Fertility of ewes inseminated intrauterinally with frozen semen using extender containing bovine serum albumin

The present study was conducted to examine the fertility of ewes inseminated intrauterinally with frozen semen using semen extender containing either egg yolk or bovine serum albumin (BSA). Sixty Suffolk and cross-bred ewes were treated with controlled internal drug release (CIDR) devices during the non-breeding season (July 2006). A CIDR was inserted into the vagina for 12 days and an intramuscular injection of 500 IU equine chorionic gonadotropin was administered one day before its removal. Ejaculates from a suffolk ram were diluted with a Tris-based extender containing either 15% (v/v) egg yolk or 10% (w/v) BSA, and the diluted semen was frozen in 0.25 ml straws. A fixed-time intrauterine artificial insemination (AI) was performed 43-47 h after CIDR removal, regardless of incidence of estrus. There was no significant difference in pregnancy rates at 60 days after AI between the extenders containing egg yolk (66.7%, 20/30 animals) or BSA (65.5%, 19/29 animals). Furthermore, there were no significant difference in the lambing rates (66.7% and 62.1%) and prolificacy (1.25 and 1.56) between the two semen extenders. The present study indicates that a semi-defined semen extender containing 10% BSA produces fertility after intrauterine AI that is similar to that achieved with semen extender containing egg yolk.     

Fate mapping of gallbladder progenitors in posteroventral foregut endoderm of mouse early somite-stage embryos

In early embryogenesis, the posteroventral foregut endoderm gives rise to the budding endodermal organs including the liver, ventral pancreas and gallbladder during early somitogenesis. Despite the detailed fate maps of the liver and pancreatic progenitors in the mouse foregut endoderm, the exact location of the gallbladder progenitors remains unclear. In this study, we performed a DiI fate-mapping analysis using whole-embryo cultures of mouse early somite-stage embryos. Here, we show that the majority of gallbladder progenitors in 9–11-somite-stage embryos are located in the lateral-most domain of the foregut endoderm at the first intersomite junction level along the anteroposterior axis. This definition of their location highlights a novel entry point to understanding of the molecular mechanisms of initial specification of the gallbladder.     

Spontaneous Rhabdomyosarcoma in a Common Marmoset ( Callithrix jacchus )

The common marmoset (Callithrix jacchus) is now widely used in various research fields, including toxicology. However, information about the background pathology of this species is scarce. Here, we report a case of rhabdomyosarcoma that spontaneously occurred in a common marmoset. A 44-month-old male common marmoset was euthanized due to bilateral hind limb paralysis. At necropsy, a 2×2×5-cm intramuscular mass was observed in the lower right back. Histologically, the mass was mainly composed of interlacing bundles of spindle-shaped tumor cells. Immunohistochemically, the tumor cells were positive for myogenin, desmin, vimentin and alpha-smooth muscle actin. Ultrastructurally, the tumor cells contained bundles of myofilaments with Z-band-like structures. Thus, the tumor was diagnosed as a rhabdomyosarcoma. To our knowledge, this is the first report of spontaneous rhabdomyosarcoma that was definitely diagnosed in the common marmoset.     

RNAi-mediated Knockdown of Xist Does Not Rescue the Impaired Development of Female Cloned Mouse Embryos

In mice, one of the major epigenetic errors associated with somatic cell nuclear transfer (SCNT) is ectopic expression of Xist during the preimplantation period in both sexes. We found that this aberrant Xist expression could be impeded by deletion of Xist from the putative active X chromosome in donor cells. In male clones, it was also found that prior injection of Xist-specific siRNA could significantly improve the postimplantation development of cloned embryos as a result of a significant repression of Xist at the morula stage. In this study, we examined whether the same knockdown strategy could work as well in female SCNT-derived embryos. Embryos were reconstructed with cumulus cell nuclei and injected with Xist-specific siRNA at 6–7 h after oocyte activation. RNA FISH analysis revealed that siRNA treatment successfully repressed Xist RNA at the morula stage, as shown by the significant decrease in the number of cloud-type Xist signals in the blastomere nuclei. However, blastomeres with different sizes (from “pinpoint” to “cloud”) and numbers of Xist RNA signals remained within single embryos. After implantation, the dysregulated Xist expression was normalized autonomously, as in male clones, to a state of monoallelic expression in both embryonic and extraembryonic tissues. However, at term there was no significant improvement in the survival of the siRNA-injected cloned embryos. Thus, siRNA injection was largely effective in repressing the Xist overexpression in female cloned embryos but failed to rescue them, probably because of an inability to mimic consistent monoallelic Xist expression in these embryos. This could only be achieved in female embryos by applying a gene knockout strategy rather than an siRNA approach.     

