Characterization, histopathology and immunogenicity of the lumpy skin disease virus isolated during 2019-20 in Bangladesh.

Introduction: Lumpy skin disease (LSD) is a highly contagious vector-borne viral disease of cattle. LSD has emerged in Bangladesh in 2019, causing significant economic losses due to its high morbidity and mortality. This research was designed to isolate, identify, and assess the immunogenicity of LSD virus (LSDV) using nodular tissue samples obtained from affected cattle during the 2019-20 outbreak across nine districts of Bangladesh. Methods: To determine the presence of LSDV in nodular tissues, we initially used iiPCR and PCR, followed by histopathological examination. 151 were positive via iiPCR and PCR among the 180 collected samples. The PCR positive 151 samples were then inoculated into 10-day-old embryonated chicken eggs via the CAM route to isolate LSDV, confirmed through PCR. Subsequently, partial sequencing and phylogenetic analysis of the P32 gene were performed to determine the origin of the circulating LSDV strain. The immunogenicity of selected LSDV strains was assessed through an ELISA test. Results: The PCR results revealed a distinct positive band at 192 bp in both the nodular tissue samples and the LSDV isolated from chicken embryo inoculations. Microscopic analysis of the nodular lesions revealed thickening of the epidermis, ballooning degeneration of keratinocytes, and proliferation of follicular epithelia. Additionally, mononuclear infiltration was observed at the demarcation line between infected and healthy tissue, with necrosis of muscular tissues beneath the epidermis. The LSDV isolate from Bangladesh exhibited a close genetic relationship with LSDV strains isolated from neighboring and other regional countries including India, Myanmar, and Mongolia. This observation strongly suggests the possibility of a transboundary spread of the LSD outbreak in Bangladesh during 2019-2020. The results of the immunogenicity test showed that the serum antibody titer remained at a protective level for up to 18 months following secondary immunization with inactivated LSDV antigen. This finding suggests that the inactivated LSDV antigen could be a potential vaccine candidate to protect cattle in Bangladesh against LSDV. Conclusion: In conclusion, our research successfully isolated, identified, and characterized LSDV in cattle nodular tissues from the 2019-20 outbreak in Bangladesh. Furthermore, it provided insights into the probable origin of the circulating strain and investigated a potential vaccine candidate to protect cattle in the region from LSDV.

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.

Exposure to Brucella spp. in Humans and Cows in a High Milk-Producing Area of Bangladesh.

Brucellosis is a zoonotic disease, caused by some species within the Brucella genus. The primary and secondary objectives of this cross-sectional study were to determine the seroprevalence of Brucella antibodies in humans and cows and identify risk factors for exposure to Brucella spp. among people in Shahjadpur sub-district, Bangladesh. Twenty-five villages were randomly selected from the 303 milk-producing villages in the sub-district. We randomly selected 5% of the total households from each village. At each household, we collected demographic information and history of potential exposure to Brucella spp. in humans. In addition, we collected serum from household participants and serum and milk from cattle and tested to detect antibodies to Brucella sp. Univariate analysis was performed to detect associations between seropositivity and demographics, risk factors, and behaviors in households. We enrolled 647 households, 1313 humans, and 698 cows. Brucella antibodies were detected in sera from 27 household participants (2.1%, 95% confidence interval [95%CI]: 1.2-2.9%). Eleven (1.6%, 95%CI 0.6-2.4%) cows had detectable Brucella antibodies in either milk or serum. About half (53%) of the 698 cows exhibited more than one reproductive problem within the past year; of these, seven (2%) had Brucella antibodies. Households with seropositive individuals more frequently reported owning cattle (78% vs. 32%, P < 0.001). Despite a low prevalence of Brucella seropositivity in the study, the public health importance of brucellosis cannot be ruled out. Further studies would help define Brucella prevalence and risk factors in this region and nationally.

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.

Epidemiology and molecular characterization of Feline panleukopenia virus from suspected domestic cats in selected Bangladesh regions.

