Dynamics and risk of transmission of bovine tuberculosis in the emerging dairy regions of Ethiopia.

The Ethiopian government has several initiatives to expand and intensify the dairy industry; however, the risk of bovine tuberculosis (bTB) spread is a challenge. To assess the rate of expansion and risk factors for transmission of bTB within-herds, we carried out a repeated cross-sectional survey at two time points, 2016/17 and 2018, in three regional cities, namely, Gondar, Hawassa and Mekelle, representing the emerging dairy belts of Ethiopia. The total number of herds involved was 128, comprising an average of 2303 cattle in each round. The Single Intradermal Comparative Cervical Tuberculin (SICCT) test was used to identify reactor status and data on herd-level risk factors were collected using a structured questionnaire. In the first survey, the apparent prevalence of bTB, as measured by the SICCT test, was 4.5% (95% CI 3.7-5.4%) at the individual animal-level and 24% (95% CI 17.5-32%) at the herd-level. There was no statistically significant change in the overall apparent prevalence or regional distribution at the second survey, consistent with the infection being endemic. The incidence rate was estimated at 3.6 (95% CI 2.8-4.5) and 6.6 (95% CI 3.0-12.6) cases/100 cattle (or herd)-years at the animal- and herd-levels, respectively. Risk factors significantly associated with the within-herd transmission of bTB were age group and within-herd apparent prevalence at the start of the observation period. We noted that farmers voluntarily took steps to remove reactor cattle from their herds as a consequence of the information shared after the first survey. Removal of reactors between surveys was associated with a reduced risk of transmission within these herds. However, with no regulatory barriers to the sale of reactor animals, such actions could potentially lead to further spread between herds. We therefore advocate the importance of setting up regulations and then establishing a systematic bTB surveillance programme to monitor the impact prior to implementing any control measures in Ethiopia.

Spoligotyping and drug sensitivity of Mycobacterium tuberculosis isolated from pulmonary tuberculosis patients in the Arsi Zone of southeastern Ethiopia.

Tuberculosis (TB) is one of the leading causes of morbidity and mortality in different zones of Ethiopia. This study was undertaken to identify the strains of Mycobacterium tuberculosis and evaluate their drug sensitivity profiles in the Arsi Zone. A total of 111 isolates of M. tuberculosis from individuals with pulmonary TB were included and speciation and strain identification were performed using Region of difference 9 and spoligotyping, respectively. The drug sensitivity patterns were assessed using Bactec MGIT 960 SIRE and GenoType MTBDRplus line probe assays. Of 111 isolates, 83% were interpretable and 56 different spoligotype patterns were identified. From these, 22 patterns were shared types while the remaining 34 were orphans. The predominant shared types were spoligotype international type (SIT) 149 and SIT53, comprising 12 and 11 isolates, respectively. Euro-American lineage was the dominant lineage followed by East-African-Indian. Phenotypically, 17.2% of tested isolates were resistant to any first-line drugs and 3.1% were multidrug-resistant. Higher (6.2%) mono-resistance was observed to streptomycin, and no resistance was observed to rifampicin or ethambutol. Genotypically, five (5.4%) isolates were resistant to isoniazid and mutated at codon S315T1 of katG. In contrast, only 1.1% of the isolates were resistant to rifampicin and were mutated at codon S531L of rpoB gene. In this study, a high proportion of orphan strains were isolated, which could suggest the presence of new strains and a high percentage of mono-resistance, warranting the need to strengthen control efforts.

Prevalence of pulmonary tuberculosis among students in three eastern Ethiopian universities.

SETTINGS: Three universities located in eastern Ethiopia: Haramaya University, Haramaya; Dire-Dawa University, Dire-Dawa; and Jigjiga University, Jigjiga. OBJECTIVE: To determine the burden of pulmonary tuberculosis (PTB) among university students and to identify risk factors for the development of TB disease. DESIGN: All full-time university students were screened for symptoms of PTB and sputum was collected for acid-fast bacilli (AFB) examination and culture for Mycobacterium tuberculosis. RESULTS: Of 35 344 students screened, we identified 153 PTB cases that occurred over the 1-year study period, or 433/100 000 students. Of these, 117 (76%) PTB cases were found through passive case finding at student health centres, while 36 (24%) previously undiagnosed patients were identified through active case finding. Sixteen cases detected using active case finding (44%) were smear-positive. Living in a dormitory with 5 students and attending university for 2 years were both significantly associated with PTB (adjusted OR 2.49 and 3.79, respectively, P < 0.001). In persons who underwent drug susceptibility testing, 11 (30.5%) had resistance to at least one first-line anti-tuberculosis drug. CONCLUSION: We found a high burden of TB among university students in eastern Ethiopia. Screening for PTB upon university admission and at regular intervals should be considered to minimise TB transmission on university campuses.