Method for estimating maximum permissible load weight for Japanese native horses using accelerometer‐based gait analysis

The aim of this study was to establish a method for estimating loading capacity for Japanese native horses by gait analysis using an accelerometer. Six mares of Japanese native horses were used. The acceleration of each horse was recorded during walking and trotting along a straight course at a sampling frequency of 200 Hz. Each horse performed 12 tests: one test with a loaded weight of 80 kg (First 80 kg) followed by 10 tests with random loaded weights between 85 kg and 130 kg and a final test with a loaded weight of 80 kg again. The time series of acceleration was subjected to fast Fourier transformation, and the autocorrelation coefficient was calculated. The first two peaks of the autocorrelation were defined as symmetry and regularity of the gait. At trot, symmetries in the 100, 110, and 125 kg tests were significantly lower than that in First 80 kg (P < 0.05, by analysis of covariance and Sidak's test). These results imply that the maximum permissible load weight is less than 100 kg, which is 29% of the body weight of Japanese native horses. Our method is a widely applicable and welfare‐friendly method for estimating maximum permissible load weights of horses.     

日本湿润的温带草地根际和微生物呼吸对土壤CO2排放的贡献及环境影响因素

Soil CO2 efflux, root mass, and root production were investigated in a humid temperate grassland of Japan over a growing season (Apr. to Sep.) of 2005 to reveal seasonal changes of soil CO2 efflux, to separate the respective contributions of root and microbial respiration to the total soil CO2 efflux, and to determine the environmental factors that control soil respiration. Minimal microbial respiration rate was estimated based on the linear regression equations between soil CO2 efflux and root mass at different experimental sites. Soil CO2 efflux, ranging from 4.99 to 16.29 μmol CO2 m-2 s-1, depended on the seasonal changes in soil temperature. The root mass at 0--10 cm soil depth was 0.82 and 1.27 kg m-2 in Apr. and Sep., respectively. The root mass at 0--10 cm soil depth comprised 60% of the total root mass at 0--50 cm soil depth. The root productivity at 0--30 cm depth varied from 8 to 180 g m-2 month-1. Microbial and root respiration rates ranged from 1.35 to 5.51 and 2.72 to 12.06 μmol CO2 m-2 s-1, respectively. The contribution of root respiration to the total soil CO2 efflux averaged 53%, ranging from 33% to 72%. The microbial respiration rate was exponentially related to soil temperature at 10 cm depth (R2 = 0.9400, P = 0.002, n = 6), and the root respiration rate was linearly related to the root production at 0--30 cm depth (R2 = 0.6561, P = 0.042, n = 6).     

Ocular lesions in Watanabe heritable hyperlipidemic rabbits

Hyperlipidemic ocular lesions are described for Watanabe heritable hyperlipidemic (WHHL) rabbits. Male WHHL rabbits 8 months old exhibited serum hyperlipidemia and ophthalmoscopically yellowish-white lesions along the corneoscleral junction and in the iris. Histopathologically, foamy macrophages aggregated in the stroma of the cornea, iris, and ciliary body were observed. These findings have been interpreted as lipid keratopathy. In addition, multiple clusters of a large number of foamy macrophages occurred throughout the choroid and sclera in association with the blood vessels. The lesions in the choroid and sclera could not be detected ophthalmoscopy, yet were much more prominent than those in the cornea, iris, and ciliary body, suggesting greater involvement and earlier onset of lipidosis at these sites associated with hyperlipidemia in WHHL rabbits.     

Hypofractionated radiation therapy in the treatment of canine thymoma: Retrospective study of eight cases

Thymomas are one of the most common tumors of the cranial mediastinum in dogs; however there is limited information available on the use of radiation therapy for treating this neoplasm. Objectives of the current retrospective observational study were to describe outcomes and side effects of a hypofractionated radiation therapy protocol in a group of dogs with confirmed thymoma. A total of eight dogs were included. To generate individualized treatment plans, we designed the planning target volume according to the limits on mean lung dose and the percentage of the total lung volume exceeding 20 Gy (V20). The total administered dose was 48–49 Gy, with one fraction per week for a total of six to seven fractions. After therapy, two dogs achieved complete responses, two achieved partial responses, and the disease remained stable in two. Two dogs died during the radiation therapy protocol and were not classified. The median mean lung dose and V20 were 6.0 Gy (range: 3.1–15.0 Gy) and 12.4% (range: 2.3–27.5%), respectively. The overall response rate was 50.0%, and the median time to response following treatment initiation was 22 days (range: 14–115 days). Acute and late side effects were common in the skin and/or lung and were self‐limiting or asymptomatic. The median survival time was not reached (range: 8–1128 days) and the 1 year survival rate was 75.0%. Hypofractionated radiation therapy was well tolerated in this sample of dogs with thymoma and may be considered when owners decline surgical treatment or the tumor is deemed unresectable.