Coprological survey of parasitic infections in pigs and cattle in slaughterhouse in Osaka, Japan.

A coprological survey was performed at a slaughterhouse in Osaka, Japan, from 2004 to 2007 on 129 pigs reared in 8 prefectures, and on 213 cattle reared in 21 prefectures. Eimeria spp., Trichuris suis, Ascaris suum and Metastrongylus spp. infections were found in 52 (40.3%), 32 (24.8%), 19 (14.7%) and 3 pigs (2.3%), respectively, while Eimeria spp., Capillaria bovis and Trichuris sp. infections were detected in 163 (76.5%), 15 (7.0%) and 8 cattle (3.8%), respectively. Our results suggest that environmentally resistant oocysts and eggs of parasites could be widespread at the farms examined.

Genetical survey of novel type of Cryptosporidium andersoni in cattle in Japan.

Previously, we reported that an isolate of novel type of Cryptosporidium andersoni detected in cattle in Japan contained Type A (identical to C. andersoni reported previously) and Type B (having a thymine nucleotide insertion unlike the Type A) genotypes in the 18S rRNA gene. Here, we conducted an extensive investigation of Cryptosporidium infections in adult cattle in Japan from 2004 to 2007. Consequently, Cryptosporidium sp. were detected in 12 of the 205 cattle examined (5.9%), and partial sequences of the Cryptosporidium oocyst wall protein (COWP) gene in all isolates were identical to those of the previously reported data for C. andersoni whereas two signals were observed in the sequence of the partial 18S rRNA gene in all the isolates. In transmission studies using five of the isolates, they all infected SCID mice. Modified multiplex PCR using DNA of a single oocyst isolated from the infected SCID mice revealed that the partial sequences in the 18S rRNA gene of 40-80% of 10 isolates were identical to the Type A genotype of C. andersoni and those of other samples were identical to the Type B genotype. These results suggested that the C. andersoni novel type is widespread in cattle throughout Japan, and have multiple copies (Types A and B) in the 18S rRNA gene.

Detection of a mixed infection of a novel Cryptosporidium andersoni and its subgenotype in Japanese cattle.

Cryptosporidium oocysts were detected in the feces of cattle in Saga, Japan. Isolates were morphologically large. We attempted to identify the species or genotypes of the isolates by analyzing the partial sequences of the 18S rRNA and Cryptosporidium oocyst wall protein (COWP) genes, and measuring the infectivity in mice. The isolates showed 100% homology with Cryptosporidium andersoni in the COWP gene sequence and it could be transmitted to mice, but in the 18S rRNA gene, there was an additional signal in the ABI sequence chromatogram. To examine the additional signal, we analyzed both the 18S rRNA and the COWP gene sequences of a single oocyst passaged from mice using a modified multiplex PCR that was able to amplify both genes. As a result, it was revealed that two distinct genotypes (Types A and B) of a novel C. andersoni type existed in the 18S rRNA gene, whereas the COWP gene sequences of both oocysts were identical to C. andersoni. Although the sequence of the 18S rRNA gene of Type A was identical to that of C. andersoni, that of Type B had a thymine insertion and was not identical to any sequence registered with GenBank. Here we report that this is a new type of C. andersoni.