Nanopore sequencing for identification and characterization of antimicrobial-resistant Escherichia coli and Salmonella spp. from tilapia and shrimp sold at wet markets in Dhaka, Bangladesh.

Wet markets in low-and middle-income countries are often reported to have inadequate sanitation resulting in fecal contamination of sold produce. Consumption of contaminated wet market-sourced foods has been linked to individual illness and disease outbreaks. This pilot study, conducted in two major wet markets in Dhaka city, Bangladesh during a 4-month period in 2021 aimed to assess the occurrence and characteristics of Escherichia coli and non-typhoidal Salmonella spp. (NTS) from tilapia (Oreochromis niloticus) and shrimp (Penaeus monodon). Fifty-four individuals of each species were collected. The identity of the bacterial isolates was confirmed by PCR and their susceptibility toward 15 antimicrobials was tested by disk diffusion. The whole genome of 15 E. coli and nine Salmonella spp. were sequenced using Oxford Nanopore Technology. E. coli was present in 60-74% of tilapia muscle tissue and 41-44% of shrimp muscle tissue. Salmonella spp. was found in skin (29%) and gills (26%) of tilapia, and occasionally in muscle and intestinal samples of shrimp. The E. coli had several Multilocus sequence typing and serotypes and limited antimicrobial resistance (AMR) determinants, such as point mutations on glpT and pmrB. One E. coli (BD17) from tilapia carried resistance genes for beta-lactams, quinolones, and tetracycline. All the E. coli belonged to commensal phylogroups B1 and A and showed no Shiga-toxin and other virulence genes, confirming their commensal non-pathogenic status. Among the Salmonella isolates, five belonged to Kentucky serovar and had similar AMR genes and phenotypic resistance patterns. Three strains of this serovar were ST198, often associated with human disease, carried the same resistance genes, and were genetically related to strains from the region. The two undetermined sequence types of S. Kentucky were distantly related and positioned in a separate phylogenetic clade. Two Brunei serovar isolates, one Augustenborg isolate, and one Hartford isolate showed different resistance profiles. This study revealed high fecal contamination levels in tilapia and shrimp sold at two main wet markets in Dhaka. Together with the occurrence of Salmonella spp., including S. Kentucky ST198, a well-known human pathogen, these results stress the need to improve hygienic practices and sanitation standards at markets to improve food safety and protect consumer health.

Animal, human, and environmental perspectives on anthrax in Bangladesh.

Background: In Bangladesh, the zoonotic transmission of anthrax from animals to humans poses substantial challenges for prevention and control programs, especially in resource-constrained settings. A comprehensive literature review was conducted focusing on anthrax infections in animals, humans, and the environment to enable better design of prevention and control strategies. Materials and methods: We followed PRISMA guidelines to collect data on anthrax infection in animals and humans from reports between 1980 and January 2023. We used a standardized data extraction template to collect data on study location, year, hosts, deaths and risk factors responsible for anthrax occurrences at the animal, human and environmental sectors. Subsequently, we conducted a thorough analysis of the data gathered to identify the factors responsible for anthrax occurrences and to propose updated strategies for anthrax prevention and control. Results: Of the 27 articles analyzed, 20 focused on animal or human anthrax, while seven addressed environmental contaminations. A total of 6354 cases of anthrax infection in animals were recorded, with 998 fatalities and an overall case fatality of 15.7 %. In humans, inadequate knowledge about anthrax and its transmission was a significant factor. Risk factors for human cutaneous anthrax included activities such as slaughtering diseased animals and contact with contaminated raw meat or blood. Risky practices such as disposal of animal carcasses in floodwaters or water bodies were observed in some areas, contributing to the persistence of the anthrax pathogen in the environment. Conclusions: Our study highlights the necessity of a multisectoral One Health approach to effectively control and prevent anthrax outbreaks in both animals and humans. This approach should include comprehensive vaccination programs, social and behavioral change activities, environmental management, and the establishment of surveillance systems. Implementing these recommendations will be crucial in addressing the complex challenges posed by anthrax in low-resource settings.

Prevalence and identification of caprine pasteurellosis in pneumonic goats in Bangladesh.

