Oocyte maturation and active motility of spermatozoa are triggered by retinoic acid in pen shell <Emphasis Type="Italic">Atrina pectinata</Emphasis>

For recovery of the declining population of pen shells in the wild, the production of pen shell juveniles for transplantation or aquaculture is underway in Japan. For more stable juvenile production, artificial fertilization methods for pen shells are needed, but methods to induce oocyte maturation (meiosis resumption) used in other bivalves, which make oocytes fertilizable, were ineffective for pen shells. Here, we report evidence showing that retinoic acid (RA) has strong activity in inducing oocyte maturation and activating sperm motility in pen shells. Treatment of fully developed oocytes with 1.0 μM all-trans-RA (at-RA) induced germinal vesicle breakdown, a typical morphological sign of oocyte maturation, but 1.0 μM at-retinol and at-retinal, 2 mM ammonia, and 1.0 μM serotonin were ineffective. Treatment with at-RA for 30 min was sufficient for oocyte maturation and was more potent than its isomers, 9-cis- and 13-cis-RA. Parallel results were obtained for sperm motility activation. Oocyte responsiveness to at-RA increased during the final stage of ovary development. Artificial fertilization was successful only with the oocytes treated with at-RA, and fertilized eggs developed to D-shaped (veliger) larvae without apparent morphological abnormalities. These results indicate the possible application of RA for the artificial fertilization of pen shells.     

Effects of VIP and GRF on the release of growth hormone in perifused bovine adenohypophysis

The effects of vasoactive intestinal polypeptide (VIP) and growth hormone releasing factor (GRF:hpGRF(1–29)-NH₂) on the release of growth hormone (GH) from anterior pituitaries from cows were examined by using an in vitro superfusion system. The pituitaries were excised randomly from cycling cows, dissected to obtain medial portions, and minced to obtain cubes with approximate dimensions of 1.5mm on a side. For each perifusion setup, 5 pieces of pituitary tissues were chambered and flushed with modified KRB solution saturated with 95% O₂-5% CO₂ at 38C. Perifusion with media containing 10⁻⁶ and 10⁻⁷M VIP for 30 min induced a significant release of GH during the treatments (P<0.05). VIP (10⁻⁸M) increased GH levels significantly (P<0.05), but to a minor degree. Perifusion with the media containing 10⁻⁶, 10⁻⁷ and 10⁻⁸M GRF for 30 min markedly increased the GH concentration and the effects continued up to 90 min after termination of the perifusion of the peptide (P<0.05, P<0.01). The GH releasing effects of GRF could be seen at doses as low as 10⁻¹¹M GRF (P<0.05, P<0.01).

These findings indicate that the GH releasing effect of VIP is less potent that that of GRF in cows.     

A Reovirus Causes Hypovirulence of Rosellinia necatrix

ABSTRACT White root rot, caused by Rosellinia necatrix, is a serious soilborne disease of fruit trees and other woody plants. R. necatrix isolate W370 contains 12 segments of double-stranded RNA (dsRNA) that is believed to represent a possible member of the family Reoviridae. W370 was weakly virulent and its hyphal-tip strains became dsRNA free and strongly virulent. The 12 segments of W370dsRNA were transmitted to hygromycin B-resistant strain RT37-1, derived from a dsRNA-free strain of W370 in all or none fashion through hyphal contact with W370. The W370dsRNA-transmitted strains were less virulent than their parent strain RT37-1 on apple seedlings, with mortality ranging between 0 to 16.7% in apple seedlings that were inoculated with the W370dsRNA-containing strains and 50 to 100% for seedlings inoculated with the dsRNA-free strains. Some W370dsRNA-containing strains killed greater than 16.7% of seedlings, but these were found to have lost the dsRNA in planta. These results indicate that W370dsRNA is a hypovirulence factor in R. necatrix. In addition, a strain lost one segment (S8) of W370dsRNA during subculture, and the S8-deficient mutant strain also exhibits hypovirulence in R. necatrix.     

