An ecological and epidemiological single-season survey of Anaplasma and Ehrlichia positive ticks in Victoria Falls National Park, Zimbabwe.

There is an urgent need for continued research on the ecology of tick-borne diseases in Africa. Our objective was to provide a preliminary description of the ecology and epidemiology of tick species, tick-borne pathogens, and animal hosts in Zimbabwe, focusing efforts at Victoria Falls National Park, for a single season. We tested the hypothesis that tick surveillance and pathogen screening data can be used to model associations among ticks, hosts, and pathogens. We collected ticks from domesticated animals and wildlife in Zimbabwe and screened the ticks for the presence of Anaplasma and Ehrlichia bacteria. Nearly 30% of the screened ticks were PCR-positive; 89% of tick species were PCR-positive, and 88% of animal species carried at least one PCR-positive tick. We sequenced a subset of amplicons that were similar to three Anaplasma species and three Ehrlichia species. The odds of a tick being PCR-positive increased when many ticks were collected from the host or the tick was collected from a cow (domesticated animal). Tick species shared host species more often than expected. We demonstrate that ticks in northwestern Zimbabwe present a One Health problem for nearby wildlife and humans.

Tuberculosis serosurveillance and management practices of captive African elephants (Loxodonta africana) in the Kavango-Zambezi Transfrontier Conservation Area.

Transfrontier conservation areas represent an international effort to encourage conservation and sustainable development. Their success faces a number of challenges, including disease management in wildlife, livestock and humans. Tuberculosis (TB) affects humans and a multitude of non-human animal species and is of particular concern in sub-Saharan Africa. The Kavango-Zambezi Transfrontier Conservation Area encompasses five countries, including Zimbabwe, and is home to the largest contiguous population of free-ranging elephants in Africa. Elephants are known to be susceptible to TB; thus, understanding TB status, exposure and transmission risks to and from elephants in this area is of interest for both conservation and human health. To assess risk factors for TB seroprevalence, a questionnaire was used to collect data regarding elephant management at four ecotourism facilities offering elephant-back tourist rides in the Victoria Falls area of Zimbabwe. Thirty-five working African elephants were screened for Mycobacterium tuberculosis complex antibodies using the ElephantTB Stat-Pak and the DPP VetTB Assay for elephants. Six of 35 elephants (17.1%) were seropositive. The risk factor most important for seropositive status was time in captivity. This is the first study to assess TB seroprevalence and risk factors in working African elephants in their home range. Our findings will provide a foundation to develop guidelines to protect the health of captive and free-ranging elephants in the southern African context, as well as elephant handlers through simple interventions. Minimizing exposure through shared feed with other wildlife, routine TB testing of elephant handlers and regular serological screening of elephants are recommended as preventive measures.

Characteristics of Foot-and-Mouth Disease Viral Strains Circulating at the Wildlife/livestock Interface of the Great Limpopo Transfrontier Conservation Area.

Foot-and-mouth disease (FMD) inflicts severe economic losses within infected countries and is arguably the most important trade-restricting livestock disease in the world. In southern Africa, infected African buffaloes (Syncerus caffer) are the major reservoir of the South African Territories (SAT) types of the virus. With the progressive expansion of transfrontier conservation areas (TFCAs), the risk of FMD outbreaks is expected to increase due to a higher probability of buffalo/livestock contacts. To investigate the dynamics of FMD within and around the Great Limpopo TFCA (GLTFCA), 5 herds of buffaloes were sampled in June 2010 to characterize circulating viruses in South Africa and Zimbabwe. Three SAT-2 and three SAT-3 viral strains were isolated in both countries, including one that was genetically linked with a recent SAT-2 outbreak in Mozambique in 2011. In addition, two groups of unvaccinated cattle (n = 192) were serologically monitored for 1 year at the wildlife/livestock interface of Gonarezhou National Park (GNP) in Zimbabwe between April 2009 and January 2010, using the liquid-phase blocking ELISA (LPBE) and a test for antibodies directed against non-structural proteins (NSP). Neither clinical signs nor vaccination of cattle were reported during the study, yet a high proportion of the monitored cattle showed antibody responses against SAT-3 and SAT-1. Antibodies against NSP were also detected in 10% of the monitored cattle. The results of this study suggest that cattle grazing in areas adjacent to the GLTFCA can be infected by buffalo or other infected livestock and that cattle trade movements can act as efficient disseminators of FMD viruses to areas several hundred kilometres from the virus source. Current methods of surveillance of FMD at the GLTFCA interface seem insufficient to control for FMD emergence and dissemination and require urgent reassessment and regional coordination.

