Occurrence of Theileria Parva and Other Haemoprotozoa in Cattle at the Edge of Hluhluwe-Imfolozi Park, Kwazulu-Natal, South Africa

Article — Artikel

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

S B A S Yusufmiaa, N E Collinsa, R Nkunaa, M Troskiea, P Van den Bosschea,b and B L Penzhorna*

South Africa occurred in the corridor ABSTRACT between the then Hluhluwe and Umfolozi Theileria parva, the most important bovine theilerial species in sub-Saharan Africa, causes Game Reserves in KwaZulu-Natal (now widespread mortality and morbidity in endemic areas. A survey was conducted using incorporated in the Hluhluwe-iMfolozi buffy-coat specimens from 60 apparently healthy adult communally herded Nguni-type Park), shortly after ECF had been eradi- cattle at the northeastern edge of the Hluhluwe-iMfolozi Park to determine, by means of cated27. The disease occurs sporadically PCR and Reverse Line Blot (RLB) hybridisation, the occurrence of Theileria and Babesia wherever cattle are in contact with infected 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 African buffaloes and in the presence of suggests that cattle in South Africa, and not only African buffaloes (Syncerus caffer), may be the vector ticks. In areas where cattle are subclinical carriers of T. parva. Other species identified were T. mutans (83.3 %), T. velifera raised, acceptance of buffaloes for conser- (70.0 %), Theileria sp. (sable) (46.8 %) and T. taurotragi (1.7 %). Two specimens (3.3 %) were vation or recreation is a major constraint 17 positive for Babesia bovis and single specimens (1.7 %) positive for B. bigemina and B. rossi, because of Corridor disease . Buffalo- respectively. Mixed infections, of up to 4 species, were common (65.0 %). Only 1 specimen derived T. parva is universally distributed was found to be positive for Trypanosoma vivax, and 2 for T. theileri, of which only the first in wild buffaloes in southern Africa, species is pathogenic. except in the Addo Elephant National Keywords: Babesia bigemina, Babesia bovis, Babesia rossi, cattle, Hluhluwe-iMfolozi Park, Park and some other game reserves in Theileria mutans, Theileria parva, Theileria taurotragi, Theileria velifera, Theileria sp. (sable), non-endemic areas where T. parva-free Trypanosoma theileri, Trypanosoma vivax, wildlife–livestock interface. animals have been translocated30.In Yusufmia SBAS,Collins N E, Nkuna R, Troskie M, Van den Bossche P, Penzhorn B L contrast to the situation in buffaloes, Occurrence of Theileria parva and other haemoprotozoa in cattle at the edge of the erythrocytic piroplasms in cattle are Hluhluwe-iMfolozi Park, KwaZulu-Natal, South Africa. Journal of the South African Veteri- usually absent or too scanty to infect ticks, nary Association (2010) 81(1): 45–49 (En.). Department of Veterinary Tropical Diseases, so the disease is usually self-limiting Faculty of Veterinary Science, University of Pretoria, Private Bag X04, Onderstepoort, 0110 in cattle. The distinct seasonality of R. South Africa. appendiculatus, which has a 2-year life cycle in southern Africa38, and transstadial transmission of T. parva also play a major INTRODUCTION to susceptible cattle by R. appendiculatus as role in this regard. The possibility of Theileriosis, a tick-transmitted proto- well as by R. zambeziensis and possibly by T. parva from buffalo transforming under zoan disease, is a major constraint for R. duttoni17. Zimbabwe theileriosis is natural conditions in South Africa to cattle production in the tropics and sub- usually a milder form of the disease, with T. parva causing ECF has always been a tropics12. Theileria parva is the most impor- transmission by R. appendiculatus between highly contentious topic, as it places the tant species in sub-Saharan Africa, while infected and susceptible cattle15. movement of buffaloes under severe T.annulata occurs further northwards and ECF was introduced to South Africa constraints36. eastwards. Three subspecies of T. parva during 1901–1903 by cattle imported In South Africa, the Kruger National were previously recognised: T. parva from Kenya and Tanzania for restocking Park and Hluhluwe-iMfolozi Park are parva, causing classical East Coast fever after the ravages of rinderpest and the regarded as endemic areas for