Since 2011, high mortality rates and symptoms consistent with vibriosis have been observed in farmed amberjack (Seriola dumerili) in Japan. To identify 41 strains isolated from diseased amberjack, a multilocus sequence analysis using nine concatenated genes (ftsZ, gapA, gyrB, mreB, pyrH, recA, rpoA, topA and 16S rRNA) was conducted. Twenty-seven strains were identified as Vibrio harveyi, suggesting an epidemic of V. harveyi infection in amberjack farms. Other strains were identified as Vibrio anguillarum, Vibrio owensii and Photobacterium damselae subsp. damselae. To develop an efficient diagnostic method for vibriosis in amberjack, a multiplex PCR system was developed to identify V. anguillarum, V. harveyi and P. damselae subsp. damselae. The method successfully discriminated between these three bacterial species, with amplification products of 350 bp for V. anguillarum, 545 bp for V. harveyi and 887 bp for P. damselae subsp. damselae and can be used for diagnosis in aquaculture farms.
Fish, like mammals, have several stages of wound healing, including inflammation, reepithelialization, proliferation, organization, and differentiation. However, little is known about the molecular mechanisms of fish skin wound healing. In the present study, 23 polymerase chain reaction (PCR) bands were identified which appeared to be specifically expressed during wound healing by a simple differential display (DD) method. Sequences of cDNAs from these 14 bands were highly homologous to known genes. However, reverse transcriptase PCR (RT-PCR) analyses of these 14 bands revealed that only one of them, the gene for matrix metalloproteinase 9 (MMP-9), was significantly expressed in wounded skin, and that the others were probably false positives. MMPs are known to have gelatinolytic activity, and film in situ zymography detected strong gelatinolytic activity in the surface and dermis of wounded skin of flounder. KEY WORDS: differential display, film in situ zymography, Japanese flounder, matrix metalloproteinase 9 (MMP-9), Paralichthys olivaceus, skin, wound healing.
During the course of our chemical analysis of the hydrophilic fractions from marine cyanobacterium Moorena producens, we have isolated natural dolapyrrolidone (Dpy, 1), a natural pyrrolidone derived from phenylalanine, for the first time as a single compound. Compound 1, with an (S)‐l absolute stereochemistry, was previously identified as a substructure that is common among several bioactive natural peptides. Surprisingly, the absolute stereochemistry of the isolated natural 1, determined through total synthesis, was (R)‐d. This result was unambiguously determined by HPLC analysis using a chiral stationary column by comparing the retention times of the natural 1 and authentic samples of synthetic enantiomers. To verify the unexpected result, the absolute stereochemistry of the natural 1 was confirmed by X‐ray crystallographic analysis of Pt‐complex derivative using the synthetic enantiomer.
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