Low level of genetic diversity and high occurrence of vector-borne protozoa in water buffaloes in Thailand based on 18S ribosomal RNA and mitochondrial cytochrome b genes

Vector-borne pathogens (VBPs) pose a great risk to ruminant production through significant economic losses. Several previous studies in Thailand have mainly been focused on the health of dairy and beef cattle. Water buffaloes are one of the important ruminants in the country, but studies on their infection with VBPs remains limited. We conducted a molecular survey on blood samples from 456 buffaloes obtained from eight provinces across different geographical locations of Thailand. The PCR diagnostics indicated that 116 (25.4%) and 59 (12.9%) of these 456 samples were positive for piroplasm and Plasmodium spp., respectively, and were found in six and all eight regions, respectively, across Thailand. Co-infections of piroplasm and Plasmodium spp. were observed in 24 cases (5.26%). Babesia spp. was not detected in any of the 12 sequenced piroplasm-positive samples in the present study. Genetic comparisons and phylogenetic analyses of within and between parasite populations, based on the 18S ribosomal (r)RNA and cytochrome b (cytb) genes for T. orientalis and P. bubalis, respectively, revealed that T. orientalis shared a high similarity within its population and could be divided into four distinct haplotypes. Haplotypes 1 and 4 were placed in the same clade with the samples previously isolated from cattle in Korea, Japan, Australia, and the USA. Haplotypes 2, and 3 were novel and were placed in a separate clade not shared with the other isolates. We also confirmed our previous investigation that at least three cytb haplotypes of P. bubalis were distributed in the country with a relatively high degree of genetic polymorphisms within its population (based on cytb sequences). Type II P. bubalis was phylogenetically closely related to P. caprae in goats in Zambia and Thailand. This study improves our current understanding on the distribution, intra- and inter-population genetic diversity, and genetic relationship of piroplasms and Plasmodium spp. in water buffaloes.