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.

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.

Gamma-interferon assay for the ancillary diagnosis of bovine tuberculosis in dairy cattle in urban and adjacent areas of Dhaka city, Bangladesh.

Background and Aim: Bovine tuberculosis (bTB) is an infectious disease of cattle, mainly caused by Mycobacterium bovis. This study aimed to evaluate the efficacy of the interferon-gamma (IFN-γ) assay and single-intradermal comparative tuberculin test (SICTT) in detecting bTB. Materials and Methods: In an earlier study, 150 positive, 83 inconclusive, and 480 negative animals from 24 cattle herds were screened using SICTT. From these groups, 125 positive, 17 inconclusive, and six negative animals were subsequently verified using the IFN-γ assay. Single-intradermal comparative tuberculin test outcomes were interpreted according to standard guidelines, whereas blood samples were collected and stimulated with purified protein derivatives. Sandwich enzyme-linked immunosorbent assay was used to measure secreted IFN-γ. Concordant and Bayesian latent class analyses were performed to evaluate test performance. Results: Results from the IFN-γ assay revealed that 83.2%, 64.7%, and 16.67% of the animals were positive in the SICTT-positive, inconclusive, and negative animal categories, respectively. Sensitivity (SE) and specificity (SP) of SICTT were 83.9% (95% confidence interval [CI]: 77.4-90.1) and 95.7% (95% CI: 86.9-99.7), respectively. Sensitivity and SP for the IFN-γ assay were 78.9% (95% CI: 71.9-85.4) and 83.9% (65.9-95.9), respectively. The use of both tests in parallel increases the SE of bTB detection (~94%), compared with SICTT alone. Conclusion: Use of the IFN-γ assay with SICTT in parallel, predominantly on cattle demonstrating an inconclusive SICTT outcome, boosts bTB detection rate in low resource settings.

Enhancing bovine tuberculosis screening at Dhaka city in Bangladesh: Integrating gamma interferon blood test as ancillary testing with tuberculin skin test.

Tuberculin skin test (TST) is the standard method for screening of bovine tuberculosis (bTB). However, gamma interferon blood test has been introduced in the bTB control program as an ancillary testing with TST in many countries of the world. The objective of this study was to recommend this screening test as an ancillary testing with TST for field application in Bangladesh. In this study 577 cattle of different age, sex and breeds from twenty nine (29) cattle herds were examined to determine skin response against bTB through single intradermal comparative tuberculin test (SICTT) that comprised of positive (n = 81), inconclusive (n = 44) and negative (n = 452) animals. Of which 74 animals that included positive (n = 63), inconclusive (n = 8) and negative (n = 3) animals were taken under this study. Blood samples were collected in heparinized tube and stimulated overnight with bovine and avian purified protein derivatives (PPDs) for the secretion of gamma interferon, and measured via sandwich ELISA. Cohen's kappa statistics was performed for the evaluation of agreement between the two tests. The agreement obtained between two tests was fair (Kappa agreement, K = 24.0%, 95% CI = 16.9-30.5%, P = 0.037). Of positive (n = 63), inconclusive (n = 8) and negative (n = 3) status of animals at SICTT, 82.54% (n = 52), 62.50% (n = 5), and 33.33% (n = 1) were found to be bTB positive respectively through this ancillary test. This test notably corroborates to TST result. A considerable number of inconclusive TB status animals were found to be positive through this gamma interferon assay. Therefore, this test could be used as an ancillary test with TST to maximize the proportion of bTB estimation in the infected cattle herd for early detection of zoonotic tuberculosis in Bangladesh before transmission at the animal-human interface.

Risk mapping and risk factors analysis of rabies in livestock in Bangladesh using national-level passive surveillance data.

