Microscopic and molecular detection of Theileria annulata in cattle from Babylon province of Iraq

Document Type : Research Paper

Authors

Department of Parasitology, College Veterinary Medicine, Al-Qasim Green University, Babylon 51013, Iraq.

10.22103/jab.2026.27095.1884

Abstract

Objective
This 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 methods
In 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.
Results
The 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.
Conclusion
PCR-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.

Keywords

References

Aboktifa, M. A., Al-ameedi, A. I., Ayad, Z. M., & Mosa, A. H. (2025). Hepatoprotective properties of piperine in experimentally hepatotoxic male rats exposed to carbon tetrachloride (CCl₄). Journal of Animal Health and Production, 13(1), 124–128. https://doi.org/10.17582/journal.jahp/2025/13.1.124.128
Ahmed, J., Yin, H., Schnittger, L., & Jongejan, F. (2002). Ticks and tick-borne diseases in Asia with special emphasis on China. Parasitology Research, 88(Suppl 1), S51–S55. https://doi.org/10.1007/s00436-001-0574-3
Ahsani, M. R., Mohammadabadi, M. R., Buchkovska, V., Ievstafiieva, Y., Kucher, D. M., Kochuk-Yashchenko, O. A., Babenko, O. I., Stavetska, R. V., Oleshko, V. P., & Kalashnyk, O. (2022). Association of stearoyl-CoA desaturase expression with cattle milk characteristics. Iranian Journal of Applied Animal Science, 12(2), 271–279. https://journals.iau.ir/article_691679.html
Al-Musawi, A. M., Awad, A. H. H., & Alkhaled, M. J. (2022). Molecular analysis of Cryptosporidium species in domestic goat in central Iraq. Iraqi Journal of Veterinary Sciences, 36(4), 1041–1045. https://doi.org/10.33899/ijvs.2022.132974.2155
Alnahass, M. G., Selim, A. M., El-Diasty, M., El-Sebaey, A. M., & Elbaz, E. (2026). Epidemiological, haematological-biochemical, and molecular investigations of bovine theileriosis with therapeutic evaluation of buparvaquone with silymarin in Dakahlia and Damietta governorates, Egypt. Veterinary Research Communications, 50(3), Article 195. https://doi.org/10.1007/s11259-026-11089-4
Badakhshan, Y., & Mohammadabadi, M. R. (2015). Thermoregulatory mechanisms of Jersey adult cattle and calves based on different body sites temperature. Iranian Journal of Applied Animal Science, 5(4), 793–798. https://journals.iau.ir/article_516394.html
Eghtedari, M., Khezri, A., Kazemi-Bonchenari, M., Yazdanyar, M., Mohammadabadi, M., Mahani, S. E., & Ghaffari, M. H. (2024). Effects of corn grain processing and phosphorus content in calf starters on intake, growth performance, nutrient digestibility, blood metabolites, and urinary purine derivatives. Journal of Dairy Science, 107(11), 9334–9346. https://doi.org/10.3168/jds.2024-25079
Ghasemi, M., Baghizadeh, A., & Mohammadabadi, M. R. (2010). Determination of genetic polymorphism in Kerman Holstein and Jersey cattle population using ISSR markers. Australian Journal of Basic and Applied Sciences, 6(12), 5758–5760. https://www.ajbasweb.com/old/ajbas_December_2010.html
Jalovecka, M., Sojka, D., Ascencio, M., & Schnittger, L. (2019). Babesia life cycle - When phylogeny meets biology. Trends in Parasitology, 35(5), 356–368. https://doi.org/10.1016/j.pt.2019.01.007
Javanmard, A., Mohammadabadi, M. R., Zarrigabayi, G. E., Gharahedaghi, A. A., Nassiry, M. R., Javadmansh, A., & Asadzadeh, N. (2008). Polymorphism within the intron region of the bovine leptin gene in Iranian Sarabi cattle (Iranian Bos taurus). Russian Journal of Genetics, 44(4), 495–497. https://doi.org/10.1134/S1022795408040169
Kagena, E. (2025). Impacts of climate change on tick-borne diseases in livestock: Case study of Theileria in cattle in New Zealand [Master’s thesis, Massey University]. https://mro.massey.ac.nz/handle/10179/74105
Kareem, S. M., Abbas, F. H., Rabeea, A. H. M. H., & Mosa, A. H. (2025). Infestation rate and molecular detection of hard ticks in pet cats from hilla city, Iraq. Journal of Animal Health and Production, 13(3), 706–710. https://doi.org/10.17582/journal.jahp/2025/13.3.706.710
Knowles, D. P., Kappmeyer, L. S., Haney, D., Herndon, D. R., Fry, L. M., Munro, J. B., Sears, K., Ueti, M. W., Wise, L. N., Silva, M., Schneider, D. A., Grause, J., White, S. N., Tretina, K., Bishop, R. P., Odongo, D. O., Pelzel-McCluskey, A. M., Scoles, G. A., Mealey, R. H., & Silva, J. C. (2018). Discovery of a novel species, Theileria haneyi n. sp., infective to equids, highlights exceptional genomic diversity within the genus Theileria: Implications for apicomplexan parasite surveillance. International Journal for Parasitology, 48(9–10), 679–690. https://doi.org/10.1016/j.ijpara.2018.03.010
Kundave, V. R., Patel, A. K., Patel, P. V., Hasnani, J. J., & Joshi, C. G. (2015). Detection of theileriosis in cattle and buffaloes by polymerase chain reaction. Journal of Parasitic Diseases, 39(3), 508–513. https://doi.org/10.1007/s12639-013-0386-2
