Comparison of cadherin and integrin localization in bovine cystic and healthy follicles

As stage progresses in the cystic follicle, granulosa cells are lost. We hypothesized that the granulosa and theca interna layers are detached in association with weakened expression of cell adhesion molecules such as cadherin (cell–cell adhesion) and integrin (cell–extracellular matrix adhesion) in cystic follicles. To elucidate this hypothesis, we immunolocalized these molecules in the granulosa and theca interna and compared them between cystic and small healthy follicles. Sections were immunostained with cadherin and integrin β₁ antibodies and their localizations were compared. Cadherin‐positive reaction was seen in the cytoplasma of all granulosa cells. No increase in the frequency of cadherin‐positive area in the granulosa layers and the intensity of cadherin immunoreaction in the theca interna was detected in cystic follicles compared with healthy ones. A dense immunoreaction product of integrin β₁ was detected in the theca interna in both cystic and healthy follicles. Intensity of integrin β₁‐immuno reaction in the granulosa layers and integrin β₁‐positive area in the theca interna was significantly lower in the cystic follicle than in the healthy follicles. These results suggest that granulosa and theca interna cells are detached while maintaining the cell–cell adhesion, resulting in the consequent loss of these layers from the cystic follicle.     

Cloning of kuruma prawn Marsupenaeus japonicus crustin-like peptide cDNA and analysis of its expression

ABSTRACT:   Antimicrobial peptides serve as an important component of the innate immune system of all species by functioning to provide a rapid first line defense against infection. Arthropod antimicrobial peptides have been well described in insects, whereas only a few molecules have been identified in crustaceans. Five variants (types 1–5) of Marsupenaeus japonicus crustin-like peptide cDNA that were obtained from a hemocyte cDNA library and polymerase chain reaction (PCR) amplification are reported here. Marsupenaeus japonicus crustin-like peptide type 1, the predominant type, has a cDNA consisting of 679 nucleotides and an open reading frame consisting of 573 base pairs coding for 191 amino acid residues. Other types contain varying glycine-rich repeats at the N-terminal amino acid sequences. The deduced amino acid sequences of these variants are highly similar to those of Litopenaeus setiferus (80% identity), Litopenaeus vannamei (80% identity) and Carcinus maenas crustins (44% identity). Expression of Marsupenaeus japonicus crustin-like peptide mRNA was detected in hemocytes, but not in the heart, hepatopancreas, gill, fore-gut, mid-gut, muscle, subcuticular epithelium or ovary. The expression level of crustin-like peptide mRNA increased significantly 1, 3 and 7 days post-peptidoglycan feeding as determined by quantitative real-time PCR. These results suggest that crustin-like peptide could have an important role in shrimp defense mechanisms.     

Effect of trehalose on the gel-forming ability,state of water and myofibril denaturation of horse mackerel <Emphasis Type="Italic">Trachurus japonicus</Emphasis> surimi during frozen storage

The cryoprotective effects of trehalose on fish myofibrillar protein were compared with those of sucrose, glucose and sorbitol. The frozen surimi with trehalose exhibited significantly higher Ca²⁺-ATPase activity through-out the storage periods, resulting in higher gel-forming ability than that of without trehalose. The amount of unfrozen water was significantly increased in the surimi upon addition of trehalose at any concentrations tested. The findings suggest that trehalose constructed bound water molecules in protein structure, consequently suppressed freeze-induced denaturation of protein and maintained gel-forming ability. An addition of 5.0% to 7.5% concentration of trehalose showed threshold behavior to increase the amount of unfrozen water and to prevent freeze-induced denaturation of protein. The effects of trehalose were almost similar to those of other sugars.     

