Comparison of Plenum and Cage-level Filter Exhaust Dust PCR Testing to Soiled Bedding Sentinel Mice (Mus musculus) on an IVC Rack.

Wendy R Williams, Shawn P Lane, Cheryl Perkins, Ken Henderson
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Abstract

The use of soiled-bedded sentinels (SBSs) has historically been the standard for colony health surveillance monitoring at our institution. With the advent of newer technologies in which dust collected from filters is tested by PCR, we compared traditional SBS with PCR testing of both exhaust air dust collected from a filter in the downstream vertical plenum (exhaust dust test [EDT]) and the SBS cage-level exhaust filter (SCEF). Our hypothesis was that both methods of filter testing would identify more pathogens than SBS testing. Twenty-five individually ventilated mouse racks that used disposable caging were sanitized and placed into rotation. Rack plenums were tested by PCR to verify negative results before the study start. Exhaust dust collection media were placed in the exhaust plenum (n = 25). SBS cages were placed on each side of the rack with 2 mice per cage (n = 42 mice), with the remaining cage slots occupied by research animals. At each triweekly cage change, the exhaust air filters were carefully removed from the cage top, placed in sterile 50-mL conical tubes, and pooled for submission. After 3mo, the SBS mice were tested via serology for bacterial and viral agents and by PCR for Helicobacter species, pinworms, and ectoparasites. In addition, the EDT filter and SCEF were collected for PCR to evaluate for the same agents. Our results indicate that the SCEF consistently detected agents more frequently than the EDT filter placed in the plenum and that the EDT filter media detected agents more frequently than did the SBS mice. Our data suggest that both PCR methods of detection are superior to SBS for individually ventilated disposable rodent cages and that the SCEF is superior to EDT. These data supported our movement of institution toward environmental monitoring as a method of rodent colony health surveillance.

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对 IVC 架上脏污垫料哨兵小鼠(Mus musculus)进行全空气和笼级过滤器排气粉尘 PCR 测试的比较。
在我们机构,使用污床哨兵(SBS)一直是菌落健康监测的标准。随着从过滤器中收集的粉尘通过 PCR 进行检测的新技术的出现,我们将传统的 SBS 与从下游垂直管道中的过滤器收集的废气粉尘(排气粉尘检测 [EDT])和 SBS 笼级排气过滤器(SCEF)的 PCR 检测进行了比较。我们的假设是,这两种过滤器检测方法都能比 SBS 检测方法识别出更多的病原体。我们对 25 个使用一次性笼具的独立通风小鼠架进行了消毒,并将其轮流放置。在研究开始前,用 PCR 对鼠架管道进行检测,以验证阴性结果。排气集尘介质被放置在排气管中(n = 25)。SBS 笼放置在机架两侧,每笼 2 只小鼠(n = 42 只),其余笼位由研究动物占据。每次三周换笼时,从笼子顶部小心取出排气过滤器,放入无菌的 50 毫升锥形试管中,然后集中送检。3 mo 后,SBS 小鼠通过血清学检测细菌和病毒,并通过 PCR 检测螺旋杆菌、蛲虫和体外寄生虫。此外,还收集了 EDT 过滤器和 SCEF 进行 PCR 检测,以评估相同的病原体。我们的结果表明,SCEF 检测到病原体的频率始终高于放置在通风孔中的 EDT 过滤器,EDT 过滤介质检测到病原体的频率高于 SBS 小鼠。我们的数据表明,对于单独通风的一次性啮齿动物笼而言,两种 PCR 检测方法都优于 SBS,而且 SCEF 优于 EDT。这些数据支持我们的机构将环境监测作为啮齿动物群落健康监测的一种方法。
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American Society of Laboratory Animal Practitioners Position Statement: Handling and Physical Restraint of Research Animals. American Society of Laboratory Animal Practitioners Position Statement: Definition of Animal Welfare. Effect of Novel High-fat Diet Feeding Methods on Food Wastage, Weight Gain, Hair Coat Grease Accumulation, and Scratching Behavior in C57BL/6NCrl Mice. Identification and Treatment of Fur Mites (Radfordia lemnina) in California Deer Mice (Peromyscus californicus) Using Selamectin. American Society of Laboratory Animal Practitioners Position Statement: Animal Care Principles.
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