GENETICS, EVOLUTION, AND PHYLOGENY - ORIGINAL PAPER Molecular identification of Eimeria hestermani and Eimeria prionotemni from a red-necked wallaby (Macropodidae; Macropus rufogriseus) in Japan Fitrine Ekawasti 1,2,3 & Kazuya Kitagawa 4 & Hiroshi Domae 4 & April Hari Wardhana 1,3,5 & Tomoyuki Shibahara 3,6 & Shigehiko Uni 7,8 & Masaharu Tokoro 9 & Kazumi Sasai 3,10 & Makoto Matsubayashi 3,5,10 Received: 25 September 2019 /Accepted: 29 January 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020 Abstract To date, more than 50 Eimeria spp. have been isolated from marsupials of the family Macropodidae. Although 18 species of Eimeria have been previously detected from multiple animal species belonging to the genus Macropus of the family, limited genetic analyses of the parasites are available, and their pathogenicity remains unclear. Here, we report the isolation of Eimeria spp. from a zoo specimen of red-necked wallaby (Macropodidae; Macropus rufogriseus). Specifically, two distinct types of Eimeria oocysts were recovered, one from the feces before treatment with an anthelmintic and the second from the intestinal contents after death of the animal. The oocysts obtained from the two sources were morphologically identified as E. hestermani and E. prionotemni, respectively. We successfully determined partial gene sequences from the two isolates, including segments of the 18S rRNA genes, and for the first time have used phylogenetic analyses of these sequences to assign the species to distinct clades. In combination with further genetic data, these results are expected to help elucidate the pathogenicity and host ranges of Eimeria spp. within the respective family and genus. Keywords Eimeria hestermani . Eimeria prionotemni . Japan . Macropodidae . Red-necked wallaby Introduction Eimeria spp. are protozoan parasites belonging to the phylum Apicomplexa that have been identified in a large number of vertebrate and invertebrate hosts. These parasites have been shown to possess strict host specificity and to develop only at specific sites, mainly the intestinal mucosa, within the host. Some species of Eimeria, especially those that infect live- stock, cause stereotypical clinical signs (known as coccidio- sis) including watery and/or bloody diarrhea, loss of appetite, All authors contributed equally in writing the manuscript. Section Editor: Berit Bangoura * Makoto Matsubayashi matsubayashi@vet.osakafu-u.ac.jp 1 Indonesian Research Center for Veterinary Science, Bogor 16114, Indonesia 2 Department of Parasitology, Veterinary Medicine, Gadjah Mada University, Yogyakarta 55281, Indonesia 3 Department of Veterinary Science, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka 598-8531, Japan 4 Ishikawa Zoo, Nomi, Ishikawa 923-1222, Japan 5 Department of Veterinary Parasitology, Faculty of Veterinary Medicine, Airlangga University, Surabaya 60115, Indonesia 6 Division of Pathology and Pathophysiology, National Institute of Animal Health, NARO, Tsukuba, Ibaraki 305-0856, Japan 7 Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia 8 Department of Public Health, Faculty of Nursing, Kobe Women’ s University, Kobe 650-0046, Japan 9 Department of Parasitology, Graduate School of Medical Sciences, Kanazawa University, Kanazawa 920-8640, Japan 10 Asian Health Science Research Institute, Osaka Prefecture University, Osaka 598-8531, Japan Parasitology Research https://doi.org/10.1007/s00436-020-06618-2