Rabies is a major zoonotic disease around the world, causing significant mortality to both humans and animals, especially in low- and middle-income countries. In Bangladesh, rabies is transmitted mostly by the bite of infected dogs and jackals to humans and domestic livestock, causing severe economic losses and public health hazards. Our study analyzed national passive surveillance data of veterinary hospital-reported rabies cases in cattle, buffalo, sheep, and goats from 2015 to 2017 in all 64 districts of Bangladesh. We used a zero-inflated negative binomial regression model to identify the main environmental and socio-economic risk factors associated with rabies occurrence in livestock, and we used model results to generate risk maps. Our study revealed that monsoon precipitation (RR=1.28, p-value=0.043) was positively associated with rabies cases in livestock, and the percentage of adults who have completed university education was also a significant predictor (RR=0.58, p-value<0.001) likely suggesting that districts with higher education levels tended to have a lower reporting of rabies cases in livestock. The standardized incidence ratio maps and predicted relative risk maps revealed a high risk of rabies cases in southeast areas in Bangladesh. We recommend implementing risk-based vaccination strategies in dogs and jackals in those high-risk areas before monsoon to reduce the burden of rabies cases in domestic ruminants and humans in Bangladesh.

Quantitative and qualitative analysis of antimicrobial usage and biosecurity on broiler and Sonali farms in Bangladesh.

Antimicrobial use (AMU) is a major contributing factor to the rising threat of antimicrobial resistance (AMR) in human and animals. To control AMR, indiscriminate antibiotic use needs to be restricted, preventive measures such as biosecurity must be prioritized and proper action plans must be implemented. This study aimed to quantify and associate AMU and biosecurity status of broiler and Sonali farms in Bangladesh. Data on all antimicrobial treatments administered during a batch production cycle and antimicrobials purchased over a year were collected from 94 conventional broiler and 51 Sonali (cross-breed) farms from the northern and southeast regions of Bangladesh. Flock-level AMU was quantified using Treatment Incidence (TI) per 100 days based on the Defined Daily Dose (TIDDDvet) expressing the number of days per 100 animal-days at risk that the flock receives a standard dose of antimicrobials. The biosecurity status (external and internal) of these farms was assessed by means of the Biocheck.UGent scoring system and the correlation between biosecurity and TIDDDvet were assessed by Pearson's correlation coefficients. Median flock TIDDDvet was 60 and 58 for broilers and Sonali flocks, indicating that the birds were treated around 60% and 58% of their lifetime with an antimicrobial dose, respectively. Minimum and maximum values of TIDDDvet ranged from 18 -188 and 31-212 in broilers and Sonali, respectively. Fluoroquinolones, sulfonamides, tetracyclines and aminopenicillins were the most frequently used antimicrobial classes. The mean external and internal biosecurity scores were 39% and 61% for broilers and 44% and 61% for Sonali, respectively. There was a statistically significant difference in the external biosecurity score in broiler farms in the two regions (p ≤ 0.001), whereas, the internal biosecurity score was borderline not statistically significantly different (p = 0.065). The biosecurity score was negatively correlated with AMU in broiler and sonali farms both for external (R2 =-0.38; -0.36) and internal biosecurity (R2 =-0.33; -0.32), respectively. As most of the farmers treated their birds with antimicrobials for a very large part of the production, it could be concluded that there is a high overuse of antimicrobials both in broiler and Sonali poultry production in Bangladesh. This study also highlighted a low level of farm biosecurity practices. Overuse of antimicrobials and low level of biosecurity practice may be due to a lack of knowledge, ignorance, avoid loss and/or lack of monitoring by governmental agencies. Therefore, urgent action is required to increase awareness and biosecurity levels and to reduce AMU in these production systems.

Investigation of Campylobacter fetus in breeding bulls of private farms in Bangladesh.