buffalo- (ECF)16; T.parva lawrencei, causing Corridor Anglo-Boer War14. It is estimated that 1.25 derived T.parva. The South African Animal disease17; and T. parva bovis, causing million of the 4 million cattle in the Disease Act (Act 35 of 1984) stipulates the Zimbabwe theileriosis or January disease15. affected area had died of ECF by 1914. The following controlled veterinary acts to This subdivision has been abandoned34. disease was eradicated from southern be performed in outbreaks of Corridor East Coast fever is an acute, usually fatal, Africa during a 50-year campaign consist- disease: Susceptible animals: Contact be- disease that is transmitted between in- ing of movement control, tick control, tween cattle and African buffaloes shall be fected cattle and susceptible cattle by the destocking of infected pastures and prevented; all cattle in a controlled area 3-host ixodid tick, Rhipicephalus appendicu- slaughtering of infected herds16. The shall be dipped or sprayed regularly by latus16. Corridor disease is an acute disease vector was not eradicated, however, and the responsible person with an efficient transmitted principally from asymptom- the original buffalo-associated T. parva remedy; no animal shall be chemothera- atic carrier African buffalo (Syncerus caffer) remained endemic in South African and peutically treated without the written Zimbabwean buffalo populations40. authorisation of the director. Contact aDepartment of Veterinary Tropical Diseases, Faculty of Corridor disease is an acute, usually animals shall be isolated and dipped or Veterinary Science, University of Pretoria, Private Bag 17 X04, Onderstepoort, 0110 South Africa. fatal disease of cattle resembling ECF .It sprayed with an efficient remedy under bDepartment of Animal Health, Institute of Tropical is caused by infection with T. parva strains the supervision of an officer or an author- Medicine, Antwerp, Belgium. from African buffaloes, the only wild ised person in the manner and at the *Author for correspondence. E-mail: [email protected] ruminant species that is a carrier of the intervals determined by the responsible 36 Received: October 2009. Accepted: February 2010. causative organism . The first outbreak in state veterinarian. Infected animals shall be

0038-2809 Jl S.Afr.vet.Ass. (2010) 81(1): 45–49 45 isolated and dipped or sprayed with an Table 1: Species-specific probes for Theileria and Babesia species. efficient remedy under the supervision of an officer or an authorised person in the Species Probe sequence (from 5’–3’)* manner and at the intervals determined B. felis TTA TGC GTT TTC CGA CTG GC by the responsible state veterinarian. In B. divergens ACT RAT GTC GAG ATT GCA C contrast to the regulations pertaining to B. microti GRC TTG GCA TCW TCT GGA ECF, slaughter of infected animals is B. bigemina CGT TTT TTC CCT TTT GTT GG therefore not enforced, on the assumption B. bovis CAG GTT TCG CCT GTA TAA TTG AG that infected animals, which may not be B. rossi CGG TTT GTT GCC TTT GTG treated, will succumb to the disease and B. canis canis TGC GTT GAC CGT TTG AC will not become subclinical carriers. B. canis vogeli AGC GTG TTC GAG TTT GCC The campaign to control the ECF epi- B. major TCC GAC TTT GGT TGG TGT demic in South Africa by intensive regula- B. bicornis TTG GTA AAT CGC CTT GGT C tory control of R. appendiculatus had a B. caballi GTT GCG TTK TTC TTG CTT TT major effect on single-host ticks, includ- Theileria/Babesia catchall TAA TGG TTA ATA GGA RCR GTT G ing Rhipicephalus (Boophilus) spp., ensur- T. parva GGA CGG AGT TCG CTT TG ing that bovine babesiosis remained of T. annulata CCT CTG GGG TCT GTG CA secondary importance for many decades. T. mutans CTT GCG TCT CCG AAT GTT In the latter part of the 20th century, relax- T. taurotragi TCT TGG CAC GTG GCT TTT ation or breakdown of tick-control mea- T. velifera CCT ATT CTC CTT TAC GAG T sures and the advent of acaricide resis- T. lestoquardi CTT GTG TCC CTC CGG G tance in Rhipicephalus (Boophilus) spp., T. equi TTC GTT GAC TGC GYT TGG caused bovine babesiosis to increase in T. buffeli GGC TTA TTT CGG WTT GAT TTT prevalence and significance to the point T. bicornis GCG TTG TGG CTT TTT TCT G where it became recognised as one of the Theileria sp. (buffalo) CAG ACG GAG TTT