Rhipicephalus zambeziensis unlikely to transmit foot-and-mouth disease virus.

The potential of the ixodid tick, Rhipicephalus zambeziensis, was investigated as a vector in the transstadial transmission of the foot-and-mouth disease virus by feeding nymphae on viraemic (log 1.0-4.0 TCID50/ml) cattle. Suspensions were prepared, at various intervals after detachment, from pools of engorged nymphae--some of which were allowed to moult first. Suspensions were inoculated into sucking mice, cell cultures and, in some cases, cattle to detect the FMD virus. Newly moulted adult ticks, derived from nymphae which had fed on viraemic cattle, were also allowed to feed on susceptible cattle. The pattern of virus detection indicated that the FMD virus was capable of surviving at least 3 d in engorged nymphae, but less than 7 d following repletion. It was concluded that R. zambeziensis is unlikely to transmit the FMD virus.

The transovarial transmission of Babesia trautmanni by Rhipicephalus simus to domestic pigs.

Rhipicephalus simus was, for the first time, experimentally proven to be a transovarial vector of Babesia trautmanni of domestic pigs. The nymphal and adult progeny of experimentally infected female ticks transmitted the infection to 2 susceptible splenectomized pigs. Features of the infection included a prepatent period of 6-8 days post-tick infestation, a febrile reaction for 3 days and a maximum parasitaemia score of 15 (more than 6 parasites per 300 red blood cells). Other clinical signs in both pigs were mild inappetence and listlessness. Both pigs recovered without any antibabesial therapy.

Transmission and diagnosis of equine babesiosis in South Africa.

The transmission and prevalence of Babesia equi and B. caballi are being studied. Rhipicephalus evertsi mimeticus an ixodid tick from Namibia was identified as a new vector of B. equi, however, R. turanicus, previously reported to be a vector, failed to transmit both B. equi and B. caballi in the laboratory. The accurate diagnosis of B. caballi is being investigated because the nature of its low level parasitaemia does not allow easy detection in thin blood smears, routinely used for diagnosis, by clinicians. Consequently its role as a pathogen remains obscure. The importance of identifying infected horses, destined for export to Babesia-free countries, is also stressed. Thick and thin blood smears, serology (IFAT) and DNA probes are currently employed to study disease prevalence. To date 293 healthy, adult, thoroughbred horses have been screened by all three methods. The percentage positives are as follows: B. equi 4.4%, 70.6%, 13% and B. caballi 0.7%, 37%, 18.4% respectively. The DNA probes were more sensitive than blood smear examination for diagnosing carrier infections but are probably not sensitive enough to identify all carrier infections. A poor correlation was found between detection of the parasites' DNA and seropositivity. However, polymerase chain reaction could be used to amplify parasite DNA in a particular sample and this could result in more accurate diagnosis.

Transmission and diagnosis of equine babesiosis in South Africa

The transmission and prevalence of Babesia equi and B. caballi are being studied. Rhipicephalus evertsi mimeticus an ixodid tick from Namibia was identified as a new vector of B. equi, however, R. turanicus, previously reported to be a vector, failed to transmit both B. equi and B. caballi in the laboratory. The accurate diagnosis of B. caballi is being investigated because the nature of its low level parasitaemia does not allow easy detection in thin blood smears, routinely used for diagnosis, by clinicians. Consequently its role as a pathogen remains obscure. The importance of identifying infected horses, destined for export to Babesia-free coutries, is also stressed. Thock and thin blood smears, serology (IFAT) and DNA probes are currently employed to study disease prevalence. To date 293 healthy, adult, throughbred horses have been screened by all three methods. The percentage positives are as follows: B. equi 4.4%, 70.6%, 13% and B. caballi 0.7%, 37%, 18.4% respectively. The DNA probes were more sensitive than blood smear examination for diagnosing carrier infections but are probably not sensitive enough to identify all carrier infections. A poor correlation was found between detection of the parasites' DNA and seropositivity. However, polymerase chain reaction could be used to amplify parasite DNA in a particular sample and its could result in more accurate diagnosis

The isolation and transmission of an unidentified Babesia sp. to cattle by Hyalomma truncatum Koch 1844.

An unidentified Babesia sp. which causes a mild disease in cattle was isolated in a splenectomized ox that received pooled blood from field cattle. That this organism is pleomorphic and resembles Babesia occultans makes it difficult to differentiate between these organisms microscopically. Initially, it was suspected that this Babesia could be B. occultans. Several attempts to transmit this parasite transovarially with Hyalomma marginatum rufipes, the vector of B. occultans, failed. Continued efforts to identify possible vectors, using Boophilus microplus, Rhipicephalus evertsi evertsi and Rhipicephalus appendiculatus, all failed. The only tick thus far identified that could have transmitted the infection transovarially in the adult stage was the two-host tick Hyalomma truncatum.

