Avian haemosporidian infections have been associated with disease outbreaks in zoos and rehabilitation centers globally. This study aimed to determine the occurrence and genetic diversity of avian haemosporidian parasites in captive birds at the National Zoological Garden in South Africa. One hundred and eighty-three blood samples from five orders and 15 species of captive flamingos, vultures, owls, ibises and parrots were analyzed for haemosporidia by nested polymerase chain reaction assays. The samples were collected as part of the zoo's studbook and archived at South African National Biodiversity Institute's Wildlife Biobank. The overall infection rate was 36.1%, and infections by Leucocytozoon spp. (33.3%) were significantly higher than Haemoproteus spp. (14.8%) (p < 0.001). Twenty-one samples (11.5%) had mixed Haemoproteus and Leucocytozoon infections. The Spotted Eagle Owl (Bubo africanus) and Barn Owl (Tyto alba) had the highest infection rates. Twenty-six sequences, similar to published sequences of Leucocytozoon spp. (lineages lCHRKLA02, lCIAE02, and lBUVIR02) and Haemoproteus spp. (lineage hTYTAL6) were obtained. Two new lineages (lBUBCAP01 and hBOSHAG02) are described in this study. This is the first molecular survey of haemosporidian parasites in captive birds of the orders Accipitriformes, Pelecaniformes, Psittaciformes, Phoenicopteriformes, and Strigiformes in South Africa. This study provides new geographical and host records of known and novel haemosporidian lineages. It highlights the need for intensive surveys of these parasites in populations of captive and free-ranging birds in South Africa, regular monitoring of infections, updated screening methods, and insect control in the zoo's animal collection to avoid disease outbreaks.
| [1] |
Alley, M. R., R. A. Fairley, D. G. Martin, L. Howe, and T. Atkinson. 2008. “An Outbreak of Avian Malaria in Captive Yellowheads/Mohua (Mohoua ochrocephala).” New Zealand Veterinary Journal 56: 247–251.
|
| [2] |
de Angeli Dutra, D., A. Fecchio, B. É. Martins, and R. Poulin. 2021. “Migratory Birds Have Higher Prevalence and Richness of Avian Haemosporidian Parasites Than Residents.” International Journal of Parasitology 51: 877–882.
|
| [3] |
Atkinson, C. T. 2005. Ecology and Diagnosis of Introduced Avian Malaria in Hawaiian Forest Birds. USGS Numbered Series, Geological Survey (U.S.).
|
| [4] |
Beadell, J. S., R. Covas, C. Gebhard, et al. 2009. “Host Associations and Evolutionary Relationships of Avian Blood Parasites from West Africa.” International Journal of Parasitology 39: 257–266.
|
| [5] |
Beadell, J. S., E. Gering, J. Austin, et al. 2004. “Prevalence and Differential Host-Specificity of Two Avian Blood Parasite Genera in the Australo-Papuan Region.” Molecular Ecology 13: 3829–3844.
|
| [6] |
Bensch, S., O. Hellgren, and J. Pérez-Tris. 2009. “MalAvi: A Public Database of Malaria Parasites and Related Haemosporidians in Avian Hosts Based on Mitochondrial Cytochrome b Lineages.” Molecular Ecology Resources 9: 1353–1358.
|
| [7] |
Bernotiene, R., V. Palinauskas, T. Iezhova, et al. 2016. “Avian Haemosporidian Parasites (Haemosporida): a Comparative Analysis of Different Polymerase Chain Reaction Assays in Detection of Mixed Infections.” Experimental Parasitolology 163: 31–37.
|
| [8] |
Botes, A., H. Thiart, N. J. Parson, and D. U. Bellstedt. 2017. “Conservation Implications of Avian Malaria: Exposure for African Penguins During Rehabilitation.” South African Journal of Science 113, no. 7/8: 2016-0244.
|
| [9] |
Chagas, C. R. F., G. Valkiūnas, G. L. De Oliveira, et al. 2017. “Diversity and Distribution of Avian Malaria and Related Haemosporidian Parasites in Captive Birds from a Brazilian Megalopolis.” Malaria Journal 16: 83.
|
| [10] |
Chaisi, M. E., S. T. Osinubi, D. L. Dalton, and E. Suleman. 2019. “Occurrence and Diversity of Avian Haemosporidia in Afrotropical Landbirds.” International Journal for Parasitology: Parasites and Wildlife 8: 36–44.
|
| [11] |
Chakarov, N., and G. Blanco. 2021. “Blood Parasites in Sympatric Vultures: Role of Nesting Habits and Effects on Body Condition.” International Journal of Environmental Research and Public Health 18: 2431.
