Developmental ability of oocytes retrieved from Meishan neonatal ovarian tissue grafted into nude mice

Ovarian xenografting makes it possible to obtain oocytes with fertilization ability from immature pigs of Western breeds. In this study, we applied these methods to the Meishan, an indigenous Chinese pig breed, and investigated the developmental competence of oocytes grown in their neonatal tissue after grafting into nude mice. First, mice harboring neonatal ovarian tissue were infused with follicle stimulating hormone (FSH) (62.5 U/ml) for 13 days starting at 10, 30, and 60 days after vaginal opening (D10‐, D30‐, and D60‐FSH groups, respectively). Development of antral follicles and their oocytes was most enhanced in the D60‐FSH group. For the next step, we examined the in vitro maturation ability of the oocytes recovered from host mice after infusion with FSH at a dose of 62.5 U/ml or 125 U/ml (FSH‐62.5 or ‐125 group) for 13 days starting at 60 days after vaginal opening. Many more oocytes with maturation ability were obtained from the FSH‐125 group. The FSH‐125 mature oocytes were fertilized in vitro, as shown by formation of male and female pronuclei, but did not reach the blastocyst stage. These results indicate that Meishan neonatal ovaries are able to produce oocytes with fertilization ability after being grafted into nude mice.     

Non-destructive Analysis of Oil-Contaminated Soil Core Samples by X-ray Computed Tomography and Low-Field Nuclear Magnetic Resonance Relaxometry: a Case Study

Non-destructive measurements of contaminated soil core samples are desirable prior to destructive measurements because they allow obtaining gross information from the core samples without touching harmful chemical species. Medical X-ray computed tomography (CT) and time-domain low-field nuclear magnetic resonance (NMR) relaxometry were applied to non-destructive measurements of sandy soil core samples from a real site contaminated with heavy oil. The medical CT visualized the spatial distribution of the bulk density averaged over the voxel of 0.31?×?0.31?×?2?mm(3). The obtained CT images clearly showed an increase in the bulk density with increasing depth. Coupled analysis with in situ time-domain reflectometry logging suggests that this increase is derived from an increase in the water volume fraction of soils with depth (i.e., unsaturated to saturated transition). This was confirmed by supplementary analysis using high-resolution micro-focus X-ray CT at a resolution of ～10?μm, which directly imaged the increase in pore water with depth. NMR transverse relaxation waveforms of protons were acquired non-destructively at 2.7?MHz by the Carr-Purcell-Meiboom-Gill (CPMG) pulse sequence. The nature of viscous petroleum molecules having short transverse relaxation times (T2) compared to water molecules enabled us to distinguish the water-saturated portion from the oil-contaminated portion in the core sample using an M(0)-T2 plot, where M(0) is the initial amplitude of the CPMG signal. The present study demonstrates that non-destructive core measurements by medical X-ray CT and low-field NMR provide information on the groundwater saturation level and oil-contaminated intervals, which is useful for constructing an adequate plan for subsequent destructive laboratory measurements of cores.     

Leydig cell hyperplasia in an ENU-induced mutant mouse with germ cell depletion

Repro22 is an N-ethyl-N-nitrosourea (ENU)-induced mutation in mice showing depletion of both male and female germ cells. In the present study, we investigated the male phenotypes of the mutant mouse at the adult stage. The repro22/repro22 homozygous mice showed reduced body weights as well as markedly reduced testis weights. Histological examination of the testes at 4 and 10 months of age showed no germ cells in the seminiferous tubules of the affected testis while a number of Sertoli cells were observed in the tubules. In addition to the germ cell depletion, the testes of the affected mouse contained expanded intertubular spaces that were filled by Leydig cell-like interstitial cells. These interstitial cells were confirmed to be Leydig cells by immunohistochmical staining using anti-3beta-HSD antibody. The estimated number of Leydig cells in the affected testes at 10 months of age increased approximately 2 fold compared with those of normal testes. Furthermore, the plasma testosterone levels of the affected mice at 10 months of age were significantly higher than those of the normal mice. These findings indicated that the repro22/repro22 mouse developed hyperplasia of Leydig cells that was presumably caused by the absence of germ cells in the seminiferous tubules.     

Experimental transmission of Bovine leukemia virus in cattle via rectal palpation

We examined whether Bovine leukemia virus (BLV) was transmitted by rectal palpation using a common sleeve between a BLV-infected cow and BLV-negative steers. Three of four steers developed antibodies against BLV as determined by agar-gel immunodiffusion (AGID) test between 7 to 10 weeks after the first rectal palpation using common sleeves from BLV-infected cow. In the steers, BLV proviral DNA were detected by PCR 1 to 5 weeks earlier than detection of the antibodies by the AGID test. Our experiments demonstrated that rectal palpation is a potential cause of BLV spread in herds and that detection of BLV proviral DNA in cattle by PCR is useful screening test for early diagnosis of BLV infection.     

Assessment of management of direct seeded rice production under different water conditions in Cambodia

In order to assess direct seeding of rice technology to cope with future agricultural labor shortage in Cambodia, agronomic experiments were conducted in 2005 and 2006 to compare direct seeding with transplanting under three water conditions (non-flooded, shallow flooded, and deep flooded conditions) with/without weed control by herbicides (bentazone and cyhalofop-butyl) for two Cambodian rice varieties (shorter stature and early maturity Sen Pidao, taller stature and longer maturity Phka Rumduol). Average rice yield in 2 years was lower in direct seeding (341 g m⁻²) than transplanting (404 g m⁻²), but interaction components with year, varieties, water conditions, and weed management were significant, and the attained maximum yield of direct seeding (510 and 464 g m⁻² for Phka Rumduol variety in shallow flooded condition with weeding in 2005 and 2006, respectively) was similar to that of transplanting. Plant length and dry weight of rice were reduced in non-flooded and deep flooded conditions compared with shallow flooded condition, and grain yield was the highest in shallow flooded condition. Yield advantage of Phka Rumduol over Sen Pidao increased under direct seeding, particularly under non-flooded conditions in 2005 because weed infestation was more suppressed in Phka Rumduol even without weeding. Increase in 100 g m⁻² of weed infestation prior to heading (dry weight basis) reduced about 20% of attainable yield with weed control. This study identified importance of stature and growth duration of rice varieties and presence of standing water as well as the weed control, in order to develop and extend direct seeding in the Cambodia.     

