Manuel Christopher A, Pugazhenthi Umarani, Leszczynski Jori K
Office of Laboratory Animal Resources, Department of Pathology, Anschutz Medical Campus, University of Colorado, Aurora, CO.
Department of Medicine, Division of Endocrinology, Metabolism, and Diabetes, Anschutz Medical Campus, University of Colorado, Aurora, CO.
J Am Assoc Lab Anim Sci. 2016 Jan;55(1):58-65.
Corynebacterium bovis causes an opportunistic infection of nude (Foxn1, nu/nu) mice, leading to nude mouse hyperkeratotic dermatitis (scaly skin disease). Enzootic in many nude mouse colonies, C. bovis spreads rapidly to naive nude mice, despite modern husbandry practices, and is very difficult to eradicate. To facilitate rapid detection in support of eradication efforts, we investigated a surveillance method based on quantitative real-time PCR (qPCR) evaluation of swabs collected from the horizontal exhaust manifold (HEM) of an IVC rack system. We first evaluated the efficacy of rack sanitation methods for removing C. bovis DNA from the HEM of racks housing endemic colonies of infected nude mice. Pressurized water used to flush the racks' air exhaust system followed by a standard rack-washer cycle was ineffective in eliminating C. bovis DNA. Only after autoclaving did all sanitized racks test negative for C. bovis DNA. We then measured the effects of stage of infection (early or established), cage density, and cage location on the rack on time-to-detection at the HEM. Stage of infection significantly affected time-to-detection, independent of cage location. Early infections required 7.3 ± 1.2 d whereas established infections required 1 ± 0 d for detection of C. bovis at the HEM. Cage density influenced the quantity of C. bovis DNA detected but not time-to-detection. The location of the cage on the rack affected the time-to-detection only during early C. bovis infections. We suggest that qPCR swabs of HEM are useful during the routine surveillance of nude mouse colonies for C. bovis infection.
牛棒状杆菌可引起无胸腺(Foxn1,nu/nu)小鼠的机会性感染,导致无胸腺小鼠发生角化过度性皮炎(鳞状皮肤病)。在许多无胸腺小鼠群体中呈地方流行性,尽管有现代饲养管理措施,牛棒状杆菌仍会迅速传播至未感染的无胸腺小鼠,且极难根除。为便于快速检测以支持根除工作,我们研究了一种基于对独立通风笼盒(IVC)饲养架水平排气歧管(HEM)采集的拭子进行定量实时PCR(qPCR)评估的监测方法。我们首先评估了从感染无胸腺小鼠地方流行群体饲养架的HEM中去除牛棒状杆菌DNA的饲养架清洁方法的效果。用于冲洗饲养架排气系统的加压水随后进行标准的饲养架清洗循环,在消除牛棒状杆菌DNA方面无效。只有在高压灭菌后,所有经过清洁的饲养架对牛棒状杆菌DNA检测均呈阴性。然后,我们测量了感染阶段(早期或已确立)、笼盒密度以及饲养架上笼盒位置对在HEM处检测到牛棒状杆菌所需时间的影响。感染阶段显著影响检测所需时间,与笼盒位置无关。早期感染在HEM处检测到牛棒状杆菌需要7.3±1.2天,而确立感染则需要1±0天。笼盒密度影响检测到的牛棒状杆菌DNA数量,但不影响检测所需时间。仅在牛棒状杆菌早期感染期间,饲养架上笼盒的位置才会影响检测所需时间。我们认为,对HEM进行qPCR拭子检测在无胸腺小鼠群体牛棒状杆菌感染的常规监测中很有用。