Abstract
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.
Highlights
In order to protect research and animal welfare[1,2], the presence of pathogens is monitored within animal facilities
The screening for bacteria in the biofilm and for P. tomentosa from the bottom of the tank can be combined by performing Polymerase Chain Reaction (PCR) for all these pathogens on the sludge sample sediment obtained after the first centrifugation
Of 115 fish tested, M. chelonae and M. haemophilum were detected in 5% and 3% of the samples, respectively
Summary
In order to protect research and animal welfare[1,2], the presence of pathogens is monitored within animal facilities. The aim is to reduce the number of fish used for health monitoring and to optimize turnover, cost, and sensitivity of the detection Such methods are an alternative to the use of animals and some techniques can be applied to screening imports in quarantine. The sentinels are in a static tank, out of the recirculating system, and their water is changed regularly, using only pre-filtration water i.e., sump water from all the systems in the room This technique is described below as a baseline for comparison with the efficacy of the environmental screening. The screening for bacteria in the biofilm and for P. tomentosa from the bottom of the tank can be combined by performing PCR for all these pathogens on the sludge sample sediment obtained after the first centrifugation. The technique is described below and some P. tomentosa eggs were detected by microscopy in this context
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