The present study was carried out to morphological investigation of ticks naturally feeding on cattle, and molecular detection of A. marginale in these ticks using the conventional polymerase chain reaction (PCR) assay. Totally 25 infested cattle with ticks were selected from rural areas in Wasit province / Iraq, during March-2020 to June-2020. The findings of clinical observation reported that ticks were distributed onto different bodily parts; udder and testis (50.3%), neck (15.98%), perineal region (28.4%), and ear (5.33%). Although hard (Ixodid) tick was the only type detected, two species of different genera were identified among infested cattle; Hyalomma anatolicum (75.15%), and Rhipicephalus sanguineus (24.85%). Regarding tick species, H. anatolicum was more prevalent (P<0.05) than R. sanguineus in neck (18.11%) region; whereas, R. sanguineus was prevalent significantly (P<0.05) in udder and testis (61.9%) compared to H. anatolicum. Regarding bodily regions, H. anatolicum and R. sanguineus were prevalent significantly (P<0.05) in udder and testis (75.15% and 61.9% respectively). Also, the range and mean number of ticks on each animal was 2-34 and 12.59 respectively. Based on the life stage of collected study ticks, 47.93%, 33.73%, and 18.34% were male, female, and nymph, respectively. Ticks of each study cattle were considered as a one sample and subjected collectively to DNA extraction. Hence, the overall findings for testing 25 samples of ticks by PCR revealed that 4 (16%) of samples were positives for msp1β gene specific to A. marginale. Additionally, male Hyalomma anatolicum ticks were the only positives for A. marginale isolates.

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