Polychromophilus spp. (Haemosporida: Plasmodiidae): First Molecular Detection in Bat Flies From Brazilian Bats

Abstract

Haemosporidian parasites exhibit a wide range of vertebrate hosts and corresponding insect vectors. Among mammals, bats host the most diverse array of haemosporidians, with seven genera identified. The genus Polychromophilus is exclusive to bats and is globally linked with hematophagous flies of the genera Basilia, Nycteribia, and Penicillidia as potential vectors. In Brazil, recent molecular studies have detected Polychromophilus in bats from the Cerrado and Atlantic Forest biomes; however, its vectors in the country remained unidentified. This study analyzed the haemosporidians infection of bat flies (24 Nycteribiidae and 43 Streblidae) collected from 13 bat species in the Legado das Águas. The bat–fly associations revealed highly specialized interactions, particularly among Basilia flies and Myotis bats. Notably, a rare interaction between Megistopoda proxima and Carollia perspicillata was also observed. Two specimens (3%) of nycteribiid flies (Basilia speiseri and Basilia lindolphoi), both collected from Myotis nigricans, tested positive for infection with Polychromophilus spp. Using cytb gene sequences, we examined the phylogenetic relationships of these Polychromophilus lineages with other global lineages. We identified two haplotypes, each clustering in distinct clades within the Polychromophilus murinus group. The presence of these parasites was further confirmed by sequencing of the clpc gene from the apicoplast genome and the nuclear asl gene. This study represents the first molecular detection of Polychromophilus spp. in a vector in Brazil, 50 years after its morphological description in the salivary glands of Basilia. These findings provide novel insights into the ecological networks in host–parasite–vector interactions in a preserved neotropical environment.

Keywords

Atlantic Forest / bat / bat fly / Brazil / haemosporidian / Polychromophilus

References

[1] Bajić, B., O. Werb, I. Budinski, et al. 2023. “Non-Invasive Investigation of Polychromophilus Parasite Infections in Bat Populations in Serbia Using Bat Flies.” Parasites Vectors 16: 170. https://doi.org/10.1186/s13071-023-05786-1.

[2] Barbier, E., M. R. André, and E. Bernard. 2024. “A Wingless Fly on a Winged Mammal: Host-Parasite Dynamics Between Basilia travassosi (Diptera: Nycteribiidae) and Myotis lavali (Chiroptera: Vespertilionidae).” Parasitology Research 123: 269. https://doi.org/10.1007/s00436-024-08294-y.

[3] Barbier, E., and E. Bernard. 2023. “Streblid Flies Parasitizing Cave Bats in Carajás, Amazonia, with a New Record for Brazil.” Acta Amazonica 53: 122–129. https://doi.org/10.1590/1809-4392202203460.

[4] Bertola, P. B., C. C. Aires, S. E. Favorito, G. Graciolli, M. Amaku, and R. Pinto-da-Rocha. 2005. “Bat Flies (Diptera: Streblidae, Nycteribiidae) Parasitic on Bats (Mammalia: Chiroptera) at Parque Estadual da Cantareira, São Paulo, Brazil: Parasitism Rates and Host-Parasite Associations.” In Memórias do Instituto Oswaldo Cruz 100, no. 1: 25–32. https://doi.org/10.1590/S0074-02762005000100005.

[5] Borner, J., C. Pick, J. Thiede, et al. 2016. “Phylogeny of Haemosporidian Blood Parasites Revealed by a Multi-Gene Approach.” Molecular Phylogenetics and Evolution 94: 221–231. https://doi.org/10.1016/j.ympev.2015.09.003.

[6] CANVA. Canva. https://www.canva.com.

