Successful treatment of babesiosis in a south-western black rhinoceros(Diceros bicornis bicornis).

Under stressful conditions, black rhinoceroses that are sub-clinical carriers of Babesia bicornis can succumb to babesiosis. After 16 days in captivity, a five-year-old female black rhino captured for relocation presented with inappetence, abdominal discomfort and constipation. After chemical immobilisation, dry faecal balls were removed from the rectum, peripheral blood smears were made and blood collected into EDTA tubes. She was treated prophylactically for colic with flunixin meglumine, penicillin and doramectin. Piroplasms were seen on fixed and stained peripheral blood smears. Overnight she developed severe haemoglobinuria, a sign consistent with babesiosis. Subsequently, DNA extracted from a blood specimen reacted with the B. bicornis probe on Reverse Line Blot (RLB) assay, confirming the diagnosis of babesiosis. Specific treatment consisted of 14 ml imidocarb dipropionate (dosage 2.4 mg/kg) administered intramuscularly by pole syringe. Fifteen days later the patient was still moderately anaemic, with the red blood cell (RBC) count, haematocrit and haemoglobin concentration within normal ranges but on microscopic examination there was a marked RBC macrocytosis and polychromasia indicative of a regenerative anaemia. DNA extracted from blood collected at that time did not react with the B. bicornis probe on RLB assay, indicating that treatment with imidocarb had been effective. Once the patient's appetite improved, she started gaining weight. After 82 days in captivity and 65 days after babesiosis had been diagnosed, she was released at the site where she had been captured.

Black-backed jackals (Canis mesomelas) from semi-arid rangelands in South Africa harbour Hepatozoon canis and a Theileria species but apparently not Babesia rossi.

Despite the importance of disease as a wildlife management challenge in South Africa, baseline data on the epidemiology of pathogens occurring in free-ranging species has received little attention to date. Black-backed jackals (Canis mesomelas) are a wide-ranging, abundant carnivore with substantial economic importance due to their role in livestock depredation. They are known reservoirs hosts of Babesia rossi, a virulent pathogen in domestic dogs in sub-Saharan Africa. We investigated the prevalence and diversity of tick-borne pathogens (TPBs) including Babesia, Theileria, Hepatozoon, Ehrlichia and Anaplasma species, together with host-attached tick diversity, in a black-backed jackal population from the semi-arid Central Karoo, a small-livestock farming region in South Africa. Using reverse line blot hybridisation, we screened 43 blood samples and sequenced the 18S rRNA gene from positive samples to confirm and characterise pathogen identity using a phylogenetic framework. Hepatozoon canis, a ubiquitous pathogen of domestic and wild canids globally, was observed in 47% of jackals, while a Theileria sp. most similar to T. ovis, a piroplasm found in small livestock, was observed in 5% of jackals. No Babesia, Ehrlichia or Anaplasma species were identified, although a Sarcocystis sp. sequence was isolated from one jackal. Host-attached ticks (n = 20) comprised three species, Amblyomma marmoreum, Haemaphysalis elliptica/zumpti and Ixodes rubicundus, commonly known ticks in the region. In summary, prevalence of TBPs in black-backed jackals from this semi-arid rangeland region was lower than in jackal populations in more mesic regions. These jackals were apparently not infected with B. rossi. While this study is one of the first investigations into the epidemiology of TBPs infecting jackals and adds to the sparse literature, further studies which span landscape uses, climate conditions and seasonality are encouraged.

Geographic range of vector-borne infections and their vectors: the role of African wildlife.

