Cornwell Emily R, Bellmund Chelsea A, Groocock Geoffrey H, Wong Po Ting, Hambury Katherine L, Getchell Rodman G, Bowser Paul R
Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853, USA.
J Vet Diagn Invest. 2013 Mar;25(2):203-9. doi: 10.1177/1040638713476865. Epub 2013 Feb 12.
Nonlethal sampling is becoming a common method to diagnose fish diseases, especially with the availability of molecular testing. Viral hemorrhagic septicemia virus (VHSV) is a viral pathogen of finfish distributed worldwide. Although VHSV has been known to occur in some parts of the world for decades, a new genotype, IVb, recently emerged in the Laurentian Great Lakes of northeastern North America. Golden shiners (Notemigonus crysoleucas; Mitchill, 1814) and fathead minnows (Pimephales promelas; Rafinesque, 1820) were exposed to VHSV-IVb doses between 10(2) and 10(6) plaque forming units per fish by intraperitoneal injection at 10°C. Both species experienced significant mortality after exposure, ranging from 38% to 52% in golden shiners and from 35% to 95% in fathead minnows. In golden shiners, a fin or gill sample was somewhat less sensitive at detecting VHSV-IVb by quantitative reverse transcription polymerase chain reaction (qRT-PCR) than a pooled organ sample (consisting of liver, anterior and posterior kidney, spleen, and heart), however the relative sensitivity increased when a fin and gill sample were tested in parallel. In fathead minnows, a fin or gill sample tested alone or in parallel was relatively more sensitive than a pooled organ sample by qRT-PCR. Specificity was 100% for all sample types in both species. The results suggest that fin and gill biopsies are useful tools to test for VHSV in live fish.
非致死性采样正成为诊断鱼类疾病的常用方法,尤其是在分子检测技术可用的情况下。病毒性出血性败血症病毒(VHSV)是一种在全世界范围内分布的硬骨鱼病毒性病原体。尽管VHSV在世界某些地区已存在数十年,但一种新的基因型IVb最近在北美东北部的五大湖出现。将金体美鳊(Notemigonus crysoleucas;米奇尔,1814年)和黑头软口鲦(Pimephales promelas;拉菲内斯克,1820年)在10℃下通过腹腔注射,使其接触每尾鱼10²至10⁶个噬斑形成单位剂量的VHSV-IVb。两种鱼在接触后均出现显著死亡率,金体美鳊的死亡率在38%至52%之间,黑头软口鲦的死亡率在35%至95%之间。在金体美鳊中,通过定量逆转录聚合酶链反应(qRT-PCR)检测VHSV-IVb时,鳍或鳃样本的敏感性略低于混合器官样本(由肝脏、前肾和后肾、脾脏和心脏组成),然而,当鳍和鳃样本同时检测时,相对敏感性会增加。在黑头软口鲦中,单独或同时检测的鳍或鳃样本通过qRT-PCR检测时相对比混合器官样本更敏感。两种鱼所有样本类型的特异性均为100%。结果表明,鳍和鳃活检是检测活鱼中VHSV的有用工具。