Heartwater, a tick-borne disease of domestic and wild ruminants, is caused by the intracellular rickettsia Ehrlichia ruminantium (previously known as Cowdria ruminantium). It is a major constraint to livestock production throughout subSaharan Africa, and it threatens to invade the Americas, yet there is no immediate prospect of an effective vaccine. A shotgun genome sequencing project was undertaken in the expectation that access to the complete protein coding repertoire of the organism will facilitate the search for vaccine candidate genes. We report here the complete 1,516,355-bp sequence of the type strain, the stock derived from the South African Welgevonden isolate. Only 62% of the genome is predicted to be coding sequence, encoding 888 proteins and 41 stable RNA species. The most striking feature is the large number of tandemly repeated and duplicated sequences, some of continuously variable copy number, which contributes to the low proportion of coding sequence. These repeats have mediated numerous translocation and inversion events that have resulted in the duplication and truncation of some genes and have also given rise to new genes. There are 32 predicted pseudogenes, most of which are truncated fragments of genes associated with repeats. Rather then being the result of the reductive evolution seen in other intracellular bacteria, these pseudogenes appear to be the product of ongoing sequence duplication events.gene duplication ͉ bacterial genome ͉ molecular sequence data ͉ intracellular adaptation E hrlichia ruminantium (previously known as Cowdria ruminantium) is an obligate intracellular bacterium in the order Rickettsiales. Species in this order cause serious diseases in man and domestic animals throughout the world. E. ruminantium is transmitted by ticks of the genus Amblyomma and causes heartwater, a fatal and economically important disease of wild and domestic ruminants. The disease occurs throughout subSaharan Africa and on several Caribbean islands, from which it threatens to invade the Americas (1), but the existing immunization procedures are rudimentary and relatively ineffective (2). E. ruminantium is a fragile bacterium with exacting culture requirements in eukaryotic cell lines; genetic manipulation has not been attempted, and little is known about its mechanisms of virulence or pathogenesis. Heartwater affects all domestic ruminants, and 80-95% of naïve animals die within 3 weeks, but those that recover have a T cell-mediated immunity to subsequent homologous challenge (3). In the absence of any directed strategy to identify T cell-stimulatory proteins we sequenced the E. ruminantium genome in the expectation that access to the complete protein-coding repertoire of the organism would facilitate the search for vaccine candidate genes.

The whitefly Bemisia tabaci cryptic species complex contains some important agricultural pest and virus vectors. Members of the complex have become serious pests in South Africa (SA) because of their feeding habit and their ability to transmit begomovirus species. Despite their economic importance, studies on the biology and distribution of B. tabaci in SA are limited. To this end, a survey was made to investigate the diversity and distribution of B. tabaci cryptic species in eight geographical locations (provinces) in SA, between 2002 and 2009, using the mitochondrial cytochrome oxidase I (mtCOI) sequences. Phylogenetic analysis revealed the presence of members from two endemic sub-Saharan Africa (SSAF) subclades coexisting with two introduced putative species. The SSAF-1 subclade includes cassava host-adapted B. tabaci populations, whereas the whiteflies collected from cassava and non-cassava hosts formed a distinct subclade, referred to as SSAF-5, and represent a new subclade among previously recognized southern Africa clades. Two introduced cryptic species, belonging to the Mediterranean and Middle East-Asia minor 1 clades, were identified and include the B and Q types. The B type showed the widest distribution, being present in five of the eight provinces explored in SA, infesting several host plants and predominating over the indigenous haplotypes. This is the first report of the occurrence of the exotic Q type in SA alongside the more widely distributed B type. Furthermore, mtCOI PCR-RFLP was developed for the SA context to allow rapid discrimination between the B, Q and SSAF putative species. The capacity to manage pests and disease effectively relies on knowledge of the identity of the agents causing the damage. Therefore, this study contributes to the understanding of South African B. tabaci species diversity, information needed for the development of knowledge-based disease management practices.

The spores of the soil-borne bacterium, Bacillus anthracis, which causes anthrax are highly resistant to adverse environmental conditions. Under ideal conditions, anthrax spores can survive for many years in the soil. Anthrax is known to be endemic in the northern part of Kruger National Park (KNP) in South Africa (SA), with occasional epidemics spreading southward. The aim of this study was to identify and map areas that are ecologically suitable for the harboring of B. anthracis spores within the KNP. Anthrax surveillance data and selected environmental variables were used as inputs to the maximum entropy (Maxent) species distribution modeling method. Anthrax positive carcasses from 1988–2011 in KNP (n = 597) and a total of 40 environmental variables were used to predict and evaluate their relative contribution to suitability for anthrax occurrence in KNP. The environmental variables that contributed the most to the occurrence of anthrax were soil type, normalized difference vegetation index (NDVI) and precipitation. Apart from the endemic Pafuri region, several other areas within KNP were classified as ecologically suitable. The outputs of this study could guide future surveillance efforts to focus on predicted suitable areas for anthrax, since the KNP currently uses passive surveillance to detect anthrax outbreaks.