The role of African wildlife in the occurrence of vector-borne infections in domestic animals has gained renewed interest as emerging and re-emerging infections occur worldwide at an increasing rate. In Africa, biodiversity conservation and the expansion of livestock production have increased the risk of transmitting vector-borne infections between wildlife and livestock. The indigenous African pathogens with transboundary potential, such as Rift Valley fever virus, African horse sickness virus, bluetongue virus, lumpy skin disease virus, African swine fever virus, and blood-borne parasites have received the most attention. There is no evidence for persistent vector-borne viral infections in African wildlife. For some viral infections, wildlife may act as a reservoir through the inter-epidemic circulation of viruses with mild or subclinical manifestations. Wildlife may also act as introductory or transporting hosts when moved to new regions, e.g. for lumpy skin disease virus, Rift Valley fever virus and West Nile virus. Wildlife may also act as amplifying hosts when exposed to viruses in the early part of the warm season when vectors are active, with spillover to domestic animals later in the season, e.g. with bluetongue and African horse sickness. Some tick species found on domestic animals are more abundant on wildlife hosts; some depend on wildlife hosts to complete their life cycle. Since the endemic stability of a disease depends on a sufficiently large tick population to ensure that domestic animals become infected at an early age, the presence of wildlife hosts that augment tick numbers may be beneficial. Many wild ungulate species are reservoirs of Anaplasma spp., while the role of wildlife in the epidemiology of heartwater (Ehrlichia ruminantium infection) has not been elucidated. Wild ungulates are not usually reservoirs of piroplasms that affect livestock; however, there are two exceptions: zebra, which are reservoirs of Babesia caballi and Theileria equi, and buffalo, which are reservoirs of Theileria parva. The latter causes Corridor disease when transmitted from buffaloto cattle, butthis appearsto be a self-limiting condition, at least in southern Africa. Wild animals are important reservoirs of tsetse-transmitted Trypanosoma spp. infection. The distribution and abundance of some tsetse species, e.g. Glossina morsitans and G. pallidipes, are closely related to the occurrence of their preferred wildlife hosts.

Hematological profile of East African short-horn zebu calves from birth to 51 weeks of age.

This paper is the first attempt to accurately describe the hematological parameters for any African breed of cattle, by capturing the changes in these parameters over the first 12 months of an animal's life using a population-based sample of calves reared under field conditions and natural disease challenge. Using a longitudinal study design, a stratified clustered random sample of newborn calves was recruited into the IDEAL study and monitored at 5-weekly intervals until 51 weeks of age. The blood cell analysis performed at each visit included: packed cell volume; red cell count; red cell distribution width; mean corpuscular volume; mean corpuscular hemoglobin concentration; hemoglobin concentration; white cell count; absolute lymphocyte, eosinophil, monocyte, and neutrophil counts; platelet count; mean platelet volume; and total serum protein. The most significant age-related change in the red cell parameters was a rise in red cell count and hemoglobin concentration during the neonatal period. This is in contrast to what is reported for other ruminants, including European cattle breeds where the neonatal period is marked by a fall in the red cell parameters. There is a need to establish breed-specific reference ranges for blood parameters for indigenous cattle breeds. The possible role of the postnatal rise in the red cell parameters in the adaptability to environmental constraints and innate disease resistance warrants further research into the dynamics of blood cell parameters of these breeds.

Origin and history to date of the World Association for the Advancement of Veterinary Parasitology (WAAVP) African Foundation.

The origin of the World Association for the Advancement of Veterinary Parasitology (WAAVP) African Foundation is described. The 16th WAAVP Conference held in South Africa in 1997 generated a surplus of ZAR 430 460 (US$ 70 116). This was invested and a foundation established to manage the fund with the intention of using it to the mutual advantage of the WAAVP and African veterinary parasitologists. To date, more than 110 scholarship applications have been screened, and 51 full and partial scholarships awarded to young African veterinary parasitologists to attend subsequent biennial WAAVP Conferences. This investment has grown into a very successful endowment currently valued at US$ 206 553. This article is written in response to many queries across the globe about the origin of this fund and how it has been invested, managed, sustained and utilised.

Piroplasm parasites of white rhinoceroses (Ceratotherium simum) in the Kruger National Park, and their relation to anaemia.

