Pedigree analysis of recaptured fish in the stock enhancement program of spotted halibut Verasper variegatus

ABSTRACT:   Spotted halibut Verasper variegatus hatchery juveniles produced in 2002 were genotyped using three microsatellite DNA markers (msDNA) and then released into natural waters. Subsequently, recaptured individuals were examined using msDNA. In order to evaluate the effectiveness of the stock enhancement program, from the genetic point of view, a pairwise F _ST test was implemented to estimate the genetic divergence between the wild captive broodstock, the hatchery offspring and the recaptured samples. The analysis showed significant differentiation between the broodstock and recaptured samples. Pedigree determination using msDNA was used to calculate the effective population size of the recaptured stock, which was found to be very low ( N _e ≈ 8). Equal family survivability was observed between the two recaptured stocks, but not between the released and recaptured stock. The number of identified families was higher and more equalized in the hatchery offspring compared to the recaptured samples, where the number of families declined. This fact was caused by an unequal family survivability just before or just after release. Separately, the number of contributing parents to the hatchery offspring was lower than the broodstock census number. Consequently, these two facts caused the genetic divergence of the recaptured stock from the broodstock.     

Evaluation of available silicon content and its determining factors of agricultural soils in Japan

To evaluate the available silicon (Si) content in agricultural soils in Japan and to investigate the determining factors of this content, we collected 180 soil samples from the surface layer of paddies and upland fields in Japan and determined their available Si contents. A phosphate buffer (PB; 0.02 M, pH 6.9) or an acetate buffer (AB; 0.1 M, pH 4.0) was used to extract available Si from the soil samples, and the Si concentrations in the extracts were determined by inductively coupled plasma-atomic emissions spectroscopy (ICP-AES). The total Si content and selected physicochemical properties were also determined for the soil samples. The median values of the available Si contents by the PB and AB methods were 48.8 and 79.7 mg kg^?1 and corresponded to 0.017% and 0.027% of the total Si content, respectively. The overall data showed log-normal distributions. The available Si content of the upland soils was significantly higher than that of the paddy soils by both the PB (p < 0.01) and AB methods (p < 0.05). The available Si contents by the PB and AB methods had a significant positive correlation (p < 0.01) and they had significant negative correlation with the total Si content (p < 0.01). The values of the available Si contents by the PB and AB methods correlated positively with the pH, total carbon (C) content, and dithionite-citrate bicarbonate extractable iron (Fed) and aluminum (Ald), acid oxalate extractable iron (Feo) and aluminum (Alo), Fed-Feo and Alo+1/2Feo values (p < 0.01). A multi-regression analysis indicated that pH, amorphous minerals and crystalline iron (Fe) oxides were the dominant determining factors of available Si in the soils, and these three variables explained approximately two thirds of the variation of available Si content in agricultural soils in Japan. In terms of soil type, Terrestrial Regosols, Dark Red soils and Andosols had relatively high available Si contents, whereas Sand-dune Regosols, Red soils and Gray Lowland soils had relatively low contents. In terms of region, the soils in the Kanto and Okinawa regions had relatively high available Si contents and those in the Kinki, Shikoku and Chugoku regions had relatively low contents. In conclusion, the available Si content and its determining factors for agricultural soils in Japan were quantitatively elucidated, and this will contribute to the establishment of rational soil management?—including the application of silicate materials, taking into account the Si-supplying power of the relevant soils—for sustainable and productive agriculture in Japan.     

Natural contamination of dietary rice straw with zearalenone and urinary zearalenone concentrations in a cattle herd

The present study was conducted to 1) identify the natural source of feed contamination by zearalenone (ZEN), which was suspected to have caused persistently increased urinary ZEN concentrations in one of our experimental cattle herds, and 2) evaluate the effects of intervention against this source of contamination. As an experimental model, a fattening Japanese Black cattle herd showing persistently increased urinary ZEN concentrations was identified. Urinary ZEN concentrations of cows fed with new rice straw (experimental group, n = 6) vs. cows that continued to feed on the old rice straw (control group, n = 4) were measured at the start (d 1) and at 2 wk (d 14) after the onset of feeding with straw. In addition, the ZEN concentration in feed and water samples was measured by using both the ELISA and HPLC methods. Furthermore, isolation and identification of fungi from rice straw and concentrate feed samples were performed. The urinary ZEN concentration [ZEN (pg/mL)/creatinine (mg/mL) = pg/mg of creatinine] of cows fed with new rice straw was significantly (P < 0.05) less (843 pg/mg of creatinine) than that of cows fed with old rice straw (15,951 pg/mg of creatinine). On both d 1 and 14, the ZEN concentrations of old rice straw were greater than those of new rice straw. In addition, fungal colonies were observed in the culture media that was obtained from the old rice straw suspected of ZEN contamination, but not in the culture media from new rice straw or other feed samples. In conclusion, our field trials clearly indicate that the rice straw fed to the cows was naturally contaminated with ZEN, and that the monitoring of urinary ZEN concentrations could prove to be a useful tool for detecting the exposure of cattle to ZEN contamination at the farm level.     