BACKGROUND: Bovine genital campylobacteriosis (BGC) is a venereal disease caused by Campylobacter fetus that has a negative impact on animal reproduction. The bull is considered to be a symptomless carrier that spreads the disease agent to breeding cows, causing infertility and sporadic abortion. AIM: The study aims to estimate the prevalence, identify risk factors of Campylobacter fetus (C. fetus) infection and antimicrobial resistance pattern of the C. fetus isolates. METHOD: A cross-sectional survey was conducted in Mymensingh district of Bangladesh. Bull smegma samples (single sample from each bull) were collected from 300 bulls from four farms and tested via culture, biochemical identification and finally 16S rRNA and cdtA gene-based molecular assays (PCR) for herd and animal-level prevalence estimation. Herd- and animal-level data on risk factors were collected from the farmers using a pretested questionnaire and analysed by univariable and multivariable logistic regression models with a p value of <0.05 was taken statistically significant for both analyses. RESULTS: Among the surveyed farms, 75% (95% CI: 19.4%-99.4%) were confirmed to have bulls infected with Campylobacter fetus at herd level. However, animal-level occurrence of C. fetus was estimated to be 8.7% (26/300) (95% CI: 5.7%-12.4%). Natural service increases the odds of campylobacteriosis 38.18 times (95% CI: 13.89-104.94) in comparison to artificial insemination for C. fetus infection in bulls. Significantly, half of the isolates (50%, n = 13) were identified to be multidrug resistant (MDR) for three to five antimicrobial agents. CONCLUSION: This study highlights the need to develop official guidelines for C. fetus control and prevention in Bangladesh including mandatory artificial insemination in reproductive cows and heifers, routine screening of breeding bulls for C. fetus free status.

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.

Dietary supplementation of Bacillus-based probiotics on the growth performance, gut morphology, intestinal microbiota and immune response in low biosecurity broiler chickens.

A feeding trial was conducted to evaluate the effects of Bacillus-based probiotics on growth performance, intestinal histo-morphology, gut microbial population and immune response in broilers. A total of 2000 Hubbard Classic day-old chicks were randomly enrolled in four experimental groups and 4 replicates of 500 birds in each group, and reared for 35 days under a low- level of biosecurity measures. The trial groups were assigned treatment-1 (T1): basal diet(control), treatment-2 (T2): basal diet plus Bacillus licheniformis (DSM17236), treatment-3 (T3): basal diet plus Bacillus subtilis (PB6), and treatment-4 (T4) basal diet plus 4% Flavomycin. All four groups were fed with maize-soybean based prepared feeds (starter, grower and finisher). Dietary inclusion of B. licheniformis significantly improved body weight gain and lessened FCR in T2 compared to other groups (p < 0.05). Probiotics increased the population of Bacillus spp. and decreased the population of Clostrium perfringens, Salmonella spp. and Escherichia coli in the jejunum and ileum in broiler birds on day 21 and 35 (p < 0.05). The highest antibody production was observed in B. licheniformis treated group (T2) compared to other probiotic treated group (T1). Taken together, the study findings suggest that B. licheniformis probiotics could be used as a feasible alternative to antimicrobials in the broiler production considering beneficial impacts at low biosecurity broiler farms.

Occurrence and multidrug resistance of Campylobacter spp. at duck farms and associated environmental and anthropogenic risk factors in Bangladesh.

BACKGROUND: The alarming rise in multi-drug resistant (MDR) zoonotic pathogens, including Campylobacter spp., has been threatening the health sector globally. In Bangladesh, despite rapid growth in poultry sector little is known about the potential risks of zoonotic pathogens in homestead duck flocks. The aim of this study was to understand the occurrence, species diversity, and multi-drug resistance in Campylobacter spp., and identify the associated risk factors in duck farms in Bangladesh. METHODS: The study involved 20 duck farms at 6 sub-districts of Mymensingh, Bangladesh. Monthly occurrence of Campylobacter spp. in potential sources at the farms during February-September, 2018, was detected by culture and PCR-based methods. Campylobacter isolates were examined for resistance to different antimicrobials. Risk factors, concerning climatic and environmental disposition, farm management, and anthropogenic practices, of Campylobacter infection were estimated by participatory epidemiological tools. RESULTS: Occurrence of Campylobacter spp. was detected in overall 36.90% (155/420) samples, more frequently in drinking water (60%, 30/50), followed by cloacal swab (37.50%, 75/200), egg surface swab (35%, 35/100) and soil of the duck resting places (30%, 15/50) but was not detected in feed samples (n = 20). PCR assays distinguished the majority (61.30%, 95/155) of the isolates as C. coli, while the rest (38.70%, 60/155) were C. jejuni. Notably, 41.7% (25/60) and 31.6% (30/95) strains of C. jejuni and C. coli, respectively, were observed to be MDR. The dynamics of Campylobacter spp., distinctly showing higher abundance during summer and late-monsoon, correlated significantly with temperature, humidity, and rainfall, while sunshine hours had a negative influence. Anthropogenic management-related factors, including, inadequate hygiene practices, use of untreated river water, wet duck shed, flock age (1-6 months), and unscrupulous use of antimicrobials were identified to enhance the risk of MDR Campylobacter infection. CONCLUSION: The present study clearly demonstrates that duck farms contribute to the enhanced occurrence and spread of potentially pathogenic and MDR C. coli and C. jejuni strains and the bacterial dynamics are governed by a combined interaction of environmental and anthropogenic factors. A long-term holistic research at the environment-animal-human interface would be integral to divulge health risk reduction approaches tackling the spread of Campylobacter spp. from duck farms.