Objective: This research aimed to assess the prevalence of caprine pasteurellosis, isolate and identify pasteurellosis (Mannheimia haemolytica and Pasteurella multocida) in pneumonic goats, and discover the main bacterial cause of pneumonia. Materials and Methods: One hundred and five samples (94 nasal swabs and 11 lung tissues) from goats suspected of having pneumonia were taken and transferred aseptically to the laboratory. Following the processing of the collected samples, Pasteurella spp. was isolated with the aid of plate culture methods. Biochemical characteristics were used to identify all bacterial isolates, which were then verified by polymerase chain reaction (PCR). Antimicrobial susceptibility testing was also carried out to evaluate the sensitivity profiles of various antibiotics. The Pasteurella haemolytica serotype-specific antigen (PHSSA) gene was used to identify isolates of M. haemolytica, and the KMT1 gene was used to identify isolates of P. multocida. Results: From the 105 clinically suspicious samples, 51 (48.57%) were identified to be Pasteurella spp. through bacteriological testing and also by PCR targeting the 16S rRNA gene. Of these, 47.87% (45/94) were nasal swabs, and 54.55% (6/11) were lung tissues. Among confirmed samples, 70.59% (36/51) were identified as M. haemolytica, and 29.41% (15/51) were identified as P. multocida. Resistance to tetracycline, streptomycin, oxytetracycline, gentamicin, and ceftriaxone was found in 50%-83% of the isolates. In addition, PCR identified the PHSSA and KMT1 genes from isolates of P. multocida and M. haemolytica, respectively. Conclusion: The present study revealed that M. haemolytica and P. multocida primarily caused pasteurellosis in pneumonic goats in Bangladesh. However, when treating these animals, the proper choice of antimicrobials should be made to control this disease.

Exploring the prevalence of antibiotic resistance patterns and drivers of antibiotics resistance of Salmonella in livestock and poultry-derived foods: a systematic review and meta-analysis in Bangladesh from 2000 to 2022.

Background: Antimicrobial resistance (AMR) is a severe public health problem that Bangladeshis are dealing with nowadays. However, we wanted to investigate the pooled prevalence of Salmonella and AMR in Salmonella strains isolated from livestock- and poultry-derived foods between 1 January 2000 and 31 August 2022. Methods: The metafor and metareg packages in the R programming language were used to conduct all analyses. We used a random-effect or fixed-effect model for pooled prevalence of Salmonella and AMR to Salmonella, depending on the heterogeneity test for each antibiotic. The heterogeneity was examined using stratified analyses, the meta-regression approach and sensitivity analysis. Results: The combined prevalence of Salmonella in livestock and poultry-derived food in Bangladesh is 37%, according to the 12-research considered (95% CI: 23%-52%). According to subgroup analysis, neomycin had the lowest prevalence of resistance (4%, 95% CI: 1%-13%), whereas tetracycline had the highest prevalence of resistance (81%, 95% CI: 53%-98%). According to univariate meta-analysis and correlation analysis, the prevalence of Salmonella increased with the study period (ß = 0.0179; 95% CI: 0.0059-0.0298, P = 0.0034; R2 = 46.11%) and without this, none of aforementioned variables was significantly associated with the detected heterogeneity and there was a positive relationship (r = 0.692, P = 0.001) between the Salmonella prevalence and study period. Conclusions: AMR is rising alarmingly in Bangladesh by livestock-derived food consumption. However, monitoring and evaluating antibiotic sensitivity trends and developing effective antibiotic regimens may improve Salmonella infection inhibition and control in Bangladesh. Policymakers should be concerned about food handling practices. Doctors should be concerned when using prescribing antibiotics.

Multidrug-resistant non-typhoidal Salmonella of public health significance recovered from migratory birds in Bangladesh.

Non-typhoidal Salmonella provides an exemplar for the One Health approach as it encompasses public and animal health, food safety, and environmental considerations. The contribution of environmental aspects is currently less well-defined. The purpose of this study was to determine the carriage occurrence of non-typhoidal Salmonella in migratory birds in Bangladesh and assess the potential significance to public and animal health. Cloacal swabs (N = 453) were collected in the years 2018-2020 from Tanguar and Hakaluki Haors, important wetland ecosystems in Northeastern Bangladesh. The prevalence of Salmonella was 13.5% (61 positive swabs). Classical serotyping identified six serovars: Salmonella enterica subsp. enterica serovars Perth, Kentucky, Albany, Infantis, Weltevreden, and Brancaster. Resistance towards 14 antimicrobials was assessed by broth microdilution minimum inhibitory concentration determination and the antimicrobial resistance (AMR) genotype established by whole-genome sequencing. S. Perth and S. Weltevreden isolates were susceptible and harbored no acquired AMR genes. Isolates from the remaining serovars were multidrug resistant, commonly possessing resistance to tetracycline, ampicillin, chloramphenicol, sulfamethoxazole, trimethoprim, and ciprofloxacin. Salmonella resistant to ciprofloxacin meets WHO criteria for priority pathogens. There was excellent concordance between resistance phenotype and the presence of corresponding AMR genes, many of which reside on Salmonella Genomic Islands. High-level ciprofloxacin resistance correlated with the presence of mutations in the chromosomal gyrB and/or parC genes. The S. Kentucky isolates were ST198, a widely distributed multidrug-resistant lineage reported in humans and animals, and constituting an ongoing risk to public health worldwide. We have demonstrated that multidrug-resistant non-typhoidal Salmonella of public health significance can be recovered from migratory birds. A potential for risk can manifest through direct interaction, transmission to food-producing livestock on farms, and dissemination via the long range migratory movements of birds. Risks can be mitigated by measures including continued surveillance and implementation of good farm biosecurity practices.