Relationship between burden of infection in ungulate populations and wildlife/livestock interfaces.

In southern African transfrontier conservation areas (TFCAs), people, livestock and wildlife share space and resources in semi-arid landscapes. One consequence of the coexistence of wild and domestic herbivores is the risk of pathogen transmission. This risk threatens local livelihoods relying on animal production, public health in the case of zoonoses, national economies in the context of transboundary animal diseases, and the success of integrated conservation and development initiatives. The level of interaction between sympatric wild and domestic hosts, defining different wildlife/livestock interfaces, characterizes opportunities of pathogen transmission between host populations. Exploring the relationship between infection burden and different types of wildlife/domestic interfaces is therefore necessary to manage the sanitary risk in animal populations through control options adapted to these multi-host systems. Here, we assessed the infection burdens of sympatric domestic cattle (Bos taurus/Bos indicus) and African buffalo (Syncerus caffer) at an unfenced interface and compared the infection burdens of cattle populations at different wildlife/livestock interfaces in the Great Limpopo TFCA. Patterns of infection in ungulate populations varied between wild and domestic hosts and between cattle populations at different wildlife/livestock interfaces. Foot-and-mouth disease, Rift Valley fever and theileriosis infections were detected in buffalo and cattle at unfenced interfaces; bovine tuberculosis was only present in buffalo; and brucellosis and lumpy skin disease only in cattle. At unfenced interfaces, cattle populations presented significantly higher Theileria parva and brucellosis prevalence. We hypothesize that cattle populations at wildlife/livestock interfaces face an increased risk of infection compared to those isolated from wildlife, and that the type of interface could influence the diversity and quantity of pathogens shared. Additional host behavioural and molecular epidemiological studies need to be conducted to support this hypothesis. If it is confirmed, the management of wildlife/livestock interfaces will need to be considered through the prism of livestock and public health.

Gousiekte in African buffalo (Syncerus caffer).

Three African buffalo (Syncerus caffer) that died after capture and translocation from Mutirikwe Recreational Park in southern Zimbabwe showed macroscopic and microscopic lesions of cardiomyopathy compatible with a diagnosis of gousiekte. The buffalo had had access to Pavetta schumanniana, a plant that is known to cause gousiekte. Death was attributed to cardiac failure as a result of previous consumption of the plant, exacerbated by the stress of translocation.

Massive outbreak of anthrax in wildlife in the Malilangwe Wildlife Reserve, Zimbabwe.

A massive outbreak of anthrax in the wildlife of the Malilangwe Wildlife Reserve in Zimbabwe between August and November 2004 resulted in the death of almost all the reserve's estimated 500 kudu (Tragelaphus strepsiceros). Other species badly affected were nyala (Tragelaphus angasi), bushbuck (Tragelaphus scriptus), waterbuck (Kobus ellipsiprymnus) and roan antelope (Hippotragus equinus), which suffered losses of approximately 68 per cent, 48 per cent, 44 per cent and 42 per cent of their populations, respectively. Buffalo (Syncerus caffer) were also badly affected and although their population suffered only a 6 per cent loss, the numbers of deaths ranked second highest after kudu. To the authors' knowledge, this is the first record of anthrax in wildlife in Zimbabwe.

On-line identification of the bioactive compounds from Blumea gariepina by HPLC-UV-MS and HPLC-UV-NMR, combined with HPLC-micro-fractionation.

The dichloromethane extract of the aerial parts of Blumea gariepina (Asteraceae) was shown to be active against the phytopathogenic fungus Cladosporium cucumerinum and to inhibit acetylcholinesterase. In order rapidly to identify the active principles, the crude extract was analysed by on-flow HPLC-1H-NMR. HPLC-micro-fractionation was performed and all peaks collected were submitted to assays against C. cucumerinum and acetylcholinesterase. By this means, the biological activities could be efficiently associated with selected HPLC peaks. Complementary on-line structural data for all peaks of interest in the crude extract were obtained from HPLC-MS and from HPLC-UV with post-column addition of UV shift reagents. This chemical screening strategy with integrated bioassays permitted the on-line identification of a number of constituents and gave useful information for an efficient isolation procedure.

Reversible anaesthesia of free-ranging lions (Panthera leo) in Zimbabwe.

