{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,27]],"date-time":"2026-03-27T19:10:51Z","timestamp":1774638651004,"version":"3.50.1"},"reference-count":45,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2023,4,19]],"date-time":"2023-04-19T00:00:00Z","timestamp":1681862400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Education, Science and Technological Development of the Republic of Serbia","award":["451-03-47\/2023-01\/ 200178"],"award-info":[{"award-number":["451-03-47\/2023-01\/ 200178"]}]},{"name":"Ministry of Education, Science and Technological Development of the Republic of Serbia","award":["30709-1"],"award-info":[{"award-number":["30709-1"]}]},{"name":"Ministry of Education, Science and Technological Development of the Republic of Serbia","award":["857251"],"award-info":[{"award-number":["857251"]}]},{"DOI":"10.13039\/100007463","name":"Rufford Small Grant Foundation","doi-asserted-by":"publisher","award":["451-03-47\/2023-01\/ 200178"],"award-info":[{"award-number":["451-03-47\/2023-01\/ 200178"]}],"id":[{"id":"10.13039\/100007463","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007463","name":"Rufford Small Grant Foundation","doi-asserted-by":"publisher","award":["30709-1"],"award-info":[{"award-number":["30709-1"]}],"id":[{"id":"10.13039\/100007463","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007463","name":"Rufford Small Grant Foundation","doi-asserted-by":"publisher","award":["857251"],"award-info":[{"award-number":["857251"]}],"id":[{"id":"10.13039\/100007463","id-type":"DOI","asserted-by":"publisher"}]},{"name":"European Union\u2019s Horizon 2020 Research and Innovation Programme","award":["451-03-47\/2023-01\/ 200178"],"award-info":[{"award-number":["451-03-47\/2023-01\/ 200178"]}]},{"name":"European Union\u2019s Horizon 2020 Research and Innovation Programme","award":["30709-1"],"award-info":[{"award-number":["30709-1"]}]},{"name":"European Union\u2019s Horizon 2020 Research and Innovation Programme","award":["857251"],"award-info":[{"award-number":["857251"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Haemogregarines (Apicomplexa: Adeleorina) are the most common and widespread reptilian blood parasites. Haemogregarina stepanowi was the first haemogregarine described from a reptile, the European pond turtle Emys orbicularis, and initial assessments indicated it was widespread across different pond turtle host species across much of Europe, the Middle East and North Africa. However, recent molecular assessments have indicated the presence of multiple genetically distinct forms in North Africa and the Iberian Peninsula, and extensive mixed infections which may be associated with a negative impact on the hosts. Here, we screened two native species, E. orbicularis and Mauremys rivulata, and the introduced Trachemys scripta from Serbia and North Macedonia for haemogregarines by amplifying and sequencing part of the 18S rRNA gene of these parasites, and used a standard DNA barcoding approach to identify leeches, the final host, attached to pond turtles. Our results again demonstrate the occurrence of considerable diversity of parasites in the analysed pond turtle species, and that T. scripta are likely infected by local haemogregarine parasites, and not those that are found in its native range. Leeches were identified as Placobdella costata, part of a lineage from Northern Europe. Mixed infections within pond turtles were again common. Current haemogregarine taxonomy does not reflect the genetic diversity identified, and a full taxonomic reassessment is needed.<\/jats:p>","DOI":"10.3390\/microorganisms11041063","type":"journal-article","created":{"date-parts":[[2023,4,19]],"date-time":"2023-04-19T04:22:51Z","timestamp":1681878171000},"page":"1063","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Molecular Screening of Haemogregarine Hemoparasites (Apicomplexa: Adeleorina: Haemogregarinidae) in Populations of Native and Introduced Pond Turtles in Eastern Europe"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7965-571X","authenticated-orcid":false,"given":"Marko","family":"Mari\u010di\u0107","sequence":"first","affiliation":[{"name":"Institute of Zoology, Faculty of Biology, University of Belgrade, Studentski trg 16, 11000 Belgrade, Serbia"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4929-2740","authenticated-orcid":false,"given":"Gorana","family":"Danon","sequence":"additional","affiliation":[{"name":"Institute of Zoology, Faculty of Biology, University of Belgrade, Studentski trg 16, 11000 Belgrade, Serbia"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6800-1243","authenticated-orcid":false,"given":"J. Filipe","family":"Faria","sequence":"additional","affiliation":[{"name":"BIOPOLIS