ACT TTG T most important livestock diseases in Theileria sp. (sable) GCT GCA TTG CCT TTT CTC C South Africa7,8. Theileria sp. (kudu) CTG CAT TGT TTC TTT CCT TTG Trypanosomosis is a complex, debilitat- ing and often fatal disease caused by *Symbols used to indicate degenerate positions: R = A/G, W = A/T, K = G/T. infection with one or more of the pathogenic tsetse fly-transmitted protozoan vicinity of the Hluhluwe-iMfolozi Park41. –20 °C until processed further at the parasites of the genus Trypanosoma5. The In a subsequent survey, blood specimens Molecular Biology Laboratory, Depart- most important species responsible for were taken randomly from healthy cattle ment of Veterinary Tropical Diseases, the disease complex, commonly known in the same area to determine the occur- Faculty of Veterinary Science, University as ‘nagana’ in livestock, include Trypano- rence of Trypanosoma species. This offered of Pretoria. soma brucei, T.congolense and T.vivax1.Two the opportunity to determine the occur- tsetse fly species, Glossina austeni and rence of other haemoparasites, especially DNA extraction G. brevipalpis, persisted in KwaZulu-Natal Theileria parva, in these cattle. The RLB DNA was extracted from 60 buffy coat despite eradication attempts13. was used for detection of Theileria species filter papers (at least 3 disks, 3 mm in A number of molecular, biochemical in these buffy-coat specimens and PCR- diameter per sample) using QIAamp and immunological tools are available RFLP was used to determine occurrence DNA Mini kit (QIAGEN®), according to that enable differentiation between Thei- of Trypanosoma species. the manufacturer’s instructions. DNA leria species4. Early molecular detection was eluted in 150 µl of AE buffer and techniques involved the use of probes to MATERIALS AND METHODS stored at 4 °C. detect repetitive regions in parasite genomic deoxyribonucleic acid (DNA). Specimen collection Polymerase Chain Reaction (PCR) for The polymerase chain reaction (PCR) Specimens were collected during Theileria and Babesia species assay has largely superseded DNA March 2006 from 60 randomly selected, Primers RLB F2 (5’-GAC ACA GGG probes. PCR has a higher sensitivity and adult, apparently healthy, communally- AGG TAG TGA CAA G-3’) and biotin- specificity for detecting parasite DNA in grazed Nguni-type cattle from various labeled RLB R2 (5’–Biotin–CTA AGA ATT blood. False positives may occur, how- herds adjacent to the northeastern edge TCA CCT CTA ACA GT-3)’ were used for ever, due to contamination. As it is not of the Hluhluwe-iMfolozi Park. The total amplification of the V4 hypervariable always possible to detect mixed infections number of cattle in the area was not region of Theileria and Babesia 18S rRNA with PCR, a reverse line blot (RLB) known. Jugular blood was collected in genes11. Reactions were performed in a hybridisation assay targeting the 18S Vacutainer® tubes coated with ethylene- final volume of 25 µ with Platinum ribosomal ribonucleic acid (rRNA) gene diamine-tetra-acetic acid (EDTA) as anti- Quantitative PCR Super mix-UDG (Invi- has been developed for simultaneous coagulant. Micro-capillary tubes were trogen), 0.25 µ of each primer (20 pmol) identification and differentiation of filled with blood from the Vacutainer and 2.5 µ of purified DNA. A touchdown distinct piroplasm species present in the tubes and centrifuged for 5 min at PCR programme was conducted in the same sample11. The RLB has proven to be 9000 r.p.m. A diamond-tipped pen was Gene Amp PCR system 9700. The cycling a very powerful tool in detecting sub- used to cut the capillary tubes immedi- conditions were as follows: 3 min at 37 °C; clinical infections. It also gives an indica- ately above the buffy coat, and the buffy 10 min at 94 °C; and 10 cycles of 94 °C for tion of new species or genotypes possibly coat was extruded onto filter paper 20 s, 67 °C for 30 s, 72 °C for 30 s, with being present in the sample32. (Whatman no. 3). The buffy-coat-spotted decreasing annealing temperature after Bovine trypanosomosis has been shown filter paper was dried, sealed in a plastic every second cycle by 2 °C for 5 times until to be prevalent in anaemic cattle in the bag containing silica gel and stored at the annealing temperature reached 57 °C.