Genotyping and molecular detection of multidrug-resistant Mycobacterium tuberculosis among tuberculosis lymphadenitis cases in Addis Ababa, Ethiopia.

Multidrug-resistant tuberculosis (MDR-TB) has emerged as a major public health problem. Drug-resistance surveillance data show that 3.9% of new and 21% of previously treated TB cases were estimated to have had rifampicin/ multidrug-resistant tuberculosis (MDR/RR-TB) in 2015. This implies that the MDR-TB is increasing alarmingly. Hence, a better understanding of drug resistance mechanisms and genotypes associated with multidrug resistance in M. tuberculosis is crucial for improving diagnostic and therapeutic methods to treat individuals with MDR-TB. The aim of this study was to analyze molecular drug resistance mutations of MDR-TB isolates from the cases of TB-lymphadenitis in relation to its genetic lineages. A cross-sectional study was conducted on culture positive cases from July to October, 2014 in Addis Ababa, Ethiopia. Sixty isolates were included to analyze drug resistance mutated gene responsible for MDR-TB in relation to its molecular genotyping. Mycobacterial culture, GenoTypeMTBDR plus and Spoligotyping were used to undertake the study. Of 60 TBLN isolates, 8.3% were identified MDR-TB cases and one isolate was isoniazid mono-resistant. Eleven isolates in T3-ETH genetic sub lineage were sensitive to both RMP and INH, while only 2 isolates were MDR-TB. Most of the RMP- resistant isolates showed mutation in codon S531L and all isolates mutated in the katG gene conferring INH resistant strains had mutations in codon of S315T1. Screening for the rpoB and katG gene mutation of tuberculosis lymphadenitis is useful in Ethiopia for an early detection and treatment of MDR-TB. Besides, there is a drug resistance variation among different lineages of Tuberculosis lymphadenitis which has important consequences for the development of efficient control strategies.

Vaccination of calves with Mycobacterium bovis Bacillus Calmette-Guerin reduces the frequency and severity of lesions of bovine tuberculosis under a natural transmission setting in Ethiopia.

Bovine tuberculosis (bTB) is highly prevalent in intensive dairy farms of the urban "milk-sheds" in Ethiopia, and vaccination could be a cost-effective disease control strategy. In the present study, the efficacy of Bacillus Calmette-Guerin (BCG) to protect against bTB was assessed in Holstein-Friesian calves in a natural transmission setting. Twenty-three 2-week-old calves were subcutaneously vaccinated with BCG Danish SSI strain 1331, and matched 26 calves were injected with placebo. Six weeks later, calves were introduced into a herd of M. bovis-infected animals (reactors) and kept in contact with them for 1 year. In vitro and in vivo immunological tests were performed to assess immune responses post-vaccination and during exposure. Successful vaccine uptake was confirmed by tuberculin skin test and IFN-γ responses in vaccinated calves. The kinetics of IFN-γ responses to early secretory antigen target 6 and culture filtrate protein 10 (ESAT6 and CFP10, respectively) and tuberculin skin test responses post-exposure suggested that the animals were infected early after being placed in contact with the infected herd as immunological signs of infection were measurable between 2 and 4 months post-initial exposure. Protection was determined by comparing gross and microscopic pathology and bacteriological burden between vaccinated and control calves. BCG vaccination reduced the proportions of tissues with visible pathology in vaccinates compared to control calves by 49% (p < .001) with 56%, 43%, 72%, and 38% reductions in the proportion of lesioned tisues in head, thoracic, abdominal lymph nodes, and lungs, respectively (p-values .029-.0001). In addition, the lesions were less severe grossly and microscopically in vaccinated calves than in non-vaccinated calves (p < .05). The reduction in the overall incidence rates of bTB was 23%, 28%, and 33% on the basis of the absence of gross pathology, M. bovis culture positivity, and histopathology, respectively, in vaccinated animals. In conclusion, BCG vaccination reduced the frequency and severity of the pathology of bTB significantly, which is likely to reduce onwards transmission of the disease.

