In a previous study, a member of Spirochaeta was presumed to cause Akoya oyster disease (AOD), and Candidatus Maribrachyspira akoyae was proposed as the putative causative agent. With the aim of performing an epidemiological study of AOD, polymerase chain reaction (PCR) assays were developed using the primers designed from shotgun metagenomic sequences identified as having high homology to Spirochaetes genes by a BLAST search. PCR assays with six primer sets produced specific target amplicons (n = 63–67) in pearl oysters collected from five AOD-endemic areas (n = 88), but did not in pearl oysters (n = 36) collected from three AOD-free areas. All individuals (n = 17) with the color index a-value above 3.0, which is an index of AOD development, were positive for all 6 primer sets in PCR. There was a positive correlation between the a-value and the results of PCR. These results show that the previous identification of Spirochaeta as an etiological agent of AOD is reasonable. Sequencing of amplification products confirmed only one nucleotide substitution in the PCR products, suggesting that the pathogenic causative agent of AOD has limited genetic diversity.
OsHV-1 国内型の病原性を種苗生産したマガキ幼生および稚貝を用いた感染実験で検討した。ウイルス源にはJPType1 が検出された死亡稚貝を用いた。ウイルス液を段階希釈して幼生の飼育海水に添加したところ,幼生に対する病原性が認められ,D型幼生のウイルス感受性は付着期幼生に比べて高かった。リアルタイムPCR法で定量したウイルスDNA量はD型幼生で最高 1.6 × 108 copies/ng(全DNA量)となった。殻高 1.8~6.1 mmの稚貝を用いた感染実験では,5日間の観察期間中に死亡は見られなかったが,ウイルスDNA量が 6.3 × 105 copies/mg(貝重量)になる群があった。また,生存していた小型稚貝の群では大型の群よりもウイルスDNA量が多い傾向が見られた。このように,OsHV-1 国内型JPType1 の病原性はマガキ幼生および稚貝に対して確認されたが,マガキの成長に伴い低下すると推察された。
The occurrence of orange/pinkish colored lesions in the adductor muscle of Yesso scallops Patinopecten yessoensis has been known for many years in Japan; however, determination of the causative agent has not been adequately investigated. Histological examination of affected scallops in southern Hokkaido typically revealed intense host responses: hemocyte infiltration, an abundance of necrotic hemocytes, lysis of muscle fibers and in some instances melanin deposits when the lesions occurred adjacent to the shell. Microbiota analysis showed that Francisella halioticida was dominant in the lesions, and in situ hybridization using F. halioticida specific probes also confirmed the presence of this bacterium within the lesions. A F. halioticida specific PCR assay detected this bacterium in the majority of scallop lesions tested. Subsequently, three bacterial isolates were obtained from scallop lesions on modified Eugon agar supplemented with antibiotics, and these bacterial isolates were found to be F. halioticida by 16S rRNA and rpoB gene sequences. These results suggest that infection with F. halioticida is the most likely cause of the adductor muscle lesions observed in Yesso scallops. Field surveys conducted in 2017 of scallops cultured in southern Hokkaido showed that the presence of adductor muscle lesions putatively caused by F. halioticida was significantly related to mortalities and poor growth of scallops.
In July 2016, there were mortalities of riverine ayu Plecoglossus altivelis in a tributary of the Tama River, Japan. A Gram-negative, motile and short rod-shaped bacterium was dominantly isolated from all examined dead fish, and identified as Aeromonas veronii biovar sobria. Biochemical characteristics and gyrB sequence of the present strains differed from those of A. veronii strains from ayu in previous years. The present strains also caused higher mortalities to ayu than A. veronii strains previously isolated. These results indicate that the present mortalities of riverine ayu in the Tama River were caused by high pathogenic A. veronii biovar sobria.
吸水過程において卵門から細菌性冷水病や細菌性腎臓病の病原細菌が卵内へ侵入する卵内感染の機序を明らかにするため,SEMを使用して,ニジマスの未受精卵,吸水卵および人為的な病原菌汚染卵における卵門および卵膜の形態を観察した。排卵された未受精卵の卵膜表面では,卵門を除き細菌が卵内に侵入する可能性のある小孔状の形態はみられなかった。卵門管の上部および管底の口径は細菌が十分に通過できるほどに大きく,吸水に伴って管内の閉塞が進む時期は実験的に卵内感染が成立した時期と一致することが確かめられた。