46 0038-2809 Tydskr.S.Afr.vet.Ver. (2010) 81(1): 45–49 Fig. 1: Occurrence of specific haemoparasites in cattle along the northeastern edge of Hluhluwe-iMfolozi Park, KwaZulu-Natal, South Africa.

Finally 40 cycles of 94 °C for 20 s, 57 °C for Fermentas, Lithuania) for fragment size Table 2: Occurrence of Theileria spp. and 30 s and 72 °C for 30 s were performed. determination. DNA fragments were Babesia spp. infections in cattle along the northeastern edge of Hluhluwe-iMfolozi Babesia bovis was used as positive control, separated by horizontal electrophoresis Park, KwaZulu-Natal, South Africa. while bovine DNA was used as negative in 0.5× TBE buffer at 100 V for 2.5 h. The control. gel was stained using Sybr Green I nucleic No. of species present No. of 10 acid gel stain (Roche diagnostics) . specimens Reverse Line Blot (RLB) hybridisation The PCR products were analysed using RESULTS 1 15 (25.0 %) the RLB hybridisation technique11,28,29.A Haemoprotozoa DNA could be extracted 2 9 (15.0 %) Theileria and Babesia genus-specific oligo- from all specimens. Occurrence of Theileria 3 26 (43.3 %) nucleotide probe and 23 species-specific spp., Babesia spp. and Trypanosoma spp. 4 4 (6.7 %) probes (Table 1) were included on the is summarised in Fig. 1. Theileria parva Theileria/Babesia genus- 6 (10.0 %) specific probe positive only membrane. occurred in 4 specimens (6.7 %). Theileria mutans (83.3 %) and T.velifera (70.0 %) were Total 60 (100 %) PCR for Trypanosoma species the most prevalent Theileria species. The primers used for the amplification These were also the only haemoparasites piroplasm stage, which is infective to of Trypanosoma species were as described10. occurring as single-species infections: ticks30. Experimental evidence suggests The first amplification was done using the 10 specimens (16.7 %) were infected with that some bovines survive the disease, forward primer 18ST nF2 (5’-CAA CGA T.mutans only, and 5 specimens (8.3 %) with however, and may serve as reservoirs of TGA CAC CCA TGA ATT GGG GA-3’) and T. velifera only. Occurrence of single- infection15. Furthermore, South Africa is 18ST nR3 (5’-TGC GCG ACC AAT AAT species and multiple-species infections is considered free of T. parva, except in TGC AAT AC-3’) as reverse primer. A shown in Table 2. Confirmed multiple- designated Corridor disease-infected semi-nested second amplification was species infections occurred in 39 (65 %) of areas including and bordering the Kruger done using 18ST nF2 with the reverse the specimens. Six specimens (10 %) were National Park and Hluhluwe-iMfolozi primer 18ST nR2 (5’GTG TCT TGT TCT positive for the Theileria/Babesia ge- Park. Therefore the national herd is es- CAC TGA CAT TGT AGT G-3’). nus-specific probe only; no hybridisation sentially naïve and completely suscepti- occurred to species-specific probes on the ble to T. parva39. RFLP for differentiation of Trypanosoma RLB membrane (Table 2). These results and those of a study on species another cattle population39, suggest that The RFLP analysis of the 18S rRNA PCR DISCUSSION cattle could be subclinical carriers of products was performed10. Briefly,a6µ Four specimens (6.7 %) were positive for T. parva parasites in South Africa. A study aliquot of amplified DNA was digested T.parva. As ECF no longer occurs in South should be carried out to determine whether with Msp1 and Eco571 restriction enzymes Africa, we assume that this result indi- these parasites could serve as a source of in buffer Y+/Tango with S-adenosyl- cates the presence of buffalo-associated infection to disease-transmitting