As part of a larger survey to map the geographical distribution of Babesia and Theileria parasites in the southern African rhinoceros population, white rhinoceroses were sampled during routine immobilisations in the Kruger National Park. Polymerase chain reaction (PCR) and reverse line blot (RLB) hybridisation assays were used to screen for the presence of piroplasms and complete blood counts were used to assess associated changes in clinical parameters. Of the 195 rhinoceroses sampled, 71 (36.4 %) tested positive for the presence of Theileria bicornis, with no significant change in the haematological parameters measured, while 18 (9.2 %) tested positive for Theileria equi. None of the rhinoceroses sampled tested positive for Babesia bicornis, a parasite associated with mortalities in black rhinoceroses.

Occurrence of blood-borne tick-transmitted parasites in common tsessebe (Damaliscus lunatus) antelope in Northern Cape Province, South Africa.

Blood samples were collected from 71 tsessebes relocated from the deproclaimed Vaalbos National Park to Mokala National Park, South Africa. DNA was extracted from the samples and the reverse line blot (RLB) hybridization technique was used to detect and identify any haemoparasites present. Six samples hybridized to the Theileria/Babesia genus-specific probe, the Theileria genus-specific probe and the Theileria sp. (sable) probe, while 3/6 also hybridized to the Theileria separata probe. Full-length 18S rRNA genes of the Theileria spp. detected were amplified, cloned and sequenced. Two novel Theileria 18S rRNA gene sequences were identified which are phylogenetically very closely related to both Theileria sp. (sable) and T. separata. All animals appeared to be in good health. It seems likely, therefore, that these Theileria spp. do not cause disease under normal circumstances. Nevertheless, care should be taken when translocating wild animals, as introduction of novel piroplasm parasites into new areas could cause clinical disease and losses in naïve wildlife and domestic animals, and new parasite species could become established in areas in which they previously did not occur.

Tick-borne blood parasites in nyala (Tragelaphus angasii, Gray 1849) from KwaZulu-Natal, South Africa.

A total of 97 blood samples of nyala (Tragelaphus angasii, Gray 1849) from South Africa were tested for the presence of tick-borne haemoparasites by means of polymerase chain reaction (PCR) and reverse line blot (RLB) hybridization. The majority of blood samples contained several different haemoparasites, often in combination. Prevalent haemoparasites were Theileria sp. (kudu), T. buffeli, Theileria sp. (sable), T. bicornis, Ehrlichia sp. Omatjenne, Anaplasma marginale and A. bovis. This serves as the first report of Theileria sp. (kudu), T. buffeli, T. bicornis, Ehrlichia sp. Omatjenne, A. marginale and A. bovis in nyala, who seem to carry multiple haemoparasites without ill effect.

Occurrence of Theileria parva and other haemoprotozoa in cattle at the edge of Hluhluwe-iMfolozi Park, KwaZulu-Natal, South Africa.

Theileria parva, the most important bovine theilerial species in sub-Saharan Africa, causes widespread mortality and morbidity in endemic areas. A survey was conducted using buffy-coat specimens from 60 apparently healthy adult communally herded Nguni-type cattle at the northeastern edge of the Hluhluwe-iMfolozi Park to determine, by means of PCR and Reverse Line Blot (RLB) hybridisation, the occurrence of Theileria and Babesia species. The presence of Trypanosoma species was determined using PCR-RFLP. Results showed that 6.7 % of the specimens were positive for Theileria parva. This significant finding suggests that cattle in South Africa, and not only African buffaloes (Syncerus caffer), may be subclinical carriers of T. parva. Other species identified were T. mutans (83.3%), T. velifera (70.0%), Theileria sp. (sable) (46.8%) and T taurotragi (1.7%). Two specimens (3.3%) were positive for Babesia bovis and single specimens (1.7%) positive for B. bigemina and B. rossi, respectively. Mixed infections, of up to 4 species, were common (65.0%). Only 1 specimen was found to be positive for Trypanosoma vivax, and 2 for T theileri, of which only the first species is pathogenic.