Program, CIBIO-InBIO, Centro de Investiga\u00e7\u00e3o em Biodiversidade e Recursos Gen\u00e9ticos, Campus de Vair\u00e3o, Universidade do Porto, 4485-661 Vair\u00e3o, Portugal"},{"name":"Departamento de Biologia, Faculdade de Ci\u00eancias, Universidade do Porto, Rua do Campo Alegre s\/n, 4169-007 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5144-2421","authenticated-orcid":false,"given":"D. James","family":"Harris","sequence":"additional","affiliation":[{"name":"BIOPOLIS Program, CIBIO-InBIO, Centro de Investiga\u00e7\u00e3o em Biodiversidade e Recursos Gen\u00e9ticos, Campus de Vair\u00e3o, Universidade do Porto, 4485-661 Vair\u00e3o, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2023,4,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"903","DOI":"10.1016\/S0020-7519(99)00045-4","article-title":"The functional importance of parasites in animal communities: Many roles at many levels?","volume":"29","author":"Poulin","year":"1999","journal-title":"Int. J. Parasitol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1016\/j.pt.2009.05.010","article-title":"Evolution of the Apicomplexa: Where are we now?","volume":"25","author":"Morrison","year":"2009","journal-title":"Trends Parasitol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"108596","DOI":"10.1016\/j.biocon.2020.108596","article-title":"A global parasite conservation plan","volume":"250","author":"Carlson","year":"2020","journal-title":"Biol. Conserv."},{"key":"ref_4","unstructured":"Telford, S.R. (2009). Hemoparasites of the Reptilia: Color Atlas and Text, CRC Press."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1111\/j.1550-7408.1995.tb01551.x","article-title":"Phylogeny of Adeleid Blood Parasites with a Partial Systematic Revision of the Haemogregarine Complex","volume":"42","author":"Siddall","year":"1995","journal-title":"J. Eukaryot. Microbiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1080\/15627020.2015.1044467","article-title":"First Hemolivia from southern Africa: Reassigning chelonian Haemogregarina parvula Dias, 1953 (Adeleorina: Haemogregarinidae) to Hemolivia (Adeleorina: Karyolysidae)","volume":"50","author":"Cook","year":"2015","journal-title":"Afr. Zool."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2773","DOI":"10.1007\/s00436-019-06430-7","article-title":"Haemogregarina daviesensis sp. nov. (Apicomplexa: Haemogregarinidae) from South American lungfish Lepidosiren paradoxa (Sarcopterygii: Lepidosirenidae) in the eastern Amazon region","volume":"118","author":"Viana","year":"2019","journal-title":"Parasitol. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.14411\/fp.2016.029","article-title":"The good, the bad and the ugly: Emys trinacris, Placobdella costata and Haemogregarina stepanowi in Sicily (Testudines, Annelida and Apicomplexa)","volume":"63","author":"Arizza","year":"2016","journal-title":"Folia Parasitol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"522","DOI":"10.1017\/S0031182013001820","article-title":"Haemogregarines from western Palaearctic freshwater turtles (genera Emys, Mauremys) are conspecific with Haemogregarina stepanowi Danilewsky, 1885","volume":"141","author":"Javanbakht","year":"2014","journal-title":"Parasitology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"547","DOI":"10.1007\/s11230-021-09995-z","article-title":"Characterization and identification of haemogregarine hemoparasites (Apicomplexa: Adeleina: Hepatozoidae) in natural populations of Mauremys leprosa leprosa and M. leprosa saharica from Morocco","volume":"98","author":"Laghzaoui","year":"2021","journal-title":"Syst. Parasitol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3315","DOI":"10.1007\/s00436-020-06821-1","article-title":"The genetic diversity of blood parasites within the freshwater turtles Mauremys leprosa and Emys orbicularis in Tunisia reveals coinfection with Haemogregarina spp.","volume":"119","author":"Achouri","year":"2020","journal-title":"Parasitol. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1007\/s11230-022-10033-9","article-title":"Blood parasite diversity (Apicomplexa: Haemogregarinidae) within the western populations of the European pond turtle Emys orbicularis","volume":"99","author":"Harris","year":"2022","journal-title":"Syst. Parasitol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"517","DOI":"10.1007\/s11230-016-9643-y","article-title":"Morphological and molecular characterisation of Haemogregarina sp. (Apicomplexa: Adeleina: Haemogregarinidae) from the blood of the Caspian freshwater turtle Mauremys caspica (Gmelin) (Geoemydidae) in Iran","volume":"93","author":"Rakhshandehroo","year":"2016","journal-title":"Syst. Parasitol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"125884","DOI":"10.1016\/j.protis.2022.125884","article-title":"Haemogregarines