[7] Ceballos-Pérez, D. F., J. Alvarez-Londoño, H. E. Ramírez-Chaves, and F. A. Rivera-Páez. 2024. “Polychromophilus (Haemosporida: Plasmodiidae): A Review of Association with Bats (Mammalia, Chiroptera) and the First Record in the Neotropical Bat, Myotis albescens (Chiroptera, Vespertilionidae) From Colombia.” International Journal for Parasitology: Parasites and Wildlife 25: 100986. https://doi.org/10.1016/j.ijppaw.2024.100986.

[8] de Oliveira Corrêa, M., A. Lazar, D. Dias, and C. R. Bonvicino. 2013. “Quirópteros Hospedeiros de Zoonoses no Brasil.” Sociedade Brasileira De Mastozoologia 67: 23–38.

[9] daSilva Biz, L., M. F. Cascaes, B. F. L. Luciano, et al. 2021. “Parasitic Interactions Between Bats (Mammalia: Chiroptera) and Flies (Insecta: Diptera) in the Intersection Area of Temperate and Tropical Climates in Brazil.” Studies on Neotropical Fauna and Environment 57: 291–300. https://doi.org/10.1080/01650521.2020.1869490.

[10] Dias, D., C. E. L. Esbérard, and R. Moratelli. 2013. “A New Species of Lonchophylla (Chiroptera, Phyllostomidae) from the Atlantic Forest of Southeastern Brazil, with Comments on L. bokermanni.” Zootaxa 3722, no. 3: 347. https://doi.org/10.11646/zootaxa.3722.3.4.

[11] Dick, C. W., and K. Dittmar. 2014. “Parasitic Bat Flies (Diptera: Streblidae and Nycteribiidae): Host Specificity and Potential as Vectors.” In Bats (Chiroptera) as Vectors of Diseases and Parasites. Parasitology Research Monographs, edited by S. Klimpel and H. Mehlhorn, Volume 5, 131–155. Springer Nature. https://doi.org/10.1007/978-3-642-39333-4_6.

[12] Dick, C. W., and J. A. Miller. 2010. “Streblidae (bat Flies).” In Manual of Central American Diptera, edited by B. V. Brown, A. Borkent, J. M. Cumming, D. M. Wood, N. E. Woodley, and M. Zumbado, Volume 2, 1249–1260. NCR Research Press.

[13] Dick, C. W., and B. D. Patterson. 2006. “Bat Flies—Obligate Ectoparasites of Bats.” In Micromammals and Macroparasites: From Evolutionary Ecology to Management, edited by S. Morand, B. R. Krasnov, and R. Poulin, 179–194. Springer.

[14] Dormann, C. F., J. Fründ, N. Blüthgen, and B. Gruber. 2009. “Indices, Graphs and Null Models: Analyzing Bipartite Ecological Networks.” The Open Ecology Journal 2: 7–24.

[15] Emmons, L. H., and F. Feer. 1990. Neotropical Rainforest Mammals: A Field Guide. The University of Chicago Press.

[16] Eriksson, A., A. Filion, M. B. Labruna, et al. 2023. “Effects of Forest Loss and Fragmentation on Bat-Ectoparasite Interactions.” Parasitology Research 122: 1391–1402. https://doi.org/10.1007/s00436-023-07839-x.

[17] FAO. 2011. Food and Agriculture Organization of the United Nations. Investigating the Role of Bats in Emerging Zoonoses: Balancing Ecology, Conservation and Public Health Interest. FAO animal production and health.

[18] Folmer, O., M. Black, W. Hoeh, R. Lutz, and R. Vrijenhoek. 1994. “DNA Primers for Amplification of Mitochondrial Cytochrome c Oxidase Subunit I from Diverse Metazoan Invertebrates.” Molecular Marine Biology and Biotechnology 3, no. 5: 294–299.

[19] França, D. A., and H. Langoni. 2025. “Prevalent Zoonoses in Sao Paulo State, Brazil: the Role of Bats and Molecular Diagnosis.” Revista Do Instituto De Medicina Tropical De São Paulo 67: e17. https://doi.org/10.1590/S1678-9946202567017.