Plant growth and nutrition in pine (Pinus thunbergii) seedlings and dehydrogenase and phosphatase activity of ectomycorrhizal root tips inoculated with seven individual ectomycorrhizal fungal species at high and low nitrogen conditions

We investigated the functions and ecological traits of seven individual ectomycorrhizal (ECM) fungal species derived from a Pinus thunbergii-dominated (nitrogen-poor) and a Robinia pseudoacacia-dominated (nitrogen-rich) area. P. thunbergii seedlings were inoculated with seven individual ECM fungal species and cultured under high nitrogen (nitrogen group) and low nitrogen (control group) conditions. Control seedlings were not inoculated with any ECM fungus. Seedlings harvested were examined for the numbers of non-mycorrhizal and ECM root tips, growth, nutrient status (nitrogen, phosphorus and carbon) and the activity of dehydrogenase and phosphomonoesterase in the root tips. The relationships among the growth, nutrient status and the enzymatic activity in the seedlings inoculated with each ECM fungal species were also analyzed by canonical discriminants. As the results, inoculation of ECM fungi made the plant growth significantly higher than those of non-inoculated seedlings. Plant growth significantly correlated with the phosphorus content, N:P ratio and phosphomonoesterase activity, especially in the nitrogen group. Dehydrogenase and phosphomonoesterase activities in the seedlings were higher when the ECM fungi from a nitrogen-rich area were inoculated than when those from a nitrogen-poor area were inoculated. The canonical discriminant analyses separated seedlings by inocula: no fungus, ECM fungi from a nitrogen-poor area, and those from a nitrogen-rich area. These results suggested the possibility that nitrogen deposition caused P shortage and the P absorbing ability was higher in the ECM fungi in nitrogen-rich forests than those in nitrogen-poor forests.     

Distribution of protein gene product 9.5-immunopositive and NADPH-diaphorase-positive neurons in the common marmoset (Callithrix jacchus) accessory olfactory bulb

The principal center of the accessory olfactory system is the accessory olfactory bulb (AOB). In primates, simians are divided into two groups, New and Old World monkeys, and the AOB is present in only New World monkeys. The common marmoset (Callithrix jacchus) is a species of New World monkey. Although the morphology of the common marmoset AOB has been demonstrated, the distribution patterns of the mitral/tufted and granule cells of the AOB remain unclear. In the present study, therefore, the distribution of the mitral/tufted and granule cells in the common marmoset AOB was examined using two histochemical markers including immuno-staining for protein gene product (PGP) 9.5 and NADPH-diaphorase staining. The vomeronasal nerves, gomeruli and mitral/tufted cells showed PGP 9.5-immunoreactivity. The mitral/tufted cells were arranged in only one or two rows along the margin of the glomerular layer to form the mitral/tufted cell layer (MTL). Since the mitral/tufted cells occurred sparsely in the common marmoset, the MTL was illegible. NADPH-diaphorase reactivity was primarily detected in the rostral and caudal areas of the AOB. In these areas, granule cells showed NADPH-diaphorase reactivity. Since the granule cells were sparse, the common marmoset AOB displayed less-developed granule cell layer. Although the functional significance of the AOB remains to be solved in the common marmoset, small-sized and less-laminated AOB may show that sexual behavior of the common marmoset has lesser dependence on the accessory olfactory system.     