Occurrence of two distinct molecular species of cathepsin B in carp Cyprinus carpio

ABSTRACT:   We purified cathepsins B₁ and B₂ from the ordinary muscle of carp Cyprinus carpio . The N-terminal amino acid sequences (12 residues) of 29 kDa bands of cathepsins B₁ and B₂ are the same and showed high homology of 75% and 83%, respectively, with the heavy chain of rat and human cathepsins B. Based on conserved sequences of other cathepsins B and the N-terminal amino acid sequences of 29 kDa bands, we cloned carp cathepsin B cDNA. The nucleotide sequence of carp cathepsin B cDNA consists of 1470 bp including a 993 bp open reading frame, encoding a deduced protein of 330 amino acids. The deduced amino acid sequence of carp cathepsin B has similarity of 80% to rainbow trout cathepsin B and of 76–78% to other vertebrate cathepsins B. The sequence of its isoform was also determined during molecular cloning, which has 94.8% similarity with first cloned cathepsin B. They are completely same in N-terminal amino acid sequence of heavy chain, active site and potential N-glycosylation site. This indicates there are at least two kinds of cathepsin B functioning in vivo in carp.     

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.     

Seasonal variation in docosahexaenoic acid content in horse mackerel caught in the East China Sea

ABSTRACT:   The fatty acid composition in horse mackerel caught off Nagasaki, off Tsushima Island, and in the middle of the East China Sea was investigated. The ratios of monoenoic and polyenoic acids to the total fatty acids in the Nagasaki and bull trawl catch specimens had a negative correlation. The docosahexaenoic acid (DHA; 22:6n-3) ratios of total fatty acids in summer-caught specimens were lower than those in winter-caught specimens, and this tendency was significantly clearer in smaller-sized fish, while DHA levels in tissues varied little throughout the year. Consistently high levels of DHA were found in various fishing areas, which suggests that horse mackerel offers a stable source of DHA.     

Chemical components and body color of horse mackerel caught in different areas

ABSTRACT: The chemical components and body color of horse mackerel caught between July 1997 and May 2000 were studied. The fish were caught with small- to medium-sized purse seine offshore from Nagasaki and from Tsushima, and with bull trawl seine and medium- to large-sized purse seine in the East China Sea. The crude lipid content of the fish caught offshore from Tsushima was higher than that of the others, and there were no significant differences among the other catches. The extractive nitrogen content of the fish caught with bull trawl seine was somewhat lower than that of the others. The body colors of the fish differed from those suggested by their common names: Kuroaji (black) , Kiaji (yellow), and Shiroaji (white). The crude lipid content and body color indicated that there was no difference in quality among the catches, with the exception of the crude lipid content of the Tsushima catch.     

Verification of Drainage Sterilization System Uses Low-Voltage Pulsed Electric Field in a Prawn Farm

It is necessary to take measures against infectious diseases in the Southeast Asian prawn farming industry. In giant tiger prawn （Penaeus monodon） aquaculture, diseases caused by viruses such as white spot syndrome virus （WSSV） and bacteria such as Vibrio have become a menace. Appropriate treatments of prawn culture pond＇s waters are advocated for preventing and controlling pathogens. The purpose of this study was to conduct an antimicrobial water treatment using a low-voltage pulsed electric field sterilization. Here we prepared a mechanical low-voltage pulsed electric field system with copper wire coiling around a titanium ring. The viability of WSSV in seawater was examined by prawn infectivity experiments. We inoculated healthy prawns （approximately 10-15 g） with the WSSV master sample that passed 0-3 times through the system. WSSV infection in prawns decreased according to the number of passes through the system. Healthy prawns survived for〉 10 days without feeding, where as prawns inoculated with the WSSV master sample showed symptoms of white spot and died in about 4 days. Two-thirds of the prawns inoculated with the WSSV master sample that was passed once through the system developed WSSV symptoms and were polymerase chain reaction （PCR） positive. However, no symptoms were observed and prawns were PCR negative when the WSSV master sample was passed three times through the system. Based on these results, we purpose that a low-voltage pulsed electric field system will serve as an efficient pond drainage sterilization system and will replace conventional treatments using chemicals such as sodium hypochlorite.