Mycobacterium tuberculosis in central Ethiopia: drug sensitivity patterns and association with genotype.

Drug resistance tuberculosis (TB) and the emergence of multidrug resistant (MDR) isolates are significant concerns regarding TB control programs in several countries. This study was undertaken to evaluate the drug sensitivity of Mycobacterium tuberculosis and to assess its association with strains and lineages of M. tuberculosis. A total of 279 M. tuberculosis strains isolated from Central Ethiopia were tested for their drug sensitivity patterns to first line TB drugs using the conventional proportion method on Löwenstein Jensen media. The association between drug sensitivity and strain type was assessed on 263 isolates of the 279 isolates. Of the 268 M. tuberculosis isolates obtained from new cases, 209 (78%) were susceptible to first line TB drugs, and 59 (22.2%) bacterial isolates were resistant to at least one of the first line drugs. The highest mono-resistance (7.5%) pertained to streptomycin (STM). Remarkably, seven of eleven isolates (63.6%) previous treatment for TB were resistant to at least one of the first line drugs. The prevalence of MDR-TB was 1.5% (4/268) for newly identified TB cases, all of which were members of the Euro-American Lineage. There was no statistically significant association (P > 0.05) between drug sensitivity, and either strains, sub-lineages or main lineages of M. tuberculosis. A significant proportion of M. tuberculosis was resistant to at least one first line anti-TB drug. Moreover, the frequencies of resistance to either isoniazid or rifampicin were high compared to data that were previously reported in some part of the country.

Molecular typing of mycobacteria isolated from tuberculous lymphadenitis cases in Addis Ababa, Ethiopia.

BACKGROUND: In Ethiopia, one of the world's 22 high tuberculosis (TB) burden countries, one third of the tuberculosis (TB) cases are attributed to tuberculosis lymphadenitis (TBLN). However, information on the molecular type of the mycobacterial species and strains that cause TBLN in the country is scarce. OBJECTIVE: To identify the species and strains of mycobacteria that cause TBLN in Ethiopia. METHODS AND RESULTS: A cross-sectional study was conducted among 206 presumed TBLN cases to characterise positive culture isolates. Mycobacterium tuberculosis complex species and strains were identified using region of difference 9 deletion and SITVIT WEB, respectively. Of the 206 fine-needle aspirate samples collected, 74 (36%) were culture-positive: 73 (98.6%) of the isolates were M. tuberculosis, and the remaining 1.4% were M. bovis. Further characterisation of the 73 M. tuberculosis isolates led to 26 distinct spoligotype international types (SITs) and 13 newly identified patterns. The most prevalent strains were SIT149, SIT53, SIT26 and SIT37 of sublineages T3-ETH, T1, CASI-DELHI and T3, respectively; these accounted for 52.6% of the total number of strains. CONCLUSION: TBLN was mainly caused by M. tuberculosis and highly clustered strains SIT149, SIT53, SIT26 and SIT37 of sublineages T3-ETH, T1, CASI-DELHI and T3, respectively.

Improved molecular detection of Ehrlichia and Anaplasma species applied to Amblyomma ticks collected from cattle and sheep in Ethiopia.

Detection of Ehrlichia and Anaplasma species in animals and tick vectors is crucial for an understanding of the epidemiology of diseases caused by these pathogens. In this study, a pair of primers designated EBR2 and EBR3 was designed from the Anaplasma 16S rDNA sequence and was used along with a previously described primer EHR 16SD for the simultaneous detection of Ehrlichia and Anaplasma species by nested PCR. The primers were used to amplify 925bp of DNA from known species of Ehrlichia and Anaplasma. Restriction with MboII and MspI enzymes allowed Ehrlichia and Anaplasma speciation. Restriction with MboII differentiated between An. marginale, Anaplasma (formerly Ehrlichia) sp. Omatjenne, and An. centrale with An. marginale and Anaplasma (formerly Ehrlichia) sp. Omatjenne yielding 2 distinct fragments each while An. centrale produced 3 distinct bands. Ehrlichia ruminantium and An. phagocytophylum remained undigested. Subsequent restriction with MspI differentiated E. ruminantium from An. phagocytophylum with 2 and 4 fragments, respectively. When used on tick samples from the field, 63 ticks (16.4%) out of 384 collected from cattle and sheep were positive for one or more species of Ehrlichia and Anaplasma. The positivity ranged from 6.3% at Andasa to 36.7% at Habernosa. Higher overall infection rates were found in Amblyomma lepidum than in Amblyomma variegatum ticks (p=0.009). Amblyomma lepidum from Habernosa were more often infected with all detected species of Anaplasma and Ehrlichia than Am. variegatum. At Bako, however, Anaplasma (formerly Ehrlichia) sp. Omatjenne was detected only in Am. variegatum. A significantly higher proportion of ticks collected from cattle (20.6%) was found positive than in those collected from sheep (3.3%) (p=0.003). Simultaneous detection of Ehrlichia and Anaplasma species and correct identification of mixed infections was possible. Since the ticks were collected from animals, the occurrence of the major species of Ehrlichia and Anaplasma in ruminants in the area is confirmed.