of the genera Haemogregarina, Hemolivia, and Hepatozoon infecting Vietnamese freshwater turtles, with additional notes on primer specificity and primer-template mismatches affecting diagnostic success","volume":"173","author":"Pribyl","year":"2022","journal-title":"Protist"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1449","DOI":"10.1017\/S0031182017000889","article-title":"Prevalence and genetic diversity of blood parasite mixed infections in Spanish terrapins, Mauremys leprosa","volume":"144","author":"Marzal","year":"2017","journal-title":"Parasitology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"85","DOI":"10.2744\/CCB-1411.1","article-title":"Identification of Blood Parasites in Individuals from Six Families of Freshwater Turtles","volume":"19","author":"Nordmeyer","year":"2020","journal-title":"Chelonian Conserv. Biol."},{"key":"ref_17","first-page":"294","article-title":"DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates","volume":"3","author":"Folmer","year":"1994","journal-title":"Mol. Mar. Biol. Biotechnol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"670","DOI":"10.1645\/GE-204R","article-title":"High Prevalence of Hepatozoon spp. (Apicomplexa, Hepatozoidae) Infection in Water Pythons (Liasis fuscus) From Tropical Australia","volume":"90","author":"Ujvari","year":"2004","journal-title":"J. Parasitol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"021","DOI":"10.14411\/fp.2021.021","article-title":"Putative intranuclear coccidium in Mauremys leprosa from Morocco","volume":"68","author":"Laghzaoui","year":"2021","journal-title":"Folia Parasitol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1016\/S0022-2836(05)80360-2","article-title":"Basic local alignment search tool","volume":"215","author":"Altschul","year":"1990","journal-title":"J. Mol. Biol."},{"key":"ref_21","first-page":"772","article-title":"PartitionFinder 2: New Methods for Selecting Partitioned Models of Evolution for Molecular and Morphological Phylogenetic Analyses","volume":"34","author":"Lanfear","year":"2017","journal-title":"Mol. Biol. Evol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1572","DOI":"10.1093\/bioinformatics\/btg180","article-title":"MrBayes 3: Bayesian phylogenetic inference under mixed models","volume":"19","author":"Ronquist","year":"2003","journal-title":"Bioinformatics"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1239","DOI":"10.1007\/s10750-021-04786-5","article-title":"Extensive sampling sheds light on species-level diversity in Palearctic Placobdella (Annelida: Clitellata: Glossiphoniiformes)","volume":"849","author":"Kvist","year":"2022","journal-title":"Hydrobiologia"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"20630","DOI":"10.1038\/s41598-022-24989-7","article-title":"Cryptic taxonomic diversity and high-latitude melanism in the glossiphoniid leech assemblage from the Eurasian Arctic","volume":"12","author":"Bolotov","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"79","DOI":"10.3391\/ai.2015.10.1.08","article-title":"Parasite host-switching from the native American red-eared slider, Trachemys scripta elegans, to the native Mediterranean pond turtle, Mauremys leprosa, in natural environments","volume":"10","author":"Meyer","year":"2015","journal-title":"Aquat. Invasions"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3467","DOI":"10.1007\/s10530-019-02076-9","article-title":"Hitch me a ride: First report of the alien leech Helobdella octatestisaca in Europe associated with freshwater turtles","volume":"21","author":"Perera","year":"2019","journal-title":"Biol. Invasions"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Kraus, F. (2009). Alien Reptiles and Amphibians: A Scientific Compendium and Analysis, Springer.","DOI":"10.1007\/978-1-4020-8946-6"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3459","DOI":"10.1007\/s00436-020-06809-x","article-title":"Host-parasite association of Placobdella costata (Glossiphoniidae: Hirudinea) and Mauremys leprosa (Geoemydidae: Testudinoidea) in aquatic ecosystems of Morocco","volume":"119","author":"Laghzaoui","year":"2020","journal-title":"Parasitol. Res."},{"key":"ref_29","unstructured":"Smit, C. (2022). Apicomplexan Diversity of Selected Southern African Snakes and Terrapins. [Master\u2019s Thesis, North-West University]."