In horse management, the alarm system with sensors in the foaling period enables the breeder can appropriately prepare the time of the parturition. It is important to prevent losses by unpredictable parturition because there are several high risks such as dystocia and the death of foals and mares during foaling. However, unlike analysis in the alarm system that detects specific motions has been widely performed, analysis of classification following specific behavior patterns or number needs to be more organized. Thus, the objective of this study is to classify signs of the specific behaviors of the mares for the prediction of pre-foaling behaviors. Five Thoroughbred mares (9-20 yrs) were randomly selected for observation of the prefoaling behaviors. The behaviors were monitored for 90 min that was divided into three different periods as 1) from -90 to -60 min, 2) from -60 to -30 min, 3) from -30 min to the time for the discharge of the amniotic fluid, respectively. The behaviors were divided into two different categories as state and frequent behaviors and each specific behavioral pattern for classification was individually described. In the state behaviors, the number of mares in the standing of the foaling group (3.17 ± 0.18 b ) at period 3 was significantly higher than the control group (1.67 ± 0.46 a ). In contrast, the number of the mares in the eating of the foaling group (1.17 ± 0.34 b ) at period 3 was significantly lower than the control group (3.33 ± 0.46 a ). In the frequent behaviors, the weaving of the foaling group was significantly higher than the control group, and looking at the belly of the foaling group was significantly lower than the control group. In period 2, defecation, weaving, and lowering the head of the foaling group were significantly higher than the control group, respectively. In period 3, sitting down and standing up, pawing, weaving, and lowering the head in the foaling group were also significantly higher than the control group. In conclusion, the behavior is significantly different in foaling periods, and the prediction of foaling may be feasible by the detection of the pre-foaling behaviors in the mares.

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Molecular markers can be used to identify and isolate specific developmental stages of germ cells and Leydig cells. Protein gene product (PGP)9.5 expression in spermatogonia and Leydig cells has been reported in several species. The stages of spermatogonia and Leydig cells expressing PGP9.5 vary depending on the species and reproductive stages. Thus, the objectives of this study were (1) to identify the localization of PGP9.5 in donkey testicular cells, and (2) to compare the expression patterns of PGP9.5 in donkey testicular cells between pre- and post-pubertal stages. Testes samples were collected following the routine field castration of six donkeys. Western blotting was performed to verify the cross-reactivity of the rabbit anti-human PGP9.5 antibody to donkey testes. Immunofluorescence was performed to investigate the expression pattern of PGP9.5 in testicular tissues at different reproductive stages. In Western blotting, the protein band of the PGP9.5 antibody appeared at approximately 27 kDa, whereas the band was not observed in the negative control treated with normal mouse IgG. In the pre-pubertal stage, the expression of deleted in azoospermia-like (DAZL) was found in some spermatogonia in pre-pubertal testicular tissues. However, the immunolabeling of PGP9.5 in testicular tissue was not observed in the seminiferous tubules. In stages 1 and 2, spermatogonia were immunolabeled with either PGP9.5 or DAZL. In contrast, PGP9.5 and DAZL were co-immunolabeled in some of the spermatogonia in stages 3 to 8. Interestingly, some Leydig cells were immunolabeled with PGP9.5 in both pre- and post-pubertal stages. In conclusion, the PGP9.5 antibody can be used as a tool to identify and isolate spermatogonia from seminiferous tubules.

Olfactory receptors (ORs) are the largest family of Gprotein-coupled receptors in the human genome. Almost 1,000 OR genes exist within the entire human genome (Firestein, 2001). Vertebrates can detect a variety of chemicals by smell, which is recognized by ORs expressed in the olfactory epithelium (OE) and pheromone receptors located in the vomeronasal organ (VNO) (Mombaerts, 1999;Fleischer et al., 2009). Although ORs are primarily expressed in chemosensory organs, many ORs have been detected and play a role in non-olfactory tissues. For ex-ample, the human OR hOR17-4 expressed in the testes mediates the chemical communication between spermatozoa and eggs (Spehr et al., 2003). In mice, MOR23 is expressed in the muscles during myogenesis and muscular regeneration (Griffin et al., 2009). ORs are also expressed in human prostate glands (Neuhaus et al., 2009), mice uterus (Kim et al., 2009), rat cardiac tissues (Drutel et al., 1995), mice autonomic nervous system ganglia (Weber et al., 2002), and rat placenta (Itakura et al., 2006). These findings suggest that the ectopic expression of ORs may play an important role in non-chemosensory tissues.Twenty genes that code for testicular ORs have been

Competing interestsNo potential conflict of interest relevant to this article was reported. Funding sourcesState funding sources (grants, funding sources, equipment, and supplies). Include name and number of grant if available.