Four p67 alleles identified in South African Theileria parva field samples

Previous studies characterizing the Theileria parva p67 gene in East Africa revealed two alleles. Cattle-derived isolates associated with East Coast fever (ECF) have a 129 bp deletion in the central region of the p67 gene (allele 1), compared to buffalo-derived isolates with no deletion (allele 2). In South Africa, Corridor disease outbreaks occur if there is contact between infected buffalo and susceptible cattle in the presence of vector ticks. Although ECF was introduced into South Africa in the early 20th century, it has been eradicated and it is thought that there has been no cattle to cattle transmission of T. parva since. The variable region of the p67 gene was amplified and the gene sequences analyzed to characterize South African T. parva parasites that occur in buffalo, in cattle from farms where Corridor disease outbreaks were diagnosed and in experimentally infected cattle. Four p67 alleles were identified, including alleles 1 and 2 previously detected in East African cattle and buffalo, respectively, as well as two novel alleles, one with a different 174 bp deletion (allele 3), the other with a similar sequence to allele 3 but with no deletion (allele 4). Sequence variants of allele 1 were obtained from field samples originating from both cattle and buffalo. Allele 1 was also obtained from a bovine that tested T. parva positive from a farm near Ladysmith in the KwaZulu-Natal Province. East Coast fever was not diagnosed on this farm, but the p67 sequence was identical to that of T. parva Muguga, an isolate that causes ECF in Kenya. Variants of allele 2 were obtained from all T. parva samples from both buffalo and cattle, except Lad 10 and Zam 5. Phylogenetic analysis revealed that alleles 3 and 4 are monophyletic and diverged early from the other alleles. These novel alleles were not identified from South African field samples collected from cattle; however allele 3, with a p67 sequence identical to those obtained in South African field samples from buffalo, was obtained from a Zambian field isolate of a naturally infected bovine diagnosed with ECF. The p67 genetic profiles appear to be more complex than previously thought and cannot be used to distinguish between cattle- and buffalo-derived T. parva isolates in South Africa. The significance of the different p67 alleles, particularly the novel variants, in the epidemiology of theileriosis in South Africa still needs to be determined.     

Prevalence and risk factors associated with <Emphasis Type="Italic">Theileria parva</Emphasis> infection in cattle in three regions of Tanzania

Ticks and tickborne diseases (TBDs) are serious constraints to cattle production in Tanzania and other tropical and subtropical countries. Among the TBDs, East Coast fever (ECF) is the most important as it causes significant economic losses to the cattle industry in Tanzania. However, control of ECF in Tanzania has continued to be a challenge due to inadequate epidemiological information. The main objective of this study was to determine the epidemiological situation of Theileria parva infections in cattle kept under pastoral and agro-pastoral farming systems in Mara, Singida, and Mbeya regions of Tanzania. Blood samples were collected from 648 cattle in the three regions. Genomic DNA was extracted and amplified in a polymerase chain reaction (PCR) using T. parva-specific primers targeting the 104-kD antigen (P104) gene. In addition, information was collected on the possible risk factors of T. parva infection (animal age, region, animal sex, tick burden, tick control method, and frequency of acaricide application). The prevalence of T. parva across the three regions was 14.2%. There was variation in prevalence among the three regions with Mara (21.8%) having a significantly higher (p = 0.001) prevalence than the other regions. Moreover, Mbeya exhibited relatively lower prevalence (7.4%) compared to the other regions. Factors found to be significantly associated with an animal being PCR positive for T. parva were region (p = 0.001) and tick burden (p = 0.003). Other factors were not found to be significant predictors of being PCR positive for T. parva. The present study showed high variation in tick burden and T. parva prevalence across the regions. Therefore, different strategic planning and cost-effective control measures for ticks and T. parva infection should be implemented region by region in order to reduce losses caused by ticks and ECF in the study area.     