[20] Galen, S. C., J. Borner, E. S. Martinsen, et al. 2018. “The Polyphyly of Plasmodium: Comprehensive Phylogenetic Analyses of the Malaria Parasites (Order Haemosporida) Reveal Widespread Taxonomic Conflict.” Royal Society Open Science 5, no. 5: 171780. https://doi.org/10.1098/rsos.171780.

[21] Gardner, R. A., and D. H. Molyneux. 1988. “Polychromophilus murinus: A Malarial Parasite of Bats: Life-History and Ultrastructural Studies.” Parasitology 96: 591–605. https://doi.org/10.1017/S0031182000080215.

[22] Garnham, P. C. C., R. Lainson, and J. J. Shaw. 1971. “A Contribution to the Study of the Haematozoon Parasites of Bats. A New Mammalian Haemoproteid, Polychromophilus deanei n. sp.” Memórias do Instituto Oswaldo Cruz 69, no. 1: 119–125. https://doi.org/10.1590/S0074-02761971000100009.

[23] Gottdenker, N. L., D. G. Streicker, C. L. Faust, and C. R. Carroll. 2014. “Anthropogenic Land Use Change and Infectious Diseases: A Review of the Evidence.” Ecohealth 11, no. 4: 619–632. https://doi.org/10.1007/s10393-014-0941-z.

[24] Graciolli, G. 2003. “Nova Espécie De Anatrichobius Wenzel, 1966 (Diptera, Streblidae) do Brasil.” Revista Brasileira De Entomologia 47: 55–58. https://doi.org/10.1590/S0085-56262003000100008.

[25] Graciolli, G. 2004. “Nycteribiidae (Diptera, Hippoboscoidea) no Sul do Brasil.” Revista Brasileira De Zoologia 21: 971–985. https://doi.org/10.1590/S0101-81752004000400035.

[26] Graciolli, G., A. G. Autino, and G. L. Claps. 2007. “Catalogue of American Nycteribiidae (Diptera, Hippoboscoidea).” Revista Brasileira De Entomologia 51, no. 2: 142–159. https://doi.org/10.1590/S0085-56262007000200004.

[27] Graciolli, G., and C. W. Dick. 2023. “Checklist of World Nycteribiidae (Diptera: Hippoboscoidea).” Accessed December 11, 2024. https://www.researchgate.net/publication/322579074_CHECKLIST_OF_WORLD_NYCTERIBIIDAE_DIPTERA_HIPPOBOSCOIDEA.

[28] Guerrero, R. 2019. Streblidae (Diptera: Pupipara) de Venezuela: Sistemática, Ecologia e Evolução. Editorial Académica Espanhola.

[29] Guillén-Rodríguez, Y. G., D. Santiago-Alarcon, L. Chapa-Vargas, M. T. Suárez-Landa, S. Albino-Miranda, and S. Ibáñez-Bernal. 2024. “Seasonal Haemosporidian Detection in Mosquitoes (Diptera: Culicidae) and Their Interactions with Vertebrate Hosts in a Mexican Cloud Forest.” Parasitology Research 123: 379. https://doi.org/10.1007/s00436-024-08387-8.

[30] Guimarães, L. R. 1966. “Nycteribiid Batfies from Panama (Diptera: Nycteribiidae).” In Ectoparasites of Panama. The Field Museum of Natural History, edited by R. L. Wenzel and V. J. Tipton, 393–404. Chicago, Field Museum of Natural History. https://doi.org/10.5962/bhl.title.2633.

[31] Guimarães, L. R., and M. A. V. D'Andretia. 1956. “Sinopse dos Nycteribiidae (Diptera) do Novo Mundo.” Arquivos De Zoologia 9: 1–175.

