Monitoring of the genetic variability of the hatchery and recaptured fish in the stock enhancement program of the rare species barfin flounder <Emphasis Type="Italic">Verasper moseri</Emphasis>

ABSTRACT:   In relation to the stock enhancement program for barfin flounder, hatchery juveniles produced in 2001 were genotyped using microsatellite DNA markers (msDNA) and then released to natural waters. Subsequently, recaptured individuals, designated as 'tentative recaptured', were examined using msDNA. In order to evaluate the effectiveness of the stock enhancement program, pairwise F _ST and genic differentiation tests were used to estimate the genetic divergence between the wild samples, the hatchery broodstock, and the tentative recaptured samples. Analysis showed significant differentiation among these three groups. Pedigree determination by msDNA was used to establish the origin of the tentative recaptured individuals, in order to elucidate whether they were hatchery produced, 'real recaptured', or wild specimens. Wild individuals were not found. The effective population size of the real recaptured stock was very low ( N _e = 16.6). Equal family survivability was observed between the released and real recaptured stocks, indicating that the genetic variability of the released stock was maintained in the natural environment. Future broodstock management, breeding designs, and family contribution equalization of the offspring to be released will be required in this rare species to avoid unintended genetic differentiation between the wild population and the hatchery broodstock.     

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.     

Complementary DNA cloning and functional analysis of lycopene β-cyclase in the brown alga Undaria pinnatifida

Fisheries Science - To date, there is no functional evidence of the enzymes catalyzing carotenogenesis in brown algae. In this study, we have investigated the activity of brown algal lycopene...     

Genetic diversity of wild mekong giant catfish Pangasianodon gigas collected from Thailand and Cambodia

The Mekong giant catfish Pangasianodon gigas is endemic to the Mekong River and is a critically endangered species. The genotypes of the microsatellite DNA (msDNA) and mitochondrial DNA (mtDNA) markers (right domain of the control region) were detected to evaluate the present status of genetic divergence of this species from the Mekong River in Thailand and Cambodia. The observed and expected heterozygosity values of Mekong giant catfish in Thailand and Cambodia were relatively low in comparison with those of other nonendangered freshwater fish species. These two populations from Thailand and Cambodia showed similar levels of genetic diversity, as evaluated by the 384 nucleotides of the mtDNA control region with 13 haplotypes. The pairwise F _ST value between the two populations based on the genotype frequencies of msDNA and mtDNA markers suggested a close genetic relationship between the populations in Thailand and Cambodia. The results of this study support the conclusion that the Mekong giant catfish is critically endangered. Care should be taken to sustain the genetic diversity of this species, as the level of genetic variability has already decreased in the wild population. This species is a target species for an ongoing stock enhancement program in the Mekong River in Thailand. It is proposed to apply these markers for proper broodstock management, such as for minimal kinship selective breeding in the hatchery.     

Allozyme variation of littleneck clam Ruditapes philippinarum and genetic mixture analysis of foreign clams in Ariake Sea and Shiranui Sea off Kyushu Island, Japan

ABSTRACT:   Allozyme variation of the littleneck clam Ruditapes philippinarum was evaluated in four samples from Nameishi and Matsuo in the Ariake Sea, Ryugatake and Ushibuka in the Shiranui Sea off Kyushu Island, Japan, and in one sample from Jinzhou, China, in the Bohai Sea. A Ruditapes bruguieri sample imported from the Korean Bay off Nampo, North Korea was also studied. Among the R. philippinarum samples, heterozygosity varied from 0.265 to 0.301 and F _IS estimates indicated significant homozygosity excess in 15 of 40 loci analyzed. Deviations from Hardy–Weinberg equilibrium were significant in all samples ( P  < 0.05). Pairwise F _ST estimates indicate that genetic differences between the Chinese and Japanese samples were very low, but significantly different from zero. Mixture proportions with 95% confidence intervals of Chinese R. philippinarum in Nameishi and Matsuo were estimated at 0.4098 [0.2512, 0.5705] and 0.4899 [0.3262, 0.6540], respectively. However, genetic invasion of stocked Chinese R. philippinarum into wild populations in the Ariake Sea remains uncertain due to the low precision of the estimates caused by the high similarity of allele frequencies between Jinzhou and the Ariake Sea.     

Persorption of luminal antigenic molecule and its specific antibody via apoptotic epithelial cells of intestinal villi and Peyer's patches into peripheral blood in rats

The possibility of persorption of bovine serum albumin (BSA) molecules from mucous epithelial cells and its mechanism were investigated in rats orally pre-immunized by BSA for 14 consecutive days. In the small and large intestines, both the BSA antigen (BSA-Ag) and its specific antibody (SpAb) were absorbed by the epithelial cells at the late apoptotic stage (ApoEp), and were subsequently transcytosed by membranes of the small vesicles. The basal cytoplasms containing highly-concentrated BSA-Ag and SpAb were occasionally fragmented into small cytoplasmic droplets that were secreted into the lamina propria. In Peyer's patches, both BSA-Ag and SpAb were more actively absorbed and transcytosed toward the dome area by the ApoEp of the dome apex than by the M cells. BSA-Ag and SpAb were finally persorbed into the portal blood and lymph, but were never secreted into the bile. They were also engulfed by macrophage-like cells in the villous lamina propria, mesenteric lymph node and spleen, and by hepatocytes in the liver. These findings suggest that sensitized soluble luminal antigens are taken up by ApoEp in the small intestine and are finally persorbed into the peripheral blood. The uptake of luminal antigen might be mediated by its luminal SpAb.     

Analysis of antibody responses by commercial western blot assay in horses with alveolar echinococcosis

Commercial western blot (WB) assay was used to detect serum antibodies specific to Echinococcus multilocularis in 23 horses in which infection was confirmed by postmortem inspection at a slaughterhouse. Livers contained from 1 to >20 nodular lesions; foci diameter ranged from 1 to 25 mm. Antibody tests of serum from all 23 animals were negative for antigen bands at 7, 16, 18, and 26-28 kDa, which show specificity in the serum of human patients. However, sera from two infected horses with the largest nodules (diameter, 25 mm) showed positive response to one of the 22-kDa and 30-kDa antigen bands. It may be possible to diagnose E. multilocularis infection in horses based on the detection of these bands on commercial WB assay.     

Morphology of the oviduct fluke,Prosthogonimus ovatus,isolated from Indonesian native chickens and histopathological observation of the infected chickens

Prosthogonimus ovatus infection was detected in 5 of 130 chickens in the oviduct and 4 chickens in the bursa of Fabricius. Scanning electron microscopy (SEM) revealed that the spines of the P. ovatus were densely distributed on the cuticula of the entire dorsal surface of body, but on the ventral surface, they were densely present to the level of ventral sucker but gradually decreased in density posteriorly, and they could not be seen in the posterior 1/3 area. The spines were finger-shaped and denticulate at the tip. Histopathological examination showed that polypous elevations, degeneration and exfoliation of the mucosal epithelium were detected in the bursa of Fabricius possibly by the suction of flukes, in addition to the stratification of the mucosal epithelium, and interstitial cell infiltration.