Genotypic diversity,a survival strategy for the apicomplexan parasite Theileria parva

The tick-borne protozoan parasite Theileria parva causes East Coast fever (ECF), a severe lymphoproliferative disease of cattle that is a major constraint to the improvement of livestock in eastern, central and southern Africa. Studies in cattle experimentally infected with T. parva have shown that the protective cytotoxic T lymphocyte (CTL) response is tightly focused, with individual animals recognizing only one or two dominant antigens, the identity of which varies with MHC class I phenotype. It is well known that cross-protection between T. parva stocks is limited, but precise evaluation of genetic diversity in field populations of the parasite has been hampered by a lack of molecular markers spanning the genome. A recently described panel of satellite markers has provided evidence for substantial genotypic diversity and recombination but does not provide cover for large segments of the genome. To address this deficiency, we undertook to identify additional polymorphic markers covering these regions and we report herein 42 newly identified PCR-RFLP markers distributed across the 4 T. parva chromosomes, as well as 19 new satellite markers for chromosomes 1 and 2. This brings the total number of available polymorphic markers to 141 for the 8.5 Mb genome. We have used these markers to characterise existing parasite stabilates and have also shown that passage of the parasite through naïve cattle and ticks can lead to substantial changes of parasite populations in resulting stabilates. These markers have also been used to show that passage of mixed parasites through an immunised calf results in the removal of the immunising genotype from the parasite population produced by ticks fed on this animal.     

Survey of ixodid ticks in domestic ruminants in Bedelle district,Southwestern Ethiopia

A survey aimed at determining the status of ticks in cattle, sheep and goats in Bedelle district (Southwestern Ethiopia) was carried out from November 2007 to April 2008. Out of the total 330 animals examined, 314 (95.2%) were found to be infested (harbouring of at least a single tick). High rates of infestations were recorded across all three host species. Factors like month of collection, age and sex of host species did not show significant association with infestation rates. A total of 5,507 ticks belonging to three genera (i.e. Amblyomma, Boophilus and Rhipicephalus) were collected during the study period of which six species were identified. The species of ticks encountered comprise of Amblyomma cohaerens (44.1%), Amblyomma variegatum (13.8%), Amblyomma lepidum (1.2%), Boophilus decoloratus (24.9%), Rhipicephalus evertsi evertsi (14.3%) and Rhipicephalus lunulatus (1.7%). As about 86.7% (4,772) of the ticks were collected from cattle, it was recognised that this could be an evidence of host preference where cattle are serving as principal host while sheep and goats serve as alternative hosts. The role of ecological factors and little attention paid by livestock owners for treatment of animals against ticks are suggested to result in the abundance of ticks. Acaricide spraying should be strategically applied to control ticks. Better result could be achieved if an emphasis is being put on spraying cattle. Studies on tick-borne diseases, involvement of wildlife species as well as related factors are recommended as they may provide a valuable basis for designing and launching an all-round control programme in the country.     

Progress in research on tick‐borne diseases: Theileriosis and heartwater

Summary

The rapid population growth in subsaharan Africa necessitates a great increase in animal production in the more humid zones. Vector‐borne diseases occurring in these zones will assume more importance, but are difficult to control. They include theileriosis and heartwater. Recent developments in research on these diseases are presented. Indigenous animal populations in endemic areas, subjected to natural selection, are far less susceptible than exotic stock.

Heartwater, caused by the rickettsia Cowdria ruminantium, transmitted by Amblyomma ticks, causes high mortality in exotic ruminants. It has received much attention in recent years, partly because the disease has been introduced from Africa into the Caribbean and threatens the American mainland. Since the recent success of in vitro culture, much progress in research has been made, but so far prevention still relies mainly on acaricidal tick control; an infection and treatment method is used on a limited scale. Antigenic diversity is a complication for immunization procedures.

Theileria parva (East Coast fever, Corridor disease and January disease) and T.annulata (Mediterranean or tropical theileriosis) are the most pathogenic of the 6 species of this protozoan genus that infect cattle. Great progress has been made in recent years in knowledge on the immunology, the epidemiology, the taxonomy and the chemotherapy of theileriosis. Intensive acaricidal tick control can now be supplemented by an attenuated schizont vaccine against T.annulata, while immunization against East Coast fever is carried out on a limited scale using virulent sporozoite infection and treatment. Research on recombinant vaccines is promising. Antigenic diversity in T.parva is a serious complication.     

