobial susceptibility to 12 antibiotics commonly used in dentistry. In addition, the isolates were analyzed molecularly for the presence of six virulence and three antibacterial resistance genes. Five pathogens were identified, of which Staphylococcus aureus (S. aureus) (49%) and S. salivarius (46%) were noticed frequently; other bacteria included S. mutans (16.8%), S. sobrinus (0.8%) and L. fermentum (13.7%). The virulence genes-clumping factor A (clfA) was detected in 62.5% isolates of S. aureus, and gelatinase enzyme E (gelE) gene was detected in 5% isolates of S. salivarius, while other virulence genes were not detected. All the tested isolates were multidrug-resistant. The overall prevalence of MDR S. aureus, Streptococcus spp. and L. fermentum was 92.2%, 95.1% and 100%, respectively. It was observed that a high proportion of isolates were found resistant to 5-8 antibiotics. A majority of S. aureus, Streptococcus spp., and L. fermentum isolates tested positive for the ß-lactamase resistance genes blaTEM and cfxA, as well as the methicillin resistance gene mecA. Phylogenetically, the resistance genes showed variable genetic character among Bangladeshi bacterial pathogens. In conclusion, S. aureus and S. salivarius were major OPD pathogens in patients attended in Mymensingh Medical College Hospital of Bangladesh, and most were Beta-lactam and methicillin resistant.

Prevalence of Dental Caries in Children in Mymensingh and Its Associated Risk Factors: A Cross-Sectional Study.

BACKGROUND: Children suffer from various oral and periodontal diseases. Dental caries is one of the most prevalent oral diseases among children in the world. This study was conducted to identify the prevalence and risk factors of dental caries in children in Mymensingh, Bangladesh. METHODS: A cross-sectional study was conducted on 362 pediatric patients who attended the Dental Unit of Mymensingh Medical College from March to September 2019. The sample size was calculated using a statistical formula and the children were selected using a systematic random sampling technique. Children and their guardians were interviewed and data were recorded using a structured questionnaire. Risk factors were analyzed using multivariate logistic regression. RESULTS: The overall prevalence of dental caries was 82.7%. The prevalence of caries was significantly higher in aged children (8-10 years) and also in rural, low-income, and illiterate families. Seven significant risk factors were identified that included residence in the rural area (OR: 7.31 [1.73-30.83]), a parental income of BDT ≤ 20,000 per month (OR: 4.75 [1.49-15.05]), reduced duration (≤1 min) of teeth cleaning (OR: 18.54 [2.05-168.17]), teeth cleaning before breakfast (OR: 93.30 [10.95-795.32]), the spoon-feeding method (OR: 12.57 [2.09-75.61]), long-term (37-48 months) breastfeeding (OR: 212.53 [8.69-5195.25]), and family oral problem (OR: 8.20 [2.57-26.16]). CONCLUSIONS: The prevalence of dental caries among the children in Mymensingh is very high and was associated with residence in rural areas, parental income, reduced duration of teeth cleaning, teeth cleaning before breakfast, the spoon-feeding method, long-term breastfeeding, and family oral problems.

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.

Metabolic profiling in periparturient dairy cows and its relation with metabolic diseases.

OBJECTIVE: Periparturient period is associated with multiple changes including serum concentration of macro minerals and drop in feed intake. Therefore, it is essential to know the actual concentrations of major macro minerals, glucose and ketone bodies in blood during the periparturient period. The objectives of the study were to study the dynamics of calcium, magnesium, phosphorus, and glucose in serum and ketone bodies in the urine of periparturient cows and to estimate the incidence of subclinical metabolic diseases. RESULTS: Results showed that all the urine samples were negative for ketone bodies. Incidence of subclinical form of hypocalcaemia, hypomagnesaemia, hypophosphatemia, and hypoglycemia in periparturient cows was 31.03%, 48.28%, 17.24% and 55.17%, respectively. Older cows with high BCS and milk yield were mostly affected with a subclinical form of hypocalcaemia and hypoglycemia. No significant differences were observed in serum level of macro minerals and glucose at different time points of periparturient period of cows fed with a balanced ration, and between two groups of cows those were given IV injection of calcium and magnesium on the day of parturition and those were not given. Age, parity, and milk yield had no significant effect on the concentration of calcium, magnesium, phosphorus, and glucose.

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.

Evaluation of sodium lauryl sulfate for the development of cow-side mastitis screening test.