Intradermal tuberculin testing of wild African lions (Panthera leo) naturally exposed to infection with Mycobacterium bovis.

African lions in the southern half of Kruger National Park (KNP) are infected with Mycobacterium bovis. Historically, reliable detection of mycobacteriosis in lions was limited to necropsy and microbiological analysis of lesion material collected from emaciated and ailing or repeat-offender lions. We report on a method of cervical intradermal tuberculin testing of lions and its interpretation capable of identifying natural exposure to M. bovis. Infected lions (n=52/95) were identified by detailed necropsy and mycobacterial culture. A large proportion of these confirmed infected lions (45/52) showed distinct responses to bovine tuberculin purified protein derivative (PPD) while responses to avian tuberculin PPD were variable and smaller. Confirmed uninfected lions from non-infected areas (n=11) responded variably to avian tuberculin PPD only. Various non-tuberculous mycobacteria (NTM) were cultured from 45/95 lions examined, of which 21/45 were co-infected with M. bovis. Co-infection with M. bovis and NTM did not influence skin reactions to bovine tuberculin PPD. Avian tuberculin PPD skin reactions were larger in M. bovis-infected lions compared to uninfected ones. Since NTM co-infections are likely to influence the outcome of skin testing, stricter test interpretation criteria were applied. When test data of bovine tuberculin PPD tests were considered on their own, as for a single skin test, sensitivity increased (80.8-86.5%) but false positive rate for true negatives (18.75%) remained unchanged. Finally, the adapted skin test procedure was shown not to be impeded by persistent Feline Immunodeficiency Virus(Ple) co-infection.

Extracts of Calpurnia aurea leaves from southern Ethiopia attract and immobilise or kill ticks.

Calpurnia aurea extracts are used in southern Ethiopia to protect stock against ticks. Acetone, hexane and water leaf extracts of C. aurea collected in southern Ethiopia were tested for repellent/attractant and acaricidal properties on unfed adult Rhipicephalus pulchellus ticks. In contrast to many other plant species evaluated, C. aurea extracts did not have repellent properties, but rather had a slight attractant capacity. With 20% and 10% acetone extracts, all ticks were either killed or their mobility severely compromised after 1microl of extract was topically applied on the abdomen. At a 5% concentration, 85% of ticks were still affected. A 10% aqueous solution also had a marked effect. The results prove the efficacy of the traditional use of this extract and may lead to a product that can be used commercially to protect animals against tick infestation, under subsistence as well as industrialized conditions.

Preliminary evaluation of the BrEMA1 gene as a tool for associating babesia rossi genotypes and clinical manifestation of canine Babesiosis.

Babesia rossi, an intraerythrocytic protozoan, causes a severe, often life-threatening disease of domestic dogs. Dogs treated early for B. rossi infection usually recover from the disease, but dogs left untreated or treated at a later stage of infection seldom survive. Dogs infected with B. rossi have varied clinical manifestations that can be categorized as uncomplicated (with a good prognosis) or complicated (with a poor prognosis). One hundred twenty-one blood samples were collected from dogs presented to the Onderstepoort Veterinary Academic Hospital and diagnosed with babesiosis by the use of a thin blood smear. An additional 20 samples were obtained from Babesia-infected dogs from private clinics around the Onderstepoort, Johannesburg, Durban, White River, and Cape Town areas. The samples were screened by PCR targeting the Babesia rossi erythrocyte membrane antigen gene (BrEMA1) and by sequencing of the polymorphic region (i.e., region with a variable number of hexapeptide repeats). Analysis of PCR products revealed 11 different gene profiles, visualized by gel electrophoresis. Twelve distinct BrEMA1 genotypes were identified by sequencing, but the numbers of hexapeptide repeats varied from 6 to 31 (classified as genotype6 to genotype31). The genotypes were retrospectively compared to the clinical case data. The most frequently encountered B. rossi parasites were those attributed to genotype19 (36.2%), genotype28 and genotype29 (20.6% each), and genotype11 (12.7%). These genotypes were also the ones associated with the poorest prognosis. This preliminary finding suggests clinically important differences between the various B. rossi genotypes identified.