The combination of medetomidine-zolazepam-tiletamine with subsequent antagonism by atipamezole was evaluated for reversible anaesthesia of free-ranging lions (Panthera leo). Twenty-one anaesthetic events of 17 free-ranging lions (5 males and 12 females, body weight 105-211 kg) were studied in Zimbabwe. Medetomidine at 0.027-0.055 mg/kg (total dose 4-11 mg) and zolazepam-tiletamine at 0.38-1.32 mg/kg (total dose 50-275 mg) were administered i.m. by dart injection. The doses were gradually decreased to improve recovery. Respiratory and heart rates, rectal temperature and relative haemoglobin oxygen saturation (SpO2) were recorded every 15 min. Arterial blood samples were collected from 5 lions for analysis of blood gases and acid-base status. For anaesthetic reversal, atipamezole was administered i.m. at 2.5 or 5 times the medetomidine dose. Induction was smooth and all lions were anaesthetised with good muscle relaxation within 3.4-9.5 min after darting. The predictable working time was a minimum of 1 h and no additional drug doses were needed. Respiratory and heart rates and SpO2 were stable throughout anaesthesia, whereas rectal temperature changed significantly over time. Atipamezole at 2.5 times the medetomidine dose was sufficient for reversal and recoveries were smooth and calm in all lions independent of the atipamezole dose. First sign of recovery was observed 3-27 min after reversal. The animals were up walking 8-26 min after reversal when zolazepam-tiletamine doses < 1 mg/kg were used. In practice, a total dose of 6 mg medetomidine and 80 mg zolazepam-tiletamine and reversal with 15 mg atipamezole can be used for either sex of an adult or subadult lion. The drugs and doses used in this study provided a reliable, safe and reversible anaesthesia protocol for free-ranging lions.

Trichinella papuae and Trichinella zimbabwensis induce infection in experimentally infected varans, caimans, pythons and turtles.

The discovery of Trichinella zimbabwensis in farm crocodiles of Zimbabwe has opened up a new frontier in the epidemiology of the Trichinella genus. The objective of the present study was to investigate the infectivity of encapsulated species (T. spiralis, T. nativa, T. britovi, T. murrelli and T. nelsoni) and non-encapsulated species (T. pseudospiralis, T. papuae and T. zimbabwensis) in caimans (Caiman crocodilus), varans (Varanus exanthematicus), pythons (Python molurus bivittatus) and turtles (Pelomedusa subrufa) raised at their natural temperature range (26-32 degrees C). Mice and chickens were used as controls. At 6 days post-infection (p.i.), adult worms were detected in the small intestine of reptiles infected with T. papuae and T. zimbabwensis, of chickens infected with T. pseudospiralis and of mice infected with all encapsulated and non-encapsulated species. At 60 days p.i., T. papuae and T. zimbabwensis adult worms were collected from the intestine of varans and caimans and larvae from muscles of the four reptile species, T. pseudospiralis larvae from muscles of chickens, and larvae of all Trichinella species from mouse muscles. The highest reproductive capacity index of both T. papuae and T. zimbabwensis was observed in varans. The results show that T. papuae and T. zimbabwensis are able to complete their entire life-cycle in both poikilothermic and homoiothermic animals.

An investigation into the source and spread of foot and mouth disease virus from a wildlife conservancy in Zimbabwe.

African buffalo were introduced into a wildlife conservancy in the southeast of Zimbabwe in an effortto increase the conservancy's economic viability, which is primarily based on eco-tourism. The buffalo were infected with SAT serotypes (SAT-1, SAT-2 and SAT-3) of foot and mouth disease (FMD) virus, and in order to isolate the conservancy and prevent the transmission of FMD to adjacent populations of domestic livestock, the conservancy was surrounded by a double-fence system, 1.8 m in height. The intention was to prevent the movement of both wildlife and domestic animals across the perimeter. However, two years after the buffalo were introduced, FMD occurred in cattle farmed just outside of the conservancy. Using serological and molecular diagnostic tests, epidemiological investigations showed that it was most likely that antelope (impala or kudu), infected through contact with the buffalo herd within the conservancy, had jumped over the fence and transmitted the virus to the cattle.

Heliotropamide, a novel oxopyrrolidine-3-carboxamide from Heliotropium ovalifolium.

Heliotropamide (1), a new alkaloid with a novel oxopyrrolidine-3-carboxamide central moiety, has been isolated as the major product of the dicholoromethane extract of Heliotropium ovalifolium aerial parts. Its structure was elucidated by spectrometric methods including ESI-HR, EI, D/CI mass spectrometry, (1)H, (13)C, and 2D NMR experiments, and chemical derivatization. Neither heliotropamide nor its acetylated derivative (1a) showed any antifungal activity against Cladosporium cucumerinum and Candida albicans, antibacterial activity against Bacillus subtilis, radical-scavenging properties in the DPPH test, or inhibitory potential toward acetylcholinesterase.