Isoniazid and rifampicin resistance mutations and their effect on second-line anti-tuberculosis treatment.

SETTING: St Peter's TB Specialized Hospital, Addis Ababa, Ethiopia. OBJECTIVE: To estimate the prevalence of mutations that cause resistance to isoniazid (INH) and rifampicin (RMP) and assess the effects of these mutations on second-line anti-tuberculosis treatment. DESIGN: GenoType(®)MTBDRplus assay results and clinical data documented at St Peter's TB Specialized Hospital over 3 years were retrospectively collected and analysed. RESULTS: The results indicated that 68.7% (n = 470) of RMP-resistant isolates had mutations at codon 531 (S531L) of the rpoB gene, while 93% (n = 481) of the INH-resistant isolates had mutations at codon 315 (S315T1) of the katG gene. The proportion of inhA mutations was 0.8% (n = 481). Treatment outcome was unfavourable in 23.7% (n = 76) of patients treated with second-line anti-tuberculosis drugs. Mutations in other codons of the rpoB gene (P > 0.05) and in the inhA promoter region (P > 0.05) were not associated with unfavourable treatment outcomes. CONCLUSION: The predominant mutations in RMP and INH resistance were observed at codons 531 and 315 in the rpoB and katG genes, respectively. Mutations in the inhA region were rare, which shows its minimal contribution to the development of resistance to ethionamide. This also suggests that treating multidrug-resistant TB patients with high doses of INH may have little effect.

Strain diversity of mycobacteria isolated from pulmonary tuberculosis patients at Debre Birhan Hospital, Ethiopia.

BACKGROUND: Ethiopia ranks seventh in the list of 22 high tuberculosis (TB) burden countries, with an incidence rate of 379 cases per 100,000 population for TB all forms. However, information on the genomic diversity of Mycobacterium tuberculosis in Ethiopia is limited. OBJECTIVE: To investigate the molecular characteristics of M. tuberculosis strains implicated in pulmonary TB in the study area. METHODS AND RESULTS: A cross-sectional study was conducted using socio-demographic, clinical and culture data combined with molecular typing analysis. The proportion of TB and M. tuberculosis isolates was not associated with risk factors (P > 0.05). Of 99 sputum samples, 80.8% were culture-positive. Speciation of isolates showed that 88.8% were M. tuberculosis. Further characterisation led to the identification of 27 different spoligotype patterns of M. tuberculosis; the most dominant shared types were SIT149, SIT53 and SIT54. Of the 27 strains, three strains were new and were reported to the SITVIT database. More than two thirds of the strains belonged to the Euro-American lineage. CONCLUSION: This study shows the presence of several clusters and new strains of M. tuberculosis circulating in pulmonary TB patients in the study area, suggesting recent transmission. Nationwide studies are recommended to map the population structure of M. tuberculosis and set control measures.

IgA response to ESAT-6/CFP-10 and Rv2031 antigens varies in patients with culture-confirmed pulmonary tuberculosis, healthy Mycobacterium tuberculosis-infected and non-infected individuals in a tuberculosis endemic setting, Ethiopia.