Tick infestation, and udder and teat damage in selected cattle herds of Matabeleland South, Zimbabwe.

A cross-sectional survey was conducted to determine tick infestation, and udder and teat damage in 286 lactating cows and heifers at six properties in the smallholder and commercial sectors in Gwanda district of Matabeleland South Province, Zimbabwe. Eight tick species were identified: Amblyomma hebraeum, Hyalomma truncatum, Hyalomma marginatum rufipes, Rhipicephalus evertsi evertsi, Rhipicephalus (Boophilus) decoloratus, Rhipicephalus appendiculatus, Rhipicephalus zambeziensis and Rhipicephalus simus. Overall, 81.5% of the cattle were tick infested; prevalence of tick-infested cattle was significantly higher on communal land (93.8%) and recently claimed land (85.1%) than on commercial farms. The mean tick load on infested cattle on communal land was significantly higher than in the other two sectors. Although 53% of the sampled cattle had some degree of udder and teat damage, very few farmers (2.6%) treated their cattle for these conditions. Udder damage was ca. two times and three times, respectively, more likely to occur in cattle on communal land compared to cattle on recently claimed land and commercial farms. The occurrence of R. appendiculatus and R. zambeziensis indicate that the cattle population in the study area is at high risk of a theileriosis outbreak, a tick-borne disease that has not been reported from this area.

Putative clinical piroplasmosis in a Burchell's zebra (Equus quagga burchelli).

A 10-year-old tame zebra gelding was presented after suffering from lethargy, nervousness, reported anaemia and icterus as well as a decreased appetite. These symptoms were seen over some months, with changing severity. The animal was immobilised, treated, and blood specimens were submitted for haematology and biochemistry. This report describes molecular characterisation of Theileria equi recovered from this animal, as well as the clinical findings, treatment and historical relevance of piroplasmosis in zebra in southern Africa.

Detection of Anaplasma antibodies in wildlife and domestic species in wildlife-livestock interface areas of Kenya by major surface protein 5 competitive inhibition enzyme-linked immunosorbent assay.

The seroprevalence of Anaplasma antibodies in wildlife (eland, blue wildebeest, kongoni, impala, Thomson's gazelle, Grant's gazelle, giraffe and plains zebra) and domestic animal (cattle, sheep and goat) populations was studied in wildlife/livestock interface areas of Kenya. Serum samples were analyzed by competitive inhibition enzyme-linked immunosorbent assay (CI-ELISA), using a recombinant antigen (MSP-5) from Anaplasma marginale surface membrane. A monoclonal antibody, FC-16, was used as the primary antibody, while anti-mouse conjugated to horseradish peroxidase was used as the secondary antibody. The results indicate a high seroprevalence in both wildlife and livestock populations, in contrast to earlier reports from Kenya, which indicated a low seroprevalence. The differences are attributed to the accurate analytical method used (CI-ELISA), as compared with agglutination techniques, clinical signs and microscopy employed by the earlier workers.

Occurrence of Theileria parva infection in cattle on a farm in the Ladysmith district, KwaZulu-Natal, South Africa.

Theileria parva causes widespread morbidity and mortality in cattle in endemic regions. An outbreak of theileriosis occurred on a farm near Ladysmith in KwaZulu-Natal, South Africa, which is not a declared Corridor disease-infected area. A survey of Red Brangus cattle from all age groups and areas of the farm was performed. Transmission of the parasite from infected animals on the farm to susceptible animals by tick transmission and tick-stabilate injection, was attempted. The survey indicated high numbers of animals with antibody titres to T. parva but only 6 infected animals, based on real-time PCR and RLB analysis. The transmission experiments failed to transmit the parasite. The study shows the difficulty in elucidating a source of infection and determining the dynamics of new infections in a herd where multiple possible sources are present and treatment with tetracyclines has taken place.