ticks. methionine according to the manufac- T. parva. Specimens were obtained from These findings would be very important, turer’s specifications (Gibco, UK) using apparently healthy cattle only, which as they could indicate that current control 0.6 U of each enzyme per µ of PCR strongly suggests that these cattle are regulations in South Africa may have to product in 15 µ total volume. The diges- subclinical carriers of buffalo-associated be revised. The assumption that only buf- tion was incubated overnight in a water T. parva. faloes are carriers of the parasite, and thus bath at 37 °C. Restriction products were Corridor disease has been regarded as merely preventing contact with buffaloes resolved on a 10 % polyacrylamide gel self-limiting in cattle because they usually is an appropriate management strategy, together with a 100 bp DNA ladder (MBI die before the parasite develops to the may no longer be tenable. Therapeutic

0038-2809 Jl S.Afr.vet.Ass. (2010) 81(1): 45–49 47 treatment of Corridor disease is not transmission could have produced a posi- ACKNOWLEDGEMENTS allowed in South Africa as recovered tive result. Theileria equi, which had not Dr Dirk Geysen is thanked for supplying intact cattle could develop a carrier status been described previously as a parasite of specimens and Ms Raksha Bhoora is and could become infective to ticks35.If canids, has been found in asymptomatic thanked for her assistance in the laboratory. carriers are present and cattle-to-cattle dogs6. It was concluded that infection of transmission occurs, it could lead to the dogs with T. equi is not unlikely to happen REFERENCES selection of a subpopulation of T. parva and that molecular methods can lead to a 1. Anene B M, Onah D N, Nawa Y 2001 Drug parasites that are better adapted to cattle correct identification of piroplasmids in resistance in pathogenic African trypano- somes: What hopes for the future? Veteri- as host. If this were to happen in South accidental hosts. Similarly, in this study, nary Parasitology 96: 83–100 Africa, as is speculated is the case in East accidental transmission of B. rossi in cattle 2. Barnett S F, Brocklesby D W 1966 The 2 Africa , it would impact negatively on the probably occurred. passage of Theileria lawrencei (Kenya) cattle industry. A fairly large percentage (10 %) of the through cattle. British Veterinary Journal 122: The high prevalence of T. mutans infec- specimens were positive for the Theileria/ 396–409 3. Brocklesby D W 1965 A new theilerial tion (83.3 %) was not really surprising, as genus-specific probe only, but no Babesia parasite of the African buffalo (Syncerus it has been shown that virtually all calves hybridisation occurred to species-specific caffer). Bulletin of Epizootic Diseases of Africa in endemic areas may be infected by the probes on the RLB membrane. This could 13: 325–330 time they are 6 months old26. African indicate that there may be novel Theileria 4. Collins N E, Allsopp MTEP,Allsopp B A buffalo also harbour T.mutans3,33 and there or Babesia species in cattle or variants of 2002 Molecular diagnosis of theileriosis and heartwater in bovines in Africa. Transactions is field and experimental evidence suggest- species present in these specimens. Alter- of the Royal Society of Tropical Medicine and ing that strains of T. mutans from buffalo natively, it could indicate the presence of Hygiene 96: 217–224 may be more pathogenic to cattle than a known species for which no probe was 5. Connor R J, Vanden Bossche P 2004 African those derived from cattle20. The prevalence included on the