Abnormal morphology in megalopae of the swimming crab,Portunus trituberculatus during seed production: causes and prevention

The occurrence of morphologically immature megalopae, which retain zoeal features such as dorsal spines and furcae of telson, is closely correlated with larval mass mortality during seed production of the swimming crab Portunus trituberculatus in Japanese hatcheries. To determine the cause of immature megalopal morphology, zoeae were reared with various supplementary schedules and density of diets (rotifer, Artemia and phytoplanktons including Chlorella vulgaris and Nannochloropsis oculata). In addition, to assess the relationship between immature morphology and endocrine control, the effect of causative dietary factor was compared with the effect of eyestalk ablation at various timing during zoeal development. Megalopal immature morphology was more distinct in Chlorella‐supplemented groups than in Nannochloropsis‐supplemented groups. High density Chlorella supplementation was associated with the highest incidence of immaturity and resulted in larval mass mortality. The premoult of the third zoeal stage was identified as a critical period at which Chlorella supplementation led to the highest incidence of immaturity. This critical period coincided with the critical period at which larval metamorphosis was regulated by the eyestalk neurosecretory system. Our results suggested that the occurrence of immature megalopal morphology under culture conditions is most likely caused by phytoplankton (especially, Chlorella) supplementation, which disrupts the endocrine regulation. On the basis of our results, we successfully prevented the occurrence of immature megalopal morphology in 500 L tanks by excluding the influence of phytoplankton before the critical period (i.e. discontinuing phytoplankton supplementation and supplying rotifer cultured with non‐phytoplankton materials).     

有机水稻杂草控制技术的研究

有效控制水田杂草的发生是有机水稻栽培关键技术。本田杂草控制技术的研究采用深水管理、施用米糠以及田面覆盖稻草、稻壳等有机物的方法进行水田杂草控制试验。结果表明,深水灌溉能够有效控制水田稗草的发生;施用米糠杂草控制效果可达到药剂控制的85%左右;田面覆盖稻草、稻壳等有机物对水田杂草均有不同程度的控制效果,可降低施用米糠成本。     

Cellular kinetics of villous epithelial cells and m cells in rabbit small intestine

The cellular kinetics of villous columnar epithelial cells and M cells in the rabbit small intestine were determined by the use of 5-bromo-2'-deoxyuridine (BrdU) as a tracer. To identify M cells, vimentin antibody was used. The BrdU-labeled nuclei of columnar epithelial cells reached the base of intestinal villi in all portions at 1 day after BrdU administration. Thereafter, BrdU-labeled cells migrated toward the villous tip, but they did not move at a uniform speed. The epithelial cells which existed in intestinal villi on circular folds moved faster than those on mucosa other than circular folds. At 7 days after BrdU administration, the leading edge of BrdU-labeled epithelial cells already disappeared from the villous tip in all portions of the small intestine. In the ileal Peyer's patch, the BrdU-labeled nuclei of microvillous epithelial cells and vimentin-positive M cells appeared near the intestinal crypt orifice at 1 day after BrdU administration, and then migrated toward the luminal surface of the follicle-associated epithelium (FAE). As they moved toward the upper portion of FAE, the number of BrdU-labeled M cells on the side of the dome decreased simultaneously. The leading edge of BrdU-labeled epithelial cells disappeared from the top of the FAE within 7 days. These results suggest that M cells may differentiate from the undifferentiated cells in intestinal crypts within 1 day and disappear from the top of the FAE after the change of their form from M cells into microvillous epithelial cells.     

Immunohistochemical and ultrastructural detection of intestinal spirochetes in Thoroughbred horses.

Studies of equine intestinal spirochetes have long focused on intestinal contents alone, but intestinal spirochetosis has been reported recently in a 21-month-old Thoroughbred colt in Japan. To define the clinical and pathological significances of intestinal spirochetosis in several horses, an epizootiologic survey with histologic, immunohistochemical, and ultrastructural methods was conducted for Brachyspira antigen-containing intestinal spirochetes in 12 diseased or injured Thoroughbred horses, aged from 35 days to 17 years. Brachyspira antigen-containing spirochetes were found in 7 of 12 horses (58.3%) and were more frequent in the cecum than in other parts of the bowel. It was not clear whether the infection was clinically related to diarrhea or dysentery, but histopathology revealed a close association between the bacterial infection and epithelial hyperplasia. Crypt epithelium consisted mainly of goblet cells and showed frequent mitosis throughout its length. Inflammatory cells and congestion were also present. There were numerous spirochetes in the crypts, and some invaded the cecal and colonic epithelia and underlying lamina propria. Ultrastructurally, the spirochetes were divided into 4 types. Three types were identified in degenerative epithelial cells or intracellularly. Brachyspira antigen-containing intestinal spirochetes invading the mucosa were capable of causing epithelial hyperplasia in the cecum and colon in the horses. The findings in this study will increase awareness of the importance of intestinal spirochetosis and may also be helpful for diagnosis and treatment of this condition.     