Little attention has been given to the role of antibodies against Mycobacterium tuberculosis (Mtb) infection. We have compared the levels of IgA and IgG against ESAT-6/CFP-10 and Rv2031c antigens in sera of patients with culture-confirmed pulmonary tuberculosis (PTB), healthy Mtb-infected and non-infected individuals in endemic TB settings. Venous blood samples were collected from 166 study participants; sera were separated and assayed by an enzyme-linked immunosorbent assay (ELISA). QuantiFERON-TB Gold In-Tube (QFTGIT) assay was used for the screening of latent TB infection. The mean optical density (OD) values of IgA against ESAT-6/CFP-10 and Rv2031 were significantly higher in sera of patients with culture-confirmed PTB compared with healthy Mtb-infected and non-infected individuals (P < 0.001). The mean OD values of IgG against ESAT-6/CFP-10 and Rv2031 were also significantly higher in sera of patients with culture-confirmed PTB compared with healthy Mtb-infected and non-infected individuals (P < 0.05). The mean OD values of IgA against both antigens were also higher in sera of healthy Mtb-infected cases compared with non-infected individuals. There were positive correlations (P < 0.05) between the level of IFN-γ induced in QFTGIT assay and the OD values of serum IgA against both antigens in healthy Mtb-infected subjects. This study shows the potential of IgA response against ESAT-6/CFP-10 and Rv2031 antigens in discriminating clinical TB from healthy Mtb-infected and non-infected cases. Nevertheless, further well-designed cohort study is needed to fully realize the full potential of this diagnostic marker.

Spoligotyping of multidrug-resistant Mycobacterium tuberculosis isolates in Ethiopia.

SETTING: St Peter Tuberculosis (TB) Specialized Hospital and the Aklilu Lemma Institute of Pathobiology, Addis Ababa, Ethiopia. OBJECTIVE: To genotype multidrug-resistant tuberculosis (MDR-TB) isolates and assess the magnitude of their clustering. DESIGN: A total of 183 consecutive MDR-TB isolates collected between September 2009 and February 2012 were characterised using molecular typing. Prior to the study, the isolates were confirmed as MDR-TB using GenoType MTBDRplus. Recent transmission index was used to analyse the clusters. RESULTS: Spoligotyping identified 43 different patterns, of which 17 consisted of at least two isolates forming clusters, while 26 had only a single isolate. The most frequent patterns were spoligo international typing (SIT) number 21 and 149. Twenty-four patterns did not match existing patterns in the SpolDB4 database. The strains belonged to three lineages, the predominant lineages being Euro-American and Indo-Oceanic, each consisting of 65 isolates. High proportions (86%) of patients were infected with clustered strains, suggesting probable recent transmission of MDR-TB in the study area. CONCLUSION: The observation of cluster formation of the spoligotype patterns of MDR-TB isolates could suggest transmission of MDR-TB strains among the population, thus warranting further attention.

Prevalence of pulmonary tuberculosis and associated risk factors in Eastern Ethiopian prisons.

OBJECTIVES: To determine the prevalence of pulmonary tuberculosis (PTB) and associated risk factors among inmates in three major prison settings of Eastern Ethiopia. DESIGN: Between July and November, 2008, 371 prisoners with a history of cough of ≥ 2 weeks were screened for PTB using direct smear microscopy and culture. Data were analysed using descriptive statistics and multivariable logistic regression. RESULTS: Of 371 PTB suspects identified by active screening, 33 (8.9%) were confirmed as smear- or culture-positive PTB. Together with the 11 PTB patients already on treatment, the point prevalence of PTB was 1913 per 100,000 (95%CI 1410-2580), about seven times higher than that of the general population. Eleven newly diagnosed PTB patients were sharing a cell with known TB patients. Factors significantly associated with PTB were young age (15-44 years of age) (OR 3.73), urban residence (OR 3.59), having a cough >4 weeks (OR 3.15), and sharing a cell with a TB patient (OR 3.39) or a prisoner with chronic cough (OR 4.5). CONCLUSIONS: The study documented a high prevalence of PTB among Ethiopian prisoners. Socio-demographic and TB management factors were identified to be underlying causes of the high transmission rate and the acquisition of new cases. Active surveillance of TB and implementing prevention and control guidelines are imperative.

Assessment of knowledge and practice about tuberculosis among eastern Ethiopian prisoners.