Trichinella zimbabwensis n.sp. (Nematoda), a new non-encapsulated species from crocodiles (Crocodylus niloticus) in Zimbabwe also infecting mammals.

Since 1995, Trichinella larvae have been detected in 39.5% of farmed crocodiles (Crocodylus niloticus) in Zimbabwe. Morphological, biological, biochemical and molecular studies carried out on one isolate from a farmed crocodile in 2001 support the conclusion that this parasite belongs to a new species, which has been named Trichinella zimbabwensis n.sp. This species, whose larvae are non-encapsulated in host muscles, infects both reptiles and mammals. The morphology of adults and larvae is similar to that of Trichinella papuae. Adults of T. zimbabwensis cross in both directions with adults of T. papuae (i.e. male of T. zimbabwensis per female of T. papuae and male of T. papuae per female of T. zimbabwensis), producing F1 offspring which produce very few and less viable F2 larvae. Muscle larvae of T. zimbabwensis, like those of T. papuae, do not infect birds. Three allozymes (of a total of 10) are diagnostic between T. zimbabwensis and T. papuae, and five are diagnostic between T. zimbabwensis and Trichinella pseudospiralis, the third non-encapsulated species. The percentage of the pairwise alignment identity between T. zimbabwensis and the other Trichinella species for the cytochrome oxidase subunit I gene, the large subunit ribosomal-DNA (mt-lsrDNA) gene and the expansion segment five, shows that T. zimbabwensis is more similar to the two non-encapsulated species T. papuae (91% for cytochrome oxidase I; 96% for mt-lsrDNA; and 88% for expansion segment five) and T. pseudospiralis (88% for cytochrome oxidase I; 90% for mt-lsrDNA; and 66-73% for expansion segment five) than to any of the encapsulated species (85-86% for cytochrome oxidase I; 88-89% for mt-lsrDNA; and 71-79% for expansion segment five). This is the first non-encapsulated species discovered in Africa. The finding of a new Trichinella species that infects both reptiles and mammals suggests that the origin of Trichinella parasites dates back further than previously believed and can contribute to understanding the phylogeny and the epidemiology of the genus Trichinella.

Vaccination to control an outbreak of Mycoplasma crocodyli infection.

Details of a severe outbreak of M. crocodyli infection in farmed crocodiles are reported. The outbreak was suspected to have been precipitated by translocation-related stress on the animals brought from a farm with a known history of M. crocodyli infection. Resorting to the use of an autogenous vaccine proved more effective in alleviating the disease manifestations than antibiotic therapy. Prospects of vaccination in the face of an outbreak are discussed.

Quinones from Heliotropium ovalifolium.

Two new benzoquinones, heliotropinones A and B, have been isolated from the aerial parts of Heliotropium ovalifolium. Their structures were elucidated by spectrometric methods including high resolution electrospray ionization (ESI-HR), EI mass spectrometry, 1H, 13C and 2D NMR experiments. The two quinones demonstrated antifungal activities against Cladosporium cucumerinum and Candida albicans as well as antibacterial activity against Bacillus subtilis.

Transmission studies on Trichinella species isolated from Crocodylus niloticus and efficacy of fenbendazole and levamisole against muscle L1 stages in Balb C mice.

Forty-four Balb C mice, aged 18 weeks were infected with crocodile (Crocodylus niloticus)-derived Trichinella species. Of the infected mice, 32 were randomly divided into two groups each containing equal numbers of males and females; levamisole treated group and fenbendazole treated group. Each group was randomly subdivided into two subgroups as follows: levamisole group (subgroup 1: treated with levamisole on day 35 post infection, and subgroup 2: treated with levamisole on days 35 and 42 post infection) and fenbendazole group (subgroup 1: treated with fenbendazole on day 35 post infection and subgroup 2: treated with fenbendazole on days 35 and 42 post infection). The first subgroups treated on day 35 post infection were slaughtered on day 42 post infection and the second subgroups were treated on day 35 and day 42 post infection and slaughtered on day 49 post infection. Two female mice were infected a day after mating and were slaughtered together with the offspring on day 64 post-infection. Ten infected control mice were given 1 ml distilled water orally as placebo, and five of these were slaughtered on day 42 post infection. The results showed that the mean reproductive capacity index of this strain (RCI) in Balb C mice was 110. There was a significant reduction (P < 0.01) in larval counts in the single treatment groups (day 35) and in the double treatment groups (days 35 and 42) for both anthelmintics when compared the number of parasites in the control groups. After a single treatment, levamisole reduced the infection by 79.9% and fenbendazole by 76.7%. Following double treatments, levamisole reduced the infection by 95.5% and fenbendazole by 99.1%. There was evidence that the infected pregnant mice transmitted the parasite to their offspring. It is not certain whether the parasite was transmitted congenitally or transmammary Alternative ways of controlling the parasite in crocodile farms in Zimbabwe are discussed.

