Expression Pattern of Vascular Endothelial Growth Factor in Canine Folliculogenesis and its Effect on the Growth and Development of Follicles after Ovarian Organ Culture

In this study, the expressions of VEGF in dog follicles were detected by immunohistochemistry and the effects of VEGF treatment on the primordial to primary follicle transition and on subsequent follicle progression were examined using a dog ovary organ culture system. The frozen‐thawed canine ovarian follicles within slices of ovarian cortical tissue were cultured for 7 and 14 days in presence or absence of VEGF. After culture, the ovaries were fixed, sectioned, stained and counted for morphologic analysis. The results showed that VEGF was expressed in the theca cells of antral follicles and in the granulosa cells nearest the oocyte in preantral follicle but not in granulosa cells of primordial and primary follicles; however, the VEGF protein was expressed in CL. After in vitro culture, VEGF caused a decrease in the number of primordial follicles and concomitant increase in the number of primary follicles that showed growth initiation and reached the secondary and preantral stages of development after 7 and 14 days. Follicular viability was also improved in the presence of VEGF after 7 and 14 days in culture. In conclusion, treatment with VEGF was found to promote the activation of primordial follicle development that could provide an alternative approach to stimulate early follicle development in dogs.     

Proposal for stock-recruitment relationship for Japanese sardine <Emphasis Type="Italic">Sardinops melanostictus</Emphasis> in North-Western Pacific

ABSTRACT:   This study assessed the stock-recruitment relationship (SRR) for the Japanese sardine Sardinops melanostictus in the North-western Pacific. Of the 20 SRR models investigated, the Akaike information criterion (AIC) was the minimum (AIC = 551.2) when the data were separated into two groups (A and B) and the log-normal distribution was applied as the error term. Group A was constructed with SRR data from 1976–1987 and 1992–2004. Group B consisted of data from 1988–1991. The AIC minimum model was R  = 22.8 S  ×  e ^ε for Group A, where R , S , and ε denote the recruitment of sardine (individual number of 0-year old fish), spawning stock biomass (SSB), and error term, respectively. This model indicated that recruitment was proportional to the SSB and that no density-dependent effect operated over the range of SSB investigated (51 000–11.3 million t). Recruitment was markedly higher (lower) when the sea surface temperature (SST) of the Kuroshio Extension area in February was low (high). The essential SRR can simply be expressed as R  = 22.8 S  ×  e ^ε with the level of recruitment deviating from the model to a greater or lesser degree depending on the environmental conditions.     

Effects of low pH on the reproductive behavior of salmonid fishes

In northern Europe and America, the acidification of lakes and rivers induced by acid precipitation has damaged fish populations. At present, rapidly expanding industrial activities in East Asia have led to continuous increases in the quantity of emission of acidic pollutants, and rain of acidic levels of pH4 has precipitated throughout Japan. In this way, the effects of acid rain on fish populations have also been manifested in Japan. In order to examine the effects of low pH on the reproductive behavior of salmonids which are known to be acid-sensitive species, changes in the frequency of upstream behavior and spawning-related female nest-digging behavior in response to changes in pH were observed in mature hime salmon (land-locked sockeye salmon) Oncorhynchus nerka, brown trout Salmo trutta and Japanese char Salvelinus leucomaenis. Digging and upstream behavior were significantly inhibited in weakly acidic water of pH 5.8–6.4 formulated using sulfuric acid. Land-locked sockeye salmon was the most sensitive to changes in pH among the three species.     

Effect of retinoic acid on gene expression profiles of bovine intramuscular preadipocytes during adipogenesis

To investigate genes involved in intramuscular adipogenesis in ruminants, 16 genes with dramatic variable expression were selected. These were selected from the differentiation‐ and proliferation‐phase libraries of our previous serial analysis of gene expression (SAGE) studies of a clonal bovine intramuscular preadipocyte (BIP) cell line. We harvested the BIP cells over 12 days after adipogenic stimulation with all‐trans retinoic acid (ATRA). Quantitative real‐time PCR confirmed the earlier SAGE study results of the expression patterns of 15 of the genes. On day 6, TG accumulation increased significantly in the BIP cells but was completely inhibited in the 3T3‐L1 cells (the monogastric reference). ATRA enhanced expression levels of six genes whereas it suppressed expression of eight genes on day 3 of adipogenesis in the BIP cells. Forty‐eight hours after transfection, the messenger RNA expression level of the adipose differentiation‐related protein (ADFP), encoded by one of the upregulated genes, in the ADFP small interference RNA (siRNA)‐transfected cells was 3.5% of that in negative control‐transfected cells. Also, 6 days after induction the TG level in the ADFP siRNA‐transfected cells was 21.8% lower than that in negative control‐transfected cells. This analysis of gene expression profiles after ATRA treatment will contribute to our understanding of the molecular mechanisms involved in bovine intramuscular adipogenesis.     

Performance of a closed recirculating system with foam separation, nitrification and denitrification units for intensive culture of eel: towards zero emission

The development of a closed recirculating aquaculture system that does not discharge effluents would reduce a large amount of pollutant load on aquatic bodies. In this study, eel were reared in a closed recirculating system, which consisted of a rearing tank, a foam separation unit, a nitrification unit and a denitrification unit. The foam separation unit has an inhalation-type aerator and supplies air bubbles to the rearing water. The growth of eel, which were fed a commercial diet, was satisfactory, with gross weight increases of up three times in 3 months. The survival rate under the congested experimental conditions was 91%. The foam separation unit maintained oxygen saturation in the rearing water at about 80%. Furthermore, fine colloidal substances were absorbed on the stable foam formed from eel mucus and were removed from the rearing water by foam separation. Ammonia oxidation and the removal of suspended solids were accomplished rapidly and simultaneously in the nitrification unit. The ammonia concentration and turbidity were kept at less than 1.2 mg of N per litre and 2.5 units, respectively. When the denitrification process was operated, nitrate that accumulated in the rearing water (151 mg of N per litre) was reduced to 40 mg of N per litre. The sludge was easily recovered from the nitrification and denitrification tanks, and the components were found suitable as compost. Based on these results, the intensive aquaculture of freshwater fish such as eel can be achieved using a closed recirculating system without emission.     

Annual dynamics of marine birnavirus (MABV) in cultured Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis> and sea water

ABSTRACT:   Seasonal changes in the infection state of marine birnavirus (MABV) in Japanese flounder Paralichthys olivaceus and in rearing sea water are described. Sea water and 10–11 healthy fish were sampled monthly from April 2002 to February 2003. The MABV genome was detected throughout the year in > 80% of the fish examined at each sampling. The virus was isolated from the liver, kidney, and spleen, but not from the brain. The detection rate in each organ increased from April to October, and then decreased. Detection of virus antigen by the indirect fluorescent antibody technique also showed that the virus was present from spring to autumn (June–September) in the liver, kidney, and spleen, but not the brain. Sequence analysis of the MABV genome at the VP2–NS region revealed two specific mutations compared to the standard yellowtail strain (Y-6). It is suggested that the infection state of MABV in Japanese flounder changes to a latent or persistent infection after autumn. MABV was detected in sea water between September and February, suggesting that virus particles in the environment are relatively higher during cool seasons.