BACKGROUND AND AIM: Subclinical mastitis (SCM) is an economically important disease for dairy cattle worldwide; therefore, regular screening is imperative to detect SCM at an early stage so as to control it. The study was conducted to compare the test characteristics of sodium lauryl sulfate (SLS) as a test reagent to detect SCM in dairy cows. MATERIALS AND METHODS: First, 106 milk samples of dairy cows were subjected to available indirect screening tests (white side test [WST], surf field mastitis test, Leucocytest, and Immucell) considering somatic cell count (SCC) as gold standard test. Then 273 milk samples were allowed to react with different concentrations of SLS with or without sodium hydroxide (NaOH) and indicators (bromothymol blue [BTB] and bromocresol purple [BCP]). RESULTS: SLS with or without NaOH yielded best reaction with the milk samples similar to Leucocytest. It was observed that the reaction of milk samples with SLS added with indicators (BTB and BCP) was easier to visualize than without indicators. SLS 3%+NaOH 1.5% with BTB and SLS 2% with BCP had high sensitivity, specificity, and substantial agreement with SCC. The area under the receiver operating characteristics curve of SLS 2% with BCP and SLS 3%+NaOH 1.5% with BTB was 0.917 and 0.875, respectively. CONCLUSION: It may be concluded that SLS 3%+NaOH 1.5% with BTB and SLS 2% with BCP may be the potential reagents for the development of an effective cow-side test to detect SCM, as the main ingredient SLS is considerably cheap and readily available in developing countries.

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.

Relationship between somatic cell counts and subclinical mastitis in lactating dairy cows.

BACKGROUND AND AIM: Subclinical mastitis (SCM) is one of the most prevalent diseases of dairy cows, and somatic cell count (SCC) is widely used to determine SCM and milk quality. However, setting the threshold of SCC is very important. This study aimed to determine the cow-level threshold of SCC to differentiate milk of SCM affected cows from normal milk in Bangladesh. MATERIALS AND METHODS: Sensitivity (Se) and specificity (Sp) along with other characteristics of different thresholds of SCC were determined considering the bacterial culture as the gold standard test. Three definitions of intramammary infection (IMI) were set based on the group of pathogens involved. Five categories of SCC thresholds were considered for analysis. RESULTS: Se and Sp of thresholds of SCC greatly varied as definitions of IMI changed. Irrespective of SCC thresholds, Se increased when isolation of major pathogens included in IMI definitions. Se decreased when SCC thresholds increased (from 100 to 300 × 103 cells/mL) for all IMI definitions and ranged from 60.6% to 20.3%. The highest Se was found at low SCC threshold (100 × 103 cells/mL), which resulted in less false-negative outcome. On the other hand, Sp increased with the increment of SCC thresholds giving rise to a less false-positive outcome. CONCLUSION: The cow-level SCC threshold of 100 × 103 cells/mL was found appropriate to identify subclinical IMI of dairy cows.

Prevalence and Multidrug Resistance Pattern of Salmonella Carrying Extended-Spectrum ß-Lactamase in Frozen Chicken Meat in Bangladesh.

ABSTRACT: Salmonella is an important foodborne pathogen that causes public health problems globally, and the increase of antimicrobial resistance in Salmonella has intensified the problem. Chicken meat is an important reservoir and disseminator of Salmonella to humans. This study aimed at estimating the burden of Salmonella carrying extended-spectrum ß-lactamase (ESBL) and their antimicrobial resistance pattern in 113 domestic frozen chicken meat samples purchased from supershops available in five divisional megacities of Bangladesh. The study also focused on the determination of ß-lactamase-, and plasmid-mediated quinolone resistance-encoding genes. All samples were analyzed for the presence of Salmonella using selective media and PCR assay. Antimicrobial susceptibility test was done by disk diffusion test, and ESBL screening was performed by double-disk synergy tests. Resistance genes were detected using multiplex PCR. Of samples, 65.5% were positive for Salmonella spp., and, of these, 58.1% isolates were ESBL producers. All the isolates were multidrug resistant (MDR): 40.5% were resistant to both three to five and six to eight antimicrobial classes; 17.6% were resistant to 9 to 11 classes, and 1.4% isolates to 12 to 15 classes. The highest rates of resistance were observed against oxytetracycline (100%), followed by trimethoprim-sulfamethoxazole (89.2%), tetracycline (86.5%), nalidixic acid (83.8%), amoxicillin (74.3%), and pefloxacin (70.3%). Notably, 48.6% of isolates demonstrated resistance to imipenem. One (1.4%) isolate was possibly extensively drug resistant. All the isolates were positive for the blaTEM gene, 2.7% were positive for blaCTX-M-1, and 20.3% for blaNDM-1. The prevalence of qnrA and qnrS genes was 4.1 and 6.8%, respectively. This study shows that ESBL-producing Salmonella are widespread in frozen chicken meat in Bangladesh, which puts greater responsibility on food processors and policy makers to ensure food safety.

Dynamics of somatic cell count and intramammary infection in lactating dairy cows.

