Use of multiplexed tandem PCR to estimate the prevalence and intensity of Theileria orientalis infections in cattle.

This study employed a semi-quantitative, multiplexed tandem PCR (MT-PCR) to assess the prevalence and infection intensity of four genotypes (buffeli, chitose, ikeda and type 5) of Theileria orientalis in cattle in Australia. Genomic DNA samples from blood samples (n=448) collected from 27 to 32 dairy cows from each of 15 dairy herds with a history of recent theileriosis outbreaks (Group 1), and from blood samples available from 24 cows with or without oriental theileriosis (Group 2) were tested using MT-PCR. Results revealed that all four genotypes were present in Group 1 cattle; genotype buffeli had the highest prevalence (80.5%), followed by genotypes ikeda (71.4%), chitose (38.6%) and type 5 (20.3%). Genotype ikeda had the highest average infection intensity in the cattle (relating to 55,277 DNA copies), followed by buffeli, chitose and type 5 (6354-51,648 copies). For Group 2, results indicated that genotype ikeda had a significantly higher average intensity of infection than buffeli in symptomatic cattle (P<0.001), and symptomatic cattle had a higher intensity of ikeda than asymptomatic cattle (P=0.004). Future studies should assess the utility of the present MT-PCR assay as a diagnostic and epidemiological tool in other parts of Australasia and the world.

Description of Cloacina atthis sp. nov. from the stomach of the euro (Macropus robustus) (Marsupialia: Macropodidae) from Western Australia based on morphological and molecular criteria.

A new species of strongyloid nematode from the genus Cloacina (Chabertiidae: Cloacininae) is described from the stomach of the hill kangaroo or euro (Macropus robustus) (Marsupialia: Macropodidae) from Western Australia. Cloacina atthis sp. nov. was found only in euros from the Pilbara region in the northwest of Western Australia, in spite of extensive collecting of the same host species from around the Australian continent. C. atthis is most closely related to Cloacina clymene, a species found in the same host species but only in the eastern half of the continent; the two species differ in minor morphological features (the shape of the wall of the buccal capsule, spicule lengths, the degree of sclerotisation of the gubernaculum and the shape of the vagina) as well as in differences in the internal transcribed spacers of ribosomal DNA. This study highlights the importance of using molecular methods when investigating the apparently disjunct distributions of strongyloid nematodes.