[32] Harl, J., T. Himmel, M. Ilgūnas, et al. 2025. “Haemoproteustartakovskyi and Plasmodium relictum (Haemosporida, Apicomplexa) Differentially Express Distinct 18S rRNA Gene Variants in Bird Hosts and Dipteran Vectors.” Parasites & Vectors 18: 63. https://doi.org/10.1186/s13071-025-06696-0.

[33] Huelsenbeck, J. P., and F. Ronquist. 2001. “MrBayes: Bayesian Inference of Phylogenetic Trees.” Bioinformatics 17, no. 8: 754–755. https://doi.org/10.1093/bioinformatics/17.8.754.

[34] Inkscape. 2023. Inkscape v1.3.2. https://inkscape.org/.

[35] Keesing, F., L. Belden, P. Daszak, et al. 2010. “Impacts of Biodiversity on the Emergence and Transmission of Infectious Diseases.” Nature 468: 647–652. https://doi.org/10.1038/nature09575.

[36] 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.

[37] Landau, I., J. M. Chavatte, and I. Beveridge. 2012a. “Johnsprentia copemani gen. nov., sp. nov. (Haemoproteidae), a Parasite of the Flying-fox, Pteropus alecto (Pteropodidae), from Queensland.” Memoirs of the Queensland Museum 56: 61–66.

[38] Landau, I., J. M. Chavatte, G. Karadjian, A. Chabaud, and I. Beveridge. 2012b. “The Haemosporidian Parasites of Bats with Description of Sprattiella alecto gen. nov., sp. nov.” Parasite 19, no. 2: 137–146. https://doi.org/10.1051/parasite/2012192137.

[39] López-Rivera, C., L. Robayo-Sánchez, A. Ramírez-Hernández, et al. 2024. “Hyperparasitism in Bat Flies (Diptera: Streblidae): New Records and Interaction Networks in the Neotropics.” Parasitology Research 123: 255. https://doi.org/10.1007/s00436-024-08221-1.

[40] Lourenço, E. C., L. A. Gomes, A. d. Viana, et al. 2020. “Co-occurrence of Ectoparasites (Insecta and Arachnida) on Bats (Chiroptera) in an Atlantic Forest Remnant, Southeastern Brazil.” Acta Parasitologica 65: 750–759. https://doi.org/10.2478/s11686-020-00224-z.

[41] 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, no. 1: 261–273. https://doi.org/10.1016/j.ympev.2007.11.012.

[42] Mathias B. D. S, G. A. Minozzo, A. W. Biondo, et al. 2023. “Molecular Investigation Confirms Myotis Genus Bats as Common Hosts of Polychromophilus in Brazil.” Microorganisms 11: 1531. https://doi.org/10.3390/microorganisms11061531.

[43] Megali, A., G. Yannic, and P. Christe. 2011. “Disease in the Dark: Molecular Characterization of Polychromophilus murinus in Temperate Zone Bats Revealed a Worldwide Distribution of this Malaria-Like Disease.” Molecular Ecology 20, no. 5: 1039–1048. https://doi.org/10.1111/j.1365-294X.2010.04905.x.

[44] Minozzo, G. A., B. da Silva Mathias, I. N. Riediger, et al. 2021. “First Molecular Detection of Polychromophilus Parasites in Brazilian Bat Species.” Microorganisms 9: 1240. https://doi.org/10.3390/microorganisms9061240.

[45] Nogueira, M. R., I. P. de Lima, R. Moratelli, V. da C. Tavares, R. Gregorin, and A. L. Peracchi. 2014. “Checklist of Brazilian Bats, with Comments on Original Records.” Check List 10, no. 4: 808–821.

[46] Obame-Nkoghe, J., N. Rahola, M. Bourgarel, et al. 2016. “Bat Flies (Diptera: Nycteribiidae and Streblidae) Infesting Cave-Dwelling Bats in Gabon: Diversity, Dynamics and Potential Role in Polychromophilus melanipherus Transmission.” Parasites Vectors 9: 333. https://doi.org/10.1186/s13071-016-1625-z.

