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.

Epidemiology ofCryptosporidiuminfection in cattle in China: a review

“…The causative agents include bacteria such as enterotoxigenic Escherichia coli (ETEC), viruses such as rotavirus, and parasites or other possible factors [10, 19]. Cryptosporidium , as an important protozoan parasite, can cause parasitic diarrhea in animals.…”

Molecular characterization of <i>Cryptosporidium parvum</i> detected in Japanese black and Holstein calves in Iwate Prefecture and Tanegashima Island, Kagoshima Prefecture, Japan

“…Infections of Cryptosporidium parvum , C. bovis , C. ryanae and C. andersoni have been reported in cattle [ 5 , 8 , 9 , 13 ]. The zoonotic subtype C. parvum [ 13 ] causes severe watery diarrhea in calves [ 3 ]. Although C. parvum has traditionally been diagnosed by the size and morphology of oocysts detected in fecal samples, the oocysts of C. parvum cannot be discriminated from those of C. bovis and C. ryanae , which are most likely asymptomatic [ 10 ].…”

“…IIa, IId and IIl subtype families have been detected in calves worldwide, with the zoonotic subtype IIaA15G2R1 of the IIa subtype family most frequently reported [ 13 ]. Previous studies on C. parvum GP60 sequences from Hokkaido [ 5 ], Gifu [ 1 ], Hyogo [ 9 ] and Okinawa [ 3 ] Prefectures, Japan, indicated that IIaA15G2R1 was the only subtype distributed in these regions. However, the number of samples and locations analyzed in those studies were insufficient to determine the genetic variation of C .…”

Direct evidence of preventive effect of severe diarrhea in calf by milk replacer-based probiotic feeding

“…Among those factors, bovine rotavirus (BRV) infection and bovine cryptosporidiosis are prevalent among cattle in Japan, and cause severe diarrhea in calf [1]. Bovine rotavirus infects cattle of all ages, but calf has a higher incidence and more severe symptoms [2]. Bovine cryptosporidiosis with diarrhea caused by Cryptosporidium parvum results in decreased appetite, weakness, and dehydration in calf [3].…”