Microbial Contamination and Antibiotic Resistance in Marketed Food in Bangladesh: Current Situation and Possible Improvements.

Antimicrobial resistance (AMR) is a public health problem worldwide. Bangladesh, like its neighboring countries, faces many public health challenges, including access to safe food, inadequate food surveillance, as well as increasing AMR. This study investigated bacterial contamination and the AMR profile of pathogens in marketed food in Bangladesh and explored barriers to reducing AMR in the country. We collected 366 tomatoes, 359 chicken and 249 fish samples from 732 vendors in traditional markets in urban, peri-urban and rural areas in Bangladesh, as well as from 121 modern retails in Dhaka capital to analyse Vibrio cholerae and Escherichia coli in fish, Salmonella in chicken, and Salmonella and E. coli in tomatoes. Antibiotic susceptibility against 11 antibiotics was tested using a disc diffusion test and interpreted by an automated zone inhibition reader. In addition, a qualitative study using key informant interviews was conducted to explore antimicrobial use and AMR reduction potential in Bangladesh. We found E. coli in 14.21% of tomatoes and 26.91% of fish samples, while 7.38% of tomatoes and 17.27% of chicken were positive for Salmonella, and 44.98% of fish were positive for Vibrio cholerae. In total 231/319 (72.4%) of all pathogens isolated were multidrug-resistant (MDR) (resistant to three or more antibiotic groups). Qualitative interviews revealed an inadequate surveillance system for antibiotic use and AMR in Bangladesh, especially in the agriculture sector. To be able to fully understand the human health risks from bacterial hazards in the food and the AMR situation in Bangladesh, a nationwide study with a one health approach should be conducted, within all sectors, including AMR testing as well as assessment of the antimicrobial use and its drivers.

A meta-analysis of COVID-19 vaccines acceptance among black/African American.

The COVID-19 pandemic had harmed Black/African Americans disproportionately. Mortality and morbidity can reduce by increasing vaccination acceptability and availability. We conducted a meta-analysis of 20 studies that show the prevalence of Black/African Americans who embrace COVID-19 vaccination between 2020 and September, 2022. Investigations conducted before and after the availability of COVID-19 vaccines found the vaccinations effective. The heterogeneity was examined using stratified analyses, the meta-regression approach, and sensitivity analysis in R programming language. This meta-analysis showed that the overall COVID-19 vaccine hesitancy among Black/African Americans is 35% (95% CI: 26%-45%). That means 65% of Black African Americans received vaccines without any hesitancy. According to correlation analysis, there was a negative relationship (r = -0.392, P = 0.021) between the prevalence of vaccine hesitancy and the survey year. Evidence suggests ethnic health disparities in Black/African Americans were for lower socioeconomic status. Some initiatives had to address health disparities, while ethnicity had not consistently been a focus. Only vaccines can prevent COVID-19 like infectious diseases. Policy makers and health educators should concern on vaccine acceptance or hesitancy related programs among Black/African American.

High prevalence of vancomycin non-susceptible and multi-drug resistant enterococci in farmed animals and fresh retail meats in Bangladesh.

The emergence of antimicrobial resistant Enterococcus spp., a main cause of untreatable nosocomial infection, in food animals and dissemination to humans is a public health risk. The study was performed to determine the prevalence and antimicrobial resistance, and virulence characteristics of Enterococcus faecalis and Enterococcus faecium in food animals and meats in Bangladesh. Enterococcus spp., were confirmed using sodA gene specific PCR, and antimicrobial resistance and virulence properties were characterized by PCR. Enterococcus spp. were recovered from 57% of the collected samples (n = 201/352). Farm samples yielded significantly higher (p ≤ 0.05) prevalence (62%) than that of retail meat samples (41%). E. faecalis (52%) is most frequently isolated species. Greater proportions of isolates exhibited resistance to tetracycline (74%), erythromycin (65%) and ciprofloxacin (34%). Fifty-one isolates are vancomycin non-susceptible enterococci (VNSE), of which forty-seven are MDR and twenty are linezolid resistant, a last line drug for VNSE. Virulence factors such as gelatinase (gelE), aggregation factor (asa1) and sex pheromone (cpd) are detected along with vancomycin resistance gene (vanA, vanB and vanC2/C3) in VNSE isolates. The high prevalence of MDR enterococci in food animals and retail meats may cause consumers infections with concomitant reduction of available therapeutic options.