Mohamadinejad, F., Mohammadabadi, M., Roudbari, Z., Eskandarynasab Siahkouhi, S., Babenko, O., Klopenko, N., Borshch, O., Starostenko, I., Kalashnyk, O., & Assadi Soumeh, E. (2024). Analysis of liver transcriptome data to identify the genes affecting lipid metabolism during the embryonic and hatching periods in ROSS breeder broilers. Journal of Livestock Science and Technologies, 12(2), 61–67. https://doi.org/10.22103/jlst.2024.23814.1554
Mohammadabadi, M. R., Torabi, A., Tahmourespoor, M., & Baghizadeh, A. (2010). Analysis of bovine growth hormone gene polymorphism of local and Holstein cattle breeds in Kerman province of Iran using polymerase chain reaction restriction fragment length. African Journal of Biotechnology, 9(41), 6848–6852.
Mohammadabadi, M., Akhtarpoor, A., Khezri, A., Babenko, O., Stavetska, R. V., Tytarenko, I., Ievstafiieva, Y., Buchkovska, V., Slynko, V., & Afanasenko, V. (2024a). The role and diverse applications of machine learning in genetics, breeding, and biotechnology of livestock and poultry. Agricultural Biotechnology Journal, 16(4), 413–442. https://doi.org/10.22103/jab.2025.24662.1644
Mohammadabadi, M., Babenko, O., Borshch, O., Kalashnyk, O., Ievstafiieva, Y., & Buchkovska, V. (2024c). Measuring the relative expression pattern of the UCP2 gene in different tissues of the Raini Cashmere goat. Agricultural Biotechnology Journal, 16(3), 317–332. https://doi.org/10.22103/jab.2024.24337.1627
Mohammadabadi, M., Golkar, A., Askari-Hesni, M., & Khezri, A. (2023). The effect of fennel (Foeniculum vulgare) on insulin-like growth factor 1 gene expression in the rumen tissue of Kermani sheep. Agricultural Biotechnology Journal, 15(4), 239–256. https://doi.org/10.22103/jab.2023.22647.1530
Mohammadabadi, M., Kheyrodin, H., Afanasenko, V., Babenko, O., Klopenko, N., Kalashnyk, O., Ievstafiieva, Y., & Buchkovska, V. (2024b). The role of artificial intelligence in genomics. Agricultural Biotechnology Journal, 16(2), 195–279. https://doi.org/10.22103/jab.2024.23558.1575
Mor, N. H., Tavera, J. V. M., Tobón, J. C., Guzmán Barragán, B. L., López, G. B., Vargas Duarte, J. J., Corredor, D. W. S., & Tafur-Gómez, G. A. (2024). Hemoparasitism in grazing cattle and risk factors associated with husbandry management in an endemic area of Eastern Colombia. Journal of Parasitic Diseases, 48(4), 924–935. https://doi.org/10.1007/s12639-024-01723-w
Nejad, F. M., Mohammadabadi, M., Roudbari, Z., Gorji, A. E., & Sadkowski, T. (2024). Network visualization of genes involved in skeletal muscle myogenesis in livestock animals. BMC Genomics, 25(1), Article 294. https://doi.org/10.1186/s12864-024-10196-3
Omer, A., Duvivier-Kali, V. F., Trivedi, N., Wilmot, K., Bonner-Weir, S., & Weir, G. C. (2003). Survival and maturation of microencapsulated porcine neonatal pancreatic cell clusters transplanted into immunocompetent diabetic mice. Diabetes, 52(1), 69–75. https://doi.org/10.2337/diabetes.52.1.69
Parveen, A., Ashraf, S., Aktas, M., Ozubek, S., & Iqbal, F. (2021). Molecular epidemiology of Theileria annulata infection of cattle in Layyah District, Pakistan. Experimental and Applied Acarology, 83(3), 461–473. https://doi.org/10.1007/s10493-021-00595-6
Patel, J., Schuett, J., & Chen, D. J. (2025). Hematology thin smears perform equally to parasitology thick and thin blood smears for the diagnosis of Plasmodium and Babesia infections in a low prevalence setting. Journal of Clinical Microbiology, 63(5), Article e0160124. https://doi.org/10.1128/jcm.01601-24
Rohallah, A., Mohammadreza, M. A., & Shahin, M. B. (2007). Kappa-casein gene study in Iranian Sistani cattle breed (Bos indicus) using PCR-RFLP. Pakistan Journal of Biological Sciences, 10(23), 4291–4294. https://doi.org/10.3923/pjbs.2007.4291.4294
Ullah, R., Shams, S., Khan, M. A., Ayaz, S., Akbar, N. U., Din, Q. U., Khan, A., Leon, R., & Zeb, J. (2021). Epidemiology and molecular characterization of Theileria annulata in cattle from central Khyber Pakhtunkhwa, Pakistan. PLOS ONE, 16(9), Article e0249417. https://doi.org/10.1371/journal.pone.0249417
Vu, Q. H., Van, H. T., Tran, V. T., Huynh, T. D. P., Nguyen, V. C., & Le, D. T. (2021). Development of a robust blood smear preparation procedure for external quality assessment. Practical Laboratory Medicine, 27, Article e00253. https://doi.org/10.1016/j.plabm.2021.e00253
Wahab, M. A., & Aziz, K. J. (2026). Prevalence of Theileria annulata and the first report of Theileria sinensis in cattle from Erbil Province, Iraq. Science Journal of University of Zakho, 14(1), 60–69. https://doi.org/10.25271/sjuoz.2026.14.1.1665
Agricultural Biotechnology Journal
Volume 18, Issue 3
June 2026
Pages 395-410

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