|
| [12] |
Ciloglu, A., A. Yildirim, O. Duzlu, Z. Onder, Z. Dogan, and A. Inci. 2016. “Investigation of Avian Haemosporidian Parasites from Raptor Birds in Turkey, with Molecular Characterisation and Microscopic Confirmation.” Folia Parasitologica63: 023.
|
| [13] |
Ciloglu, A., V. A. Ellis, R. Bernotienė, G. Valkiūnas, and S. Bensch. 2019. “A New One-Step Multiplex PCR Assay for Simultaneous Detection and Identification of Avian Haemosporidian Parasites.” Parasitology Research 118: 191–201.
|
| [14] |
Edgar, R. C. 2004. “MUSCLE: Multiple Sequence Alignment with High Accuracy and High Throughput.” Nucleic Acids Research 32: 1792–1797.
|
| [15] |
Ejiri, H., Y. Sato, K. S. Kim, et al. 2011. “Entomological Study on Transmission of Avian Malaria Parasites in a Zoological Garden in Japan: Bloodmeal Identification and Detection of Avian Malaria Parasite DNA from Blood-Fed Mosquitoes.” Journal Medical Entomology 48: 600–607.
|
| [16] |
Fecchio, A., C. R. F. Chagas, J. A. Bell, and K. Kirchgatter. 2019. “Evolutionary Ecology, Taxonomy, and Systematics of Avian Malaria and Related Parasites.” Acta Tropica 204: 105364.
|
| [17] |
Ferrell, S. T., K. Snowden, A. B. Marlar, M. Garner, and N. P. Lung. 2007. “Fatal Hemoprotozoal Infections in Multiple Avian Species in a Zoological Park.” Journal of Zoo and Wildlife Medicine 38: 309–316.
|
| [18] |
Galosi, L., F. E. Scaglione, G. E. Magi, et al. 2019. “Fatal Leucocytozoon Infection in a Captive Grey-Headed Parrot (Poicephalus robustus suahelicus).” Journal of Avian Medicine and Surgery 33: 179–183.
|
| [19] |
Graczyk, T. K., J. F. Cockrem, M. R. Cranfield, J. T. Darby, and P. Moore. 1995. “Avian Malaria Seroprevalence in Wild New Zealand Penguins.” Parasite 2: 401–405.
|
| [20] |
Grilo, M. L., R. E. T. Vanstreels, R. Wallace, et al. 2016. “Malaria in Penguins—Current Perceptions.” Avian Pathology 45: 393–407.
|
| [21] |
Gutiérrez-López, R., L. Gangoso, J. Martínez-de la Puente, et al. 2015. “Low Prevalence of Blood Parasites in a Long-Distance Migratory Raptor: The Importance of Host Habitat.” Parasites & Vectors 8: 189.
|
| [22] |
Harl, J., T. Himmel, G. Valkiūnas, M. Ilgūnas, et al. 2020. “Geographic and Host Distribution of Haemosporidian Parasite Lineages from Birds of the Family Turdidae.” Malaria Journal 19: 335.
|
| [23] |
Harl, J., T. Himmel, G. Valkiūnas, M. Ilgūnas, et al. 2022. “Avian Haemosporidian Parasites of Accipitriform Raptors.” Malaria Journal 21: 14.
|
| [24] |
Hellgren, O., J. Waldenström, and S. Bensch. 2004. “A New PCR Assay for Simultaneous Studies of Leucocytozoon, Plasmodium, and Haemoproteus from Avian Blood.” Journal of Parasitology 90: 797–802.
|
| [25] |
Himmel, T., J. Harl, A. Kübber-Heiss, et al. 2019. “Molecular Probes for the Identification of Avian Haemoproteus and Leucocytozoon Parasites in Tissue Sections by Chromogenic In Situ Hybridization.” Parasites Vectors 12: 282.
|
| [26] |
Himmel, T., J. Harl, J. Matt, and H. Weissenbock. 2021. “A Citizen Science-Based Survey of Avian Mortality Focusing on Haemosporidian Infections in Wild Passerine Birds.” Malaria Journal 20: 417.
|
| [27] |
Huang, X., D. Huang, Y. Liang, et al. 2020. “A New Protocol for Absolute Quantification of Haemosporidian Parasites in Raptors and Comparison with Current Assays.” Parasites Vectors 13: 354.
|
| [28] |
Huelsenbeck, J. P., and F. Ronquist. 2001. “MRBAYES: Bayesian Inference of Phylogenetic Trees.” Bioinformatics 17: 754–755.
|
| [29] |
Jia, T., X. Huang, and C. Zhang. 2018. “Malaria Parasites and Related Haemosporidians Cause Mortality in Cranes: A Study on the Parasites Diversity, Prevalence and Distribution in Beijing Zoo.” Malaria Journal 17: 234.