Feline panleukopenia (FPL) is a highly contagious cat disease and is endemic in Bangladesh. The study aims to describe the epidemiology and molecular characterization of the Feline panleukopenia virus from the suspected domestic cats in selected Bangladesh regions. Randomly, 161 rectal swabs were collected from the pet hospitals between July 2021 and December 2022. A structured questionnaire was administered through face-to-face interviews with cat owners in order to collect data on potential risk factors for FPL, such as age, sex, sharing litter boxes and every day utensils in multicat households, vaccination history, hospital visits for other diseases, and season. The rectal swabs were tested by PCR targeting the VP2 capsid protein gene, and six PCR-positive samples were further sequenced for molecular characterizations. The risk factors for FPLV were identified using multivariable logistic regression analysis. The overall prevalence of FPL among suspects was 22.9%. The mortality and case fatality were 10.6%, and 45.9%, respectively. However, mortality in kittens was significantly higher (16.4%) than younger cats. The odds of FPL were 8.83 times (95% CI: 3.14-24.85) higher among unvaccinated cats than vaccinated cats. The winter season had almost six times (95% CI: 1.38-24.40) higher odds of FPL than rainy season. In a multicat house, the odds of FPL was about five times (95% CI: 1.93-13.45) higher for cats that shared a litter box and food utensils compared to those that did not engage in such sharing. Visiting hospitals for other reasons nearly triples the odds of FPL (OR: 2.80, 95% CI: 1.04-7.54) compared to cats that do not visit hospitals. Analysis of partial sequence of the VP2 gene revealed genetic variations among the isolates from different regions. Among these isolates, four were identical to FPLV isolates from South Korea and China, while one showed complete homology with FPLV isolates from Thailand. In contrast, the remaining one was 100% identical to Carnivore protoparvovirus-1 isolated from a feline sample in Italy. Our isolates were classified into three distinct clades alongside Feline panleukopenia virus and Carnivore protoparvovirus-1. One in every three suspected cats was infected with Feline panleukopenia. Regular vaccination of the cats, especially those that share common litter box and food utensils and visit hospitals for other purposes, will help reduce the prevalence of FPL in Bangladesh. Besides, it is worth emphasizing the existence of genetic diversity among the circulating Feline panleukopenia viruses in Bangladesh.

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.

Molecular characterization and antibiotic resistance profile of ESBL-producing Escherichia coli isolated from healthy cow raw milk in smallholder dairy farms in Bangladesh.

Background and Aim: The emergence of antimicrobial-resistant bacteria, such as Escherichia coli in milk, is a serious public health concern as milk is considered a complete food and an important part of daily human diet worldwide, including in Bangladesh. However, there have been no reports on the molecular characterization and antibiotic resistance profile of extended-spectrum beta-lactamase (ESBL)-producing E. coli from milk of healthy cows in Bangladesh. Therefore, this study aimed to detect and characterize ESBL-producing E. coli (ESBL-Ec) in milk samples from healthy cows in smallholder dairy farms in Mymensingh district, Bangladesh, and assess the potential risk of consuming this milk. Materials and Methods: A total of 100 milk samples were collected from apparently healthy cows on smallholder dairy farms. Escherichia coli was isolated from the collected samples using standard methods. The detection of ESBL-Ec was performed phenotypically using cultural methods and genotypically by ESBL genetic determinants using multiplex polymerase chain reaction. Antimicrobial susceptibility testing of the ESBL-Ec isolates was performed using the disk diffusion method with 15 common antimicrobials. Results: In this study, out of the 100 samples tested, 70 (70%) were found to be positive for E. coli. Among these, 41 (58.6%) strains were identified as ESBL-producing, both phenotypically and genotypically, with the presence of blaCTX-M, blaTEM, and blaSHV individually or combined (blaCTX-M plus blaTEM plus blaSHV). The antibiogram of these ESBL-positive isolates revealed high resistance against commonly used antibiotics, such as ampicillin, cefotaxime, and gentamicin (100%), azithromycin (88%), oxytetracycline (27%), nalidixic acid, cotrimoxazole/trimethoprim (24%), and streptomycin (22%). In addition, one isolate showed resistance to 4th generation of cephalosporin (cefepime). Most importantly, extensive multidrug resistance was found in many ESBL-Ec isolates. However, the isolates were highly sensitive to drugs such as ceftriaxone (100%) and imipenem (100%). This is the first study to detect ESBL-Ec in raw milk from healthy cows on smallholder dairy farms in Bangladesh. Conclusion: More than 58% of the E. coli isolated from raw milk of healthy cows tested positive for ESBL production and showed resistance to most commonly used antimicrobials which may be alarming for human health. A limitation of our study is that we had a small size of sample collected from one district in Bangladesh. Therefore, a larger sample size covering a wider geographic area, and using multi-locus sequence typing and whole genome sequencing could provide a more comprehensive understanding of the prevalence and characteristics of ESBL-Ec in Bangladesh.