OBJECTIVE: To assess the level of knowledge and practice related to TB and identify predictive factors in three prisons in eastern Ethiopia: Dire Dawa, Jijiga and Harar. DESIGN: Between July and November 2008, 382 TB patients and suspects were interviewed using a structured questionnaire. Data were analysed using descriptive statistics and ordinal logistic regression. RESULTS: Only six (1.6%) prisoners described the cause of TB as being bacterial, while a wind locally known as 'nefas' was frequently mentioned (36.1%); nearly 75% of the prisoners correctly described breath as a mode of TB transmission; 116 (30.7%) did not know any measures for TB prevention and control; and half of the participants did not know that anti-tuberculosis drugs were provided free of charge. Significant predictors of TB knowledge were: incarceration in the Jijiga (OR 9.62, P < 0.001) and Dire Dawa (OR 2.14, P = 0.016) prisons, those who did not consult and receive treatment for TB symptoms (OR 2.46, P < 0.001), and prisoners without a past history of TB (OR 2.72, P = 0.002). CONCLUSION: The study demonstrates that prisoners have a modest level of biomedical knowledge. As part of the National TB Programme, health education programmes need to be implemented to enhance prisoners' knowledge of TB.

Molecular typing of Mycobacterium bovis isolated from tuberculosis lesions of cattle in north eastern Ethiopia.

Mycobacterium bovis strains isolated from tuberculosis (TB) lesions from 1138 cattle slaughtered at Kombolcha abattoir in north eastern Ethiopia were characterised. Detailed postmortem examination, bacteriological culturing, regions of difference PCR and spoligotyping were used. At least one TB lesion was observed in 57 (5 per cent) of the cattle, of which 27 (47 per cent) yielded mycobacteria isolates. Of the 27 isolates, 25 were identified as M bovis and two as Mycobacterium tuberculosis. The M bovis isolates were grouped into six clusters of strains, and the M tuberculosis isolates were typified to one strain only with the reference SIT262. Three clusters of previously unreported M bovis strains were identified and reported to the Mycobacterium bovis spoligotype database. They were designated the reference numbers SB1490, SB1491 and SB1492.

Zoonotic tuberculosis and brucellosis in Africa: neglected zoonoses or minor public-health issues? The outcomes of a multi-disciplinary workshop.

Late in 2007, veterinary, medical and anthropological professionals from Europe and Africa met in a 2-day workshop in Pretoria, South Africa, to evaluate the burden, surveillance and control of zoonotic tuberculosis and brucellosis in sub-Saharan Africa. Keynote presentations reviewed the burden of these diseases on human and livestock health, the existing diagnostic tools, and the available control methods. These presentations were followed by group discussions and the formulation of recommendations. The presence of Mycobacterium bovis and Brucella spp. in livestock was considered to be a serious threat to public health, since livestock and animal products are the only source of such infections in human beings. The impact of these pathogens on human health appears to be relatively marginal, however, when compared with Mycobacterium tuberculosis infections and drug resistance, HIV and malaria. Appropriate diagnostic tools are needed to improve the detection of M. bovis and Brucella spp. in humans. In livestock, the 'test-and-slaughter' approach and the pasteurization of milk, which have been used successfully in industrialized countries, might not be the optimal control tools in Africa. Control strategies should fit the needs and perceptions of local communities. Improved intersectoral and international collaboration in surveillance, diagnosis and control, and in the education of medical and veterinary personnel, are advocated.

Effect of skin testing and segregation on the prevalence of bovine tuberculosis, and molecular typing of Mycobacterium bovis, in Ethiopia.

In 2002, the prevalence of bovine tuberculosis (tb) among 500 cattle on Holeta Farm, near Addis Ababa, Ethiopia, was 48 per cent, and the farm was divided into positive and negative herds. After three consecutive rounds of skin testing and segregation of skin test-positive and -negative animals, the prevalence of bovine tb was reduced from 14 per cent to 1 per cent in the negative herd in a year. Spoligotyping of 41 isolates from 17 cows gave an identical and unique spoligotype pattern, which can be represented as the binary number 1100000101111110111111100010000000000100000, where 1 indicates the presence of a spacer and 0 represents a loss. This spoligotype pattern had not previously been reported on the Mycobacterium bovis spoligotype database, and it was therefore designated SB1176, Ethiopian M bovis strain 1 (EMbs1). The variable number tandem repeat (VNTR) profile of the strain was 5254(*)33.1, which differed from the VNTR profile of strains reported in Great Britain.

Bovine tuberculosis on small-scale dairy farms in Adama Town, central Ethiopia, and farmer awareness of the disease.

