Glycoprotein hormone receptor family in medaka (Oryzias latipes)

Four types of GPHR cDNAs have been cloned from ovary and testis of medaka (Oryzias latipes) and their gene constructions have been determined. Two of them are closely related to known fish receptors for FSH and LH, respectively. Changes in their mRNA levels were examined during the course of oogenesis. FSH receptor mRNA could not be detected from 20 h before ovulation, whereas LH mRNA remained 5 h before ovulation.     

Isolation of heat-tolerant myoglobin from Asian swamp eel Monopterus albus

Myoglobin from Asian swamp eel Monopterus albus was purified from fish muscle using salt fractionation followed by column chromatography and molecular filtration. The purified Mb of 0.68?mg/g wet weight of muscle was determined for its molecular mass by MALDI-TOF-MS to be 15,525.18?Da. Using isoelectric focusing technique, the purified Mb showed two derivatives with pI of 6.40 and 7.12. Six peptide fragments of this protein identified by LC-MS/MS were homologous to Mbs of sea raven Hemitripterus americanus, yellowfin tuna Thunnus albacores, blue marlin Makaira nigicans, common carp Cyprinus carpio, and goldfish Carassius auratus. According to the Mb denaturation, the swamp eel Mb had thermal stability higher than walking catfish Clarias batrachus Mb and striped catfish Pangasius hypophthalmus Mb, between 30 and 60?(°)C. For the thermal stability of Mb, the swamp eel Mb showed a biphasic behavior due to the O(2) dissociation and the heme orientation disorder, with the lowest increase in both Kd(f) and Kd(s). The thermal sensitivity of swamp eel Mb was lower than those of the other Mbs for both of fast and slow reaction stages. These results suggest that the swamp eel Mb globin structure is thermally stable, which is consistent with heat-tolerant behavior of the swamp eel particularly in drought habitat.     

Feeding events of Japanese lates <Emphasis Type="Italic">Lates japonicus</Emphasis> detected by a high-speed video camera and three-axis micro-acceleration data-logger

The feeding events of wild animals have been widely researched in recent decades. The use of invasive methods to determine stomach contents in which the fish is killed can, however, be considered to be unjustifiable in the case of an endangered fish, such as the Japanese late. Here, we report on a method that can be used to verify feeding events of a fish species without the need for killing the fish through the use of a high-speed video camera and a three-axis micro-acceleration data-logger. High-quality images obtained by the video showed that the fish opens its mouth with a quick downward movement of the mandibles and ingests prey via suction feeding. The movement of the mandibles was also simultaneously recorded by changes in the three-axis accelerations obtained by the logger attached to the dorsum. We attached the logger with an automatic release system to the dorsum of one captive Japanese lates and two wild fish together with an acoustic transmitter and then released the fish in the Shimanto River in Japan. After retrieving the logger, the movement records of the fish obtained by the three-axis accelerometer showed the same pattern of feeding events as those in captivity. A total of 13 feeding events at night and five during the daytime were obtained for the three fish during a total 129.7 h of recording.     

Identification of Antiviral Agents Targeting Hepatitis B Virus Promoter from Extracts of Indonesian Marine Organisms by a Novel Cell-Based Screening Assay

The current treatments of chronic hepatitis B (CHB) face a limited choice of vaccine, antibody and antiviral agents. The development of additional antiviral agents is still needed for improvement of CHB therapy. In this study, we established a screening system in order to identify compounds inhibiting the core promoter activity of hepatitis B virus (HBV). We prepared 80 extracts of marine organisms from the coral reefs of Indonesia and screened them by using this system. Eventually, two extracts showed high inhibitory activity (>95%) and low cytotoxicity (66% to 77%). Solvent fractionation, column chromatography and NMR analysis revealed that 3,5-dibromo-2-(2,4-dibromophenoxy)-phenol (compound 1) and 3,4,5-tribromo-2-(2,4-dibromophenoxy)-phenol (compound 2), which are classified as polybrominated diphenyl ethers (PBDEs), were identified as anti-HBV agents in the extracts. Compounds 1 and 2 inhibited HBV core promoter activity as well as HBV production from HepG2.2.15.7 cells in a dose-dependent manner. The EC₅₀ values of compounds 1 and 2 were 0.23 and 0.80 µM, respectively, while selectivity indexes of compound 1 and 2 were 18.2 and 12.8, respectively. These results suggest that our cell-based HBV core promoter assay system is useful to determine anti-HBV compounds, and that two PBDE compounds are expected to be candidates of lead compounds for the development of anti-HBV drugs.     

