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.

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.

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.

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.