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Al-Quraishy, S., Abdel-Ghaffar, F., Dkhil, M.A., and Abdel-Gaber, R. (2021). Haemogregarines and Criteria for Identification. Animals, 11.","DOI":"10.3390\/ani11010170"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1645\/GE-2470.1","article-title":"Molecular Characterization of Hepatozoon Species in Reptiles from the Seychelles","volume":"97","author":"Harris","year":"2011","journal-title":"J. Parasitol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"292","DOI":"10.1654\/4594.1","article-title":"Molecular Detection of Hemolivia (Apicomplexa: Haemogregarinidae) from Ticks of North African Testudo graeca (Testudines: Testudinidae) and an Estimation of Their Phylogenetic Relationships Using 18S rRNA Sequences","volume":"80","author":"Harris","year":"2013","journal-title":"Comp. Parasitol."},{"key":"ref_33","first-page":"71","article-title":"Effect of Haemogregarina stepanovi Danilewsky, 1885 (Apicomplexa: Haemogregarinidae) on erythrocyte morphology in the European Pond Turtle, Emys orbicularis (Linnaeus, 1758) Blanford, 1876 (Testudines: Emydidae)","volume":"5","author":"Mihalca","year":"2004","journal-title":"Rev. Sci. Parasitol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"443","DOI":"10.1515\/acve-2015-0037","article-title":"Cytological and molecular identification of Haemogregarina stepanowi in blood samples of the European pond turtle (Emys orbicularis) from quarantine at Belgrade zoo","volume":"65","year":"2015","journal-title":"Acta Vet."},{"key":"ref_35","first-page":"22","article-title":"Haemoparasites of the genus Haemogregarina in a population of european pond turtles (Emys orbicularis) from Dr\u0103g\u0103\u015fani, V\u00e2lcea county, Romania","volume":"2","author":"Mihalca","year":"2002","journal-title":"Sci. Parasitol."},{"key":"ref_36","unstructured":"Thrall, M.A., Dale, C., Baker, E., and Lassen, E.D. (2004). Veterinary Hematology and Clinical Chemistry, Lippincott Williams & Wilkins."},{"key":"ref_37","first-page":"155","article-title":"Prevalence and intensity of Haemogregarina stepanowi (Apicomplexa: Haemogregarinidae) in two species of freshwater turtles (Mauremys caspica and Emys orbicularis) in Iran","volume":"2","author":"Javanbakht","year":"2014","journal-title":"J. Entomol. Zool. Stud."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"22","DOI":"10.4274\/tpd.galenos.2022.38257","article-title":"Prevalence of Haemogregarina stepanowi and Assessment of Some Risk Factors in Mauremys rivulata (Valenciennes, 1833) Freshwater Turtles (Testudines: Geoemydidae)","volume":"47","author":"Ceylan","year":"2023","journal-title":"Turk. Parazitolojii Derg."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"731","DOI":"10.2478\/s11756-012-0069-y","article-title":"Coexistence of Placobdella costata (Fr. M\u00fcller, 1846) (Hirudinida: Glossiphoniidae) and mud turtle Emys orbicularis","volume":"67","author":"Bielecki","year":"2012","journal-title":"Biologia"},{"key":"ref_40","unstructured":"Lukin, E.I. Pijavki presnych i solonovatich vodojemov. In Fauna USSR. Pijavki; Academy of Science of the USSR, Leningrad, UK, 1976; 484p."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"259","DOI":"10.2478\/s11686-014-0232-4","article-title":"Presence of the leech Placobdella costata in the south of the Iberian Peninsula","volume":"59","author":"Romero","year":"2014","journal-title":"Acta Parasitol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"506","DOI":"10.4172\/2155-9546.1000506","article-title":"Placobdella costata an Ectoparasite for Mauremys caspica in North of Iran","volume":"8","author":"Bashirichelkasari","year":"2017","journal-title":"J. Aquac. Res. Dev."},{"key":"ref_43","first-page":"949","article-title":"First record of Mauremys sinensis (Gray, 1834) and its natural overwintering in Central Europe","volume":"11","author":"Jablonski","year":"2018","journal-title":"Herpetol. Notes"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1654\/1525-2647-85.1.48","article-title":"The Occurrence of Protozoan Parasites (Schellackia sp. Reichenow, 1919, Tritrichomonas sp. Kofoid, 1920, and Proteromonas sp. Kunstler, 1883) in Lacertid Lizards from Selected Localities in Europe","volume":"85","year":"2018","journal-title":"Comp. Parasitol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"337","DOI":"10.2478\/s11686-012-0048-z","article-title":"Apicomplexa primers amplify Proteromonas (Stramenopiles, Slopalinida, Proteromonadidae) in tissue and blood samples from lizards","volume":"57","author":"Maia","year":"2012","journal-title":"Acta Parasitol."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/11\/4\/1063\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:18:48Z","timestamp":1760123928000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/11\/4\/1063"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,4,19]]},"references-count":45,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2023,4]]}},"alternative-id":["microorganisms11041063"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms11041063","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,4,19]]}}}