blot. Not all probes that animal trypanosomoses. In CoetzerJAW, of T. velifera, which is also known to infect are specific for all the Theileria and Babesia Tustin R C (eds) Infectious diseases of livestock 37 (2nd edn). Oxford University Press Southern buffalo , was also quite high (70.0 %). species that could occur in cattle were Africa, Cape Town: 251–296 This piroplasm is generally regarded as used in this study; for example, B. occultans 6. Criado-Fornelio A, Martinez-Marcos A, 21 apathogenic in cattle . Theileria taurotragi, was not included. Alternatively, the con- Buling-Saraña A, Barba-Carretero J C 2003 which is mildly pathogenic in cattle centration of species-specific DNA may Molecular studies on Babesia, Theileria and but has been associated with cerebral have been too low to be detected in the Hepatozoon in southern Europe. Part 1. Epizootiological aspects. Veterinary Parasi- theileriosis (so-called turning disease) hybridisation reaction. Sequence analysis tology 113: 189–201 was initially described from eland could be carried out on these specimens 7. de Vos A J 1979 Epidemiology and control of 9,18,19,22 Taurotragus oryx . The high preva- that hybridised only to the Theileria and bovine babesiosis in South Africa. Journal of lence of Theileria sp. (sable) (46.8 %) is Babesia genus-specific probe to determine the South African Veterinary Association 50: interesting. This species, which has been the nature of these specimens. 357–362 8. de Vos A J, de Waal D T, Jackson L A 2004 reported from cattle in Tanzania, is Two of the 60 specimens were positive Bovine babesiosis. In CoetzerJAW,Tustin incriminated in causing mortalities in for Trypanosoma theileri and 1 was positive R C (eds) Infectious diseases of livestock (2nd sable antelope (Hippotragus niger) and for T. vivax. Results of a previous survey edn). Oxford University Press Southern possibly roan antelope (Hippotragus of the same area showed that bovine Africa, Cape Town: 406–424 equinus)29, neither of which occurs in trypanosomosis is prevalent in the vicinity 9. Flanagan H O, le Roux J M W 1957 Bovine 41 cerebral theileriosis: a report on two cases Hluhluwe-iMfolozi Park. It has also been of the Hluhluwe-iMfolozi Park . Accord- occurring in the Union. Onderstepoort reported from African buffalo, blue wilde- ing to that survey, in which anaemic Journal of Veterinary Research 27: 453–461 beest (Connochaetes taurinus), klipspringer animals had been selected, 60.5 % of the 10. Geysen D, Delespaux V, Geerts S 2003 (Oreotragus oreotragus) and reedbuck specimens were positive for Trypanosoma PCR-RFLP using SSU-rDNA amplification (Redunca arundinum)29, all of which occur congolense41. Trypanosoma vivax is one of as an easy method for species-specific diagnosis of Trypanosoma species in cattle. in Hluhluwe-iMfolozi Park. A closely the species responsible for causing nagana, Veterinary Parasitology 110: 171–180 related Theileria species has been reported while Trypanosoma theileri (subgenus 11. Gubbels J M, de Vos A P,van der Weide M, from dogs in KwaZulu-Natal25. Megatrypanum) is generally regarded as Viseras J, Schouls LM,deVries E, Jongejan Two of the specimens were positive for non-pathogenic. F 1999 Simultaneous detection of bovine B. bovis, indicating a degree of the addi- In summary, we determined occurrence Theileria and Babesia species by reverse line blot hybridization. Journal of Clinical Micro- tional challenge the herd may be exposed of haemoprotozoa in 60 cattle at the wild- biology 37: 1782–1789 to. A third sample was positive for B. life/livestock