Microscopy and Molecular Detection of Haemoparasites in Birds in Ibadan, Nigeria
Main Article Content
Abstract
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.
Article Details

This work is licensed under a Creative Commons Attribution 4.0 International License.
References
Banda PC. Agriculture: The backbone of the African economy. Int J Adv Res Social Sci Environ Stud Technol. 2022; 7(1): 58-61. DOI: 10.48028/iiprds/ijarssest.v7.i1.05
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
Liverpool-Tasie LS, Sanou A, and Tambo JA. Climate change adaptation among poultry farmers: Evidence from Nigeria. Clim Change. 2019; 157(3): 527-544. DOI: 10.1007/s10584-019-02574-8
Mlondo S, Tembe D, Malatji MP, Khumalo ZT, and Mukaratirwa S. Molecular identification of helminth parasites of the Heterakidae and Ascarididae families of free-ranging chickens from selected rural communities of KwaZulu-Natal province of South Africa. Poult Sci. 2022; 101(8): 101979. DOI: 10.1016/j.psj.2022.101979
Tembe D, Malatji MP, and Mukaratirwa S. Occurrence, prevalence, and distribution of haemoparasites of poultry in sub-Saharan Africa: A scoping review. Pathogens. 2023; 12(7): 945. DOI: 10.3390/pathogens12070945
Lawal JR, Bello AM, Balami SY, Dauda J, Malgwi KD, Ezema KU, et al. Prevalence of haemoparasites in village chickens (Gallus gallus domesticus) slaughtered at poultry markets in Maiduguri, Northeastern Nigeria. J Anim Sci Vet Med. 2016; 1(2): 39-45. DOI: 10.31248/JASVM2016.013
Attaran H, Luo J, Bo W, Nabavi R, and He HX. Haemosporidian blood parasites in nestling birds of prey in Mongolia. bioRxiv. 2021. DOI: 10.1101/2021.06.26.450033
Chagas CRF, Harl J, Preikša V, Bukauskaitė D, Ilgūnas M, Weissenböck H, et al. Lankesterella (Apicomplexa, Lankesterellidae) blood parasites of passeriform birds: Prevalence, molecular and morphological characterization, with notes on sporozoite persistence in vivo and development in vitro. Animals. 2021; 11(5): 1451. DOI: 10.3390/ani11051451
Nourani L, Djadid ND, Rabiee K, Mezerji MS, Shakiba M, Bakhshi H, et al. Detection of haemosporidian parasites in wild and domestic birds in northern and central provinces of Iran: Introduction of new lineages and hosts. Int J Parasitol Parasites Wildl. 2020; 13(1): 203-212. DOI: 10.1016/j.ijppaw.2020.10.001
El-Ghany WAA. Avian haemosporidian parasites: An updated review. Arch Vet Sci. 2023; 28(2): 1-17. DOI: 10.5380/avs.v28i2.91244
Adejuwon JO. Trends and periodicities of annual rainfall over Ibadan region, Nigeria. World Water Policy. 2022; 8(2): 132-141. DOI: 10.1002/wwp2.12081
Hellgren O, Waldenström J, and Bensch S. A new PCR assay for simultaneous studies of Leucocytozoon, Plasmodium, and Haemoproteus from avian blood. J Parasitol. 2004; 90(4): 797-802. DOI: 10.1645/GE-184R1
Saitou N, and Nei M. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Mol Biol Evol. 1987; 4(4): 406-425. DOI: 10.1093/oxfordjournals.molbev.a040454
Felsenstein J. Confidence limits on phylogenies: An approach using the bootstrap. Evolution. 1985; 39(4): 783-791. DOI: 10.1111/j.1558-5646.1985.tb00420.x
Jukes TH, and Cantor CR. Evolution of protein molecules. In: Munro HN, editor. Mammalian protein metabolism. Academic Press, 1969. p. 21-132. Available at: https://evoluscope.fr/phylographe/biblio/JukesCantor1969.pdf
Tamura K, Stecher G, and Kumar S. MEGA 11: Molecular evolutionary genetics analysis version 11. Mol Biol Evol. 2021; 38(7): 3022-3027. DOI: 10.1093/molbev/msab120
Scaglione FE, Pregel P, Cannizzo FT, Pérez-Rodríguez AD, Ferroglio E, and Bollo E. Prevalence of new and known species of haemoparasites in feral pigeons in northwest Italy. Malar J. 2015; 14(1): 99. DOI: 10.1186/s12936-015-0617-3
Lawal JR, Ibrahim UI, Biu AA, and Musa HI. Natural infections of avian haemosporidian parasites among free-range chickens (Gallus gallus domesticus) in Yamaltu Deba, Gombe State, Nigeria. Acta Sci Vet Sci. 2020; 2(4): 12-20. Available at: https://actascientific.com/ASVS/pdf/ASVS-02-0058.pdf
Ciloglu A, Ellis VA, Bernotienė R, Valkiūnas G, and Bensch S. A new one-step multiplex PCR assay for simultaneous detection and identification of avian haemosporidian parasites. Parasitol Res. 2019; 118(1): 191-201. DOI: 10.1007/s00436-018-6153-7
Ogbaje CI, Okpe JA, and Oke P. Haemoparasites and haematological parameters of Nigerian indigenous (local) and exotic (broiler) chickens slaughtered in Makurdi major markets, Benue State, Nigeria. Alex J Vet Sci. 2019; 63(2): 90-98. DOI: 10.5455/ajvs.53637
Lawal JR, Ibrahim UI, Biu AA, and Musa HI. Molecular detection of avian haemosporidian parasites in village chickens (Gallus gallus domesticus) in Gombe State, Nigeria. J Vet Med Anim Sci. 2022; 5(1): 1095. Available at: https://meddocsonline.org/journal-of-veterinary-medicine-and-animal-sciences/Molecular-detection-of-avian-haemosporidian-parasite-%20in-village-chickens-gallus-gallus-domesticus-in-gombe-state-nigeria.pdf
Sol D, Jovani R, and Torres J. Parasite mediated mortality and host immune response explain age-related differences in blood parasitism in birds. Oecologia. 2003; 135(4): 542-547. DOI: 10.1007/s00442-003-1223-6
LaPointe DA, Atkinson CT, and Samuel MD. Ecology and conservation biology of avian malaria. Ann NY Acad Sci. 2012; 1249(1): 211-226. DOI: 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
Musa S, Mackenstedt U, Woog F, and Dinkel A. Untangling the actual infection status: Detection of avian haemosporidian parasites of three Malagasy bird species using microscopy, multiplex PCR, and nested PCR methods. Parasitol Res. 2022; 121(10): 2817-2829. DOI: 10.1007/s00436-022-07606-4
Valkiūnas G, Iezhova TA, Križanauskienė A, Palinauskas V, Sehgal RN, and Bensch S. A comparative analysis of microscopy and PCR-based detection methods for blood parasites. J Parasitol. 2008; 94(6): 1395-1401. DOI: 10.1645/GE-1570.1