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Long-chain (≥ C20) polyunsaturated fatty acids (LC-PUFA), such as eicosapentaenoic acid (20:5n-3, EPA) and docosahexaenoic acid (22:6n-3, DHA), are necessary for human health and are obtained from marine fish-derived oils. Marine fish are LC-PUFA-rich animals; however, many of them require LC-PUFA for growth. Therefore, it is suggested that they do not have sufficient ability to biosynthesize LC-PUFA. To evaluate in vivo LC-PUFA synthetic activity in fish cells, fish-derived cell lines from red sea bream (Pagrus major, PMS and PMF), Japanese flounder (Paralichthys olivaceus, HINAE), and zebrafish (Danio rerio, BRF41) were incubated with n-3 fatty acids labeled by radioisotopes or stable isotopes, and then, n-3 PUFA were analyzed by thin-layer chromatography or liquid chromatography-mass spectrometry. Labeled EPA and DHA were biosynthesized from labeled α-linolenic acid (18:3n-3) in BRF41, whereas they were not detected in PMS, PMF, or HINAE cells. We next cloned the fatty acid desaturase 2 (Fads2) cDNAs from PMF cells and zebrafish, expressed in budding yeasts, and then analyzed the substrate specificities of enzymes. As a result, we found that Fads2 from PMF cells was a ∆6/∆8 desaturase. Collectively, our study indicates that cell lines from red sea bream and Japanese flounder were not able to synthesize EPA or DHA by themselves, possibly due to the lack of ∆5 desaturase activity. Furthermore, this study provides a sensitive and reproducible non-radioactive method for evaluating LC-PUFA synthesis in fish cells using a stable isotope and liquid chromatography-mass spectrometry.

The protective efficacy of intranasal (IN) administration of inactivated feline calicivirus (FCV) vaccine against homologous or heterologous FCV infection was investigated. Groups of cats immunized with the experimental inactivated, non-adjuvanted FCV vaccine via either the IN or subcutaneous (SC) route were exposed to homologous or highly heterologous FCV. Both the IN and SC immunization protocols established robust protection against homologous FCV infection. Although neither immunization regimen conferred protection against the heterologous strain, clinical scores and virus titres of oral swabs were lower in cats in the IN group compared to those in the SC group, accompanying a faster neutralizing antibody response against the heterologous virus in cats in the IN group. The IN group secreted more IgA specific to FCV proteins in oral washes (lavage fluids from the oral cavity) than the SC group. IN immunization with an inactivated whole FCV particle, which protects cats from homologous virus exposure and shortens the period of heterologous virus shedding, may serve as a better platform for anti-FCV vaccine.

Chattonella antiqua isolated in 2010 showed extremely more potent fish-killing activities against red sea bream, Japanese horse mackerel, and blue damselfish than those of Chattonella marina isolated in 1985. Chemiluminescence and electron spin resonance (ESR) analyses suggested greater reactive oxygen species (ROS)-producing activity of C. antiqua than that of C. marina. Sodium benzoate, a hydroxyl radical scavenger, significantly suppressed the fish-killing activity of C. antiqua on blue damselfish. The chlorophyll level in the gill tissue of blue damselfish exposed to flagellate cells increased along with the exposure time, and the cell count of gill-associated C. antiqua estimated with chlorophyll level was higher than that of C. marina. These results suggest that the ROS-producing activity and affinity of Chattonella cells to the gill surface may be important factors influencing the fish-killing activity of Chattonella species.

A number of artificial earthquake ground motions compatible with time-frequency characteristics of recorded actual earthquake ground motions as well as the given target response spectrum are generated using wavelet transform. The coefficient of variation (C. O. V.) of maximum displacement on elasto-plastic SDOF systems excited by these artificial ground motions are numerically evaluated.