interface in northern Kwa- 12. Jongejan F, Uilenberg G 1994 Ticks and bigemina. This extremely low prevalence Zulu-Natal. The most important finding control methods. Revue Scientifique et Tech- of Babesia spp. indicates effective tick con- was that 4 cattle (6.7 %) were infected nique, Office International des Épizooties 13: trol in the area. with T. parva. The presence of subclinical 1201–1226 13. Kappmeier K, Nevill E M, Bagnall R J 1998 One of the specimens was positive for T. parva carriers, which were also recently 39 Review of tsetse and trypanosomosis in B. rossi. This parasite had previously only found in another cattle population in South Africa. Onderstepoort Journal of Veteri- been found in canids23,24,31 and thus this KwaZulu-Natal, would suggest that the nary Research 65: 195–203 result may have been due to contamina- premises on which current Corridor- 14. Lawrence J A 1992 History of bovine tion. With PCR product contamination, disease-control strategies in South Africa theileriosis in southern Africa. In Norval R A I, Perry B D, YoungA S (eds) The epidemi- however, more than 1 false positive result are based may have to be reassessed. ology of theileriosis in Africa. Academic Press, would have been expected. Accidental This paper emanates from project London: 1–31 transmission of parasites to aberrant V015/08, approved by the Research 15. Lawrence J A, de Vos A J, Irvin A D 1994 hosts is possible. These parasites may sur- Committee of the Faculty of Veterinary Zimbabwe theileriosis. In Coetzer J A W, vive and be detected, although it is un- Science and the Animal Use and Care Thompson G R, Tustin R C (eds) Infectious diseases of livestock with special reference to likely that the parasite would be able to Committee of the University of Pretoria southern Africa, Vol. 1. Oxford University reproduce and cause disease in the acci- and funded by National Research Foun- Press, Cape Town: 326–328 dental host. It is possible that accidental dation grant GUN 44403 toBLPenzhorn. 16. Lawrence J A, Perry B D, Williamson S M

48 0038-2809 Tydskr.S.Afr.vet.Ver. (2010) 81(1): 45–49 2004 East Coast fever. In CoetzerJAW, Veterinary Parasitology 155: 152–157 buffalo, and transmission by Amblyomma Tustin R C (eds) Infectious diseases of livestock 25. Matjila P T,Leisewitz A L, Oosthuizen M C, gemma. Veterinary Parasitology 8: 31–37 (2nd edn). Oxford University Press South- Jongejan F, Penzhorn B L 2008 Detection of 34. Perry B D, Young A S 1993 The naming ern Africa, Cape Town: 448–463 a Theileria species in dogs in South Africa. game: the changing fortunes of East Coast 17. Lawrence J A, Perry B D, Williamson S M Veterinary Parasitology 157: 34–40 fever and Theileria parva. Veterinary Record 2004 Corridor disease. In CoetzerJAW, 26. Moll G, Lohding A, Young A S 1984 Epide- 133: 613–616 Tustin R C (eds) Infectious diseases of livestock miology of theileriosis in the Trans-Mara 35. Potgieter F T, Roos J A, de Vos A J 1985 (2nd edn). Oxford University Press Southern Division, Kenya. Husbandry and disease Implications of chemotherapy of Theileria Africa, Cape Town: 468–471 background and preliminary investigations lawrencei infections (Corridor disease) in 18. Lawrence J A, Williamson S M 2004 Turning on theileriosis in calves. Preventive Veteri- cattle. South African Journal of Science 81: 44 sickness. In Coetzer J A W, Tustin R C (eds) nary Medicine 2: 801–832 36. Potgieter F T, Stoltsz W H, Blouin E F, Roos Infectious diseases of livestock (2nd edn). 