Electrical conductivity and total dissolved solid of raw milk for the detection of bovine subclinical mastitis.

Background and Aim: Bovine subclinical mastitis (SCM) is highly prevalent among dairy cattle. A cross-sectional study was conducted in Bangladesh to evaluate the performance of electric conductivity (EC) and total dissolved solids (TDS) tests for the detection of SCM. Materials and Methods: We randomly selected 108 milk samples from cows of different breeds in the primary milk-producing region of Pabna and Sirajgonj districts of Bangladesh. Samples were subjected to the California mastitis test (CMT), white side test (WST), electric conductivity (EC), TDS, and culture. A cow was considered positive for SCM if it tested positive in CMT, WST, and culture, whereas a cow was considered negative for SCM if it tested negative in all three methods. These gold standards have been used to evaluate the performance of the EC and TDS tests. The optimal EC and TDS cutoff values for the detection of SCM were determined using the "optimal cutoff" function in R version 4.3.1. Results: The optimal EC cutoff value for SCM detection was found to be 6159 µS/cm or 6.16 mS/cm. A positive likelihood ratio (LR+) of 31.2 and an area under the curve (AUC) of 0.905 were obtained for this cutoff value. The optimal cutoff value for TDS was 3100 mg/L of milk, which resulted in a positive LR+ of 45.5 and an AUC of 0.924. Conclusion: To the best of our knowledge, this is the first study to evaluate the performance of EC and TDS tests in detecting SCM in Bangladesh. These results suggest that EC and TDS tests, which are inexpensive, rapid, and easy to conduct, can effectively detect SCM at the farm level.

Epidemiological investigation of lumpy skin disease outbreaks in Bangladeshi cattle during 2019-2020.

Lumpy skin disease (LSD) is an acute infectious viral disease of cattle with a wide distribution that emerged in Bangladesh in 2019, causing huge economic losses. This study was undertaken to investigate the epidemiological features of LSD emergence in nine districts of Bangladesh between December 2019 and December 2020. A total of 8215 cattle from 603 herds were investigated and LSD was diagnosed based on the characteristic clinical findings. A standard questionnaire was administered to collect herd-level data including location, herd size, number of LSD-infected cattle, number died due to LSD, farm type, season, house type, vector presence, sanitation and fly repellent use. Similarly, data on clinical signs, sex, age, animal class and breed of the LSD-infected cattle were also recorded. The herd-level attack risk (%) and mortality risk (%) were calculated based on the number of infected and dead cattle, respectively, as a proportion of total cattle. The herd-level risk factors for LSD were identified using a multivariable Poisson regression model. The most common clinical signs were skin nodules (100%), fever (97.9%) and depression with anorexia and weight loss (97.9%). Crossbred (84.9%) and female (72.2%) cattle were mostly affected by LSD. The overall LSD attack risk, mortality risk and case fatality were 26.5%, 0.26% and 0.97%, respectively. The LSD attack risk was significantly higher in small herds (risk ratio: [RR] 1.39; 95% CI: 1.27; 1.53) than large herds. In addition, significantly higher LSD attack risk was observed in semi-intensive management systems (RR = 1.29; 95% CI: 1.01; 1.64) than intensive management systems. Moreover, it was also significantly higher in hut (RR = 1.81; 95% CI: 1.12; 2.92), temporary (RR = 1.62; 95% CI: 1.21; 2.17) and tin-shed houses (RR = 1.29; 95% CI: 1.11; 1.51) than in semi-building houses. To the best of our knowledge, this is the first detailed epidemiological study of LSD emergence in South Asia. Female crossbred cattle in small herds under semi-intensive management should be prioritized for LSD surveillance and vaccination to prevent further outbreaks and control the impact of the disease in Bangladesh.

Hierarchical true prevalence, risk factors and clinical symptoms of tuberculosis among suspects in Bangladesh.

