Draghi Andrew, Bebak Julie, Daniels Stephen, Tulman Edan R, Geary Steven J, West A Brian, Popov Vsevolod L, Frasca Salvatore
Department of Pathobiology and Veterinary Science, University of Connecticut, Storrs, Connecticut 06269, USA.
Dis Aquat Organ. 2010 Feb 24;89(1):39-49. doi: 10.3354/dao02171.
Arctic charr Salvelinus alpinus production facilities, nonproduction water sources and effluents in the United States and Canada were sampled to determine if chlamydiae associated with epitheliocystis were present in water and were associated with inclusions of epitheliocystis in gill tissue. Gills from 607 fish from 13 sites were processed for histopathologic examination and DNA extraction. Water was collected from 21 locations for DNA testing. Eighteen fish from one location had inclusions of epitheliocystis with proliferative and inflammatory gill lesions. Inclusions were stained using the Gimenez technique and, at the ultrastructural level, consisted of intracytoplasmic membrane-bound vacuoles containing reticulate and intermediate bodies in a fibrillar matrix. PCR using Order Chlamydiales-specific primers performed on DNA extracts from 12 of 13 infected fish yielded amplicons that were identical to (GQ302988) or differed at one base from (GQ302987) the 16S ribosomal RNA gene signature sequence of 'Candidatus Piscichlamydia salmonis', which is the chlamydia that was previously identified in epitheliocystis inclusions of farmed Atlantic salmon. In situ hybridization using a approximately 1.5 kb riboprobe corresponding to the 'Candidatus Piscichlamydia salmonis' 16S rRNA genetic sequence (AY462244) confirmed its presence within Arctic charr gill inclusions. DNA isolated from water samples was tested by Chlamydiales-specific PCR and yielded 54 partial 16S rRNA genetic sequences spanning the signature region; however, no 16S rRNA genetic sequences associated with epitheliocystis were identified. This is the first report of 'Candidatus Piscichlamydia salmonis' associated with epitheliocystis in Arctic charr, the first identification of 'Candidatus Piscichlamydia salmonis' from a freshwater production location, and the first reported occurrence in North America.
对美国和加拿大的北极红点鲑（Salvelinus alpinus）养殖设施、非养殖水源及废水进行采样，以确定与上皮囊肿相关的衣原体是否存在于水中，以及是否与鳃组织中的上皮囊肿包涵体有关。对来自13个地点的607条鱼的鳃进行处理，用于组织病理学检查和DNA提取。从21个地点采集水样进行DNA检测。来自一个地点的18条鱼有上皮囊肿包涵体，伴有鳃的增殖性和炎性病变。使用吉姆萨技术对包涵体进行染色，在超微结构水平上，包涵体由胞质内膜结合的液泡组成，液泡内含有在纤维状基质中的网状体和中间体。对13条受感染鱼中的12条的DNA提取物进行衣原体属特异性引物PCR，得到的扩增产物与“鲑鱼假衣原体（Candidatus Piscichlamydia salmonis）”的16S核糖体RNA基因特征序列（GQ302988）相同，或与该序列在一个碱基上不同（GQ302987），“鲑鱼假衣原体”是先前在养殖大西洋鲑的上皮囊肿包涵体中鉴定出的衣原体。使用对应于“鲑鱼假衣原体”16S rRNA基因序列（AY462244）的约1.5 kb核糖探针进行原位杂交，证实其存在于北极红点鲑鳃包涵体内。对从水样中分离的DNA进行衣原体属特异性PCR检测，得到54个跨越特征区域的部分16S rRNA基因序列；然而，未鉴定出与上皮囊肿相关的16S rRNA基因序列。这是关于“鲑鱼假衣原体”与北极红点鲑上皮囊肿相关的首次报告，是首次从淡水养殖地点鉴定出“鲑鱼假衣原体”，也是在北美首次报告其出现。
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