East Coast fever and multiple El Niño Southern oscillation ranks

East Coast fever (ECF), a tick-borne disease of cattle, is a major constraint to livestock development in Africa in general and southern Zambia in particular. Understanding the transmission patterns of this disease complex is very difficult as shown by previous studies in southern and eastern Zambia due to the interplay of risk factors. In this long-term study, we investigated whether global weather changes had any influence on disease transmission in traditionally kept cattle in southern Zambia. The results from this study show a strong association between increased Theileriaparva contacts in cattle and the presence of El Niño, clearly linking a simple climatic index to disease outbreaks. We therefore propose that in southern Zambia, the simple and readily available multiple El Niño Southern oscillation index (MEI) ranks be used in planning ECF control programmes and early warning.     

East coast fever: 4. Further studies on the protection of cattle immunized with a combination of theilerial strains

Four experiments were carried out in which cattle immunized with three cryopreserved stabilates, Theileria parva (Muguga), T. parva (Kiambu 5) and T. lawrencei (Serengeti transformed) and simultaneously treated with a single dose of long acting tetracycline were challenged with field isolates of T. parva. Three of the challenge isolates were from Tanzania and one from Kenya. The immunized cattle showed either mild or inapparent diease reations while susceptible control cattle suffered severe disease and 72% died. These experiments further demonstrated the efficacy of the immunization method against T. parva isolates from a wide distribution within East Africa. The value of such laboratory screening of field isolates prior to field exposures is discussed.     

East coast fever: 3. Chemoprophylactic immunization of cattle using oxytetracycline and a combination of theilerial strains

In the first of two experiments performed, 29 animals were immunized by chemoprophylaxis against either a combination of three theilerial strains, Theileria parva (Muguga), T. parva (Kiambu 5) and T. lawrencei (Serengeti transformed), or against one or two elements of the combination separately, and then challenged with either T. parva (Kiambu 1) or T. lawrencei (Solio KB1), both of which were known to be heterologous to T. parva (Muguga). The animals immunized against the combination had mild or inapparent reactions to challenge, whilst the susceptible control cattle and many of those immunized against one or two strains either had severe reactions of died.In the second experiment, 17 cattle immunized by chemoprophylaxis against the combination were challenged with lethal doses of three theilerial strains, T. parva (Entebbe 1), T. parva (Entebbe 2) and T. parva (Ukunda), recently isolated from two widely separated areas of East Africa. They had mild or inapparent reactions whilst 13 of 15 susceptible control cattle died.Chemoprophylaxis using a combination of theilerial strains and a limited oxytetracycline regimen was shown to be an effective means of immunizing cattle against East African theilerioses. The use of this technique as a method of immunizing cattle before exposure in the field is proposed.     

Tick communities at the expanding wildlife/cattle interface in the Eastern Cape Province, South Africa: implications for Corridor disease

Corridor disease, transmitted by the brown ear tick (Rhipicephalus appendiculatus), is one of Africa's most pathogenic tick-borne diseases for cattle. With a focus on this species, we investigated the community parameters (richness, diversity and abundance) of ticks in the Eastern Cape, South Africa, and how this may be linked to the increasing wildlife/cattle interface in the region. There were significantly more ticks of a greater diversity and richness at sites positioned at the wildlife/cattle interface ('treatment sites') compared to sites where wildlife was absent (controls). Significantly, R. appendiculatus was only found at the treatment sites. Therefore, it is believed that the wildlife/cattle interface may be playing a crucial role in increasing the occurrence, abundance and distribution of R. appendiculatus in the Eastern Cape. The implications of a Corridor disease outbreak in the region are discussed.     

Re-infection of wildlife populations with rinderpest virus on the periphery of the Somali ecosystem in East Africa

We report surveillance for rinderpest virus in wildlife populations in three major ecosystems of East Africa: Great Rift Valley, Somali and Tsavo from 1994 to 2003. Three hundred and eighty wild animals were sampled for detection of rinderpest virus, antigen or genome and 1133 sampled for antibody in sera from Kenya, Uganda, Ethiopia and Tanzania from 20 species. This was done modifying for wildlife the internationally recommended standards for rinderpest investigation and diagnosis in livestock. The animals were selected according to susceptibility and preference given to gregarious species, and populations were selected according to abundance, availability and association with livestock. Rinderpest virus, antigen and/or genome were detected in Kenya; within Tsavo, Nairobi and Meru National Parks. Serological results from 864 animals (of which 65% were buffalo) from the region were selected as unequivocal; showing the temporal and spatial aspects of past epidemics. Recent infection has been only in or peripheral to the Somali ecosystem (in Kenya). Our evidence supports the hypothesis that wildlife is not important in the long-term maintenance of rinderpest and that wildlife are infected sporadically most likely from a cattle source, although this needs to be proven in the Somali ecosystem. Wildlife will continue to be a key to monitoring the remaining virus circulation in Africa.     