An attempt to improve the immunization of sheep against heart-water by using different combinations of 3 stocks of Cowdria ruminantium.

Neither sheep immune to the Ball 3, the Kwanyanga or the Mara stocks of Cowdria ruminantium nor those immunized with combinations of these 3 stocks were protected against challenge with the Mali stock. Against challenge with the Welgevonden stock, however, immunization with each of the 3 combinations and with single stocks effected a protective immunity.

Corridor disease in South Africa: a review of the current status.

The African buffalo (Syncerus caffer) is a reservoir host of Theileria parva lawrencei the causative organism of Corridor or buffalo disease of cattle. This organism can apparently transform its behaviour when tick-passaged through cattle to resemble that of Theileria parva parva, causing classic East Coast fever (ECF). These are major considerations for the strict quarantine measures imposed on buffaloes from Corridor disease endemic areas in South Africa. Results of continuing studies on diagnosis, chemotherapy, transmission and attempts to transform the behaviour of T.p. lawrencei in the laboratory, are discussed.

Parasites of domestic and wild animals in South Africa. XXIII. Helminth and arthropod parasites of warthogs, Phacochoerus aethiopicus, in the eastern Transvaal Lowveld.

A total of 69 warthogs, Phacochoerus aethiopicus, were collected from 4 localities within the Kruger National Park, eastern Transvaal Lowveld. These animals harboured 16 nematode species, 2 trematodes, 1 or 2 species of adult cestodes and the larval stages of 4 cestodes. No pattern of seasonal abundance could be determined for any of the helminths. The warthogs were also infested with 3 flea species, 1 louse species, 8 ixodid tick species, 1 argasid tick and the nymphae of a pentastomid. The seasonal abundance of fleas of the genus Echidnophaga, of the sucking louse Haematopinus phacochoeri and the ixodid ticks Amblyomma hebraeum, Rhipicephalus appendiculatus, Rhipicephalus simus and Rhipicephalus zambeziensis was determined.

Freeze-drying of Aegyptianella pullorum.

Heparinized whole blood, parasitized with Aegyptianella pullorum, was collected from 2 fowls. Buffered lactose peptone (BLP) was added v/v as a stabilizer and the mixture lyophilized in 2 ml aliquots after rapid or slow freezing. At different stages during the freeze-drying process, as well as after lyophilization and reconstitution with 1.8 ml of sterile water, samples were taken and injected into pullets. Infectivity was maintained throughout. However, the prepatent period was lengthened after freezing and particularly after lyophilization when there was some loss of viability.

Isolation of pure Babesia equi and Babesia caballi organisms in splenectomized horses from endemic areas in South Africa.

Both Babesia equi and Babesia caballi are endemic in large parts of South Africa. Attempts were made to obtain pure local isolates of both B. equi and B. caballi for the purpose of developing serological tests to study the epidemiology of equine babesiosis in this country. The indirect fluorescent antibody test was used to screen horses for B. equi and B. caballi in an endemic area. Seven horses and 3 donkeys between 3 and 36 months of age that tested negative were subsequently splenectomized. The splenectomy operation was performed through the abdominal approach. A 100% survival rate was achieved through this method, probably because it reduced the risk involved in the operation. Blood collected from naturally infected horses and passaged in fully susceptible splenectomized horses and a donkey, under laboratory conditions, produced 2 isolates of Babesia caballi and 1 of B. equi. Microscopical and serological examinations confirmed that these were pure isolates.

Besnoitia besnoiti: studies on the definitive host and experimental infections in cattle.

Domestic cats, 11 other species of carnivorous mammals, 6 species of snakes, and white-backed vultures were tested for their possible role as definitive hosts of Benoitia besnoiti by feeding with cystic material from chronically infected bovines. None of the species tested is a definitive host; hence, the life cycle of this parasite remains obscure. In attempts to produce clinical cases of besnoitiosis by experimental infection, bovines were inoculated IV, SC, and IP with cystozoites or tachyzoites. Immunosuppression of the animals was essential for the development of severe cases and skin lesions; cystozoites proved to be more pathogenic than tachyzoites.

The persistence of colostral Anaplasma antibodies and incidence of in utero transmission of Anaplasma infections in calves under laboratory conditions.