BACKGROUND: The study was aimed to estimate the true prevalence of human tuberculosis (TB); identify risk factors and clinical symptoms of TB; and detect rifampicin (RIF) sensitivity in three study areas of Bangladesh. METHODS: The cross-sectional study was conducted in three Bangladesh districts during 2018. Potential risk factors, clinical symptoms, and comorbidities were collected from 684 TB suspects. Sputum specimens were examined by LED microscopy. TB hierarchical true prevalence, risk factors and clinical symptoms were estimated and identified using a Bayesian analysis framework. Rifampicin sensitivity of M. tuberculosis (MTB) was detected by GeneXpert MTB/RIF assay. RESULTS: The median TB true prevalence was 14.2% (3.8; 34.5). Although overall clustering of prevalence was not found, several DOTS centers were identified with high prevalence (22.3% to 43.7%). Risk factors for TB identified (odds ratio) were age (> 25 to 45 years 2.67 (1.09; 6.99), > 45 to 60 years 3.43 (1.38; 9.19) and individuals in families/neighborhoods where a TB patient(s) has (ve) already been present (12.31 (6.79; 22.60)). Fatigue, night sweat, fever and hemoptysis were identified as important clinical symptoms. Seven of the GeneXpert MTB/RIF positive sputum specimens (65) were resistant to rifampicin. CONCLUSIONS: About one in every seven TB suspects was affected with TB. A number of the TB patients carry multi drug resistant MTB. Hierarchical true prevalence estimation allowed identifying DOTS centers with high TB burden. Insights from this study will enable more efficient use of DOTScenters-based TB surveillance to end the TB epidemic in Bangladesh by 2035.

Risk factors for bovine rotavirus infection and genotyping of bovine rotavirus in diarrheic calves in Bangladesh.

Bovine rotavirus (BRV) is considered the leading cause of calf diarrhea worldwide, including Bangladesh. In this study we aimed to identify risk factors for BRV infection and determine the G and P genotypes of BRV strains in diarrheic calves. Fecal samples were collected from 200 diarrheic calves in three districts between January 2014 and October 2015. These samples were screened to detect the presence of BRV using rapid test-strips BIO K 152 (RTSBK). The RTSBK positive samples were further tested by polyacrylamide gel electrophoresis and the silver staining technique to detect rotavirus dsRNA. Risk factors were identified by multivariable logistic regression analysis. The G and P genotypes of BRV were determined by RT-PCR and sequencing. A phylogenetic tree was constructed based on the neighbor-joining method using CLC sequence viewer 8.0. About 23% of the diarrheic calves were BRV positive. The odds of BRV infection were 3.8- (95% confidence interval [95% CI]: 1.0-14.7) and 3.9-times (95% CI:1.1-14.2) higher in Barisal and Madaripur districts, respectively, than Sirrajganj. The risk of BRV infection was 3.1-times (95% CI: 1.5-6.5) higher in calves aged ≤ 5 weeks than those aged >5 weeks. Moreover, the risk of BRV infection was 2.6-times (95% CI:1.1-5.8) higher in crossbred (Holstein Friesian, Shahiwal) than indigenous calves. G6P[11] was the predominant genotype (94.4%), followed by G10P[11] (5.6%). The BRV G6 strains were found to be closest (98.9-99.9%) to Indian strains, and BRV G10 strains showed 99.9% identities with Indian strain. The VP4 gene of all P[11] strains showed >90% identities to each other and also with Indian strains. The most frequently identified BRV genotype was G6P[11]. About 23% of calf diarrhea cases were associated with BRV. To control disease, high-risk areas and younger crossbred calves should be targeted for surveillance and management. The predominant genotype could be utilized as the future vaccine candidate or vaccines with the dominant genotype should be used to control BRV diarrhea in Bangladesh.

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.

Risk factors and true prevalence of bovine tuberculosis in Bangladesh.

Bovine tuberculosis (bTb) is endemic in Bangladesh but the true prevalence has not yet been reported. Our objectives for this study were to determine the true prevalence and identify risk factors for bTb at the animal- and herd-level in Bangladesh. A total of 510 cows were randomly selected during January 2018 to December 2018. Caudal fold (CFT) and comparative cervical tuberculin tests (CCT) were serially interpreted. Animal- and herd-level risk factor data were collected using a pre-tested questionnaire. The hierarchical true prevalence of bTb was estimated within a Bayesian framework. The herd- and animal-level risk factors were identified using mixed effects logistic regression. The apparent prevalence of bTb was 20.6% [95% Confidence Interval (CI): 17.3; 24.3] based on CFT. The animal-level true prevalence of bTb was 21.9 (13.0; 32.4). The herd-level true prevalence in different regions varied from 41.9% to 88.8%. The region-level true prevalence was 49.9 (13.8; 91.2). There is a 100% certainty that herds from Bhaluka and Mymensingh Sadar upazilas are not free from bTb. The odds of bTb were 3.9 times (1.2; 12.6) higher in herds having more than four cows than those with ≤ 4 cows. On the other hand, the risk of bTb was 3.3 times higher (1.0; 10.5) in non-grazing cows than grazing cows. Crossbred cows were 2.9 times (1.5; 5.9) more likely to be infected with bTb than indigenous cows. The risk of bTb in animals with cough was 2.3 times (1.2; 4.3) higher than those without cough. Crossbred, non-grazing cows with cough should be targeted for bTb surveillance. Herds of the Mymensingh, Sadar and Bhaluka regions should be emphasized for bTb control programs. Estimation of Bayesian hierarchical true prevalence facilitates identification of areas with higher prevalence and can be used to indicate regions that where true prevalence exceeds a pre-specified critical threshold.

