title: Molecular characterization of Cryptosporidium parvum from two different Japanese prefectures, Okinawa and Hokkaido.
abstract: Infectious diarrhea is the most frequent cause of morbidity and mortality in neonatal calves. Cryptosporidium parvum is one of the main pathogens associated with calf diarrhea. Although diarrhea is a symptom of infection with various pathogens, investigations to detect the types of pathogens have never been performed in Japan. This study investigated the prevalence of four major diarrhea-causing pathogens in calves: C. parvum, rotavirus, coronavirus, and enterotoxigenic Escherichia coli (E. coli K99). Commercial immunochromatography testing of all four pathogens and molecular analysis of C. parvum with diarrhea in calves from southernmost Okinawa and northernmost Hokkaido, Japan, were conducted. The frequencies of C. parvum, rotavirus, coronavirus, and E. coli (K99) in Okinawa were 50%, 28%, 2.3%, and 4.7%, respectively. Watery fecal stools were significantly correlated with C. parvum (p<0.05). In oocyst calculations for C. parvum, no significant difference was observed between the single-infection cases and the mixed-infection cases with rotavirus. Interestingly, molecular analyses targeting small subunit ribosomal RNA as well as glycoprotein 60 (GP60) genes revealed that the C. parvum nucleotide sequences from the two prefectures were identical, indicating that C. parvum with a uniform characteristic is distributed throughout Japan. GP60 subtyping analysis identified C. parvum from Okinawa and Hokkaido as belonging to the IIaA15G2R1 subtype, a known zoonotic subtype. Hence, control of cryptosporidiosis is important not only for pre-weaned calves, but also for human health.
authors.affiliation: Laboratory of Veterinary Parasitology, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka 020-8550, Japan; National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-13 Inada-cho, Obihiro 080-8555, Japan.
authors.forename: Madoka
authors.initials: M
authors.lastname: Ichikawa-Seki
authors.affiliation: Laboratory of Veterinary Parasitology, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka 020-8550, Japan.
authors.forename: Junya
authors.initials: J
authors.lastname: Aita
authors.affiliation: Transboundary Animal Diseases Center, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan; National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-13 Inada-cho, Obihiro 080-8555, Japan.
authors.forename: Tatsunori
authors.initials: T
authors.lastname: Masatani
authors.affiliation: Yaeyama Livestock Hygiene Service Center, 1-2 Miyara, Ishigaki, Okinawa 907-0243, Japan.
authors.forename: Moemi
authors.initials: M
authors.lastname: Suzuki
authors.affiliation: Yaeyama Livestock Hygiene Service Center, 1-2 Miyara, Ishigaki, Okinawa 907-0243, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Nitta
authors.affiliation: Tamayose Veterinary Hospital, 204-332 Maezato, Ishigaki, Okinawa 907-0002, Japan.
authors.forename: Genta
authors.initials: G
authors.lastname: Tamayose
authors.affiliation: Laboratory of Veterinary Parasitology, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka 020-8550, Japan.
authors.forename: Takehiro
authors.initials: T
authors.lastname: Iso
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-13 Inada-cho, Obihiro 080-8555, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Suganuma
authors.affiliation: Honbetsu Veterinary Clinical Center, Tokachi Agricultural Mutual Aid Association, Honbetsu 089-3324, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Fujiwara
authors.affiliation: Hokubu Veterinary Clinical Center, Tokachi Agricultural Mutual Aid Association, Asyoro 089-3708, Japan.
authors.forename: Keita
authors.initials: K
authors.lastname: Matsuyama
authors.affiliation: Hokubu Veterinary Clinical Center, Tokachi Agricultural Mutual Aid Association, Asyoro 089-3708, Japan.
authors.forename: Tadamasa
authors.initials: T
authors.lastname: Niikura
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-13 Inada-cho, Obihiro 080-8555, Japan.
authors.forename: Naoaki
authors.initials: N
authors.lastname: Yokoyama
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-13 Inada-cho, Obihiro 080-8555, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Suzuki
authors.affiliation: Yubetsu Herd Management Service, Baro, Yubetsu-cho, Hokkaido 093-0731, Japan.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Yamakawa
authors.affiliation: Department of Clinical Veterinary Science, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-11 Inada-cho, Obihiro 080-8555, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Inokuma
authors.affiliation: Laboratory of Veterinary Parasitology, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka 020-8550, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Itagaki
authors.affiliation: Yaeyama Livestock Hygiene Service Center, 1-2 Miyara, Ishigaki, Okinawa 907-0243, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Zakimi
authors.affiliation: National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Nishi 2-13 Inada-cho, Obihiro 080-8555, Japan. Electronic address: nisikawa@obihiro.ac.jp.
authors.forename: Yoshifumi
authors.initials: Y
authors.lastname: Nishikawa
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000453:epidemiology / Q000469:parasitology*; D003457:Cryptosporidiosis / Q000453:epidemiology / Q000469:parasitology*; D016785:Cryptosporidium parvum / Q000235:genetics*; D016054:DNA, Protozoan / Q000235:genetics; D003967:Diarrhea / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary; D007564:Japan / Q000453:epidemiology; D017720:Molecular Epidemiology; D010802:Phylogeny
chemical_list: D016054:DNA, Protozoan
keywords: Cryptosporidium parvum; GP60; Immunochromatography test; Japan; SSUrRNA