Effects of tick infestation on Boran (Bos indicus) cattle immunised against theileriosis in an endemic area of Kenya

Boran (Bos indicus) heifers were immunised by infection with local Theileria parva parva, T p lawrencei and T mutans stocks and treated with parvaquone and later exposed to natural tick and tick-borne disease challenge in the Trans-Mara Division of Kenya. The Theileria species parasites in the challenge were maintained in African buffalo and cattle and the tick vectors were supported by several species of wild Bovidae and domestic livestock present in the area. Thirty immune cattle were observed for 30 weeks while grazing on a ranch in the Trans-Mara Division. Of these, 15 were immersed in toxaphene at weekly or twice-weekly intervals while 15 cattle remained without acaricide application. Cattle which became pregnant were withdrawn from the experiment. There was no evidence of any clinical tick-borne disease in either group of cattle during the experiment. Five species of ixodid ticks infested the cattle during the experiment and cattle which were not treated with acaricide had far more ticks. Rhipicephalus appendiculatus was the most abundant tick species, with a mean infestation of 46 adults per animal in the undipped cattle. This tick also appeared to be the cause of the observed reduction in weight gains. Major haematological parameters did not differ significantly between the groups. Behavioural studies showed that the undipped cattle spent less time grazing and ruminating. This study has shown that, at the expense of some loss in productivity, zebu cattle, immunised against ticks and theileriosis, can be kept despite tick infestation.     

Sequence variation identified in the 18S rRNA gene of Theileria mutans and Theileria velifera from the African buffalo (Syncerus caffer)

The African buffalo (Syncerus caffer) is a natural reservoir host for both pathogenic and non-pathogenic Theileria species. These often occur naturally as mixed infections in buffalo. Although the benign and mildly pathogenic forms do not have any significant economic importance, their presence could complicate the interpretation of diagnostic test results aimed at the specific diagnosis of the pathogenic Theileria parva in cattle and buffalo in South Africa. The 18S rRNA gene has been used as the target in a quantitative real-time PCR (qPCR) assay for the detection of T. parva infections. However, the extent of sequence variation within this gene in the non-pathogenic Theileria spp. of the Africa buffalo is not well known. The aim of this study was, therefore, to characterise the full-length 18S rRNA genes of Theileria mutans, Theileria sp. (strain MSD) and T. velifera and to determine the possible influence of any sequence variation on the specific detection of T. parva using the 18S rRNA qPCR. The reverse line blot (RLB) hybridization assay was used to select samples which either tested positive for several different Theileria spp., or which hybridised only with the Babesia/Theileria genus-specific probe and not with any of the Babesia or Theileria species-specific probes. The full-length 18S rRNA genes from 14 samples, originating from 13 buffalo and one bovine from different localities in South Africa, were amplified, cloned and the resulting recombinants sequenced. Variations in the 18S rRNA gene sequences were identified in T. mutans, Theileria sp. (strain MSD) and T. velifera, with the greatest diversity observed amongst the T. mutans variants. This variation possibly explained why the RLB hybridization assay failed to detect T. mutans and T. velifera in some of the analysed samples.     

Transformation of Theileria parva derived from African buffalo (Syncerus caffer) by tick passage in cattle and its use in infection and treatment immunization.

A sporozoite stabilate (St. 199) of Theileria parva was obtained by feeding nymphal Rhipicephalus appendiculatus on an African buffalo (Syncerus caffer) and was used to immunize cattle by the infection and treatment method. Nymphal ticks were applied to one of the steers 90 days later and it was shown that the resultant adult tick had become infected. Using tick/cattle passage, two passage lines of T. parva were established. By the fifth tick/cattle passage, the parasite stocks had changed their behaviour to that of T. parva derived from cattle as the parasite produced relatively high schizont parasitosis and piroplasm parasitaemia in cattle, and had become highly infective to ticks. At various passage levels the parasite populations were characterized by behaviour and by monoclonal antibodies against T. parva schizonts using infected cell culture isolates from cattle during acute infections. The monoclonal antibody profile showed little evidence of antigen change of the parasite during passage through cattle, which was confirmed in a two-way cross-immunity experiment using sporozoite stabilate derived from ticks obtained from the buffalo and fourth passage in cattle. The implication of these results, particularly in relationship to immunization of cattle against T. parva derived from buffalo, is discussed.     