Cultivar and anatomical analysis of corolla enlargement of petunia (Petunia hybrida Vilm.) by cytokinin application

We found that the corolla of petunia (Petunia hybrida Vilm.) could be conspicuously enlarged by the separate application of three cytokinins: forchlorfenuron (CPPU), N⁶-benzylaminopurine (BA), and zeatin. To obtain the same enlargement as that achieved by CPPU, approximately 30 and 900 times the concentration of BA and zeatin, respectively, were required. CPPU at 3.2 μmol/L increased the limb area of the corollas of 15 cultivars to between 1.3 and 2.4 times (1.8 times on average) the size of the control area. The increase was negatively correlated (R = 0.58) with the “genetic” limb area (i.e., that of the untreated plant). The enlargement of the corolla caused by cytokinin application was mainly attributed to an increase in cell number in most cultivars. This increase resulted from a high rate of cell proliferation and from prolongation of the cell proliferation phase during corolla development. This anatomical change caused by cytokinin application was similar to the anatomical difference among cultivars because genetic differences in limb area resulted mainly from differences in cell number.     

Metabolism of glucose,fructose, and sucrose in tobacco leaves during maturity

Phosphorus sorption and desorption processes in selected soils were investigated to evaluate the usefulness of phosphorus sorption isotherms in the estimation of the phosphorus supplying capacity of soils. There was a distinct hysteresis in the phosphorus sorption and desorption isotherms, resulting in an overestimation of the replenishing ability of soils to supply phosphorus to the soil solution, when phosphorus sorption isotherms were used for the estimations.

To attain a value of 0.2 ppm P in the soil solution, 115 µmol P/g soil are required in the Kuromatsunai Ando soil (Ochric Andosols) based on the estimation by the P desorption isotherms; this value is 29% higher than that indicated by the P sorption isotherm. Also 42 µmol P/g soil are required in the Mikatagahara Yellow soil (Helvic Acrisols)—which corresponds to a value 23% higher than that indicated by the P sorption isotherm.

Two types of hysteresis subloops were observed: A reversible type and an irreversible type. The former was found typically in the highly weathered Mikatahagara Yellow soil while the latter was observed in the Kuromatsunai Ando soil.     

The assessment of genetic diversity within and among the eight subpopulations of Japanese Black cattle using 52 microsatellite markers

Japanese Black cattle are at risk for genetic homogeneity due to intensive use of a few sires. Therefore, assessment of the actual genetic diversity of this breed is important for future breeding plans. In the present study, we investigated the genetic diversity within and among eight subpopulations of Japanese Black cattle using 52 microsatellite markers. The parameters for genetic diversity of Japanese Black cattle were comparable to those of other cattle breeds, suggesting that the relatively high genetic diversity of the breed. However, upon comparison among the eight subpopulations, the Hyogo subpopulation showed markedly low genetic diversity. The results of the pairwise F_ST values, phylogenetic network and structure analysis indicated that the Hyogo population has remarkably high level of genetic differentiation from other populations, while Yamagata, Niigata, Hiroshima and Kagawa populations have low levels of genetic differentiation. Furthermore, multidimensional scaling plots indicated that individuals in some subpopulations were separated from individuals in the other subpopulations. We conclude that while the overall genetic diversity of Japanese Black cattle is still maintained at a relatively high level, that of a particular subpopulation is significantly reduced, and therefore the effective population size of the breed needs to be controlled by correct mating strategies.     

Xanthan gum and locust bean gum substrate improves bovine embryo development

One of the major difference between the in vivo and in vitro embryonic environments is the stiffness of the culture substrate. Xanthan gum (XG) and locust bean gum (LBG) are natural materials that are safe, inexpensive and easy to handle. In this study, we investigated the effects of using a polysaccharide culture substrate made from 1% XG and 1% LBG (XG-LBG gel) on bovine embryonic development. Oocytes collected from bovine ovaries were subjected to maturation, and fertilization to generate embryos at an early developmental stage (>4 cell stage). Cleaved embryos were further cultured in a well of 96-well cell culture plate coated with or without XG-LBG gel for 5 days. While the developmental rate up to the blastocyst stage did not differ between the two culture systems (control, 38.0 vs. gel, 38.6%), blastocysts developed on the XG-LBG gel produced significantly high cell numbers and ATP content. Embryos cultured on XG-LBG gels for 24 hr had high expression levels of F-actin and a highly even distribution of E-cadherin. In addition, embryos developed on XG-LBG gel demonstrated increased translocation of YAP to the nucleus and increased connective tissue growth factor (CTGF) protein levels (downstream of Hippo signalling). These findings suggest that soft culture substrates improve embryonic development by enhancing mechanotransduction, including YAP-CTGF signalling.     

Changes in growth,higher salinity tolerance and behavioral traits in Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> after repeated cross-breeding

ABSTRACT:     To improve stock enhancement strategies, the influences of repeated cross-breeding on several traits were studied in hatchery-reared juveniles of the Japanese flounder. Ten families (seven caught in the wild and three from cross-bred brood fish) were produced by artificial fertilization. Ninety-four days after hatching, we investigated settlement, burying in the sand and escape behavioral traits, and then measured tolerance of high salinity and six morphological traits. Juveniles from the repeatedly cross-bred brood fish showed significantly higher growth and tolerance than those from the wild-caught brood fish. Among the behavioral traits, a significantly lower settling speed and incidence of burying in the sand were observed in the juveniles from the repeatedly cross-bred families. Heritability ( h ²) of growth traits was more than 0.2 in both origins, while that of other traits was close to zero. These results suggest that repeated cross-breeding of Japanese flounder under hatchery conditions improves growth performance and leads to the loss of certain behavioral traits that are considered essential to survival in wild environments.