Knowledge, Attitudes and Practices Regarding Antibiotic Use and Resistance among Veterinary Students in Bangladesh.

The use of antibiotics in animals for both therapeutic and non-therapeutic purposes is a major driver of the emergence and spread of antimicrobial resistance (AMR). While several studies have investigated prescription and consumption patterns in humans, little attention has been paid to the veterinary sector. A cross-sectional study was conducted in 3002 veterinary students (VS) and non-medical students (NMS) from 12 universities in Bangladesh to explore their knowledge, attitudes and practices (KAP) about antibiotics and AMR using a self-administered questionnaire, and assess the influence of the veterinary curriculum. KAP regarding antibiotic use and AMR was significantly higher in veterinary than non-medical students, and in first-year than final-year students. However, gaps in knowledge and practices were highlighted, suggesting deficiencies in training. Moreover, final-year veterinary students were found to be more likely than first-year students to use antibiotics without instructions, which could indicate deficiencies in their curriculum. Although the study suggested a positive impact of the veterinary curriculum on KAP about antibiotics and AMR in Bangladesh, critical gaps remain that are likely to contribute to inadequate use in their future practice. Therefore, there is scope for improving educational programs on AMR in professional curricula.

Virulence and Antimicrobial Resistance Profiles of Salmonella enterica Serovars Isolated from Chicken at Wet Markets in Dhaka, Bangladesh.

Virulent and multi drug resistant (MDR) Salmonellaenterica is a foremost cause of foodborne diseases and had serious public health concern globally. The present study was undertaken to identify the pathogenicity and antimicrobial resistance (AMR) profiles of Salmonellaenterica serovars recovered from chicken at wet markets in Dhaka, Bangladesh. A total of 870 cecal contents of broiler, sonali, and native chickens were collected from 29 wet markets. The overall prevalence of S. Typhimurium, S. Enteritidis, and untyped Salmonella spp., were found to be 3.67%, 0.57%, and 1.95% respectively. All isolates were screened by polymerase chain reaction (PCR) for eight virulence genes, namely invA, agfA, IpfA, hilA, sivH, sefA, sopE, and spvC. S. Enteritidis isolates carried all virulence genes whilst S. Typhimurium isolates carried six virulence genes except sefA and spvC. A diverse phenotypic and genotypic AMR pattern was found. Harmonic descending trends of resistance patterns were observed among the broiler, sonali, and native chickens. Interestingly, virulent and MDR Salmonella enterica serovars were found in native chicken, although antimicrobials were not used in their production cycle. The research findings anticipate that virulent and MDR Salmonella enterica are roaming in the wet markets which can easily anchor to the vendor, consumers, and in the food chain.

Behavioral preventive measures and the use of medicines and herbal products among the public in response to Covid-19 in Bangladesh: A cross-sectional study.

The present study was conducted to assess the behavioral preventive measures and the use of medicines and herbal foods/products among the public in response to Covid-19. A cross-sectional survey comprised of 1222 participants was conducted from 27 June to 20 July 2020. Kruskal-Wallis test was used to identify the differences in behavioral preventive practices across different demographic categories. To identify the factors associated with the use of preventive medicines and herbal foods/products, multivariable logistic regression was performed. Most participants adopted the recommended preventive practices such as washing hands more frequently (87.5%), staying home more often (85.5%), avoiding crowds (86%), and wearing masks (91.6%). About half of the smokers reported a decreased rate of smoking during the pandemic. Also, 14.8% took medicines, 57.6% took herbal foods/products, and 11.2% took both medicines and herbal foods/products as preventive measure against Covid-19. Arsenicum album, vitamin supplements, and zinc supplements were the most commonly used preventive medicines. Gender, age, and fear of Covid-19 were significantly associated with the use of both preventive medicines and herbal foods/products. For the management of Covid-19 related symptoms, paracetamol, antihistamines, antibiotics, and mineral (zinc and calcium) supplements were used most often. Most participants sought information from non-medical sources while using medicines and herbal products. Moreover, potentially inappropriate and unnecessary use of certain drugs was identified.