Contacts between domestic livestock and wildlife at the Kruger National Park Interface of the Republic of South Africa

One of the most important transboundary animal diseases (TADs) in the southern African region is foot-and-mouth disease (FMD). In this region, a pathway for spread of FMD virus is contacts between cattle and certain species of wildlife. The objective of this study was to evaluate contacts between cattle and wildlife in the Kruger National Park (KNP) and the adjacent Limpopo province for the time periods October 2006 to March 2007 and April to September 2007. In this study, 87 livestock owners and 57 KNP field rangers were interviewed. Fifteen (17%) livestock owners reported contacts between wildlife and cattle. More livestock owners reported observing contacts between cattle and all wildlife species during October-March than April-September (p = 0.012). However, no difference was found between these periods for contacts between cattle and individual wildlife species. A total of 18 (32%) field rangers reported contacts between cattle and wildlife. The most common species-specific contacts were between cattle and buffalo (63/year), cattle and impala (17/year) and cattle and lion (10/year). There were no significant differences in rangers reporting observed contacts between cattle and wildlife during October-March versus April-September or between rangers reporting observed contacts outside versus within the KNP. Overall, there was no evidence of higher contact rates between cattle and wildlife in the study area during October-March compared to April-September. Contact data collected in this study can be used to better understand the transmission of FMD virus in this region.     

East coast fever: 1. Chemoprophylactic immunization of cattle against Theileria parva (Muguga) and five theilerial strains

In one experiment, groups of cattle were immunized by chemophylaxis or therapy against Theileria parva (Muguga), and were challenged with five recently-isolated strains of Theileria. Two of the strains caused mild or inapparent reactions in the immunized animals, and three caused significant reactions. None of 25 immunized animals died on challenge, whereas 22 of 25 susceptible (non-immunized) animals died.In a second experiment, groups of cattle were immunized against five field strains using chemoprophylaxis. They had minimal reactions and were solidly immune both to isologous challenge and to subsequent challenge with T. parva (Muguga). This demonstrated that cattle could be effectively immunized against lethal challenge with several theilerial strains by inoculating them with 1 ml of an infective stabilate of the appropriate strain and the concomitant daily inoculation of 5 mg/kg of a formulation of oxytetracycline on 4 consecutive days starting on the day of infection.It is suggested that T. parva (Muguga) may be a valuable component of a polyvalent East Coast fever (ECF) vaccine in East Africa.     

Acaricide resistance profiles of single- and multi-host ticks from communal and commercial farming areas in the Eastern Cape and North-West Provinces of South Africa

A field study (February 2000 to August 2001) was conducted on communal and commercial farms in the Eastern Cape and North-West Provinces of South Africa to detect the levels of tick resistance to commonly used acaricides. The larvae obtained from engorged females of the one-host tick Boophilus decoloratus, the two-host tick Rhipicepalus evertsi evertsi and the three-host ticks Amblyomma hebraeum and Rhipicephalus appendiculatus were tested against various concentrations of amitraz, chlorfenvinphos and cypermethrin using the Shaw Larval Immersion Test method. Ticks from the communal farms showed higher levels of resistance to cypermethrin and some resistance to chlorfenvinphos whilst no resistance was detected against amitraz. However, ticks from commercial farms were equally resistant to amitraz, chlorfenvinphos and cypermethrin. The B. decoloratus populations tested were considerably more resistant to all the acaricides tested than the R. evertsi evertsi, A. hebraeum and R. appendiculatus populations. This supports the hypothesis that single-host ticks develop resistance faster than multi-host ticks. This trend was recorded on most of the farms where single- and multi-host ticks co-existed. It was concluded that the use of acaricides at high frequencies and high concentrations was one of the main causes of tick resistance in the study areas. Possible factors which caused the resistance problems are discussed and acaricide management strategies recommended.     

