James Oloya

African 1, an Epidemiologically Important Clonal Complex of Mycobacterium bovis Dominant in Mali, Nigeria, Cameroon, and Chad

We have identified a clonal complex of Mycobacterium bovis present at high frequency in cattle in population samples from several sub-Saharan west-central African countries. This closely related group of bacteria is defined by a specific chromosomal deletion (RDAf1) and can be identified by the absence of spacer 30 in the standard spoligotype typing scheme. We have named this group of strains the African 1 (Af1) clonal complex and have defined the spoligotype signature of this clonal complex as being the same as the M. bovis BCG vaccine strain but with the deletion of spacer 30. Strains of the Af1 clonal complex were found at high frequency in population samples of M. bovis from cattle in Mali, Cameroon, Nigeria, and Chad, and using a combination of variable-number tandem repeat typing and spoligotyping, we show that the population of M. bovis in each of these countries is distinct, suggesting that the recent mixing of strains between countries is not common in this area of Africa. Strains with the Af1-specific deletion (RDAf1) were not identified in M. bovis isolates from Algeria, Burundi, Ethiopia, Madagascar, Mozambique, South Africa, Tanzania, and Uganda. Furthermore, the spoligotype signature of the Af1 clonal complex has not been identified in population samples of bovine tuberculosis from Europe, Iran, and South America. These observations suggest that the Af1 clonal complex is geographically localized, albeit to several African countries, and we suggest that the dominance of the clonal complex in this region is the result of an original introduction into cows naïve to bovine tuberculosis.

Isolation of non-tuberculous mycobacteria from pastoral ecosystems of Uganda: public health significance

BackgroundThe importance of non-tuberculous mycobacteria (NTM) infections in humans and animals in sub-Saharan Africa at the human-environment-livestock-wildlife interface has recently received increased attention. NTM are environmental opportunistic pathogens of humans and animals. Recent studies in pastoral ecosystems of Uganda detected NTM in humans with cervical lymphadenitis and cattle with lesions compatible with bovine tuberculosis. However, little is known about the source of these mycobacteria in Uganda. The aim of this study was to isolate and identify NTM in the environment of pastoral communities in Uganda, as well as assess the potential risk factors and the public health significance of NTM in these ecosystems.MethodA total of 310 samples (soil, water and faecal from cattle and pigs) were examined for mycobacteria. Isolates were identified by the INNO-Lipa test and by 16S rDNA sequencing. Additionally, a questionnaire survey involving 231 pastoralists was conducted during sample collection. Data were analysed using descriptive statistics followed by a multivariable logistic regression analysis.ResultsForty-eight isolates of NTM were detected; 25.3% of soil samples, 11.8% of water and 9.1% from animal faecal samples contained mycobacteria. Soils around water sources were the most contaminated with NTM (29.8%). Of these samples, M. fortuitum-peregrinum complex, M. avium complex, M. gordonae, and M. nonchromogenicum were the most frequently detected mycobacteria. Drinking untreated compared to treated water (OR = 33), use of valley dam versus stream water for drinking and other domestic use (OR = 20), sharing of water sources with wild primates compared to antelopes (OR = 4.6), sharing of water sources with domestic animals (OR = 5.3), and close contact with cattle or other domestic animals (OR = 13.8) were the most plausible risk factors for humans to come in contact with NTM in the environment.ConclusionsThe study detected a wide range of potentially pathogenic NTM from the environment around the pastoral communities in Uganda. Drinking untreated water and living in close contact with cattle or other domestic animals may be risk factors associated with the possibility of humans and animals acquiring NTM infections from these ecosystems.

European 1: a globally important clonal complex of Mycobacterium bovis.