[47] Pacheco, M. A., and A. A. Escalante. 2023. “Origin and Diversity of Malaria Parasites and Other Haemosporida.” Trends Parasitology 39: 501–516. https://doi.org/10.1016/j.pt.2023.04.004.

[48] Paglia, A. P., G. A. B. Fonseca, A. B. Rylands, et al. 2012. “Lista Anotada dos Mamíferos do Brasil.” Occasional Papers in Conservation Biology 6: 76.

[49] Peracchi, A. L., I. P. Lima, N. R. Reis, M. R. Nogueira, and H. Ortêncio-Filho. 2006. “Ordem Chiroptera.” In Mamíferos do Brasil, edited by N. R. Reis, A. L. Peracchi, W. A. Pedro, and I. P. Lima, 153–230. Editora UEL, Londrina.

[50] Perkins, S. L., and J. Schaer. 2016. “A Modern Menagerie of Mammalian Malaria.” Trends in Parasitology 32: 772–782. https://doi.org/10.1016/j.pt.2016.06.001.

[51] Perkins, S. L., and J. J. Schall. 2002. “A Molecular Phylogeny of Malarial Parasites Recovered From Cytochrome b Gene Sequences.” Journal of Parasitology 88: 972–978. https://doi.org/10.1645/0022-3395(2002)088[0972:AMPOMP]2.0.CO;2.

[52] Ramasindrazana, B., S. M. Goodman, N. Dsouli, et al. 2018. “Polychromophilus spp. (Haemosporida) In Malagasy Bats: Host Specificity and Insights on Invertebrate Vectors.” Malaria Journal 17: 318. https://doi.org/10.1186/s12936-018-2461-8.

[53] Rambaut, A. 2010. FigTree: Tree Figure Drawing Tool Version 1.4.0, Institute of Evolutionary Biology. University of Edinburgh.

[54] Ramírez-Martínez, M. M., and L Tlapaya-Romero. 2023. “Association of Bat Flies (Diptera: Streblidae) and Bats: Richness and Host Specificity in Western Mexico.” International Journal for Parasitology: Parasites and Wildlife 21: 160–167. https://doi.org/10.1016/j.ijppaw.2023.05.001.

[55] Reis, N. R. 1981. Estudos ecológicos dos quirópteros de matas primárias e capoeiras da região de Manaus, Amazonas. 242 f. Tese (Doutorado)—Universidade do Amazonas, INPA, Manaus.

[56] Reis, N. R., M. N. Fregonezi, A. L. Peracchi, and O. A. Shibatta (Orgs.). 2013. Morcegos do Brasil: Guia de Campo. 1a Edição. Technical Books Editora (Série Manuais & guias TB).

[57] Reis, N. R., A. L. Peracchi, C. B. Batista, I. P. Lima, and A. D. Pereira (Orgs.). 2017. História Natural dos Morcegos Brasileiros: Chave de Identificação de Espécies. 1a edição. Technical Books Editora.

[58] Reis, N. R., A. L. Peracchi, and G. Santos. 2008. Ecologia de Morcegos. Technical Books Editora; Universidade Estadual de Londrina, Programa de Pós-Graduação Ciências Biológicas.

[59] Rosyadi, I., H. Shimoda, A. Takano, T. Yanagida, and H. Sato. 2022. “Isolation and Molecular Characterization of Polychromophilus spp. (Haemosporida: Plasmodiidae) from the Asian Long-Fingered Bat (Miniopterus fuliginosus) and Japanese Large-Footed Bat (Myotis macrodactylus) in Japan.” Parasitology Research 121: 2547–2559. https://doi.org/10.1007/s00436-022-07592-7.

[60] Ruiz, F., Y.-M. Linton, D. J. Ponsonby, et al. 2010. “Molecular Comparison of Topotypic Specimens Confirms Anopheles (Nyssorhynchus) dunhami Causey (Diptera: Culicidae) in the Colombian Amazon.” Memórias do Instituto Oswaldo Cruz 105: 899–903. https://doi.org/10.1590/S0074-02762010000700010.