|
| [30] |
Kearse, M., R. Moir, A. Wilson, et al. 2012. “Geneious Basic: An Integrated and Extendable Desktop Software Platform for the Organization and Analysis of Sequence Data.” Bioinformatics 28: 1647–1649.
|
| [31] |
Krone, O., J. Waldenström, G. Valkiūnas, et al. 2008. “Haemosporidian Blood Parasites in European Birds of Prey and Owls.” Journal of Parasitology 94: 709–715.
|
| [32] |
Kumar, S., G. Stecher, M. Li, C. Knyaz, and K. Tamura. 2018. “MEGA X: Molecular Evolutionary Genetics Analysis Across Computing Platforms.” Molecular Biology and Evolution 35: 1547–1549.
|
| [33] |
Labuschagne, K., L. Gerber, I. Espie, and S. Carpenter. 2008. “Culicoides Biting Midges at the National Zoological Gardens of South Africa.” Onderstepoort Journal of Veterinary Research 74: 343–347.
|
| [34] |
LaPointe, D. A., C. T. Atkinson, and M. D. Samuel. 2012. “Ecology and Conservation Biology of Avian Malaria.” Annals of the New York Academy of Sciences 1249: 211–226.
|
| [35] |
Lertwatcharasarakul, P., C. Salakij, P. Prasopsom, et al. 2021. “Molecular and Morphological Analyses of Leucocytozoon Parasites (Haemosporida: Leucocytozoidae) in Raptors from Thailand.” Acta Parasitologica 66: 1406–1416.
|
| [36] |
Levine, N. D. 1988. The Protozoan Phylum Apicomplexa: Volume 2. 1st ed. CRC Press.
|
| [37] |
Lutz, H. L., W. M. Hochachka, J. I. Engel, et al. 2015. “Parasite Prevalence Corresponds to Host Life History in a Diverse Assemblage of Afrotropical Birds and Haemosporidian Parasites.” PLoS ONE 10: e0121254.
|
| [38] |
Martinsen, E. S., S. L. Perkins, and J. J. Schall. 2008. “A Three-Genome Phylogeny of Malaria Parasites (Plasmodium and Closely Related Genera): Evolution of Life-History Traits and Host Switches.” Molecular Phylogenetics and Evolution 47: 261–273.
|
| [39] |
Martinsen, E. S., I. Paperna, and J. J. Schall. 2006. “Morphological Versus Molecular Identification of Avian Haemosporidia: An Exploration of Three Species Concepts.” Parasitology 133: 279–288.
|
| [40] |
Meister, S. L., F. Wyss, C. Wenker, S. Hoby, and W. U. Basso. 2023. “Avian Haemosporidian Parasites in Captive and Free-Ranging, Wild Birds from Zoological Institutions in Switzerland: Molecular Characterization and Clinical Importance.” International Journal for Parasitology: Parasites and Wildlife 20: 46–55.
|
| [41] |
Niebuhr, C. N., R. Poulin, and D. M. Tompkins. 2016. “Is Avian Malaria Playing a Role in Native Bird Declines in New Zealand? Testing Hypotheses Along an Elevational Gradient.” PLoS ONE 11: e0165918.
|
| [42] |
Okanga, S., G. S. Cumming, and P. A. R. Hockey. 2013. “Avian Malaria Prevalence and Mosquito Abundance in the Western Cape, South Africa.” Malaria Journal 12: 370.
|
| [43] |
Pérez-Rodríguez, A., I. de la Hera, S. Fernández-González, and J. Pérez-Tris. 2014. “Global Warming Will Reshuffle the Areas of High Prevalence and Richness of Three Genera of Avian Blood Parasites.” Global Change Biology 20: 2406–2416.
|
| [44] |
Pérez-Rodríguez, A., J. de la Puente, A. Onrubia, and J. Pérez-Tris. 2013. “Molecular Characterization of Haemosporidian Parasites from Kites of the Genus Milvus (Aves: Accipitridae).” International Journal for Parasitology 43: 381–387.
|
| [45] |
Palinauskas, V., R. Žiegytė, T. A. Iezhova, M. Ilgūnas, R. Bernotienė, and G. Valkiūnas. 2016. “Description, Molecular Characterisation, Diagnostics and Life Cycle of Plasmodium Elongatum (lineage pERIRUB01), the Virulent Avian Malaria Parasite.” International Journal for Parasitology 46: 697–707.
|
| [46] |
Paperna, I., and P. Martelli. 2008. “Haemosporidian Infections in Captive Exotic Glossy Starling Lamprotornis chalybaeus in Hong Kong.” Folia Parasitologica 55: 7–12.
