Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 18 3 2026 08 23 Microscopic and molecular detection of Theileria annulata in cattle from Babylon province of Iraq Microscopic and molecular detection of Theileria annulata in cattle from Babylon province of Iraq 395 410 5444 10.22103/jab.2026.27095.1884 FA Ghefran Shiaa Mazhar Department of Parasitology, College Veterinary Medicine, Al-Qasim Green University, Babylon 51013, Iraq. 0009-0004-1442-0599 Safaa M Kareem Department of Parasitology, College Veterinary Medicine, Al-Qasim Green University, Babylon 51013, Iraq. 0000-0003-3885-5891 Journal Article 2026 05 03 ObjectiveThis study aimed to investigate the detection of theileriosis caused by Theileria annulata in cattle applying microscopic and molecular methods. The other goal of the study was to evaluate the effects of geographical region, age, and sex on infection rates. Materials and methodsIn this study, 300 bovine blood samples were collected from different areas of Babil province, Iraq. The study was conducted between September 2025 and March 2026. Giemsa-stained blood smears were prepared for 200 samples for microscopic examination. 100 samples were analyzed by PCR to target the mitochondrial cytochrome b gene of T. annulata. Chi-square test was used in SPSS version 27 for statistical analysis. ResultsThe results showed that 66 (33%) of 200 bovine blood samples examined microscopically were positive for Theileria annulata. In the PCR method, 54 (54%) of 100 samples that studied the mitochondrial cytochrome b gene were positive. This could indicate a higher sensitivity of PCR than the microscopic method. The prevalence of infection was 53.3% in women and 55% in men, which was not significant. Also, the age of the individuals did not report a significant difference in the infection rate (χ² = 0.34, p = 0.84). However, the geographical regions showed a significant difference. So that the highest infection rate was related to Al-Shumali (73.9%) and the lowest rate was related to Al-Qasim (35%) (χ² = 8.922, p < 0.01). These results indicate that there is a significant relationship between the region and the infection rate. Phylogenetic analysis of ten T. annulata isolates (PX849363–PX849372) confirmed their genetic relationship with reference strains available in GenBank. ConclusionPCR-based molecular detection demonstrated higher sensitivity than microscopy for identifying T. annulata infection in subclinical infections that may contribute to parasite transmission within herds. These findings highlight the need for integrated control strategies, including regular tick management, molecular surveillance, and improved animal husbandry, to decrease the impact of bovine theileriosis and enhance cattle health and productivity in endemic areas. ObjectiveThis study aimed to investigate the detection of theileriosis caused by Theileria annulata in cattle applying microscopic and molecular methods. The other goal of the study was to evaluate the effects of geographical region, age, and sex on infection rates. Materials and methodsIn this study, 300 bovine blood samples were collected from different areas of Babil province, Iraq. The study was conducted between September 2025 and March 2026. Giemsa-stained blood smears were prepared for 200 samples for microscopic examination. 100 samples were analyzed by PCR to target the mitochondrial cytochrome b gene of T. annulata. Chi-square test was used in SPSS version 27 for statistical analysis. ResultsThe results showed that 66 (33%) of 200 bovine blood samples examined microscopically were positive for Theileria annulata. In the PCR method, 54 (54%) of 100 samples that studied the mitochondrial cytochrome b gene were positive. This could indicate a higher sensitivity of PCR than the microscopic method. The prevalence of infection was 53.3% in women and 55% in men, which was not significant. Also, the age of the individuals did not report a significant difference in the infection rate (χ² = 0.34, p = 0.84). However, the geographical regions showed a significant difference. So that the highest infection rate was related to Al-Shumali (73.9%) and the lowest rate was related to Al-Qasim (35%) (χ² = 8.922, p < 0.01). These results indicate that there is a significant relationship between the region and the infection rate. Phylogenetic analysis of ten T. annulata isolates (PX849363–PX849372) confirmed their genetic relationship with reference strains available in GenBank. ConclusionPCR-based molecular detection demonstrated higher sensitivity than microscopy for identifying T. annulata infection in subclinical infections that may contribute to parasite transmission within herds. These findings highlight the need for integrated control strategies, including regular tick management, molecular surveillance, and improved animal husbandry, to decrease the impact of bovine theileriosis and enhance cattle health and productivity in endemic areas. https://jab.uk.ac.ir/article_5444_587b7b833034299fdd5f4b10e7dc9fca.pdf