Teruyoshi Narita

Motility of spermatozoa obtained from testes of Japanese pearl oyster Pinctada fucata martensii

Effects of NH3 concentration in sea water and pH of sea water on the motility of spermatozoa obtained from testes were examined in the Japanese pearl oyster Percent motility at 30 s after dilution increased with increasing NH3 concentration in sea water from 0.75–2.0 mM. When spermatozoa were diluted with sea water containing 0.75 mM NH3, which is widely used as the insemination fluid in the hatchery of this species, the percent motility increased with time elapsed after dilution, and peaked at 5 min. For spermatozoa diluted with sea water containing 2.0 mM NH3, the percent motility increased rapidly and peaked at 30 s. The pH of sea water increased with increasing NH3 concentration from 8.2 (0 mM NH3) to 9.9 (5.0 mM NH3). When spermatozoa were diluted with artificial sea water at various pH (buffered without NH3 at 6.0–10.0), only spermatozoa diluted with artificial sea water of pH 10.0 were motile, and the percent was considerably lower than those in ammonical sea water. These results indicate that sea water containing 2.0 mM NH3 is a suitable solution for evaluating sperm motility, and that NH3 and/or ammonium ions may activate sperm motility in this species.

Effects of cryopreservation on sperm structure in Japanese pearl osyter Pinctada fucata martensii

To clarify factors reducing the motility and fertility of cryopreserved spermatozoa of the Japanese pearl oyster Pinctada fucata martensii, the structure of spermatozoa before and after cryopreservation was observed by scanning electron microscopy. Testicular spermatozoa were diluted with cryopreservation diluent (10% methanol+18% fetal bovine serum+72% sea water), and dispensed into 0.25-mL straws. The straws were cooled at a rate of approximately −20 °C/min to −50°C, and subsequently immersed in liquid nitrogen. Percentage motility of spermatozoa before cryopreservation was 69.9±4.2%, and that of cryopreserved spermatozoa was 24.0±1.8%, respectively. In cryopreserved spermatozoa, the percentage that lacked or had a deformed flagellum was 56.6±3.9%, while in fresh spermatozoa this was 8.7±2.0%. In cryopreserved spermatozoa, the percentage of deformed acrosomes was 76.6±5.2%, while in fresh spermatozoa this was only 0.9±0.3%. Cryopreserved spermatozoa with a normal acrosome and flagellum were only 15.4±3.5% of those in fresh spermatozoa. These results indicate that lesion of the flagellum and deformation of the acrosome occurred through the cryopreservation procedure, and both types of damage lead to loss of the motility and fertility in thawed spermatozoa.

Population dynamics of portunid crab Charybdis bimaculata in Ise Bay, central Japan

The present study dealt with seasonal and interannual variations in the abundance and biomass, and spatio-temporal distributions of the portunid crab Charybdis bimaculata dominant in Ise Bay, central Japan. The abundance and biomass of the crab decreased in summer when the oxygen-poor water developed in central or inner parts of the bay, and then increased through new recruits from autumn (October–November) to the following spring (March–May) when the oxygen-poor water disappeared. Berried females were collected mainly from spring to autumn. Recruits were collected in any season. Particularly in winter, most recruits were located in the innermost part of the bay. According to the cohort separation based on size frequency distribution in carapace width of the crab specimens, the cohorts that were derived from spawning in spring to summer largely contributed to establishing and maintaining the benthic populations for the following year in the bay, whereas those from other seasons failed to recruit because of serious damage caused by the oxygen-poor water. Most crab individuals one year post hatch contributed to spawning and then died by the winter of the same year.