Initial vegetation recovery following a blowdown of a conifer plantation in monsoonal East Asia: Impacts of legacy retention, salvaging, site preparation, and weeding

All or a part of a sequence of forest practices (i.e., salvage logging, site preparation, planting crop trees, and weeding) has been implemented after natural disturbances for the rapid re-establishment of tree cover. Forest policies in Japan have recently changed from monocultural planting of coniferous crop trees to planting native broadleaved trees to restore forests and nurture local biodiversity following large windthrows. However, the effects of this new practice on preserving biodiversity, as well as the effects of legacy retention, have never been verified in Asia. Thus, the objective of our research was to compare the effects of legacy retention with plantation after salvaging on the initial stage of vegetation recovery in a blowdown area, specifically focusing on plant species diversity, the occurrence of alien species, and the composition of plant species. Following the analysis of our results, we finally describe appropriate practices to alter disturbed coniferous plantations to bring the species composition closer to that of the original natural mixed forests.A control (A, legacy retention) and three experimental treatment sites (B, salvage logged, site prepared, and Quercus crispula seedlings planted; C, same as B, but weeded once during the summer; and D, residual rows that emerged after establishing sites for planting) were prepared, and quadrats were set. Eleven indicators of the ground condition and the number of vascular plant species, including ferns, were quantified, and the number and abundance of residual and newly colonized plants of the main woody species were estimated.Our main findings were as follows: (1) in unsalvaged sites and residual rows, the diversity of plant species was poor, but a variety of plant species compositions were observed due to the heterogeneous conditions of the ground and ample residual plants; (2) in the planting site, many species appeared, but little variety of the species composition was observed due to the homogeneous condition of the ground and the destruction of residual plants; (3) a large number of alien species emerged in broad, unvegetated areas; (4) the impact of site preparation overwhelmed the impact of salvage logging on the initial recovery of plant species; and (5) to restore a natural mixed forest, a combination of legacy retention and plantation after salvaging would be the most appropriate.     

Where water is oxidized to dioxygen: structure of the photosynthetic Mn4Ca cluster

The oxidation of water to dioxygen is catalyzed within photosystem II (PSII) by a Mn(4)Ca cluster, the structure of which remains elusive. Polarized extended x-ray absorption fine structure (EXAFS) measurements on PSII single crystals constrain the Mn(4)Ca cluster geometry to a set of three similar high-resolution structures. Combining polarized EXAFS and x-ray diffraction data, the cluster was placed within PSII, taking into account the overall trend of the electron density of the metal site and the putative ligands. The structure of the cluster from the present study is unlike either the 3.0 or 3.5 angstrom-resolution x-ray structures or other previously proposed models.     

Blood biochemical values in Japanese Black breeding cows in Kagoshima Prefecture,Japan

To obtain blood biochemical basic data of Japanese Black breeding cows in Kagoshima Prefecture, Japan, blood samples were obtained from 857 clinically healthy multiparous cows on 71 farms. Cows were divided into three stages: the prepartum stage (between 20 and 80 days before parturition, n=290), lactation stage (between 20 and 80 days after parturition, n=283) and maintenance stage (between 120 and 200 days after parturition, n=284). The mean concentration of total protein and total cholesterol, and the mean activities of aspartate aminotransferase in the lactation stage tended to be higher than those in the prepartum and maintenance stages. The mean concentration of glucose in the prepartum stage tended to be lower than that in the lactation and the maintenance stages. The mean concentration of nonesterified fatty acids and beta-hydroxybutyrate in the prepartum stage tended to be higher than those in the lactation and maintenance stages. The mean concentration of serum retinol was approximately 30 µg/dl in all stages. These results suggest that the blood biochemical values of multiparous Japanese Black cows vary with breeding stages, and it is considered that the blood parameters obtained in this study are useful as indices for health management of Japanese Black breeding cows.     

Intestinal Tritrichomonas suis (=T. foetus) infection in Japanese cats

Tritrichomonas suis (=T. foetus) has recently been reported to be a causative agent of chronic large-bowel diarrhea in cats. While the disease was previously attributed to Pentatrichomonas hominis, the etiologic agent for feline trichomonal diarrhea was identified as T. suis. Although feline trichomonosis due to T. suis has been reported at prevalences ranging from 14 to 31% in Europe and the U.S., no reports of the pathogen have been published to date in Japan. In 2008, however, we encountered a case of feline trichomonosis at the Veterinary Teaching Hospital of Hokkaido University. The parasite was identified as T. suis by nested PCR amplification of partial internal transcribed spacer region 1 and 5.8S ribosomal RNA gene sequences with T. suis-specific primers and DNA sequencing of the amplified products. We then conducted surveys for feline trichomonosis in three different animal hospitals using either cultivation and/or PCR-based assays. The results revealed that 13 of 147 samples (8.8%) were positive for T. suis, and that 5 of the 13 infected cats, which ranged between 1 month and 7.5 years-old, showed chronic diarrhea. Seven of the infected cats were purebred and 6 were mixed breed. These findings suggested that feline trichomonosis is prevalent in Japan, and that T. suis may play a role as a causative agent of feline chronic diarrhea.