Ostland V E, Stannard J A, Creek J J, Hedrick R P, Ferguson H W, Carlberg J M, Westerman M E
Kent SeaTech Corporation, 11125 Flintkote Ave., Suite J, San Diego, California 92121, USA.
Dis Aquat Organ. 2006 Oct 17;72(2):135-45. doi: 10.3354/dao072135.
The present study identifies an emerging disease associated with an aquatic Francisella-like bacterium that can cause mortality in hybrid striped bass Morone chrysops x M. saxatilis reared intensively in freshwater. Clinically affected fish were lethargic, had scattered haemorrhagic cutaneous lesions and diffuse gill pallor. The head kidney and spleen were markedly swollen and contained numerous interstitial granulomas; histological examination revealed small, pleomorphic Gram-negative coccobacilli within vacuolated cells. The bacterium could not be cultured from head kidney homogenates either with standard or enriched microbiological media or following inoculation of a Chinook salmon embryo (CHSE)-214 cell line. No amplification product was obtained from head kidney DNA by polymerase chain reaction (PCR) assay using Piscirickettsia salmonis-specific primers. PCR analysis of infected head kidney homogenate with primers designed for the eubacterial 16S rRNA produced a single amplicon. Phylogenetic analysis of this DNA sequence demonstrated that the sequence aligned most closely with members of the genus Francisella, identified from tilapia Oreochromis spp. in Taiwan and an aquatic Francisella species that was recently isolated from the three-line grunt Parapristipoma trilineatum in Japan. This Francisella-like disease was transmitted to naive hybrid striped bass fingerlings by intraperitoneal injection of tissue homogenates prepared from a natural outbreak. All fish developed gross and histological lesions identical to those from natural outbreaks. Intracellular Gram-negative bacteria were observed within the cytoplasm of cells (presumably macrophages) within the granulomas, but bacteria were not recovered. The 16S DNA sequence of the bacterium obtained from tissues of experimentally infected fish was identical to that obtained from the fish used as infected donor tissue.
本研究鉴定出一种与水生弗朗西斯菌属样细菌相关的新出现疾病,该细菌可导致在淡水中集约化养殖的杂交条纹鲈(条纹鲈♀×白鲈♂)死亡。临床受影响的鱼表现为嗜睡、体表有散在出血性病变且鳃弥漫性苍白。头肾和脾脏明显肿大,含有大量间质肉芽肿;组织学检查显示空泡化细胞内有小的、多形性革兰氏阴性球杆菌。无论是使用标准或富集微生物培养基,还是接种奇努克鲑胚胎(CHSE)-214细胞系后,均无法从受感染鱼的头肾匀浆中培养出该细菌。使用鲑鱼立克次氏体特异性引物通过聚合酶链反应(PCR)检测头肾DNA,未获得扩增产物。用针对真细菌16S rRNA设计的引物对受感染的头肾匀浆进行PCR分析,产生了一个单一扩增子。对该DNA序列进行系统发育分析表明,该序列与在台湾罗非鱼中鉴定出的弗朗西斯菌属成员以及最近从日本三线矶鲈中分离出的一种水生弗朗西斯菌物种的序列最为接近。通过腹腔注射由自然暴发制备的组织匀浆,将这种弗朗西斯菌属样疾病传播给未感染的杂交条纹鲈幼鱼。所有鱼都出现了与自然暴发相同的大体和组织学病变。在肉芽肿内的细胞(可能是巨噬细胞)胞质内观察到细胞内革兰氏阴性细菌,但未分离到细菌。从实验感染鱼组织中获得的细菌16S DNA序列与用作感染供体组织的鱼中获得的序列相同。