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The diagnostic sensitivity (DSe) of the Rose Bengal test (RBT), the complement fixation test (CFT), the serum agglutination test (SAT), the competitive enzyme-linked immunosorbent assay (cELISA) and the indirect ELISA (iELISA) were determined in naturally infected cattle in KwaZulu-Natal province of South Africa with known infectious status from culture (gold standard). Natural brucellosis infection status of animals was determined by culturing and identification of Brucella abortus biovar 1 from abomasal fluid, milk, hygroma fluid, lymph nodes or uterine discharges samples. The diagnostic specificity (DSp) of the tests mentioned above was determined using samples from known negative herds. There was no statistically significant difference between the tests in their ability to diagnose brucellosis. The RBT and iELISA had the highest DSe of 95.8%, whereas RBT and CFT had the highest DSp of 100%. In South African laboratories, the RBT and CFT serological tests are used, because of the cost efficacy of CFT when compared to the less labour intensive but more expensive iELISA.

In a search for tools to distinguish antigenic variants of Ehrlichia ruminantium, we sequenced the major antigenic protein genes (map1 genes) of 21 different isolates and found that the sequence polymorphisms were too great to permit the design of probes which could be used as markers for immunogenicity. Phylogenetic comparison of the 21 deduced MAP1 sequences plus another 9 sequences which had been previously published did not reveal any geographic clustering among the isolates. Maximum likelihood analysis of codon and amino acid changes over the phylogeny provided no statistical evidence that the gene is under positive selection pressure, suggesting that it may not be important for the evasion of host immune responses.

BackgroundBrucellosis is an infectious and contagious zoonotic bacterial disease of both humans and animals. In developing countries where brucellosis is endemic, baseline data on the prevalence of brucellosis, using abattoir facilities, is important.ObjectivesThe aim of this study was to determine the seroprevalence of antibodies against Brucella in slaughter cattle at Gauteng province, South Africa and to characterize isolates of Brucella spp.MethodsIn this cross‐sectional study, un‐clotted blood samples with corresponding organ tissue samples were collected from slaughtered cattle. Serological [Rose Bengal test (RBT), complement fixation test (CFT) and indirect ELISA (iELISA)], molecular (PCR) and bacteriological methods were used to detect Brucella antibodies and Brucella spp. from 200 slaughtered cattle in 14 abattoirs.ResultsThe RBT revealed a seroprevalence of brucellosis as 11.0% (22 of 200) and iELISA confirmed 5.5% (11 of 200). The estimated seroprevalence from RBT and iELISA was 5.5% while RBT and CFT was 2.0% (4 of 200). Brucella melitensis (n = 6) and B. abortus (n = 5) were isolated from 11 cattle tissues (5.5%) as confirmed to species level with AMOS PCR and differentiated from vaccine strains with Bruce‐ladder PCR. Seven of the 11 isolates originated from seropositive cattle of which five were biotyped as B. abortus bv 1 (n = 2) and B. melitensis bv 2 (n = 1) and B. melitensis bv 3 (n = 2).ConclusionsThis is the first documentation of B. melitensis in cattle in South Africa. The zoonotic risk of brucellosis posed by Brucella‐infected slaughter cattle to abattoir workers and consumers of improperly cooked beef cannot be ignored.

Bovine brucellosis is endemic in Rwanda; however, little information is available on seroprevalence and risk factors. Therefore, a cross-sectional study was conducted among cattle farmed at the wildlife-livestock-human interface (n = 1691) in five districts and one peri-urban district (n = 216). Cattle were screened using the Rose Bengal test, then the results were confirmed by indirect enzyme-linked immunesorbent assay. Potential risk factors were determined with a questionnaire and analyzed for their association with seropositivity. In all districts, the animal and herd-level seroprevalence was 7.4% (141/1907) and 28.9% (61/212), respectively, 8.3% (141/1691) and 30.9% (61/198) at the interface, and 0.0% (0/216) in peri-urban areas. Among the potential risk factors, old age (≥5 years), cattle farmed close to wildlife, herds of cattle and small ruminants, history of abortions, and replacement animals were significantly associated with brucellosis (p < 0.05). Low awareness of zoonotic brucellosis, assisting calving without biosafety protection, drinking raw milk, and manual milking were each observed in more than 21.7% of cattle keepers whose herds were seropositive. This study confirmed brucellosis endemicity in cattle farmed close to wildlife in Rwanda, suggesting the need to focus control efforts in these areas. Educated farmers with a high awareness of zoonotic brucellosis had low bovine brucellosis seropositivity, which emphasizes the importance of education.