Molecular study of Anaplasma spp. in horses, sheep, and goats with phylogenetic analysis in northwest Iran

Anaplasmosis is a tick-borne disease with worldwide distribution, affecting ruminants, equines, carnivores, and humans. This study aimed to investigate Anaplasma phagocytophilum (A. phagocytophilum) in horses from Ardabil province and Anaplasma ovis (A. ovis) in small ruminants from East Azerbaijan province using the Nested PCR method. Blood samples were collected from the jugular vein of ۱۰۰ healthy horses in the Ardabil province and ۱۵۶ healthy sheep and goats (۱۱۶ sheep and ۴۰ goats) in the East Azerbaijan Province during the spring and summer seasons of ۲۰۱۶ in northwest Iran. The gathered blood samples were stored at -۲۰°C until the molecular experiments were conducted. Nested PCR was employed to detect A. phagocytophilum in horses and A. ovis in small ruminants using extracted DNA and amplifying ۱۶S rRNA and msp۴ genes. The Chi-square test of independence was used to determine the relationship between Anaplasma spp., infection, and independent variables, including age, gender, animal species, and sampling location. None of the ۱۰۰ samples collected from horses in the Ardabil province were positive for A. phagocytophilum. In the East Azerbaijan province, ۱۱ out of the ۱۵۶ (۷.۰۵%) blood samples collected from sheep and goats tested positive for A. ovis. In addition, A. ovis infection was not significantly related to the independent variables. Phylogenetic analysis showed that the sequence obtained in the present study (MH۷۹۰۲۷۳) has ۱۰۰% homology with the sequence obtained from sheep infected with Anaplasma in Ahvaz (JQ۶۲۱۹۰۳.۱). The study’s findings can aid in preventing and controlling anaplasmosis in farm animals in northwestern Iran.

Keywords:

Anaplasma phagocytophilum , Anaplasma ovis , Nested PCR , Sequencing