We have identified a globally important clonal complex of Mycobacterium bovis by deletion analysis of over one thousand strains from over 30 countries. We initially show that over 99% of the strains of M. bovis, the cause of bovine tuberculosis, isolated from cattle in the Republic of Ireland and the UK are closely related and are members of a single clonal complex marked by the deletion of chromosomal region RDEu1 and we named this clonal complex European 1 (Eu1). Eu1 strains were present at less than 14% of French, Portuguese and Spanish isolates of M. bovis but are rare in other mainland European countries and Iran. However, strains of the Eu1 clonal complex were found at high frequency in former trading partners of the UK (USA, South Africa, New Zealand, Australia and Canada). The Americas, with the exception of Brazil, are dominated by the Eu1 clonal complex which was at high frequency in Argentina, Chile, Ecuador and Mexico as well as North America. Eu1 was rare or absent in the African countries surveyed except South Africa. A small sample of strains from Taiwan were non-Eu1 but, surprisingly, isolates from Korea and Kazakhstan were members of the Eu1 clonal complex. The simplest explanation for much of the current distribution of the Eu1 clonal complex is that it was spread in infected cattle, such as Herefords, from the UK to former trading partners, although there is evidence of secondary dispersion since. This is the first identification of a globally dispersed clonal complex M. bovis and indicates that much of the current global distribution of this important veterinary pathogen has resulted from relatively recent International trade in cattle.

Characterisation of mycobacteria isolated from slaughter cattle in pastoral regions of Uganda

BackgroundBovine tuberculosis is a zoonotic problem in pastoral cattle and communities in Uganda. Tuberculin tests in pastoral cattle had shown a high herd but low animal prevalence, with a high proportion of avian reactors. No work had been done to identify the mycobacterial species involved. The objective of the study was to isolate and characterise Mycobacterial species causing tuberculous lesions in slaughtered animals. Lesioned organs compatible with bovine tuberculosis in slaughtered cattle from pastoral areas in Uganda were collected and cultured to isolate mycobacteria. AccuProbe culture identification kits for the Mycobacterium tuberculosis complex, M. avium complex and M. avium were used to identify the isolates. Spoligotyping and Insertion Sequence (IS) 1311 and IS1245 Restriction Fragment Length Polymorphism analysis (RFLP) were used to further characterise the isolates.ResultsOf the 61 lesioned organs and tissues cultured, 19 isolates were identified as M. bovis, 3 as M. avium subsp.hominissuis, 1 as M. intracellulare, 1 as a mixed culture of M. bovis and M. avium sp. and 1 as M. avium sp. and unidentified mycobacteria. Eleven other mycobacteria outside the tuberculosis and avium complex groups were also isolated. Ten new spoligopatterns grouped into three clusters were identified from M. bovis isolates. Two of the three M. avium subsp.hominissuis isolates showed similar patterns on the IS1311 RFLP but all were different on the IS1245 RFLP.ConclusionThe isolation of M. bovis confirms the ongoing infection with spoligotypes unique to Uganda. Isolation of environmental mycobacteria could explain the high avian or non specific tuberculin reactor patterns commonly observed in pastoral cattle and suggests their pathogenic or opportunistic role in the infection of cattle with disseminated bovine tuberculous lesions.

A cross-sectional study of bovine tuberculosis in the transhumant and agro-pastoral cattle herds in the border areas of Katakwi and Moroto districts, Uganda

A study to determine the prevalence of bovine tuberculosis in the transhumant and agro-pastoral cattle herds in the border areas of Katakwi and Moroto districts in Uganda was carried out from July 2006 to January 2007 using comparative intradermal tuberculin test containing bovine and avian PPDs. A total of 1470 animals, 612 (41.6%) males and 858 (58.4%) females, 883 (60%) young, 555 (37.8%) adult and 32 (2.2%) old animals were included. The study involved a cross-sectional multistage sampling technique with random selection of individual animals from a herd. The results revealed a 1.3% overall prevalence of bovine tuberculosis in cattle herds in the study area, with a marked variation between sub-counties. The highest recorded prevalence was 6.0% in Kapujan, while no cases were recorded in Ongogonja, Magoro and Katakwi sub-counties. Distinctly different patterns in the avian-bovine reactions were also found in different sub-counties. A multivariate logistic regression showed more positive reactions (OR = 6.3; 95%CI (1.4–26.34) in females than males. BTB prevalence did not differ significantly between cattle maintained in pastoral and agro pastoral production systems. The study demonstrated a relatively low prevalence of bovine tuberculosis in local zebu cattle reared under traditional husbandry systems in Uganda, suggesting low infectiousness of the disease under such mode of production. The risk associated with the consumption of raw milk among the pastoral communities and that, the pooling of milk together from different animals is a common practice, warrants more investigation into the zoonotic transmission of tuberculosis within these communities.

