Veterinary Medicine International collected aseptically from the lung lobes (left apical, left diaphragmatic, right apical, right cardiac, right diaphrag- matic, and right accessory), lymph nodes of the head (retropharyngeal and mandibular), lymph nodes of lungs (mediastinal and bronchial), and mesenteric lymph nodes. Data were collected on the presence, size, and distribution of visible lesions in each carcass. Samples from tissues con- taining visible lesions were collected and placed into sterile universal bottles containing 5 mL of 0.9% saline solution(pH 7.2) and kept at −20◦C at Semera Regional Animal Health Laboratory until they were transported to ALIPB laboratory under cold chain for isolation of the causative agents. 2.7. Isolation of Mycobacteria from Tissue Samples. Isolation of mycobacteria from tissues was done in accordance with OIE protocols [31]. Brieﬂy, the specimens were sectioned into pieces using sterile blades and then homogenized by pestle and mortar. The homogenate was decontaminated by adding an equal volume of 4% NaOH followed by centrifugation at 1000 g for 15 minutes. The supernatant was discarded, while the sediment was neutralized by 1% (0.1N) HCl using phenol red as an indicator. Neutralization was achieved when the colour of the solution changed from purple to yellow. Thereafter, 0.1 mL of suspension from each sample was spread onto a slope of Lo¨wenstein-Jensen (LJ) medium. Duplicates of LJ media were used; one enriched with sodium pyruvate, while the other was enriched with glycerol. Cultures were incubated at 37◦C in a slant position for one week and in upright position for 11 weeks with weekly observation for mycobacterial growth. Whenever, colonies were seen, subculturing and Ziehl-Neelsen staining were performed to conﬁrm the presence of acid fast bacilli. Positive colonies were preserved with freezing media, and some portions of the colonies were heat-killed in water bath at 80◦C for 45 minutes. The frozen and heat killed isolates were stored at −20◦C for future mycobacteriology and further molecular typing analysis. 2.8. Molecular Characterization of Mycobacterial Isolates. Mycobacterium genus typing was conducted as described previously [32], and spoligotyping of Mycobacterium tuber- culosis complex isolate from goat was performed as pre- viously described by Kamerbeek and coauthors [33]. Both methods were described in detail in previous publication [22]. 2.9. Data Management and Analysis. Data were classiﬁed, ﬁltered, coded using EpiData software and Microsoft Excel sheet, and was transferred and analyzed using STATA version 11 (Stata Corp., Collage station, TX). Pearson chi-square was used to evaluate the statistical signiﬁcance of the associations of diﬀerent categorical variables with skin test results. Bivariate and multivariable logistic regression anal- yses were performed to quantify crude and adjusted eﬀects of prespeciﬁed risk factors on tuberculin reactivity. P values less than 5% were considered statistically signiﬁcant. In cases of estimating the eﬀect of diﬀerent risk factors in terms of OR with corresponding 95% conﬁdence interval, statistically 3 signiﬁcance was assumed if the conﬁdence interval did not include one among its values. 3. Results 3.1. Animal Prevalence. On the basis of CIDT test, the animal prevalence of TB was 0.5% (10/2231) at a cutoﬀ ≥4 mm and 3.8% (86/2231) at a cutoﬀ ≥ 2 mm. At ≥2 mm cut-oﬀ point, there were signiﬁcant diﬀerences in proportions of reactors among the four districts (χ2 = 26.385, P = 0.000), between sheep and goat (χ2 = 6.46, P = 0.011) and between pregnant and nonpregnant females (χ2 = 5.342, P = 0.021) (Table 1). Multivariable logistic regression analysis at ≥2 mm cut-oﬀ point showed that older small ruminants (5 years and above) had 13 times the odds of being tuberculin reactors compared with age category less than 2 years old (adjusted OR = 13.79; CI: 2.22–85.55). Female small ruminants with parity number greater than 4 had 0.05 odds of being bovine tuberculin positivity in relative to those with less than 2 parity numbers (adjusted OR = 0.05; CI: 0.01–0.31) (Table 2). At ≥4 mm cut-oﬀ points, there was no statistical signiﬁcance diﬀerence in the proportion of bovine tuberculin positivity between groups in relation to the diﬀerent variables considered. 3.2. Herd Prevalence. The herd prevalence was 20% (95% CI: 12–28%) and 47% (95% CI: 37–56%) at ≥4 mm and ≥2 mm cut-oﬀ points, respectively. In multivariable logistic regression analysis, no signiﬁcant association was found in herd positivity between groups in relation to district of origin, herd size category, and production system at ≥2 mm cut-oﬀ point (Table 3). 3.3. Prevalence of Mycobacterium Avium Complex Infection. According to the CIDT test result of the avian tuberculin skin reaction, the overall animal prevalence of Mycobacterium avium complex infection was 2.8% (95% CI: 2.1–3.5%) and 6.8% (95% CI: 5.8–7.9%) at ≥4 mm and ≥2 mm cut-oﬀ points, respectively. 3.4. Postmortem Lesions in Tuberculin Reactor Goats. Five goats which were positive to bovine TB in CIDT test at cutoﬀ ≥ 4 mm were purchased, slaughtered, and investigated for gross tuberculous lesions. Tuberculous lesions were detected in diﬀerent organs (left diaphragmatic lung, retropharyngeal lymph node, parotid lymph node, right bronchial lymph node, mesenteric lymph node, intestinal wall, and mesen- tery). Two of them had partially disseminated TB lesions which involved lung, intestine and the lymph nodes of thoracic and abdominal cavities. Upon incision of the lung, lesions showed a yellowish caseous material indicating a characteristic of tuberculous lesion (Figure 1). In mesen- tery and mesenteric lymph nodes, greenish discharge was observed in the lesions. 3.5. Molecular Typing of the Isolates from Goats. All tis- sue samples obtained from slaughtered tuberculin reactor goats were positive for mycobacterial growth on LJ culture medium. Further molecular characterization indicated that