[61] Sándor, A. D., A. Péter, A. Corduneanu, et al. 2021. “Wide Distribution and Diversity of Malaria-Related Haemosporidian Parasites (Polychromophilus spp.) in Bats and Their Ectoparasites in Eastern Europe.” Microorganisms 9, no. 2: 230. https://doi.org/10.3390/microorganisms9020230.

[62] Schaer, J., S. L. Perkins, J. Decher, et al. 2013. “High Diversity of West African Bat Malaria Parasites and a Tight Link with Rodent Plasmodium Taxa.” Proceedings of the National Academy of Sciences 110: 17415–17419. https://doi.org/10.1073/pnas.1311016110.

[63] Schaer, J., D. M. Reeder, M. E. Vodzak, et al. 2015. “Nycteria Parasites of Afrotropical Insectivorous Bats.” International Journal for Parasitology 45: 375–384. https://doi.org/10.1016/j.ijpara.2015.01.008.

[64] Simmons, N. B. 2005. “Order Chiroptera.” In . Mammal Species of the World: A Taxonomic and Geographic Reference, edited by D. E. Wilson and D. M. Reeder, 3rd ed., 312–529. Johns Hopkins University Press.

[65] Szentiványi, T., W. Markotter, M. Dietrich, et al. 2020. “Host Conservation Through Their Parasites: Molecular Surveillance of Vector-Borne Microorganisms in Bats Using Ectoparasitic Bat Flies.” Parasite 27: 72. https://doi.org/10.1051/parasite/2020069.

[66] Taylor, M., and M. D. Turtle. 2019. Bats: An Illustrated Guide to All Species. Ivy Press.

[67] Thompson, J. D., D. G. Higgins, and T. J. Gibson. 1994. “Clustal W: Improving the Sensitivity of Progressive Multiple Sequence Alignment Through Sequence Weighting, Position-Specific Gap Penalties and Weight Matrix Choice.” Nucleic Acids Research 22: 4673–4680.

[68] Timm, L., S. P. Rosskopf, O. Werb, J. van Schaik, and J. Schaer. 2025. “Europe-Wide Distribution and Bat-Host Specific Lineages in the Malarial Parasite Polychromophilus murinus Revealed Through Genetic Screening of Bat Flies.” Infection, Genetics and Evolution 127: 105707. https://doi.org/10.1016/j.meegid.2024.105707.

[69] Urbieta, G. L., G. Graciolli, and J. Vizentin-Bugoni. 2021. “Modularity and Specialization in Bat–Fly Interaction Networks Are Remarkably Consistent Across Patches Within Urbanized Landscapes and Spatial Scales.” Current Zoology 67, no. 4: 403–410. https://doi.org/10.1093/cz/zoaa072.

[70] Valkiūnas, G. 2005. Avian Malaria Parasites and Other Haemosporidia. CRC Press.

[71] Vizotto, L. D., and V. A. Taddei. 1973. Chave para Determinação de Quirópteros Brasileiros. São José do Rio Preto: Gráfica Francal.

[72] Witsenburg, F., L. Clément, A. López-Baucells, et al. 2015. “How a Haemosporidian Parasite of Bats Gets Around: The Genetic Structure of a Parasite, Vector and Host Compared.” Molecular Ecology 24, no. 4: 926–940. https://doi.org/10.1111/mec.13071.

[73] Witsenburg, F., N. Salamin, and P. Christe. 2012. “The Evolutionary Host Switches of Polychromophilus: A Multi-Gene Phylogeny of the Bat Malaria Genus Suggests a Second Invasion of Mammals by a Haemosporidian Parasite.” Malaria Journal 11: 53. https://doi.org/10.1186/1475-2875-11-53.

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.