Antibiotic-resistant Escherichia coli in deer and nearby water sources at Safari parks in Bangladesh.

BACKGROUND AND AIM: The emergence and rapid dissemination of multidrug-resistant (MDR) bacteria in different ecosystems is a growing concern to human health, animal health, and the environment in recent years. The study aimed to determine the antibiotic resistance in Escherichia coli from deer and nearby water sources at two different Safari parks in Bangladesh. MATERIALS AND METHODS: A number of 55 fresh fecal samples of deer and six water samples from nearby lakes were collected from two Safari parks. Samples were processed, cultured, and carried out biochemical tests for E. coli. The antibiotic susceptibility was determined by disk diffusion method. To identify the resistance genes, polymerase chain reaction was performed. RESULTS: A total of 32 E. coli isolates from 55 fecal samples and 6 of 6 E. coli isolates from lake water were isolated. From fecal E. coli isolates, ampicillin and sulfamethoxazole were 90.63% (n=29/32) resistant and 87.5% (n=28/32) were resistant to tetracycline and nalidixic acid. High resistance was also observed to other antibiotics. On the contrary, all E. coli isolates from water sources were 100% (n=6/6) resistant to ampicillin, tetracycline, sulfamethoxazole, and nalidixic acid. MDR was revealed in all water samples, whereas 96.88% (n=31/32) was found in fecal isolates. A number of bla TEM, tet A, and Sul2 genes were detected from both isolates. CONCLUSION: This study for the 1st time highlights, a significant proportion of E. coli isolates in wildlife deer and nearby water sources were MDR in Bangladesh.

Antibiotic resistance of Escherichia coli isolated from broilers sold at live bird markets in Chattogram, Bangladesh.

OBJECTIVE: The present study was carried out to appraise the antibiotic resistance and to detect some of the target resistant genes in Escherichia coli (E. coli) isolated from apparently healthy broilers. MATERIALS AND METHODS: Cloacal swab samples (n = 60) were collected from apparently healthy broilers (n = 60) sold at two different live bird markets (LBMs) of Chattogram, Bangladesh. Isolation and identification of the Escherichia coli was done by the following standard bacteriological techniques followed by biochemical tests. The antibiotic susceptibility of E. coli isolates was determined by the disk diffusion method. The antibiotic resistant genes were detected by polymerase chain reaction (PCR) using specific primers. RESULTS: The overall prevalence of E. coli in broilers was 61.67% (n = 37/60) (95% CI = 49-72.93). The antibiogram study showed that the isolates were 100% resistant to ampicillin and tetracycline followed by sulfomethoxazole-trimethoprim (94.59%, n = 35/37) and nalidixic acid (91.89%, n = 34/37). To the contrary, 56.76% (n = 21/37) isolates were sensitive to both ceftriaxone and gentamicin followed by colistin (48.65%, n = 18/37). All of E. coli isolates were multidrug resistant (MDR) and carried bla TEM, tetA, and Sul2 genes. CONCLUSION: The presence of MDR genes in E. coli isolates in broilers could pose a serious public health threat.

Antibiotic resistance of Escherichia coli isolated from captive Bengal tigers at Safari parks in Bangladesh.

OBJECTIVES: The present study was carried out to assess the antibiotic resistance and to identify the resistance genes in Escherichia coli from captive Bengal tigers at two Safari parks in Bangladesh. MATERIALS AND METHODS: A number of 24 environmental fecal swab samples of Bengal tigers were collected from two different Safari parks in Bangladesh. For the isolation of E. coli, samples were submitted to a number of bacteriological screening and biochemical tests. The antibiotic susceptibility of E. coli isolates was determined by disk diffusion method. RESULTS: Results demonstrated that 18 environmental fecal samples were positive to E. coli in bacteriological screening and biochemical test. The overall prevalence of E. coli in Bengal tiger was 75% (n = 18/24). The antibiogram study unveiled that all the isolates were resistant to ampicillin. Sulfamethoxazole-trimethoprim, nalidixic acid, and tetracycline were 89% (n = 16/18) resistant. On the contrary, 100% (n = 18/18) of the isolates were sensitive to colistin sulfate. bla TEM was detected in 78% (n = 14/18) ampicillin-resistant isolates, whereas sul2 was found in 31% (n = 5/16) of the sulfamethoxazole-trimethoprim-resistant isolates. CONCLUSION: This study, first time in Bangladesh, highlights a significant proportion of environmental fecal samples from captive Bengal tigers at Safari parks harboring antibiotic resistant E. coli. Transmission of resistant E. coli from Bengal tigers to humans and the environment could pose a public health risk at Safari parks in Bangladesh.