Mocho Jean-Philippe, Martin Darren J, Millington Mollie E, Saavedra Torres Yolanda
Biological Research Facility, The Francis Crick Institute;
Biological Research Facility, The Francis Crick Institute.
J Vis Exp. 2017 Dec 8(130):55306. doi: 10.3791/55306.
Health monitoring systems are developed and used in zebrafish research facilities because pathogens of Danio rerio such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa have the potential to impair animal welfare and research. The fish are typically analyzed post mortem to detect microbes. The use of sentinels is a suggested way to improve the sensitivity of the surveillance and to reduce the number of animals to sample. The setting of a pre-filtration sentinel tank out of a recirculating system is described. The technique is developed to prevent water pollution and to represent the fish population by a careful selection of age, gender, and strains. In order to use the minimum number of animals, techniques to screen the environment are also detailed. Polymerase Chain Reaction (PCR) on surface sump swabs is used to significantly improve the detection of some prevalent and pathogenic mycobacterial species such as Mycobacterium fortuitum, Mycobacterium haemophilum, and Mycobacterium chelonae. Another environmental method consists of processing the sludge at the bottom of a holding tank or sump to look for P. tomentosa eggs. This is a cheap and fast technique that can be applied in quarantine where a breeding device is submerged into the holding tank of imported animals. Finally, PCR is applied to the sludge sample and A. hydrophila is detected at the sump's bottom and surface. Generally, these environmental screening techniques applied to these specific pathogens have led to an increased sensitivity compared to the testing of pre-filtration sentinels.
健康监测系统在斑马鱼研究设施中得到开发和应用,因为斑马鱼的病原体,如嗜水气单胞菌、分枝杆菌属和绒毛伪毛细线虫,有可能损害动物福利和研究。通常对鱼进行死后分析以检测微生物。使用哨兵鱼是提高监测灵敏度和减少采样动物数量的一种建议方法。本文描述了在循环系统外设置预过滤哨兵鱼缸的方法。该技术旨在防止水污染,并通过仔细选择年龄、性别和品系来代表鱼群。为了使用最少数量的动物,还详细介绍了环境筛查技术。对表面集水池拭子进行聚合酶链反应(PCR),可显著提高对一些常见致病分枝杆菌物种的检测,如偶然分枝杆菌、嗜血性分枝杆菌和龟分枝杆菌。另一种环境检测方法是处理养殖池或集水池底部的污泥,以寻找绒毛伪毛细线虫卵。这是一种廉价且快速的技术,可应用于检疫,即将繁殖装置浸入进口动物的养殖池中。最后,将PCR应用于污泥样本,在集水池底部和表面检测到嗜水气单胞菌。一般来说,与预过滤哨兵鱼检测相比,这些针对特定病原体的环境筛查技术提高了检测灵敏度。