Seroprevalence and risk factors for bovine brucellosis in the Chittagong Metropolitan Area of Bangladesh.

Brucellosis is a neglected endemic zoonosis in Bangladesh and has a significant impact on public health and animal welfare of dairy farming as well as dairy farm economics. A cross-sectional study was conducted to evaluate the seroprevalence of and risk factors for brucellosis in dairy cattle in the Chittagong metropolitan area (CMA) of Chittagong, Bangladesh. We collected serum samples (n = 158) from six randomly selected dairy farms in the CMA between February and November, 2015. The Rose Bengal Plate Test (RBPT) and a competitive ELISA (cELISA) were used as the screening and confirmatory tests respectively. Farm level and animal level demographic and risk factor data were collected using a questionnaire. The risk factors were analysed using a multivariable logistic regression with random effects. The overall seroprevalences of antibodies against brucellosis in cattle were 21.5% (34/158) and 7.6% (12/158) based on parallel and serial interpretation of the two tests respectively. Our results revealed that 20.3% (32/158) samples were positive using the RBPT and 8.9% (14/158) were positive using the cELISA. The within-herd seroprevalence ranged from 10% to 26.3% and 5 to 20.7% using the RBPT and cELISA tests respectively. The odds of seropositivity were significantly higher in lactating cows (OR: 2.59; 95% CI: 1.02-6.55), cows producing less than 2 litres of milk (OR: 29.6; 95% CI: 4.3-353.8), cow producing 2-12 litres of milk (OR: 4.8; 95% CI: 1.1-33.4) and cows with reproductive disorders (OR: 3.2; 95% CI: 1.2-10.1). About 7.6% (12/158) and 1.3% (2/158) of cattle were found to be infected with acute and chronic brucellosis respectively. Based on these results, we suggest that cows that have reproductive disorders and are producing little milk should be prioritized for brucellosis screening in CMA. The screening tests should be used to control brucellosis in cattle in order to protect animal welfare, human health and to minimize the economic losses.

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

A cross-sectional survey was conducted in selected districts of Bangladesh to estimate the prevalence of bovine tuberculosis (bTB), and to identify the risk factors for bTB. We included 1865 farmed cattle from 79 herds randomly selected from five districts. Herd and animal level data were collected using semi-structured interviews with cattle herd owners. The single intradermal comparative tuberculin test (SICTT) was used to estimate the prevalence of bTB. The risk factors were identified using mixed-effect multiple logistic regression analyses. The overall herd and animal level prevalences of bTB were estimated to be 45.6% (95% Confidence Interval [CI] = 34.3-57.2%) and 11.3 (95% CI = 9.9-12.8%), respectively, using the OIE recommended >4 mm cut-off. The true animal level prevalence of bTB was estimated to be 11.8 (95% Credible Interval = 2.1-20.3%). At the herd level, farm size, bTB history of the farm and type of husbandry were significantly associated with bTB status in univariable analysis. Similarly, age group, sex, pregnancy status and parity were significantly associated with bTB at cattle level. However, in multivariable analysis only herd size at the herd level and age group and pregnancy status at the cattle level were significant. Compared to a herd size of 1-10, the odds of bTB were 22.8 (95% CI: 5.2-100.9) and 45.6 times (95% CI: 5.0-417.7) greater in herd sizes of >20-50 and >50, respectively. The odds of bTB were 2.2 (95% CI: 1.0-4.5) and 2.5 times (95% CI: 1.1-5.4) higher in cattle aged >3-6 years and > 6 years, compared to cattle aged ≤1 year. Pregnancy increased the odds of bTB by 1.7 times (95% CI: 1.2-2.4) compared to non-pregnant cattle. Taken together, the results suggest high herd and animal level prevalence of bTB in these 5 districts, with the greatest risk of bTB in older and pregnant cattle within large herds (>20), and highlight an urgent need for continued surveillance and implementation of bTB control programs in Bangladesh.

