_1776544821 20260418204021000 Rovedar Daryoushbabazadeh@gmail.com Rovedar Journal of World’s Poultry Science J. World's Poult. Sci. 2980-7999 03 05 2026 5 1 Diano Oumar Mamadou Diop https://orcid.org/0009-0004-7140-9337 Abdulrauf Adekunle Usman https://orcid.org/0009-0007-6614-6343 Theophilus Aghogho Jarikre https://orcid.org/0000-0001-8222-198X Oluwole Oyetunde Oni https://orcid.org/0000-0001-7013-1282 Introduction: Avian haemoparasites pose significant threats to poultry health and productivity. While microscopy is traditionally used for parasite identification, molecular methods, such as polymerase chain reaction (PCR), offer enhanced sensitivity and specificity. The present study aimed to determine the prevalence of haemoparasites in poultry species in Ibadan, Nigeria, and to compare the diagnostic performance of microscopy and nested PCR for detection and identification.Materials and methods: Blood samples were obtained from 390 healthy birds, including commercial layers chickens (153), turkeys (75), broiler chickens (69), local chickens (60), pigeons (30), and ducks (3), randomly selected from 10 farms in Ibadan, Nigeria. Thin blood smears were Giemsa-stained and examined microscopically. Genomic DNA was extracted and subjected to nested PCR targeting the mitochondrial cytochrome b gene of haemosporidian parasites. Positive amplicons were sequenced and phylogenetically analyzed. Results: The haemoparasites were microscopically detected in 44.6% (174/390) of samples, including Plasmodium (P.) spp. (18.2%), Haemoproteus spp. (15.9%), Leucocytozoon spp. (5.6%), Babesia spp. (2.8%), and microfilariae (2.1%). The PCR detected infections in 53.3% (208/390), confirming P. gallinaceum (12.3%), Haemoproteus spp. (19%), Leucocytozoon spp. (9%), Babesia spp. (3.3%), and additional unidentified haemosporidian lineages (8.7%). The PCR demonstrated significantly greater sensitivity than microscopic analysis. Infections were more prevalent among females (60.3%), adult birds (55.2%), and during the rainy season (54%). Sequencing confirmed the presence of P. gallinaceum as the most prevalent pathogen (97.87-97.94%). Phylogenetic analysis supported the molecular identification and revealed evolutionary relationships among detected lineages. Conclusion: The present study confirmed a high prevalence of haemoparasites in poultry in Ibadan, Nigeria, and underscored the superior sensitivity of PCR over microscopy for detection. The integration of molecular and morphological approaches enhanced diagnostic accuracy and provided deeper insights into parasite diversity and epidemiology. 03 05 2026 1 8 https://creativecommons.org/licenses/by/4.0 10.58803/jwps.v5i1.102 https://jwps.rovedar.com/index.php/JWPS/article/view/102 https://jwps.rovedar.com/index.php/JWPS/article/download/102/119 https://jwps.rovedar.com/index.php/JWPS/article/download/102/119 10.48028/iiprds/ijarssest.v7.i1.06 Heise H, Crisan A, and Theuvsen L. The poultry market in Nigeria: Market structures and potential for investment in the market. Int Food Agribus Manag Rev. 2015; 18(A): 197-222. DOI: 10.22004/ag.econ.207011 10.1007/s10584-019-02574-8 10.1016/j.psj.2022.101979 10.3390/pathogens12070945 10.31248/JASVM2016.013 10.1101/2021.06.26.450033 10.3390/ani11051451 10.1016/j.ijppaw.2020.10.001 10.5380/avs.v28i2.91244 10.1002/wwp2.12081 10.1645/GE-184R1 10.1093/oxfordjournals.molbev.a040454 10.1111/j.1558-5646.1985.tb00420.x 10.1016/B978-1-4832-3211-9.50009-7 10.1093/molbev/msab120 10.1186/s12936-015-0617-3 10.31080/ASVS.2020.02.0058 10.1007/s00436-018-6153-7 10.5455/ajvs.53637 10.30564/vsr.v3i2.4127 10.1007/s00442-003-1223-6 10.1111/j.1749-6632.2011.06431.x Rukambile E, Chengula A, Swai E, and Jongejan F. Poultry ecto-, endo- and haemoparasites in Tanzania: A Review. Austin J Vet Sci Anim Husb. 2020; 7(1): 1066. https://austinpublishinggroup.com/veterinary-science-research/fulltext/avsah-v7-id1066.pdf 10.1007/s00436-022-07606-4 10.1645/GE-1570.1