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.

Correction: Bovine tuberculosis prevalence and risk factors in selected districts of Bangladesh.

[This corrects the article DOI: 10.1371/journal.pone.0241717.].

Zoonotic tuberculosis knowledge and practices among cattle handlers in selected districts of Bangladesh.

We assessed zoonotic tuberculosis (zTB) knowledge and prevention and control practices of 404 cattle handlers via a survey in three dairy-intensive districts of Bangladesh. Most respondents were aged 30-49 (52%) and male (95%). Almost all (99%) recognized the important public health burden of tuberculosis in Bangladesh, however, most (58%) had inadequate knowledge about zTB transmission to humans. Inappropriate practices such as: not using protective equipment (98%); smoking, drinking or eating food whilst working with cattle (69%); and sharing the same premises with animals (83%) were identified. Cattle handlers educated at secondary or higher levels were 2.82- (95% CI: 1.59-5.10) and 5.15 times (95% CI: 1.74-15.20) more likely to have adequate knowledge of control and prevention activities compared to those with no formal education. Those who had reared animals for 1-5 years were 2.67 times (95% CI: 1.44-4.91) more likely to have adequate knowledge, compared to those who reared animals for >15 years. Cattle handlers with a monthly incomes of 10,000-20,000 taka were significantly (Odds Ratio = 0.36, 95% CI: 0.14-0.92) less likely to have adequate knowledge compared to those with monthly incomes <10,000 taka. Cattle handlers with high school or higher education were 6.98 times (95% CI: 2.47-19.71) more likely to use appropriate zTB control and prevention practices compared to those without formal education. Those who had reared animals for 1-5 years, 6-10 years and 11-15 years were 2.72- (95% CI: 1.42-5.24), 2.49- (95% CI: 1.29-4.77) and 2.86 times (95% CI: 1.13-7.23) more likely to apply appropriate practices compared to those who reared animals for >15 years. Overall, education, duration of cattle rearing and monthly income predicted zTB knowledge and practices. There is an urgent need to educate those at high-risk of zTB transmission on issues including the handling of infected animals, and general hygiene. A One Health approach, to support the Sustainable Development Goals and the End TB strategy, appears to be the way forward.

Qualitative risk assessment of transmission pathways of highly pathogenic avian influenza (HPAI) virus at live poultry markets in Dhaka city, Bangladesh.