Bio-economic analysis of super-intensive closed shrimp farming and improvement of management plans: a case study in Japan

Fisheries Science - Crustacean aquaculture is a multibillion-dollar industry worldwide that continues to show significant growth. Shrimp farming has been intensified for decades, and...     

Carbon nanotube enhanced label-free immunosensor for amperometric determination of oocyte maturation-inducing hormone in fish

Maintaining high-quality fish eggs stably and efficiently is important for aquaculture. We developed a label-free immunosensor system for measuring 17,20β-dihydroxy-4-pregnen-3-one (DHP). DHP is suddenly secreted before ovulation as a maturation-inducing hormone in fish, and therefore, DHP levels are an indicator for predicting ovulation. The method is based on immunologic reactions and amperometric measurement using cyclic voltammetry (CV). For biomolecular immobilization on the surface of sensing electrode, Au electrode, we used self-assembled monolayers of thiol-containing compounds to fix anti-DHP immunoglobulin. In addition, we used a single-walled carbon nanotube to improve sensitivity. Using this electrode, we were able to determine the CV signal change caused by the antigen–antibody complex. The proposed immunosensor system showed a linear correlation (correlation coefficient: 0.9827) between the anodic peak current of the CV and the DHP level in range from 15.6 to 50,000 pg ml^?1. The sensor system was then applied to monitor DHP of goldfish (Carassius auratus). Blood plasma of fish was collected every 3 h after administering a DHP inducer. In the measurement, the anodic peak current of the CV showed distinct changes depending on DHP levels in the blood plasma. A good relationship was observed between DHP levels determined by our proposed system and the conventional method (correlation coefficient: 0.9351).     

Construction of a linkage map for watermelon (Citrullus lanatus (Thunb.) Matsum & Nakai) using random amplified polymorphic DNA (RAPD)

Summary A linkage map for watermelon (Citrullus lanatus) was constructed on the basis of RADP, ribosomal DNA restriction fragment length polymorphism (RFLP), isozyme, and morphological markers using F₁BC1. A segregating population of 78 individuals was the result of a backcross of a cultivated inbred line (H-7; Citrullus lanatus; 2n=22) and a wild form (SA-1; C. lanatus; 2n=22), in which the latter was the recurrent (male) parent. A total of 69 RAPD, one RFLP, one isozyme, and three morphological markers was found to segregate in the BC1 population. Linkage analysis revealed that 62 loci could be mapped to 11 linkage groups that extended more than 524 centimorgans (cM), while 12 loci segregated independently of all other markers. The locus for exocarp color was linked to two RAPD markers within a region of 5 cM on linkage group 4. The locus for flesh color was linked to a RAPD marker within a region of 30 cM on linkage group 6. The isozyme marker GOT was located on the linkage group 1. Linkage group 2 contained a locus for ribosomal DNA within 5 cM of a RAPD marker. Half of the RAPD markers on the linkage group 7 displayed severely distorted segregation. The construction of linkage map using molecular markers is necessary for the breeding of watermelon to introduce useful gene of wild watermelon efficiently. However the linkage map that was constructed for the most part on the basis of RAPD markers could not cover significant parts of the genome, the linkage map provides breeders of watermelons the possibility of tagging useful agronomic traits, as well as the gene for exocarp color.Abbreviations RAPD random amplified polymorphic DNA - RFLP restriction fragment length polymorphism - GOT glutamate oxaloacetate transaminase - MDH malate dehydrogenase - ACP acid phosphatase - 6PGH 6-phosphogluconate dehydrogenase     

One-year stress relaxation of timber joints assembled with pretensioned bolts

In our previous study, great increases of hysteretic damping and initial slip resistance of timber joints were attained by applying axial pretension to the steel fasteners. To evaluate the effectiveness of this method, 1-year stress-relaxation measurement was carried out. Nine prestressed joints were prepared and three of them were restressed after 3 and then 6 months after the initial prestressing. All joints were exposed to indoor conditions, and relaxation of the pretension was regularly measured from time-dependent decreases of axial strain of the bolts. After measurement, the joints were subjected to cyclic and monotonic loading tests until failure. The average ratio of residual stress to the initial prestress after 1 year was about 0.23 and 0.66, respectively, for joints without restressing and those with restressing. A simulated stress-relaxation curve developed from the four-element relaxation model predicted 3% of the initial stress after 5 years. Without a regular restressing program, the initial prestressing effect therefore must be considered negligible. However, about 20% of the pre-stress level can be reasonably assumed if restressing is carried out annually. This small residual stress was found to introduce suffi cient frictional damping to signifi cantly increase the equivalent viscous damping ratio of the joints. Part of this study was presented at the 10th World Conference on Timber Engineering, Miyazaki, June 2008