Tick infestations on livestock in the yemen arab republic and their potential as vectors of livestock diseases

A survey of ticks on cattle, camels, sheep, goats and donkeys in four different geographical locations of the Yemen Arab Republic (YAR) was carried out to provide more information on the possible risk of tick-borne diseases to imported exotic cattle included in the YAR's plans for livestock development. The most abundant ticks were Hyalomma spp. particularly on camels. Ticks found on cattle included Hyalomma spp., Amblyomma variegatum, Boophilus annulatus and Rhipicephalus spp. In general with the exception of camels tick burdens on all species of domestic livestock were very low. Two hundred and ninety eight serum samples from miscellaneous adult cattle throughout the country were negative to a test for Anaplasma marginale antibodies. It is speculated that tick burdens in the YAR are too low for significant disease transmission and the implications of the findings are discussed.     

Cyhalothrin—a novel acaricidal and insecticidal synthetic pyrethroid for the control of the cattle tick (Boophilus microplus) and the buffalo fly (Haematobia irritans exigua)

SUMMARY Cyhalothrin, a novel synthetic pyrethroid, was evaluated for control of the major resistant strains of the cattle tick (Boophilus microplus) and for control of the buffalo fly (Haematobia irritans exigua) on cattle. In regulated treatment trials with 0.007% cyhalothrin, greater than 99% control of the Biarra, Mackay, Mt Alford, DDT resistant and Ulam cattle tick strains was obtained. Protective-period trials were conducted in which animals which had been sprayed with 0.007% cyhalothrin then received a continuing heavy challenge of the organophosphate-resistant Biarra tick strain. The first semi-engorged adult ticks appeared no earlier than 27 days after treatment, which corresponds to a minimum protective period against reinfestation of 7 days. Protective-periods ranging from 7 to 15 days were obtained in trials that were conducted.     

New data regarding distribution of cattle ticks in the south-western Indian Ocean islands

Recent studies have produced new insight into the origin and distribution of some cattle ticks in the south-western Indian Ocean islands. Rhipicephalus appendiculatus, introduced from Tanzania in 2002, is now well established on Grande Comore but has not yet reached the other islands of the archipelago (Mohéli, Anjouan and Mayotte). Only one of the two clades identified in Africa has settled so far. Amblyomma variegatum, which was not supposed to be able to persist in the Antananarivo region (1300 m) nor in other Malagasy regions of high altitude without regular introductions of ticks by infested cattle, is now endemic as a general rule up to 1600 m although other regions of lower altitude (1400 m) are still free of the tick. This species remains confined in a small area of the west coast on La Reunion Island. On the contrary, Hyalomma dromedarii could not settle on Madagascar where it was introduced in 2008 and Rhipicephalus evertsi evertsi is not yet present in Grande Comore despite regular introductions by infested cattle from Tanzania. A phylogeographic approach has been carried out at an intra-specific level for A. variegatum. This study has led to the identification of two main lineages, one covering all species distribution and one restricted to East Africa and the Indian Ocean area. These two lineages are in sympatry in Madagascar where a high genetic diversity has been described, whereas a lower genetic diversity is observed on other islands. These results seem to agree with the historical data concerning the introduction of the tick in the Indian Ocean area.     

Options for the control of tsetse-transmitted livestock trypanosomosis. An epidemiological perspective

Tsetse-transmitted livestock trypanosomosis affects livestock in large parts of sub-Saharan Africa. In southern Africa two epidemiological situations can be distinguished. The disease can have an endemic nature with high morbidity and low mortality in the livestock population. Endemic livestock trypanosomosis is found mainly in areas where cattle constitute the main host of tsetse and reservoirs of trypanosomes. Epidemic trypanosomosis, with high morbidity and high mortality is found in areas where wildlife persist as main reservoir and where livestock come into contact with tsetse flies transmitting trypanosomes from the sylvatic reservoir. Based on the differences in impact of the disease on livestock health in these two epidemiological settings, the appropriateness of the available trypanosomosis control tools differs. In trypanosomosis endemic areas, trypanocidal drug use could be the most suitable approach. Possible problems associated with the development of resistance in trypanosomes to the drugs need to be investigated further. In epidemic situations, vector control seems the most appropriate long-term solution.