Analysis of environmental samples obtained from the Live Poultry Markets (LPMs) of Dhaka City, Bangladesh, has revealed that the highest degree of prevalence of highly pathogenic avian influenza A (HPAI, H5N1), besides other subtypes of the LPAI virus, poses the plausible risk of transmission of these viruses between human and poultry species. The present study was conducted using the OIE risk analysis framework to assess the risk level of each pathway successively. The estimated risk parameters were integrated towards to obtain the overall risk level for each specific HPAI transmission pathway using the matrix adapted by Cristobel Zepeda accompanying other expert consultations. The relevant data obtained from published and unpublished sources, together with survey data of field observations, were used to formulate and confirm the risk pathways and their associated risks. The results revealed that the risk of the release of the HPAI virus was medium when exposure was high. Additionally, the consequence would be considered very high with a medium degree of uncertainty for all parameters. Ultimately, the overall risk for transmission was estimated as medium with a medium degree of uncertainty. The findings of this study reveal that there is a significant threat that HPAI virus transmission could occur among poultry and humans and effectively sustain within the environment of the LPMs. Our findings are primarily focused on public health considerations, the hygienic slaughter of poultry and the relevant cleaning and sanitation practices conducted in the LPMs to support evidence-based decision-making processes. The findings of the study have the potential to be used to formulate effective risk reduction measures and can be further adapted in low-resource settings without major infrastructural changes required of the LPMs. All of which would reduce the risk of HPAI virus release and further lessen the degree of exposure and transmission in established LPMs.

Identification and characterization of Salmonella spp. from samples of broiler farms in selected districts of Bangladesh.

BACKGROUND AND AIM: Salmonella spp. are an important group of pathogens responsible for human and animal diseases. This study aimed to estimate the prevalence and identify and characterize of Salmonella spp. isolated from broiler farms of Gazipur, Tangail, and Dhaka districts of Bangladesh. This study also evaluated the difference of Salmonella positivity status between two groups of farms, good practices adapted in broiler rearing at the project intervened farms, and non-project intervened traditional farms. MATERIALS AND METHODS: A total of 352 samples including 128 cloacal swabs, 32 whole carcasses, 64 feed, 64 water, and 64 attendants' hand rinses were collected through convenient sampling technique from 16 poultry food safety project of Food and Agricultural Organization of United Nations Bangladesh intervened farms and other 16 non-project intervened farms in the same location. Various cultural based techniques and biochemical methods were employed for the estimation of prevalence, isolation, and identification of Salmonella spp. which was further evaluated by polymerase chain reaction. Antimicrobial susceptibility test using disk diffusion methods and serogrouping by slide agglutination test was accomplished for additional characterization. RESULTS: Among the samples, an overall prevalence of Salmonella spp. was 31.25% (110/352) (95% confidence interval [CI]=26.44-36.38%). However, the prevalence of Salmonella spp. was 24.43% (43/176) (95% CI=18.28-31.47) in project intervened farms and 38.07% (67/176) (95% CI=30.87-45.68%) in non-intervened farms. Among the 110 isolates, 31.82% (35/110) were fitted under serogroup B, and the rest of the isolates 75 (68.18%) under serogroup D. Of 110 isolates, 82.72%, 77.27%, 81.82%, and 79.09% were susceptible to ciprofloxacin, gentamycin, norfloxacin, and streptomycin, respectively. In addition, 81.82% and 80% isolates were resistant to erythromycin and tetracycline, respectively. Isolated Salmonella spp. presented moderate resistance to both amoxicillin and azithromycin. Alarmingly, 80.91% (89/110) isolates were shown to be multidrug-resistant Salmonella spp. CONCLUSION: The study has presented a significant variation of the prevalence of Salmonella spp. between project intervened and non-project intervened farms, and this indicates project intervened farms are comparatively safer than the non-intervened farms considering public health and food safety grounds. This research outcome also has highlighted a substantial proportion of poultry origin multidrug resistance Salmonella spp. is a potential source of public health hazards. In this regard, proper awareness creation and motivational activities on good agriculture practices in poultry rearing and maintaining good personal hygiene at the farmers' level are warranted through participatory training.

A further outbreak of classical swine fever in indigenous pigs in Kurigram district, Bangladesh.

