The Miyako and Yonaguni horses are native horses in Okinawa. Here, we evaluated their genetic relationship using microsatellite data and Kiso horses, which have four subpopulations, as a reference population for evaluating this relationship. Microsatellite data from 35 Miyako, 78 Yonaguni, and 172 Kiso horses were evaluated using the STRUCTURE software for analyzing multilocus genotype data to investigate the population structures and their underlying relationship. The results of the STRUCTURE analysis were stable when ΔK was 2, suggesting that the Okinawan horses are different from the Kiso horses. Moreover, the results were also stable when ΔK was 6; the sample was then divided into four subpopulations of the Kiso horses and two Okinawan horse breeds. However, the diagrams from the STRUCTURE analysis were unstable when ΔK was 3. These results suggest that the genetic relationship of the Okinawan horse breeds may be close, similar to that among the subpopulations of the Kiso horses.
The Iranian Turkoman horse is considered one of the major categories of Iranian native horses. The aim of this research was to study the genetic and nongenetic factors affecting body conformation traits in Iranian Turkoman horses. For this, measurements were performed on body conformation traits of 121 horses. To study the effect of nongenetic factors, the Lsmeans procedure was used. Variance components and heritability were estimated by restricted maximum likelihood method (REML) and AI-REML convergence algorithm. The standard deviation was estimated to be 7.04 cm for croup depth and 2.08 cm for chest width, and the coefficient of variation was estimated to be 11.27% for croup length and 2.58% for withers height. Sex had a significant effect on head-neck circumference (P<0.05), withers height (P<0.05), chest width (P<0.05), and croup height (P<0.05). Province had a significant effect on withers height (P<0.05) and croup depth (P<0.05). The lowest and highest heritability estimates were for head-neck circumference (0.12 ± 0.06) and neck-body circumference (0.33 ± 0.09), respectively. The lowest and highest additive genetic variance estimates were for head length (0.64) and pelvis width (18.34), respectively. Generally, the medium to high estimated heritability for the traits in this study indicate that genetic improvement would be possible in these traits.
In order to promote conservation of the traditional Tokara horse in its remaining three breeding areas in Japan (Nakanoshima, Kaimondake, and Iriki), we genotyped 123 horses using 31 microsatellite markers and determined their genetic diversity. On average, the number of alleles (NA), observed heterozygosity (HO), expected heterozygosity (HE), and inbreeding coefficient (FIS) among all horses were 3.0, 0.424, 0.481, and 0.108, respectively. Compared with other endangered horse breeds, we found that, even though the size of the Tokara horse population has recently increased, the NA, HO, and HE of Tokara horses are still notably lower than those of other breeds. Neighbor-joining tree and STRUCTURE analysis showed that the current population of Tokara horses is divided into three subpopulations, corresponding to their respective feeding and breeding areas: Nakanoshima, Kaimondake, and Iriki. This subdivision was also reflected in the NA of microsatellite DNAs, with four, three, and four different loci showing single alleles in Nakanoshima, Kaimondake, and Iriki horses, respectively. These alleles are considered to have become fixed as a consequence of breeding within the limited number of horses in each area. Since Tokara horses are currently strongly divided into subpopulations, it is vitally important to exchange several horses among their different breeding units in order to maintain the genetic diversity of the Tokara horse as a unique breed. The data obtained in this study contribute toward explaining the history of Tokara horses and also provide important information for future monitoring of population diversity and guiding conservation measures for this endangered breed.
Equine scintigraphy has been legally permitted in Japan since 2009; however, it has not yet been a routine modality for horses. One reason is the legal regulations concerning the disposal of contaminated bedding. However, overseas, the bedding after scintigraphy can be disposed following radioactivity decay, but this is not allowed in Japan. Therefore, beddings are required to stored permanently in a controlled area, implying that large amounts of beddings such as straw would be kept untreated, which is quite unpractical. This may cause a hospital owner to hesitate to construct an equine scintigraphy facility. Therefore, it is proposed that water-dispersed paper bedding is disposed as aqueous waste after radioactivity decay. The purpose of this study was to check the availability of bedding, thus radioisotopes were not used in this study. Three horses were housed individually in stalls covered with water-dispersed paper bedding for 48 hr. Physical condition, including body weight, was monitored, and a complete blood cell count and biochemical analysis were conducted. The results revealed that physical conditions and results of blood analysis were all stable within the normal range, and the veterinarian did not find any specific abnormality in any of the three horses. No marked changes in the levels of blood cortisol were observed before and after stalling, suggesting almost no stress for the horses. Because the water-dispersed paper bedding did not negatively affect the horses, it can be used as a substitute for conventional straw bedding.
In order to establish an efficient system for serological diagnosis of equine viral arteritis in Japan, we compared enzyme-linked immunosorbent assays (ELISAs) provided by two manufacturers (Nisseiken Co., Ltd., Tokyo, Japan, and VMRD Inc., Pullman, WA, U.S.A.) by testing a series of horse sera. The results revealed that 159 of 160 virus-neutralizing (VN) antibody-positive serum samples were positive in both the Nisseiken-ELISA and VMRD-ELISA. Of the VN-negative sera (n=157), 134 and 154 samples were negative in the Nisseiken-ELISA and VMRD-ELISA, respectively. Sensitivity was 99.4% for both the Nisseiken-ELISA and VMRD-ELISA. The specificity of the VMRD-ELISA (98.1%) was significantly higher than that of the Nisseiken-ELISA (85.4%, P<0.05). The diagnostic performance of the VMRD-ELISA was superior to that of the Nisseiken-ELISA because of this greater specificity.
Synovium-derived mesenchymal stromal cells (SM-MSCs) from seven Thoroughbreds with naturally occurring intra-articular fracture proliferated to over ten million cells by the second passage. Using three experimental Thoroughbreds, columnar osteochondral defects were made arthroscopically at the bilateral distal radius. Five million allogenic SM-MSCs were implanted into the right defect, and another five million were injected into the right radio-carpal joint (implantation site). No SM-MSCs were implanted into the left defect or the same joint (control site). At 3 and 6 weeks after surgery, ten million autologous SM-MSCs were injected into the right joints. Radiolucent volumes of defects calculated by analysis of postmortem CT images 9 weeks after surgery were decreased in implanted sites compared with control sites in all horses. The average scores for ICRS gross and histopathological grading scales in implanted sites were equal to or higher than those of the controls. These results suggest that allogenic implantation and subsequent autologous injection of SM-MSCs might not obstruct subchondral bone formation in defects.