Prevalence of Bovine tuberculosis in Ethiopian slaughter cattle based on post-mortem examination

A study aimed at describing the magnitude and distribution of gross lesions compatible with bovine tuberculosis (BTB) in Ethiiopian slaughter cattle in five abattoirs (four municipal and one export) located in various cattle husbandry systems in Ethiopia was carried out from July 2006 to January 2007 using detailed meat inspection procedure. Five representative abattoirs (four municipal and one export) located in distinct livestock management systems were selected. A total of 3322 cattle; 2876 (86.6%) male, 446 (13.4%) female; 3094 (93.1%) indigenous zebu, 140 (4.2%) crossbred and 88 (2.7%) pure exotic cattle were included in the study. A nine-year meat inspection record was also analyzed to elucidate the trend of BTB in the local cattle population.Of the carcasses inspected, 337 (10.2%, 95%CI= [9.2–11.2]) had lesions suggestive of tuberculosis, 69 (20.5%) generalized and 268 (79.5%) localized.TB prevalence showed a marked variation between categories of age, breed, class of animals, abattoir, geographic origin and husbandry system. It was higher in old and young animals than middle age group; in exotic than local breed; in calves than other classes of animals. The highest and lowest prevalence of TB was recorded in Adama (24.7%, 95%CI= [21.1–28.7]) and Yabello abattoirs (4.2%, 95%CI= [2.6–6.6]), respectively. Cattle whose origin was from Addis Ababa and its surrounding areas had higher prevalence of TB infection (23.9%, 95%CI= [17.6–31.5]).Cattle maintained in dairy farms had high degree of exposure (23.9%, 95%CI= [16.7–32.9]) to the infection than those maintained in other types of management system. Analysis of meat inspection records also revealed an increasing incidence of TB over the years. Our study demonstrated a high prevalence of tuberculosis in Ethiopian slaughter cattle and this could infer to similar scenario in a reference cattle population in the country. In view of Ethiopia’s increasing involvement in livestock export trade, the reported high prevalence of tuberculosis could be a major obstacle, particularly at this moment when sanitary requirements from importing countries are so much strict. Furthermore, the growing concern over increasing incidence of tuberculosis/HIV/AIDS co-infection, the high incidence of extra- pulmonary tuberculosis and a high risk of acquiring zoonotic tuberculosis among the majority of the population emphasize the need for paying the necessary attention towards the control of bovine tuberculosis.

Responses to tuberculin among Zebu cattle in the transhumance regions of Karamoja and Nakasongola district of Uganda

Responses to tuberculin in Zebu cattle of the transhumant pastoral farming system in Karamoja region and Nakasongola district in the north-eastern and mid-central regions in Uganda, respectively, were investigated using a comparative intradermal tuberculin skin test. Of the 1864 cattle tested from 30 large units (superherds) in Karamoja and 7 herds in Nakasongola, a total of 28 animals from 19 herds (51.4%) tested positive. Inter-district tuberculin reactor prevalence variations seemed to be influenced by climate, with impact on both the management patterns and transmissibility of agent. High herd tuberculin reactor prevalence (51.4%) was attributed to widespread contacts and mixing of animals between herds. Low individual animal tuberculin test positivity (mean = 1.4%) was attributed to low transmissibility of the agent under the Karamoja climate, which is semi-arid, and to increased resistance due to non-specific response to environmental mycobacteria and natural selection, since there was no active control against bovine tuberculosis. Owing to similarities in management practices in Karamoja and widespread risk factors, it was difficult to identify which were more important, but variations in sources of drinking water pointed to provision of lake and borehole water during dry season as reducing the risk. Positive bovine tuberculin reactor prevalence and skin reactor status were related to age.