Deaths of native scavenging pigs were reported in mid-November 2015 at Nageswari sub-district, Kurigram district of Bangladesh. The investigation for a suspected classical swine fever (CSF) outbreak was accomplished via a joint outbreak investigation team from Department of Livestock Services (DLS) and Food and Agriculture Organization, Emergency Center for Transboundary Animal Disease (FAO-ECTAD), Bangladesh. Out of 592 pigs, 396 were infected and among them 263 died. The attack rate and case fatality rate were 66.9% and 66.4%, respectively. The epidemic curve constructed using the data captured from the CSF outbreak site was nearly bell-shaped, indicating a point source epidemic. The basic reproduction numbers (R0) were estimated to be 1.6 (95% Confidence interval [CI]: 1.5-1.7) and 1.5 (95% CI: 1.3-1.7) based on attack rate and exponential growth rate methods, respectively. Adult pigs showed signs of high fever, staggering gait and depression, whereas piglets either died without any premonitory signs or purulent exudates in the eyes were observed. Post-mortem examination was carried out on a 6-month-old piglet. The necropsy findings included were swollen lymph nodes deep red in colour, and haemorrhages on serous and mucous membranes of the intestinal organs together with button-like ulceration in the intestines. Nasal swabs and tissue samples (spleen, lung and liver) were tested using real-time reverse transcriptase polymerase chain reaction (RT-qPCR) and found to be positive for CSF virus. One-step RT-PCR was used to amplify 1,148 base pair of E2 gene in extracted RNA and was sequenced using standard Sanger's sequencing. Phylogenetic analysis revealed the virus as genotype 2.2 that clustered with CSF virus sequences from Bangladesh and India. This is only the second report of a CSF outbreak in Bangladesh. CSF appears to be an emerging transboundary disease in this country. A special programme for controlling swine diseases is needed since pigs are being reared by marginalized communities of Bangladesh to provide livelihoods and food security.

Peste des Petits Ruminants Risk Factors and Space-Time Clusters in Bangladesh.

Peste des Petits Ruminants (PPR) is endemic in Bangladesh, but its spatial distribution and risk factors have not yet been reported. Using four years of national-level, passive surveillance data (2014 to 2017), in this study we aimed to identify risk factors, create PPR risk maps and describe PPR time-space clusters. We selected PPR case records-mainly based on presumptive diagnosis of small ruminants in subdistrict veterinary hospitals-and sheep and goat population data from all 64 districts of Bangladesh. Peste des Petits Ruminants cumulative incidence per 10,000 animals at risk per district was used to conduct cluster and hotspot analysis and create predictive maps for each year and all 4 years combined. The association between PPR cumulative incidence and hypothesized risk factors-including climatic variables, elevation, road length, river length, railroad length, land cover, and water bodies-was analyzed using a geographically weighted regression model. The total number of PPR cases reported during the study period was 5.2 million. We found that most PPR cases (27.6%) were reported in the monsoon season. The highest and lowest proportions of cases were reported from Rajshahi (36.1%) and Barisal divisions (2.1%), respectively. We identified five space-time clusters, 9 high-high clusters, and 9 hotspots. The predicted cumulative incidences of PPR were persistently higher in north-east, north-west, and south-east parts of Bangladesh. Road length (P = 0.03) was positively associated with PPR incidence in Bangladesh. Results suggest that movement of animals (road length) plays an important role in the epidemiology of PPR in Bangladesh. Along with restriction of animal movement, hotspots and high-high clusters should be targeted first for immunization coverage in Bangladesh and similar PPR endemic countries to achieve eradication.

A Decade of Avian Influenza in Bangladesh: Where Are We Now?

Highly pathogenic avian influenza (HPAI) has been a public health threat in Bangladesh since the first reported outbreak in poultry in 2007. The country has undertaken numerous efforts to detect, track, and combat avian influenza viruses (AIVs). The predominant genotype of the H5N1 viruses is clade 2.3.2.1a. The persistent changing of clades of the circulating H5N1 strains suggests probable mutations that might have been occurring over time. Surveillance has provided evidence that the virus has persistently prevailed in all sectors and caused discontinuous infections. The presence of AIV in live bird markets has been detected persistently. Weak biosecurity in the poultry sector is linked with resource limitation, low risk perception, and short-term sporadic interventions. Controlling avian influenza necessitates a concerted multi-sector 'One Health' approach that includes the government and key stakeholders.