Detection of a Theileria species in dogs in South Africa.

A Theileria species was detected by PCR in blood samples collected from dogs in the Pietermaritzburg area and was also found in dogs presented at the Outpatients Clinic of the Onderstepoort Veterinary Academic Hospital (OVAH), in the Pretoria area, South Africa. In the Pietermaritzburg area, 79 of the 192 samples were positive, while 3 out of 1137 of the Onderstepoort samples were positive. Three positive samples from Pietermaritzburg were co-infected with Ehrlichia canis. PCR positive samples were further analysed by the Reverse Line Blot (RLB) and sequence analysis. Phylogenetic analysis of the 18S rRNA full-length gene sequences of one sample (VT12) from Pietermaritzburg and two samples from OVAH (BC281 and BC295) revealed a close relationship with sequences of Theileria species (sable). Clinical signs of the dogs that were examined at Pietermaritzburg and OVAH included an immune-mediated condition with severe thrombocytopenia. These findings identify a Theileria sp. in dogs for the first time in South Africa and add yet another microorganism to the growing list of haemoprotozoan parasites infecting dogs worldwide. The clinical significance of this infection in dogs is poorly resolved.

Molecular characterisation of Babesia gibsoni infection from a pit-bull terrier pup recently imported into South Africa.

Canine babesiosis caused by Babesia gibsoni was diagnosed in a 3-month-old Pit-bull pup during a routine clinical examination. Diagnosis was confirmed by way of smear examination, PCR, Reverse Line Blot (RLB) and sequence analysis which showed 100% homology with B. gibsoni (Japan AB118032) and Babesia sp. (Oklahoma) (AF205636). Haematology showed moderate anaemia and severe thrombocytopenia. Treatment was initiated with diminazene aceturate (Berenil RTU) followed by 2 doses of imidocarb diproprionate (Forray-65) 3 days and 14 days later, respectively. Babesia gibsoni DNA was still detectable 2 weeks post-treatment on the PCR/RLB test. A 10-day course of combination drug therapy using atovaquone and azithromycin was initiated. Blood samples taken on Day 1 and Day 40 after completion of treatment were negative for B. gibsoni DNA on PCR/RLB test. The implications of a possible introduction of B. gibsoni into South Africa are discussed.

Molecular evidence for transplacental transmission of Theileria equi from carrier mares to their apparently healthy foals.

The intra-erythrocytic parasite Theileria equi is one of two tick-transmitted causative agents of equine piroplasmosis. Piroplasms of T. equi can be transmitted across the equine placenta and once a horse is infected, it appears to remain a lifelong carrier, since anti-theilerial drugs suppress but do not eliminate the parasite. Carrier mares may transmit the organism to their offspring and this may result in abortion or neonatal piroplasmosis, but observations by some researchers suggest that foals may be born as carriers yet remain apparently healthy. Using a T. equi-specific oligonucleotide probe, we have determined that transplacental transmission occurs early in equine foetal development and that carrier mares may give birth to healthy carrier foals. Investigation of parasite levels and the effect of maternal colostrum on the newborn suggests that colostral T. equi antibody may act to suppress parasitaemia in the newborn, reducing the incidence of clinical neonatal piroplasmosis.

Occurrence of Babesia felis and Babesia leo in various wild felid species and domestic cats in Southern Africa, based on reverse line blot analysis.

Reverse line blot (RLB) is a hybridization assay that can be used to detect various blood parasites and differentiate between them. Results, using the RLB, showed that Babesia felis and Babesia leo occurred as single or mixed infections in various felid species, but most frequently in domestic cats and lions, respectively. Prevalence of infection in free-ranging cheetahs in Namibia was low (7, 5%), whereas 50% of free-ranging lions in South Africa and Swaziland were infected. A large number (52, 9%) of samples tested positive only for Babesia, neither B. felis nor B. leo. This could be an indication of at least one further, as yet undescribed, Babesia species in felids.