Development of a deep amplicon sequencing method to determine the proportional species composition of piroplasm haemoprotozoa as an aid in their control (27th Conference of WAAVP Madison USA)

Umer Naveed Chaudhry (Speaker)

Activity: Academic talk or presentation typesOral presentation

Description

Piroplasmosis is caused by tick-borne haemoprotozoa of the genera Theileria and Babesia, that impose a serious impact on animal production and human health. While multiple haemoprotozoan species can infect a single host, there is a lack of reliable molecular diagnostic tools with which to understand the composition of these complex parasite communities. Haemoprotozoa vary in their epidemiology, drug sensitivity, pathogenicity and interaction of co-infected species in the host, while common features include that animals become the persistent carriers of these pathogens after primary recovery from infection and can play a significant roles as a reservoir hosts. In the present study, we developed for the first time, the use of deep amplicon sequencing using an Illumina Mi-seq platform to identify haemoprotozoan communities and to establish the concept of a “protobiome”. Haemoprotozoa of ruminants were used to develop the concept, because mixed species infections are common. First, four phenotypically verified species of Theileria and Babesia were used to prepare mock pools with random number of parasites, and amplified with four different numbers of PCR cycles to assess the species representation bias. Second, we evaluated the threshold of the deep amplicon sequencing assay for each of the four species present at different levels of parasitemia and to confirm the accurate relative quantification of all four species. Finally, we applied the assay to the field samples to afford insight of the species composition of haemoprotozoan communities in small and large ruminants in the Punjab province of Pakistan. The “protobiome” concept has a wide range of potential applications in veterinary and human research including responses to drug treatment, parasite epidemiology/ecology, parasite interactions during mixed infections and parasite control strategies.
Period2019
Event titleWAAVP USA
Event typeConference