Thomas-Jinu S, Goodwin A E
Aquaculture/Fisheries Center, University of Arkansas at Pine Bluff, AR 71601, USA.
J Fish Dis. 2004 Jan;27(1):29-35. doi: 10.1046/j.1365-2761.2003.00507.x.
Variability in pathogenicity of Flavobacterium columnare makes disease treatment difficult because there is currently no way to easily recognize those strains that warrant aggressive treatments. In order to identify suitable virulence markers, 17 isolates of F. columnare were cultured from six different fish species. The DNA from all isolates was analysed using random amplified polymorphic DNA (RAPD). Bootstrap analysis of the RAPD data produced a tree with three major groups supported by bootstrap scores of 80-100%. Virulence of the isolates was determined by bath exposure of channel catfish, Ictaluruspunctatus (Rafinesque), and golden shiners, Notemigonus crysoleucas (Mitchill), to broth cultures of F. columnare. In channel catfish, 13 of 17 isolates produced 100% mortality within 48 h post-exposure. All isolates of cyprinid fish origin clustered in a single RAPD group. At least two of the four isolates that were not virulent in channel catfish were of cyprinid fish origin. There was a wide variation in cell morphology between isolates with lengths of cells or cell chains ranging from 3 to 11 microm, even under identical culture conditions. Most of the shorter or single cell isolates fell into a single RAPD group. No clear association was identified between virulence and any other characteristic, including RAPD group.
柱状黄杆菌致病性的变异性使得疾病治疗变得困难,因为目前尚无简便方法来识别那些需要积极治疗的菌株。为了确定合适的毒力标记,从六种不同鱼类中培养出17株柱状黄杆菌分离株。使用随机扩增多态性DNA(RAPD)对所有分离株的DNA进行分析。对RAPD数据的自展分析产生了一棵树,其中三个主要组得到了80%-100%的自展分数支持。通过将斑点叉尾鮰(Ictalurus punctatus,Rafinesque)和美国拟鲤(Notemigonus crysoleucas,Mitchill)置于柱状黄杆菌肉汤培养物中进行浸浴暴露,来确定分离株的毒力。在斑点叉尾鮰中,17株分离株中有13株在暴露后48小时内导致100%的死亡率。所有鲤科鱼类来源的分离株都聚集在一个RAPD组中。在斑点叉尾鮰中无毒性的4株分离株中,至少有两株来自鲤科鱼类。即使在相同培养条件下,分离株之间的细胞形态也存在很大差异,细胞或细胞链长度在3至11微米之间。大多数较短或单细胞的分离株属于一个单一的RAPD组。未发现毒力与任何其他特征(包括RAPD组)之间存在明确关联。