A cross-sectional study was conducted on 378 households, owning 524 head of cattle, to determine the prevalence of bovine tuberculosis (TB) and assess its public health implications. Stratified systematic sampling and a questionnaire were used, in addition to the comparative intradermal tuberculin test, mycobacterial culture and biochemical and drug susceptibility tests. Using a confidence interval (CI) of 95%, the herd and animal prevalences were 15% (11.4%, 18.6%) and 11% (8.32%, 13.68%), respectively. A significant variation in prevalence (P) was observed in regard to herd size (chi2 = 32.3, P < 0.0001); age group (chi2 = 11.47, P < 0.05); and reproductive status (chi2 = 10.9, P < 0.05). The herd prevalence was significantly associated with feeding practices (chi2 = 6.6, P < 0.05) and respiratory signs (chi2 = 8.7, P < 0.01). A significant (chi2 = 8.4, P < 0.01) association was recorded between the presence of reactor cattle and human TB cases in a household. Thirty-five percent (132/378) of the respondents knew about bovine TB while only 32% (121/378) were aware it could be transmitted from cattle to humans. Mycobacterium bovis and M. tuberculosis were isolated from 18.2% and 11.4% (n = 44) of the milk of reactor cows, respectively. In conclusion, the herd and animal prevalences of bovine TB were moderate. The fact that human TB patients and reactor cattle were present in the same household suggests the transmission of mycobacterial species between cattle and their owners, while the awareness of these farmers about bovine TB and its transmission was generally poor.

Kinetics of interferon-gamma (IFN-gamma) release in the peripheral blood of calves vaccinated with BCG.

A study was conducted on 13 Friesian-Zebu crossbred calves, the ages of which lie between 6 and 18 months, to investigate the kinetics of IFN-gamma release in the peripheral blood following Bacille Calmete Guerine (BCG) vaccination. After being screened for bovine tuberculosis (BTB), the calves were vaccinated with 1 mL inoculums containing 6 x 10(6) CFU of BCG. The level of IFN-gamma in the peripheral blood was measured two times before vaccination and seven times after vaccination. using a sandwich ELISA. The kinetics of IFN-gamma post vaccination presented itself in three phases: rising, falling, and steady phases. The concentration of IFN-gamma, before and after vaccination, both in stimulated and non-stimulated samples, was statistically significant (P < 0.011). Strong positive correlation (r = 0.86) was recorded between the levels of IFN-gamma release in avian PPD- and bovine PPD-stimulated samples. Of the total 13 calves, 11 (84.6%) reacted positively to tuberculin inoculation 15 weeks post vaccination. It is concluded that the IFN-gamma rises immediately after BCG vaccination, reaching its peak two weeks post vaccination, and then declines gradually in the following weeks. The strong positive reaction of calves to tuberculin inoculation 15 weeks post vaccination showed the capability of BCG in causing the release of IFN-gamma in the peripheral blood, indicating its role in protection against infection with Mycobacterium bovis in calves.

Comparison between comparative tuberculin and gamma-interferon tests for the diagnosis of bovine tuberculosis in Ethiopia.

A study to determine and compare the sensitivities and specificities of the comparative cervical tuberculin (CCT) and gamma-interferon (IFN-gamma) tests for the diagnosis of bovine tuberculosis was conducted on 30 zebu oxen. The results of the tests were compared with the presence of acid-fast bacilli found by bacteriological culturing and histopathological examinations. The sensitivity and specificity of CCT test were found to be 90.9% and 100%, respectively. Those of the commercial IFN-gamma test were determined to be 95.5% and 87.7%, respectively. No significant differences were found between the sensitivities (Yates' corrected chi 2 = 0.32; p = 0.57) or the specificities (Yates' corrected chi 2 = 2.54; p = 0.11) of the two tests. Furthermore, a positive correlation (r = 0.76) was recorded between the increase in skin thickness following injection of bovine purified protein derivative (PPD) and the optical density in the gamma-interferon assay with bovine PPD. On the other hand, the correlation (r = 0.47) between the change in skin thickness following injection of avian PPD and the optical density in the gamma-interferon assay with avian PPD was relatively weak. On the basis of this preliminary investigation, it was concluded that the choice between the two tests depends on their cost and simplicity and on livestock management and time factors rather than on their respective diagnostic value.