Recently, single nucleotide polymorphisms (SNPs) have been used to identify genes or genomic regions responsible for economic traits, including genetic diseases in domestic animals, and to examine genetic diversity of populations. In this study, we genotyped 70 chicken autosomal SNPs using DigiTag2 assay to understand the genetic structure of the Japanese native chicken breeds Satsumadori and Ingie, and the relationship of these breeds with other established breeds, Rhode Island Red (RIR), commercial broiler and layer. Five breeds, each consisting of approximately 20 chickens, were subjected to the assay, revealing the following: Average expected heterozygosities of broiler, Satsumadori, RIR, layer and Ingie were 0.265, 0.254, 0.244, 0.179 and 0.176, respectively. Phylogenetic analysis using the concatenated 70 autosomal SNP genotypes distinguished all chickens and formed clusters of chickens belonging to the respective breeds. In addition, the 2-D scatter plot of the first two principal components was consistent with the phylogenic tree. Taken together with the pairwise F(st) distances, broiler and RIR were closely positioned near each other, while Ingie was positioned far from the other breeds. Structure analysis revealed that the probable number of genetic clusters (K) was six and four with maximum likelihood and ΔK values, respectively. The clustering with maximum likelihood revealed that, in addition to the clustering of the other five breeds, the Satsumadori was subdivided into two genetic clusters. The clustering with ΔK value indicated that the broiler and Rhode Island Red were assigned to the same genetic cluster.
We surveyed reservoir animals of leptospires in the northern part of Miyazaki Prefecture, where a cluster of human leptospirosis had occurred during the summer of 2006. Leptospira was isolated from 6 of 57 large Japanese field mice (Apodemus speciosus). The serogroups of the isolates were Autumnalis (5 strains) and Hebdomadis (1 strain) and the partial nucleotide sequences of their flaB genes suggested that the isolates belonged to L. interrogans. The human patient sera reacted specifically with the Leptospira strain isolated from the mice captured around the area where each patient occurred, suggesting that mice are the source of human infection. We also detected leptospiral DNAs by flaB-polymerase chain reaction in the kidneys of large feral animals; wild boars (positive ratio 10.3%; 4 of 39) and deer (19.2%; 10 of 52). The Leptospira spp. harbored by these animals were deduced to be L. interrogans (in 5 animals) and L. borgpetersenii (in 9 animals) by the nucleotide sequences of the amplicons. Anti-Leptospira antibodies were also detected among symptomatic hound dogs. These results suggest that these feral animals may cause leptospirosis and pose a potential risk to hunters and workers in the meat processing industry.
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