Mechanical transfer of Theileria orientalis: possible roles of biting arthropods, colostrum and husbandry practices in disease transmission.

BACKGROUND: The intracellular protozoal parasite Theileria orientalis has rapidly spread across South-eastern Australia, substantially impacting local cattle industries since 2006. Haemaphysalis longicornis appears to be a biological vector in the endemic regions. Mechanical transfer of blood by biting arthropods, in colostrum or iatrogenic transmission though husbandry procedures is another possible mode of transmission. This study assesses the risk of these mechanical modes of transmission. METHODS: Blood was collected from a T. orientalis Ikeda positive Angus steer, and was inoculated into the jugular vein of 9 calves in 3 treatment groups, each with 3 animals. Calves in Group 1 received 10 ml of cryopreserved blood, while those in Groups 2 and 3 received 1 ml (fresh blood) and 0.1 ml (cryopreserved), respectively. An additional three animals remained as negative controls and the donor calf was also followed as a positive control. Blood was collected over 3 months, and analysed via qPCR for the presence of the parasite. Samples of the sucking louse Linognathus vituli were collected opportunistically from calves 5 months after inoculation and tested for T. orientalis. For the colostral transmission study, 30 samples of blood and colostrum were collected from cows at calving in an endemic herd. These samples along with blood from their calves were tested by qPCR for T. orientalis and for antibodies to the major piroplasm surface protein (MPSP). RESULTS: Eight of the nine inoculated calves became positive for T. orientalis. The prepatent period of these infections was inversely correlated with inoculation dose. All negative control calves remained negative and the positive control calf remained positive. Samples of L. vituli tested positive for T. orientalis Ikeda, while some samples of colostrum were also shown to be qPCR and anti-MPSP positive. All calves in the colostral study tested qPCR negative although one was antibody-positive. CONCLUSIONS: T. orientalis is capable of being mechanically transferred by intravenous inoculation with small volumes of blood and is detectable up to 5 months post-infection. Animals infected by this means may play a significant role in the transmission of the disease by acting as asymptomatic carriers. Other modes of blood transfer, including biting arthropods and colostral transfer are also possible modes of disease transmission.

Detection of Theileria orientalis genotypes in Haemaphysalis longicornis ticks from southern Australia.

BACKGROUND: Theileria are blood-borne intracellular protozoal parasites belonging to the phylum Apicomplexa. Previously considered a benign parasite in Australia, outbreaks of clinical disease resulting from Theileria orientalis genotypes have been reported in Australia since 2006. Since this time, outbreaks have become widespread in south-eastern Australia, resulting in significant adverse impacts on local dairy and beef industries. This paper provides the first investigation into the possible biological and mechanical vectors involved in the rapid spread of the parasite. METHODS: To identify possible vectors for disease, ticks, biting flies and mosquitoes were collected within active outbreak regions of Gippsland, Victoria. Ticks were collected from cattle and wildlife, and mosquitoes and biting flies were collected in traps in close proximity to outbreak herds. Ticks were identified via DNA barcoding of the mitochondrial cytochrome oxidase I gene. Barcoded ticks were pooled according to species or phylogenetic group and tested for the presence of T. orientalis and the genotypes Ikeda, Chitose and Buffeli using real-time PCR. RESULTS: DNA barcoding and phylogenetic analysis identified ticks from the following species: Haemaphysalis longicornis, Ixodes holocyclus, Ixodes cornuatus, Ixodes hirsti, and Bothriocroton concolor. Additional Haemaphysalis, Ixodes and Bothriocroton spp. were also identified. Of the ticks investigated, only H. longicornis ticks from cattle carried theilerial DNA, with the genotypes Ikeda, Chitose and Buffeli represented. Mosquitoes collected in close proximity to outbreak herds included; Aedes camptorhynchus, Aedes notoscriptus, Coquillettidia linealis, Culex australicus, and Culex molestus. Low levels of T. orientalis Buffeli genotype were detected in some mosquitoes. The haematophagous flies tested negative. CONCLUSIONS: This is the first demonstration of a potential vector for T. orientalis in the current Australasian disease outbreak.