Foot-and-Mouth Disease Space-Time Clusters and Risk Factors in Cattle and Buffalo in Bangladesh.

Foot-and-mouth disease (FMD) is highly endemic in Bangladesh. Using passive surveillance data (case records from all 64 districts of Bangladesh, 2014-2017) and district domestic ruminant population estimates, we calculated FMD cumulative incidence per 10,000 animals at risk per district, conducted cluster (Moran's spatial autocorrelation and scan statistics) and hotspot analysis (local indicator of spatial association statistic), created predictive maps and identified risk factors using a geographically weighted regression model. A total of 548,817 FMD cases in cattle and buffalo were reported during the four-year study period. The highest proportion (31.5%) of cases were reported during the post-monsoon season, and from Chattogram (29.2%) division. Five space-time clusters, 9 local clusters, and 14 hotspots were identified. Overall, higher cumulative incidences of FMD were consistently predicted in eastern parts of Bangladesh. The precipitation in the pre-monsoon season (p = 0.0008) was positively associated with FMD in Bangladesh. Results suggest climate plays an important role in the epidemiology of FMD in Bangladesh, and high risk zones exist. In a resource limited-setting, hotspots and clusters should be prioritized for vaccination coverage, and surveillance for FMD should be targeted in eastern areas of Bangladesh and during the post-monsoon season.

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.

Molecular detection of Toxoplasma gondii from aborted fetuses of sheep, goats and cattle in Bangladesh.

The study was planned to apply the PCR method for detection of T. gondii infection in sheep, goats and cattle aborted fetuses from Mymensingh, Bangladesh. A total of 58 fetal tissue samples (brain, liver, heart, skeletal muscle and placenta) of sheep (5), goats (5) and cattle (2) were selected for study. Aborted fetuses were taken from serologically positive mothers by indirect ELISA. Among them 24 and 34 samples were subjected for PCR assay by using TgB1 and TgTox4 primers respectively. DNA fragments were visualized under UV illumination after gel run. The results demonstrated 15.52% tissue samples from sheep and goat aborted fetuses were positive for T. gondii parasite. Among different tissue samples, brain, liver and heart showed presence of T. gondii parasite. None of tissue samples showed positive in case of cattle. The results of the PCR exhibited that T. gondii infection might be considered as one of the major causative agents for abortion in ewes and does. Further studies are needed to improve our knowledge on different genotypes of T. gondii that infect sheep, goat and cattle population in Bangladesh.

Seroprevalence of Toxoplasma gondii infection in ruminants in selected districts in Bangladesh.

To estimate the seroprevalence of toxoplasmosis and risk factors for seropositivity in sheep, goats and cattle in Dhaka, Mymensingh, Sirajganj and Chittagong districts in Bangladesh, 1104 sera samples (552 sheep, 300 goats and 252 cattle) were randomly selected and tested by an indirect enzyme linked immunosorbent assay (iELISA). The overall seroprevalence was 12.2% (135/1104), and was significantly (P=0.008) higher in goats (16.0%) than cattle (8.3%). The odds of seropositivity was 2.09 times (95% confidence interval [CI]: 1.23-3.67) higher in goats than cattle. In sheep, herd type, district and pregnancy status were significant risk factors. The odds of seropositivity was 2.1 (95% CI: 1.00-4.84), 7.29 (95% CI: 12.71-423.41) and 10.88 (95% CI: 5.42-23.41 times higher in sheep-only flocks, in Mymensingh district and in pregnant sheep than in mixed flocks, Chittagong district and non-pregnant sheep, respectively. In cattle, breeds and district were significant risk factors. The odds of seropositivity was 5.79 (95% CI: 1.13-24.62) and 4.29 (95% CI: 1.38-16.34) times higher in Holstein Friesian cross cattle and in Mymensingh district than in indigenous cattle and Chittagong district, respectively. This study indicates that exposure of sheep, goats and cattle to oocysts of T. gondii is widespread, suggesting that the consumption of raw and undercooked meat of these animals might be a source of human toxoplasmosis. Risk factor information can be used to design control programs to reduce exposure.