Twenty-six calves, born from 25 Anaplasma-infected, intact and splenectomized cows, from a herd kept under strict tick-free laboratory conditions, were monitored for the presence of Anaplasma antibodies, using the rapid card agglutination test. Serum was collected at birth, weekly for 12 weeks, and then monthly for approximately 6 months. Specific antibodies passively acquired could be detected in calf sera for an average period of 8 weeks after birth. Calves that remained positive for longer than 12 weeks were suspected of having contracted in utero infections. Infection of the calves was confirmed by splenectomy. It was concluded that 4 calves in Group I contracted in utero infections. Two of the dams were chronically infected, whilst the other 2 underwent acute primary reactions during the 1st and 2nd trimesters of gestation, respectively. Subsequently all calves born from infected cows in this tick-free herd were serologically screened before being splenectomized at an average age of 8 months. Out of 50 cows, 8 in utero infected calves were identified serologically and this finding was confirmed through splenectomy or subinoculation of blood. Both Anaplasma centrale and Anaplasma marginale were carried transplacentally. Splenectomized and intact cows, chronically infected or undergoing primary reactions during the 1st, 2nd or 3rd trimester of gestation, produced infected calves. A 15,6% incidence of in utero transmitted infections were observed amongst 77 calves under these conditions. None of the 13 splenectomized cows, undergoing primary A. centrale infections during gestation, aborted. Clinical signs of disease were not observed in any of the 12 in utero infected calves prior to splenectomy. The implications of these findings are discussed.

An investigation into the clinical pathological changes and serological response in horses experimentally infected with Babesia equi and Babesia caballi.

Serologically negative horses, as determined with the indirect fluorescent antibody test (IFA), were infected with Babesia equi and 60 days later with Babesia caballi. The only clinical signs of disease observed in these animals were a febrile reaction and slight icterus. Haematological changes included a drop in haematocrit and haemoglobin concentration, as well as lowered platelet counts. The serum concentrations of albumin, iron and phosphorus were lowered. Mildly elevated serum bilirubin and fibrinogen concentrations were observed. Antibody titres were determined with the IFA and complement fixation (CF) tests. Antibodies to B. equi were first detected between Days 10-19 and 12-38 with the IFA and CF test, respectively, while the corresponding IFA periods for B. caballi were 6-8 days after infection. The parasitaemia of both B. equi and B. caballi infections never reached the 1% level.

The transstadial transmission of Babesia caballi by Rhipicephalus evertsi evertsi.

Rhipicephalus evertsi evertsi larvae were fed on the ears of rabbits. Seven days after larval infestation, unfed, newly moulted nymphae were manually removed to infest a splenectomized donkey showing a patent Babesia caballi infection. Engorged nymphae were collected from the donkey and the ensuing adult ticks were placed on a susceptible horse. The horse contracted a B. caballi infection showing a prepatent period of 19 days after tick infestation. A very low parasitaemia, (highest score 2), which was patent for only 10 days, was recorded. The lowest packed cell volume recorded was 16%.

Chronic splenitis as a possible complicating factor in calves inoculated with an attenuated redwater vaccine.

In two consecutive years a total of 3810 calves were inoculated with the Onderstepoort redwater live-blood vaccine, containing attenuated strains of Babesia bigemina and Babesia bovis. Twenty calves died 8-13 days later due to B. bigemina-infection. Necropsies were performed on 19 of these animals. In 11 calves the spleen was distinctly smaller than normal and its morphology was distorted, and showed changes of chronic splenitis. Histological lesions in the spleens of 6 of the 11 calves are described. The potentiating effect of the splenic lesions to reduce the ability to contain the vaccine reaction, is discussed.

Tick transmission of Anaplasma centrale.

Anaplasma centrale was isolated from a field collection of Rhipicephalus simus. Transstadial transmission of A. centrale with adult ticks was demonstrated, but the infection was not carried transovarially. Ticks from this collection were subsequently reared as a non-infected, laboratory strain. It was proved that the Onderstepoort live blood vaccine strain of A. centrale, isolated by Theiler in 1911, is still tick transmissible after more than 75 years of needle passage through cattle in the laboratory. Attempts to demonstrate transstadial transmission of the vaccine strain with Boophilus decoloratus and Boophilus microplus failed.

The genome of anaplasma: DNA base composition and DNA/DNA hybridization.

The Tm value of DNA from Anaplasma centrale and Anaplasma marginale was found to be 87.1 degrees C and 89.3 degrees C, respectively. The G + C content, calculated from the Tm, was 45.1% for A. centrale and 48.5% for A. marginale. Identical hybridization patterns were obtained when the DNA from one species was hybridized to restriction endonuclease-digested DNA from the other species.

The inability of a South African Babesia bovis vaccine strain to infect Boophilus microplus.

A strain of Babesia bovis that had been attenuated by rapid syringe passage through a series of 23 splenectomized calves was unable to infect its vector Boophilus microplus. An attempt to transmit the attenuated Australian Babesia bigemina G strain with a South African strain of B. microplus was likewise unsuccessful. The epidemiological implication of these observations in terms of babesiosis control is discussed.

A column purification procedure for the removal of leucocytes from parasite-infected bovine blood.

A cellulose column procedure is described which removes white cells from bovine blood infected with Babesia and Anaplasma. The efficiency of this method was confirmed by the absence of white blood cell DNA in lysates from column-filtered infected as well as non-infected blood.