The foot-and-mouth disease risk posed by African buffalo within wildlife conservancies to the cattle industry of Zimbabwe.

Quantification of the risk that African buffalo (Syncerus caffer) (isolated within wildlife conservancies in Zimbabwe by a double fencing system) would infect cattle outside the conservancies with foot-and-mouth disease (FMD) virus was assessed by scenario-pathway analysis. Of the five scenarios considered, the greatest annual risk (1:5000) for cattle would be from antelope jumping over the outer perimeter fence of the conservancy and infecting cattle on the outside. The other transmission scenarios (including air-borne transmission) had a FMD risk that was low to very low. Risk management would include means to prevent the escape of antelope from the conservancies and restriction of cattle density in the proximity of the perimeter fence.

The epidemiology of rabies in Zimbabwe. 1. Rabies in dogs (Canis familiaris).

The epidemiology of rabies in dogs in Zimbabwe is described using data from 1950, when rabies was re-introduced after a 37-year absence, to 1996. Dogs constituted 45.7% of all laboratory-confirmed rabies cases and were the species most frequently diagnosed with the disease. Slightly more cases were diagnosed from June to November than in other months. From 1950 to the early 1980s, most dog cases were recorded from commercial farming areas, but since the early 1980s most have been recorded from communal (subsistence farming) areas. This change appears to be due to improved surveillance in communal areas and not to any change in the prevalence of rabies. Dog rabies therefore appears to be maintained mainly in communal area dog populations, particularly the large communal area blocks. Urban rabies was not important except in the city of Mutare. Where dog rabies prevalence was high, the disease was cyclic with periods between peak prevalence ranging from 4-7 years. Dog rabies cases were, on the whole, independent of jackal rabies and rabies in other carnivores. There was a significant negative relationship between the annual number of rabies vaccine doses administered nationally to dogs and the annual number of dog rabies cases lagged by one year, indicating that the past levels of immunisation coverage have had a significant effect on the number of rabies cases. However, dog vaccination coverage has clearly not been adequate to prevent the regular occurrence of rabies in dogs.

The epidemiology of rabies in Zimbabwe. 2. Rabies in jackals (Canis adustus and Canis mesomelas).

The epidemiology of rabies in Canis adustus (the side-striped jackal) and Canis mesomelas (the black-backed jackal) in Zimbabwe is described using data collected from 1950-1996. Cases in the two species made up 25.2% of all confirmed cases, second only to domestic dogs. Since the species of jackal cases was not recorded on rabies submission forms, the country was divided into areas according to species dominance and jackal cases were assigned to either C. adustus or C. mesomelas dominant zones or a sympatric zone where the relative status of the species is not known. Jackal rabies in both species is maintained in the commercial farming sector. Jackal rabies in the C. adustus zone occurs as dense epidemics, which begin at a single focus and spread centrifugally. The foci were initiated by rabid dogs, but once initiated the epidemic is maintained by C. adustus independently of other species. The extent of outbreaks in the C. adustus zone was limited by geographical (landuse type and jackal species interface) boundaries. Jackal rabies in C. adustus zones showed two seasonal peaks with the main peak occurring during late summer and the second peak during winter. In the C. mesomelas zone jackal rabies was more sparse but it occurred during most years. C. mesomelas is also able to maintain rabies independently of other species, although the epidemiology of the disease in this species is unclear. Transmission of rabies cycles between the two jackal species zones does not appear to occur as epidemics terminate when crossing the C. adustus and C. mesomelas interface boundaries.

Infectivity of Trichinella sp. isolated from Crocodylus niloticus to the indigenous Zimbabwean pig (Mukota).

An experimental infection of the indigenous Zimbabwean pig (Mukota) with a Trichinella sp. derived from crocodile (Crocodylus niloticus) was performed. The same larval isolates of Trichinella were infected to rats as a control. The muscles of both pigs and rats were found to be heavily infected with the first-stage larvae. The present study constitutes the first report of a successful experimental infection of the pig with Trichinella sp. originating from crocodile.