OBJECTIVE: The influence of intramammary infection (IMI) and types of bacteria was assessed on somatic cell count (SCC) in dairy cows' milk with respect to breed, age, parity, stage of lactation, milk production, and mammary quarter location. MATERIALS AND METHODS: After recording data in a structured questionnaire, 360 samples of quarter milk were collected. The samples were subjected to SCC and isolation and identification of bacteria. The data were analyzed to find out the significant influence of independent factors on SCC and IMI. RESULTS: The infected quarters had a significantly higher mean SCC (210.52 × 103 cells/ml) compared to uninfected ones (32.72 × 103 cells/ml). The mean SCC was the highest for IMI with Enterobacter spp. (338.00 × 103 cells/ml) followed by Bacillus spp. (319.20 × 103 cells/ml), coagulase-negative Staphylococci (CNS) (268.17 × 103 cells/ml), Staphylococcus aureus (218.31 × 103 cells/ml), and Escherichia coli (200.75 × 103 cells/ml) and the lowest for Pseudomonas aeruginosa (66.33 × 103 cells/ml). Milk of rear quarters had a significantly higher SCC than the front quarters. SCC increased with increasing age, parity, and lactation stage regardless of whether cows are infected or not. The IMI was more prevalent in rear quarters (42.2%) and cows at early (≤7 days) lactation (100.0%). Cows having a parity of ≥5 and crossbred and high yielding (>5 l) cows had also a higher rate of IMI of 38.2%, 36.7%, and 38.2%, respectively. CONCLUSION: The IMI and type of bacteria were the principal factors for SCC variation. Besides, mammary quarter location, age, and parity should be taken into consideration during the interpretation of SCC.

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.

Microbial risk assessment of ready-to-eat mixed vegetable salads from different restaurants of Bangladesh Agricultural University campus.

OBJECTIVE: The study was aimed to analyze the microbiological quality of mixed vegetable salads and to understand the risk related with its consumption from different restaurants around Bangladesh Agricultural University campus in Mymensingh. MATERIALS AND METHODS: Sixty (60) samples of mixed vegetable salads were taken from twelve (12) different restaurants in five different time points from each restaurant. In parallel, restaurant workers were asked for handling practices while the consumers were interviewed about their salad consumption pattern and whether they had experienced any health-related problems. Microbial risk assessment of Staphylococcus spp., Salmonella spp., and Escherichia coli was estimated by Monte Carlo simulation (10,000 iterations), an exponential model. RESULTS: Aerobic plate count was ranged from 7.73 ± 0.61 to 9.04 ± 0.26 log cfu/gm, Staphylococcus spp. from 4.64 ± 0.61 to 6.42 ± 0.53 log cfu/gm, Salmonella spp. from 4.75 ± 0.08 to 5.27 ± 0.53 log cfu/gm, and E. coli from 4.98 ± 0.20 to 6.66 ± 0.80 log cfu/gm. From the survey, it was found that total consumers had 18% chances where the male had 13% and the female had 30% chances of being infected with salads. Again frequent, average, and occasional consumers had 31%, 13%, and 0% chances, respectively, of being infected with those salads. From the Monte Carlo simulation, the calculated mean annual risks of Staphylococcus spp., Salmonella spp., and E. coli infection for the three exposure scenarios were found to be about 100%. CONCLUSION: The study actually revealed the potential microbial contamination in mixed vegetable salads which may impact on food safety and human health. So, the findings suggest that following hygienic measures during processing and handling the microbiological quality of mixed vegetables salads can be improved.

Prevalence and risk factors of subclinical mastitis in lactating dairy cows in north and south regions of Bangladesh.

The purpose of the study was to identify the potential risk factors for subclinical mastitis (SCM) in lactating dairy cows in Bangladesh. A cross-sectional study was carried out on randomly selected 212 smallholder dairy farms of Sadar upazilas of Rangpur, Mymensingh, and Satkhira districts of Bangladesh during January to October 2011. The direct interview using a structured questionnaire and physical examination of the cows were done to collect data on 15 variables. Milk samples collected from study cows were subjected to California Mastitis Test (CMT). The diagnosis of SCM was based on the results of CMT and physical examination of udder and milk. The bivariable followed by multivariable analysis was done using SPSS 17.0. Of the total cows examined, 20.2 % had subclinical mastitis. In bivariable analysis, eight risk factors were identified. However, in the final model of multivariable analysis, four potential risk factors were identified. These were history of previous clinical mastitis (odds ratio (OR) 10.51, p<0.001), pendulous type of udder (OR 2.26, p=0.008), no grass feeding (OR 1.84, p=0.039), and body condition score (BCS) 2.5 or less (OR 7.25, p=0.054). Four different factors were significantly associated with the occurrence of subclinical mastitis, which need to be considered in the control of the disease. However, particular emphasis should be given on grass feeding and BCS because these traits can be modified or improved to allow prevention of SCM.