Prevalence of zoonotic Mycobacterium bovis (bTB) disease in human population is underreported from the North of Pakistan. Here, we report on the proportion of human bTB disease among the overall TB patients, drug resistance pattern of bTB isolates, and knowledge, attitude, and practices (KAP)-based analysis of bTB disease. For this purpose, sputum samples from a total of 300 clinically diagnosed TB patients and 100 randomly selected school children suspected of pulmonary TB were processed by culture as well as polymerase chain reaction (PCR) for isolation, identification, and confirmation of Mycobacterium tuberculosis (mTB) and bTB species. Isolates of bTB were processed for drug susceptibility tests. Data on KAP regarding TB were obtained on a pretested questionnaire. Sputum-based PCR results indicated that 288/300 (96%) were confirmed as mTB, while 12/300 (4%) were found as bTB diseases. Interestingly, none of the school child was declared positive for either mTB or bTB. Notably, 274/300 (91.3%) positively cultured samples were identified as mTB, 13/300 (4.3%) as bTB, while 5/300 (1.7%) as mixed containing both. Importantly, except one, all of the bTB isolates were found resistant to pyrazinamide. Surprisingly, most of the bTB isolates (~70%) were found resistant to a broad range of first- and second-line anti-TB drugs. SplitsTree and recombination analysis indicated no evidence of intergenic recombination. Finally, residence, occupation, presence of animals at home, and sleeping alongside animals were found significantly associated with occurrence of bTB disease. To the best of our knowledge, we report for the first time on the high (4%) burden of bTB disease in human TB patients in Peshawar, Pakistan.

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The dietary supplementation of trace minerals and probiotics has long been practiced to reduce the harmful effects of high environmental temperature in poultry. The study aimed to assess the effect of single or combined supplementation of ZnSO 4 •H 2 O and probiotic (Protexin ® ) on the growth performance, gut biometry and immune organs histomorphology in broilers reared under cyclic heat stress. For this purpose, 280 day-old broiler chicks were randomly divided into seven groups designated as control (basal diet), HS (heat stress + basal diet), HS-Zn30 (HS + 30 mg ZnSO 4 •H 2 O/kg feed), HS-Zn60 (HS + 60 mg ZnSO 4 •H 2 O/kg feed), HS-Pro (HS + 0.1 g Protexin ® /kg feed), HS-Zn30-Pro (HS + 30 mg ZnSO 4 •H 2 O/kg feed + 0.1 g Protexin ® /kg feed), and HS-Zn60-Pro (HS + 60 mg ZnSO 4 •H 2 O/kg feed + 0.1 g Protexin ® /kg feed). Each group consisted of five replicates (8 birds per replicate). The birds were fed on starter (0 -21days) and grower (22 -42 days) diets. From day 22 onwards, the birds were exposed to cyclic heat stress (temperature and relative humidity at 35 ± 1 °C and 75 ± 5% respectively) for 8 hours. The results revealed that birds exposed to heat stress showed significant decrease in body weight (BW) and increase in feed conversion ratio (FCR) compared to the control group. The dietary supplementation of ZnSO 4 •H 2 O and probiotic either singly or in combination improved BW and FCR throughout the experimental period in heat-stressed broilers. Area of lymphatic nodule and lymphoid follicle in caecal tonsils and bursa of Fabricius increased in HS-Pro, HS-Zn30-Pro and HS-Zn60-Pro groups compared to HS group. In conclusion, zinc and probiotics combination partially improved performance and immune organs histomorphology in heat-stressed broilers.

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