Artificial Infection of Rosellinia necatrix with Purified Viral Particles of a Member of the Genus Mycoreovirus Reveals Its Uneven Distribution in Single Colonies

ABSTRACT Rosellinia necatrix mycoreovirus 3 (W370) (RnMYRV-3/W370, described as RnMYRV-3 in this paper), a member of the newly established genus Mycoreovirus within the family Reoviridae, is the hypovirulence factor of the white root rot fungus, Rosellinia necatrix. Two virus-free fungal isolates (W37 and W97) that were somatically incompatible with the virus-harboring field isolate (W370) were transfected with purified RnMYRV-3 particles. Virus infection was confirmed by electrophoresis and northern hybridization of viral double-stranded RNA. RnMYRV-3 was transmissible from transfected strains to their respective, virus-free counterparts via hyphal anastomosis. Virus-transfected strains produced smaller lesions on apple fruits than did their virus-free counterparts. Virus-cured strains were indistinguishable from wild-type strains in culture morphology and displayed approximately the same virulence level on apples. Virus-transfected strains had "mosaic" colony portions consisting of thin, fast-growing and dense, slow-growing mycelia, and grew more slowly as a whole than their virus-free, parental strains. The level of virus accumulation varied among virus-transfected subcultures and within its single colonies. Virus-transfected strains were occasionally cured, as was W370. Such a phenomenon may be ascribed to uneven viral distribution in single colonies and the difficulty in viral transmission to virus-free hyphae.     

Histopathology of red crown rot of soybean

Histopathological analyses of red crown rot (RCR) of soybean were performed using field-grown and indoor-grown soybean plants. These analyses revealed that soybean plants formed a periderm to prevent pathogen invasion. When a break formed in the periderm, an additional periderm newly differentiated into the inner cells and prevented pathogen invasion. If the pathogen reached the intravascular tissues, some xylem parenchyma cells died, and the vessels were occluded by gum. Periderm formation and vessel occlusion were observed not only in slightly diseased soybean plants, but also in seriously diseased plants, indicating that both reactions function as part of the defense response against RCR.     

Horizontal and vertical variations in survival rates of juvenile bivalves,Anadara kagoshimensis (Tokunaga, 1906), Ruditapes philippinarum (Adams & Reeve, 1850) and Atrina sp., experimentally caged in the inner area of the Sea of Ariake,Japan

International Aquatic Research - In the inner area of the Sea of Ariake, rearing experiments were conducted to examine how the survival rates of juveniles of three bivalve species (Anadara...     

Effect of vasoactive intestinal peptide on the release of growth hormone in perifused pituitary and hypothalamus of the goat

The effect of vasoactive intestinal peptide (VIP) on the release of growth hormone (GH) from the adenohypophysis of the goat was studied in vitro using the perifusion system. Two perifusion systems were employed to differentiate potential direct effects of VIP on the pituitary from indirect effects mediated through the hypothalamus. The first perifusion system used single chambers housing only pituitary fragments. The second system used two chambers in tandem, one containing hypothalamus and the second the pituitary fragments, the flow passing through the hypothalamic chamber first. VIP (10(-6), 10(-7), 10(-8)M) stimulated significant GH release in both perifusion systems (P less than 0.05, P less than 0.01) in a concentration related fashion. The quantity of GH release induced by the 10(-9)M and 10(-10)M VIP groups were not significant. Further, the GH released from the system that perifused the pituitary alone (10(-8)M VIP) was significantly higher than that containing both the hypothalamus and the pituitary fragments (P less than 0.05). The relative lower GH secretory response to the same dose of VIP applied to the hypothalamus-pituitary suggests that the perifused caprine hypothalamus may release an inhibitory factor, such as somatostatin in vitro. These results suggest that VIP stimulates GH release by acting directly on the adenohypophysis of the goat.     

Effect of iron-quercetin complex on reduction of nitrite in in vitro and in vivo systems

This study investigated whether reducing agents such as quercetin and iron(II) facilitate formation of nitric oxide (NO) gas from orally ingested nitrite in an vivo study. When 3 mg/kg Na (15)NO2 was orally administered to rats with or without iron(II) or quercetin, Hb (15)NO, which is indicative of systemic (15)NO, was detected in the blood, with the maximum blood concentration of Hb (15)NO at 15 min after nitrite or nitrite plus quercetin treatment, whereas after administration of nitrite plus iron(II) or nitrite plus iron(II) and quercetin, the time was shortened to 10 min. Interestingly, iron(II), quercetin, or iron(II) plus quercetin did not affect the total amount of Hb (15)NO generated from orally administered Na (15)NO2. However, the systemic nitrite concentration was significantly decreased in the presence of iron(II) or iron(II) plus quercetin. These results may indicate that iron(II) is critical to the generation of NO gas from nitrite, whereas quercetin contributed little under the in vivo experimental conditions.     

Survival time of juvenile pen shell Atrina pectinata (Bivalvia: Pinnidae) in hyposaline water

International Aquatic Research - Survival time of juvenile pen shell Atrina pectinata (shell length&nbsp;≈&nbsp;34&nbsp;mm) exposed to hyposaline water without sediments was...