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These parasites are transmitted by several black fly species, but knowledge of the factors determining the diversity and prevalence in these vectors, which is crucial for fully understanding disease epidemiology, is largely unexplored. In this study, we investigated factors associated with the prevalence and diversity of <jats:italic>Leucocytozoon<\/jats:italic> species in black flies from Thailand.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>Adults of two black fly taxa (<jats:italic>Simulium asakoae<\/jats:italic> Takaoka and Davies complex and <jats:italic>S. khelangense<\/jats:italic> Takaoka, Srisuka and Saeung) were collected using sweep nets at nine locations in northern and northeastern regions of Thailand. Specimens were identified morphologically and the results corroborated by DNA barcoding. Molecular methods using specific primers for amplification of the mitochondrial cytochrome<jats:italic> b<\/jats:italic> (cyt <jats:italic>b<\/jats:italic>) gene of <jats:italic>Leucocytozoon<\/jats:italic> were used to detect the parasite in black flies. Species and lineages of <jats:italic>Leucocytozoon<\/jats:italic> were determined using the MalAvi database of malaria parasites and related haemosporidians in avian hosts. Regression analysis was used to examine relationships between <jats:italic>Leucocytozoon<\/jats:italic> diversity and prevalence, black fly abundance and habitat characteristics.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>A total of 11,718 adult black flies were collected, of which 4367 were members of the <jats:italic>S. asakoae<\/jats:italic> complex and 7351 were <jats:italic>S. khelangense<\/jats:italic>. For molecular detection of <jats:italic>Leucocytozoon<\/jats:italic>, we randomly selected 300 individual female black flies of the <jats:italic>S<\/jats:italic>. <jats:italic>asakoae<\/jats:italic> complex and 850 females of <jats:italic>S<\/jats:italic>. <jats:italic>khelangense<\/jats:italic> pooled into groups of five individuals (= 170 pools).  A total of 34 of the 300 specimens of the <jats:italic>S. asakoae<\/jats:italic> complex and 118 of the 170 pools of <jats:italic>S<\/jats:italic>. <jats:italic>khelangense<\/jats:italic> were positive for <jats:italic>Leucocytozoon<\/jats:italic>. Fifty-four lineages (haplotypes) were identified, all of which belonged to those reported in domestic chickens, <jats:italic>Gallus gallus<\/jats:italic>, with one exception that was identified in <jats:italic>S. khelangense<\/jats:italic> and found to be closely related to the <jats:italic>Leucocytozoon<\/jats:italic> lineages reported in owls; this is the first record of the latter lineage in Asian black flies. Among these haplotypes, nine and 45 were exclusively found in the <jats:italic>S. asakoae<\/jats:italic> complex and <jats:italic>S. khelangense<\/jats:italic>, respectively. No lineage was shared between these black fly taxa. Analysis of similarity (ANOSIM) revealed significant <jats:italic>Leucocytozoon<\/jats:italic> lineage composition between the two black flies. Phylogenetic analysis found that <jats:italic>Leucocytozoon<\/jats:italic> lineages in the <jats:italic>S. asakoae<\/jats:italic> complex and <jats:italic>S. khelangense<\/jats:italic> are largely isolated, agreeing with the ANOSIM result. The overall prevalence of <jats:italic>Leucocytozoon<\/jats:italic> in the <jats:italic>S. asakoae<\/jats:italic> complex was 11.3% and ranged from 9% to 13% in each collection. <jats:italic>Leucocytozoon<\/jats:italic> prevalence in <jats:italic>S. khelangense<\/jats:italic> was 21%, varying from 13% to 37% in each collection. The Shannon H\u2032 index indicated greater <jats:italic>Leucocytozoon<\/jats:italic> diversity in <jats:italic>S. khelangense<\/jats:italic> (H\u2032\u2009=\u20093.044) than in the <jats:italic>S. asakoae<\/jats:italic> complex (H\u2032\u2009=\u20091.920). Regression analysis revealed that <jats:italic>Leucocytozoon<\/jats:italic> diversity was positively related to black fly abundance and negatively related to maximum air temperature.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>The results of this study show that the prevalence and diversity of <jats:italic>Leucocytozoon<\/jats:italic> lineages in the <jats:italic>S<\/jats:italic>. <jats:italic>asakoae<\/jats:italic> complex and <jats:italic>S. khelangense<\/jats:italic> from Thailand were associated with the abundance of these black flies and with air temperature. The <jats:italic>Leucocytozoon<\/jats:italic> lineages identified also showed some degree of black fly taxon specificity, possibly related to different abundance peaks of these vectors. The environmental conditions that favor the development of black flies are possibly a driver of <jats:italic>Leucocytozoon<\/jats:italic> prevalence, diversity and vector\u2013parasite co-evolution.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Graphical Abstract<\/jats:title>\n                \n              <\/jats:sec>","DOI":"10.1186\/s13071-024-06567-0","type":"journal-article","created":{"date-parts":[[2024,11,19]],"date-time":"2024-11-19T09:24:23Z","timestamp":1732008263000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Diversity and prevalence of Leucocytozoon in black flies (Diptera: Simuliidae) of Thailand"],"prefix":"10.1186","volume":"17","author":[{"given":"Waraporn","family":"Jumpato","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wannachai","family":"Wannasingha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chavanut","family":"Jaroenchaiwattanachote","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronnalit","family":"Mintara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Komgrit","family":"Wongpakam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter H.","family":"Adler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pairot","family":"Pramual","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,11,19]]},"reference":[{"key":"6567_CR1","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1002\/9780813804620.ch4","volume-title":"Parasitic diseases of wild birds","author":"DJ Forrester","year":"2008","unstructured":"Forrester DJ, Greiner EC. 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