Genetic diversity of wild soybean (<Emphasis Type="Italic">Glycine soja</Emphasis> Sieb. et Zucc.) and Japanese cultivated soybeans [<Emphasis Type="Italic">G. max</Emphasis> (L.) Merr.] based on microsatellite (SSR) analysis and the selection of a core collection

Wild soybeans, Glycine soja, are a source of genetic variation to improve soybeans. To improve the efficiency evaluation of conserved germplasm a core or mini-core collection approach that maximizes allelic diversity in a proportion of the whole collection has frequently been advocated. The genetic diversity of a wild soybean collection (1,305 accessions) plus Japanese cultivated soybeans (53 accessions) were analyzed at 20 SSR marker loci. Higher levels of allelic diversity were found in wild soybeans (28 alleles per locus) than Japanese cultivated soybean (five alleles per locus). The genetic distance between wild soybeans from different regions reflected their proximity. Accessions from Russia consisted of a diverse array of alleles resulting in accessions being spread further apart in a PCA plot than accessions from other regions. Accessions of wild soybean from Korea included many rare alleles and thus had a high representation in the core collection. The two core collections developed here, traditional and mini, consisted of 192 accessions with 97% of the allelic diversity (14% of the whole collection) and 53 accessions with 62.4% of the allelic diversity (5% of the whole collection), respectively.     

Development of a method for forest cafeteria tests for evaluating chemical deterrents against bark stripping by sika deer (Cervus nippon yesoensis)

Damage caused by sika deer (Cervus nippon yesoensis) is a serious problem in commercial and environmental (non-harvested) forests in Hokkaido, northern Japan. Cafeteria tests in forests may be useful for evaluating the efficacy of chemical deterrents against bark stripping by deer. To develop a method for forest cafeteria tests in the continuous snow cover period, two experiments were carried out. In the experiments, logs were produced from tree trunks, and used as carriers of chemical deterrents. Carriers were installed in forests and fed to deer. The first experiment was to find suitable sites and installation methods for carriers. Criteria for the local suitability and the installation methods were as follows: a) Sites where deer are active should be selected; b) Carriers should be installed along actively used deer trails; c) Installation sites of carriers should be changed in response to deer movement; d) Carriers should be produced from tree species that deer naturally prefer; and e) Each carrier should be partially buried in the snow. The second experiment evaluated the feasibility of a cafeteria test method based on the results of the first experiment. The method was used for 13 sets of the cafeteria test, in which the deterrent effectiveness of 5 chemicals (wood tar, rosin, wood vinegar, and 2 pyroligneous liquors) was examined. We obtained results from all the sets. The chemicals tested did not deter bark stripping by deer. Nevertheless, the method used in the present study was practical for the cafeteria tests.     

Effects of feeding polyphenol‐rich winery wastes on digestibility,nitrogen utilization,ruminal fermentation,antioxidant status and oxidative stress in wethers

Four wethers were used in a 4 × 4 Latin square design experiment to evaluate the availability of two types of winery wastes, winery sediment and grape pomace, as ruminant feeds possessing antioxidant activities. Each wether was assigned to one of the following four treatments: (i) 75 g/kg winery sediment (WS) on a dry matter (DM) basis; (ii) 166 g/kg DM winery grape pomace (WP); (iii) control diet (CD; 17 g/kg DM soybean meal);and (iv) only tall fescue hay (TFH; no additive). Winery sediment and grape pomace had high levels of polyphenols and of radical scavenging activities. Feeding with winery sediment and grape pomace did not negatively affect the intake, but it depressed crude protein (CP) digestibility compared with CD (P = 0.052 and P < 0.01 for WS and WP, respectively). Polyphenols in winery wastes decreased ruminal ammonia production (P = 0.089 and P < 0.05), likely due to their inhibitive effect on microbial activities in the rumen. The addition of winery sediment and grape pomace decreased urinary 8‐hydroxy‐2'‐deoxyguanosine (8‐OHdG; an index of oxidative damages) excretion per day (P < 0.05 and P = 0.059). The results indicated that winery sediment and grape pomace could alter nitrogen metabolism and/or act as new antioxidants for ruminants.     

Transportation of freeze-dried mouse spermatozoa under different preservation conditions

Freeze-dried mouse spermatozoa can be used for normal embryonic development after injection into oocytes, thus indicating that freeze-drying is a useful method for the storage and transportation of genetic materials from animals. We recently reported that storage of freeze-dried mouse spermatozoa requires maintenance at temperatures lower than -80 C for long-term preservation and a pressure of 0.37 mbar at primary drying and that these conditions significantly improve the developmental rate to the blastocyst stage. In this study, we examined the influence of transportation and preservation conditions on freeze-dried spermatozoa. Freeze-dried spermatozoa stored for 2 or 2.5 years at 4 or -80 C were transported round trip overland between Shizuoka and Hokkaido prefectures in Japan or by air between Japan and Belgium. The freeze-drying conditions consisted of primary drying at pressures of 0.04, 0.37 and 1.03 mbar and secondary drying at a pressure of 0.001 mbar. Embryos (2-cell stage) from freeze-dried spermatozoa dried at 0.04 mbar and stored at 4 C for 2 years with and without overland transportation did not develop to term. The development rates of embryos from spermatozoa stored at -80 C for up to 2 years and transported overland, by air and without transportation were 8, 1 and 28%, respectively. The development rates of embryos from spermatozoa without transportation were significantly higher than with transportation (P<0.05). These data indicate that freeze-dried spermatozoa stored at -80 C with and without transportation can retain their ability to generate viable offspring after storage for up to 2 years. However, there are limitations to be considered in the transportation of freeze-dried spermatozoa at ambient temperature.