Molecular characterization of Mycobacterium bovis isolates from Ethiopian cattle

BackgroundBovine Tuberculosis (BTB) is a widespread and endemic disease of cattle in Ethiopia. Information relating to genotypic characteristics of Mycobacterium bovis strains affecting the cattle population in Ethiopia is limited. We carried out molecular characterization of M. bovis strains isolated from BTB infected cattle using the spoligotyping technique. The relationship between distribution of spoligotypes and recorded variables was also investigated. A new approach that can numerically reflect the degree of genetic polymorphism in a M. bovis population was also developed. The study was conducted from July 2006 to January 2007 in cattle slaughtered at five representative abattoirs in Ethiopia.ResultsForty-five M. bovis isolates were obtained from 406 pathologic tissue specimens collected from 337 carcasses with lesions compatible with BTB. Twelve spoligotypes were identified from 34 distinct strains; with SB1176 as a dominant spoligotype (41.2% of the isolates) followed by SB0133 (14.7%). Comparison of spoligotypes with an M. bovis global database http://www.mbovis.org revealed six new spoligotypes which were subsequently registered in the database with international identification codes of SB1517, SB1518, SB1519, SB1520, SB1521 and SB1522. The majority of strains were obtained from cattle slaughtered at Addis Ababa abattoir. On the basis of the Spoligotype Evolutionary Index, SEI (a numeric expression approach to make standardized comparison of spoligotype evolution), M. bovis isolates from Ethiopia were relatively more heterogeneous (SEI = 3.2) compared to isolates from other countries. This might be attributed to extensive livestock movement linked to trading or seasonal migration, high degree of livestock mingling, and also diversities of the countrys agricultural and livestock ecosystems, in addition to lack of disease control measures that led to high infection prevalence. Multiple spoligotype infection was recorded in nine (50%) of infected carcasses and this may indicate the prevailing high degree of super infection.ConclusionsThis study provided molecular evidence for the widespread distribution of M. bovis in the cattle population in Ethiopia. It also demonstrated a relatively high degree of genetic polymorphism of the isolates. Further molecular investigation of M. bovis strains in humans and other domestic animals is recommended in order to elucidate the zoonotic importance as well as reservoirs and pattern of transmission among various hosts.

Distribution and intensity of Echinococcus granulosus infections in dogs in Moroto District, Uganda

This study was carried out during August 2007–March 2008 in pastoral areas of Moroto District, Uganda. It investigated the distribution and infection intensity of Echinococcus granulosus in the dog population and involved the postmortem examination of 327 dogs (106 domesticated; 80 semi-domesticated, and 141 strays; comprised of 163 females and 164 males). The overall prevalence of E. granulosus was 66.3% (95% CI = 60.8–71.4) with parasite burdens of 6–5,213 among the infected dogs. The prevalence of E. granulosus was primarily associated with the season. While the dogs were more likely to have high parasite burdens during the rainy as opposed to the dry season, the parasite burden of E. granulosus infection was also highly associated with age and husbandry. Young dogs were at greater risk of carrying a heavy burden of E. granulosus infection than adults. Likewise, stray dogs were more likely to have a heavy parasite burden. As the study documented a high prevalence and intensity of E. granulosus infection in the dog population in Moroto District of Uganda, further studies need to be carried out in human and intermediate hosts to elucidate the cycle of transmission that could help to design appropriate controlling measures.

African swine fever among slaughter pigs in Mubende district, Uganda

Owing to frequent reports of suspected outbreaks and the presence of reservoir hosts and vectors (warthogs, bushpigs and O. moubata ticks), African swine fever (ASF) is believed to be an endemic disease in Uganda. There have, however, been very few studies carried out to confirm its existence in Uganda. This study was carried out to describe the prevalence of ASF based on pathologic lesions and analysis of serum samples from slaughtered pigs during a suspected outbreak in the Mubende district of Uganda. The study was based on visits to 22 slaughterhouses where individual pigs were randomly selected for a detailed ante-mortem and post-mortem inspections. Sera were also collected for laboratory analysis. A total of 997 pigs (53.7% male and 46.3% female) were examined for lesions suggestive of ASF and sero-positivity of sera for ASF antibodies. The sera were tested using enzyme-linked immunosorbent assay (ELISA) and positive samples were further confirmed with an immunoblot assay. The results showed that 3.8% (38/997) of the pigs examined had clinical signs and post-mortem lesions suggestive of ASF. Two of 997 (0.2%) sera analysed were positive for ASF antibodies. Of the sub-counties investigated, Bagezza (12%) and Kiyuni (11%) had the highest prevalence of lesions suggestive of ASF based on ante- and post-mortem examination results, while Mubende town council (1.7%) had the lowest. This study found a low number of pigs (3.8%) with lesions suggestive of ASF at slaughter and an even lower number of pigs (0.2%) that were seropositive at slaughter, however a significantly higher number of pigs were slaughtered during the outbreak as a strategy for farmers to avoid losses associated with mortality.