27. Neitz W O 1955 Corridor disease: a fatal J A 1988 Corridor disease in South Africa: A Oxford University Press Southern Africa, form of bovine theileriosis encountered in review of the current status. Journal of the Cape Town: 475–477 Zululand. Bulletin of Epizootic Diseases of South African Veterinary Association 59: 19. Lawrence J A, Williamson S M 2004 Theileria Africa 3: 121–123 155–160 taurotragi infection. In Coetzer J A W,Tustin 28. Nijhof A M, Penzhorn B L, Lynen G, Mollel J 37. Schreuder B E C, Uilenberg G, Tondeur W R C (eds) Infectious diseases of livestock (2nd O, Bekker C, Jongejan F 2003 Babesia bicornis 1977 Studies on Theileriidae (Sporozoa) in edn). Oxford University Press Southern sp. n. and Theileria bicornis sp. n.: tick-borne Tanzania. VIII. Experiments with African Africa, Cape Town: 478–479 parasites associated with mortality in the buffalo (Syncerus caffer). Tropenmedizin und 20. Lawrence J A, Williamson S M 2004 Theileria black rhinoceros (Diceros bicornis). Journal of Parasitologie 28: 367–371 mutans infection. In CoetzerJAW,Tustin Clinical Microbiology 41: 2249–2254 38. Short N J, NorvalRAI1981 Seasonal activ- R C (eds) Infectious diseases of livestock (2nd 29. Nijhof A M, Pillay V, Steyl J, Prozesky L, ity of Rhipicephalus appendiculatus Neu- edn). Oxford University Press Southern Stoltsz W H, Lawrence J A, Penzhorn B L, mann 1901 (Acari: Ixodidae) in the highveld Africa, Cape Town: 480–482 Jongejan F 2005 Molecular characterization of Zimbabwe Rhodesia. Journal of Parasitol- 21. Lawrence J A, Williamson S M 2004 Theileria of Theileria species associated with mortality ogy 67: 77–84 velifera infection. In CoetzerJAW,Tustin R in four species of African antelopes. Journal 39. Thompson B E, Latif A A, Oosthuizen M C, C (eds) Infectious diseases of livestock (2nd of Clinical Microbiology 43: 5907–5911 Troskie M, Penzhorn B L 2008 Occurrence edn). Oxford University Press Southern 30.NorvalRAI,PerryBD,Young A S 1992 The of Theileria parva infection in cattle on a farm Africa, Cape Town: 483–484 epidemiology of theileriosis in Africa. Academic in the Ladysmith district, KwaZulu-Natal, 22. Martin H, Brocklesby D W 1960 A new Press, London South Africa. Journal of the South African parasite of the eland. Veterinary Record 72: 31. Nuttall G F H 1910 On haematozoa occur- Veterinary Association 79: 31–35 331–332 ring in wild animals in Africa. 1. Piroplasma 40. Uilenberg G 1999 Immunization against 23. Matjila P T, Leisewitz A L, Jongejan F, rossi N. Sp. and Haemogregarinia canis adusti diseases caused by Theileria parva: a review. Bertschinger H J, Penzhorn B L 2008 n. Sp. found in the jackal. Parasitology 3: Tropical Medicine and International Health Molecular detection of Babesia rossi and 108–116 4(9): A12–A20 Hepatozoon sp. in African wild dogs (Lycaon 32.OuraCAL,Bishop R P, Wampande E M, 41. Van den Bossche P,Esterhuizen J, Nkuna R, pictus) in South Africa. Veterinary Parasitology Lubega G W, Tait A 2004 Application of a Matjila T, Penzhorn B, Geerts S, Marcotty T 157: 123–127 reverse line blot assay to the study of 2006 An update of the bovine trypano- 24. Matjila P T, Leisewitz A L, Jongejan F, haemoparasites in cattle in Uganda. Inter- somosis situation at the edge of the Penzhorn B L 2008 Molecular detection national Journal of Parasitology 34: 603–613 Hluhluwe-iMfolozi Park, KwaZulu-Natal of tick-borne protozoal and ehrlichial 33. Paling R W,Grootenhuis J G, Young A S 1981 Province, South Africa. Onderstepoort infections in domestic dogs in South Africa. Isolation of Theileria mutans from Kenyan Journal of Veterinary Research 73: 77–79

0038-2809 Jl S.Afr.vet.Ass. (2010) 81(1): 45–49 49