Use of a rigid-type polyethylene T-cannula,silicone tubing,and adhesion netting for surgical cannulation of ileum of the horse

A rigid-type of polyethylene T-cannula was fitted into the anterior ileum of six horses in order to improve the cannulation techniques. A piece of polyethylene net was fastened onto the intestinal wall around the cannula to prevent dislodgment of the cannula by promoting a secure adhesion between the ileum and the abdominal wall. The cannula barrel sheathed with silicone tubing was exteriorized through a stab incision at the lateral ventral wall on the transverse line of the second lumber vertebra, and a flange was screwed onto the barrel. The feeding regime gradually increased concentrate without roughage prevented any colic signs. The use of these techniques succeeded in the ileal cannulation with no leakage of digesta.     

Three-dimensional common-feature hypotheses of inhibitors of calling behaviour and in vitro [14C]acetate incorporation by pheromone glands of Plodia interpunctella

Some octopamine (OA) agonists were found to suppress the calling behaviour and pheromone biosynthesis in vitro of the Indian meal moth, Plodia interpunctella (Hübner), a stored-product pest. Compounds were screened using a calling behaviour bioassay of female P interpunctella. Three active derivatives, with activity at the nanomolar level, were identified. In order of decreasing pheromonostatic activity these were: 2-(2-ethyl-6-methylanilino)oxazolidine > 2-(2,6-diethylanilino)thiazolidine > 2-(2,6-diethylanilino)oxazolidine. These compounds showed also in vitro inhibitory activities in de novo pheromone biosynthesis. Three-dimensional pharmacophore hypotheses were built from a set of 19 compounds. Among the ten common-featured models generated by the program Catalyst/HipHop, a hypothesis including a ring aromatic group (RA), a positive ionizable group (PI) and two hydrophobic aliphatic (HpA1) features was considered to be essential for inhibitory activity in the calling behaviour and pheromone biosynthesis in vitro. Active compounds mapped well onto all the RA, PI and HpA1 features of the hypothesis. Less-active compounds were shown not to achieve the energetically favourable conformation which was found in the active molecules in order to fit the 3-D common-feature pharmacophore models. The present studies demonstrate that inhibition of calling behaviour and PBAN-stimulated incorporation of radioactivity is by OA-agonistic activity.     

Quantitative structure—activity studies of some octopaminergic agonists against Periplaneta americana

The quantitative relationship between the structure of 2-(arylimino) thiazolidines (AITs), arylethanolamines (AEAs) and 2-(arylalkylamino)-2-thi-azolines (AATs) and their octopamine (OA)-agonist activities against the ventral nerve cord of American cockroach, Periplaneta americana L., was analysed using reported physicochemical parameters and regression analysis. The electronic nature of a substituent was the most important factor for AAT, followed by the hydrophobic effects: the more electron-donating and the more hydrophobic the substituent, the greater the activity. The hydrophobic nature of a substituent in AEA was the most important factor, followed by the steric effects: the more hydrophobic the substituent, the greater the activity. The more electron-donating the substituent of AIT, the greater the activity.     

Meiotic Competence of Canine Oocytes Embedded in Collagen Gel

The present study was conducted to examine the meiotic competence of canine oocytes embedded in collagen gel, and to investigate the effects of timed exposure of the oocytes embedded in collagen gel to gonadotrophins during maturation culture, on their nuclear maturation. Cumulus-oocyte complexes (COCs) were collected from bitches at the anoestrous and dioestrous stages of the reproductive cycle. In the first experiment, half of the COCs were embedded in collagen gels. The COCs with or without collagen-gel embedding were cultured in a TCM-199 medium supplemented with 0.1 IU/ml human menopausal gonadotropin (hMG) and 10 IU/ml human chorionic gonadotropin (hCG) for 72 h. In the second experiment, the COCs embedded in collagen gels were cultured in TCM-199 medium with gonadotrophins (hMG and hCG) for various periods (0, 24, 48 and 72 h) and then cultured in the medium without gonadotrophins until reaching total culture period (72 h). The percentage of the oocytes reaching metaphase I and metaphase II (MI/MII) was significantly higher (p < 0.05) in COCs with collagen-gel embedding than in COCs without collagen-gel embedding. The percentage of oocytes that were arrested at the germinal vesicle stage was significantly lower (p < 0.05) in oocytes cultured with gonadotrophins than in oocytes cultured without gonadotrophins. However, there were no significant differences in the percentages of oocytes that reached each stage of meiosis among the groups, irrespective of the duration of exposure to gonadotrophins. These observations indicate that embedding of COCs by collagen gel enhances the meiotic competence of canine oocytes, but removal of hormone supplement from maturation medium does not improve the ability of the oocytes to reach MII stage.