|
| [47] |
Parsons, N. J., and L. G. Underhill. 2005. “Oiled and Injured African Penguins Spheniscus demersus and Other Seabirds Admitted for Rehabilitation in the Western Cape, South Africa, 2001 and 2002.” African Journal of Marine Science 27: 289–296.
|
| [48] |
Parsons, N. J., N. M. Voogt, A. M. Schaefer, M. A. Peirce, and R. E. T. Vanstreels. 2017. “Occurrence of Blood Parasites in Seabirds Admitted for Rehabilitation in the Western Cape, South Africa, 2001–2013.” Veterinary Parasitology 233: 52–61.
|
| [49] |
Pornpanom, P., C. R. Chagas, P. Lertwatcharasarakul, C. Kasorndorkbua, G. Valkiūnas, and C. Salakij. 2019. “Molecular Prevalence and Phylogenetic Relationship of Haemoproteus and Plasmodium Parasites of Owls in Thailand: Data from a Rehabilitation Centre.” International Journal for Parasitology: Parasites and Wildlife 9: 248–257.
|
| [50] |
Pornpanom, P., C. Kasorndorkbua, P. Lertwatcharasarakul, and C. Salakij. 2021. “Prevalence and Genetic Diversity of Haemoproteus and Plasmodium in Raptors from Thailand: Data from Rehabilitation Center.” International Journal for Parasitology: Parasites and Wildlife 16: 75–82.
|
| [51] |
Pulgarin, P., C. Gómez, N. Bayly, et al. 2018. “Migratory Birds as Vehicles for Parasite Dispersal? Infection by Avian Haemosporidians over the Year and Throughout the Range of a Long-Distance Migrant.” Journal of Biogeography 46: 83–96.
|
| [52] |
Ranford-Cartwright, L. C. 2023. “Special Issue: Avian Malaria.” Parasitology 150: 1263–1265.
|
| [53] |
Richard, F. A., R. N. M. Sehgal, H. I. Jones, and T. B. Smith. 2002. “A Comparative Analysis of PCR-Based Detection Methods for Avian Malaria.” Journal of Parasitology 88: 819–822.
|
| [54] |
Schultz, A., and P. Whittington. 2005. “High Prevalence of Avian Malaria Infection to Avifauna at Cape Receife, Eastern Cape, South Africa.” Ostrich 76: 56–60.
|
| [55] |
Silva-Iturriza, A., V. Ketmaier, and R. Tiedemann. 2012. “Prevalence of Avian Haemosporidian Parasites and Their Host Fidelity in the Central Philippine Islands.” Parasitology International 61: 650–657.
|
| [56] |
Taioe, M. O., M. C. Mnisi, M. Chaisi, et al. 2020. “Mosquito Identification and Haemosporidian Parasites Detection in the Enclosure of the African Penguins (Spheniscus demersus) at the SANBI Zoological Garden.” International Journal for Parasitology: Parasites and Wildlife 13: 98–105.
|
| [57] |
Valkiūnas, G. 2005. Avian Malaria Parasites and Other Haemosporidia. CRC Press. Raton, Florida.
|
| [58] |
Valkiūnas, G., T. A. Iezhova, C. Loiseau, A. Chasar, T. B. Smith, and R. N. M. Sehgal. 2008. “New Species of Haemosporidian Parasites (Haemosporida) from African Rainforest Birds, with Remarks on Their Classification.” Parasitology Research 103: 1213–1228.
|
| [59] |
Vanstreels, R., R. da Silva-Filho, C. Kolesnikovas, et al. 2015. “Epidemiology and Pathology of Avian Malaria in Penguins Undergoing Rehabilitation in Brazil.” Veterinary Research 46: 30.
|
| [60] |
Waldenström, J., S. Bensch, D. Hasselquist, and Ö. Östman. 2004. “A New Nested Polymerase Chain Reaction Method Very Efficient in Detecting Plasmodium and Haemoproteus Infections from Avian Blood.” Journal of Parasitology 90: 191–194.
|
| [61] |
Wardjomto, M. B., M. Ndlovu, A. Pérez-Rodríguez, T. Pori, and T. Nangammbi. 2021. “Avian Haemosporidia in Native and Invasive Sparrows at an Afrotropical Region.” Parasitology Research 120: 2631–2640.
|
| [62] |
Zagalska-Neubauer, M., and S. Bensch. 2016. “High Prevalence of Leucocytozoon Parasites in Freshwater Breeding Gulls.” Journal of Ornithology 157: 525–532.
|
RIGHTS & PERMISSIONS
2025 The Author(s). Integrative Zoology published by International Society of Zoological Sciences, Institute of Zoology/Chinese Academy of Sciences and John Wiley & Sons Australia, Ltd.