Ragland Natalie H, Miedel Emily L, Engelman Robert W
Department of Comparative Medicine, H Lee Moffitt Cancer Center and Research Institute, University of South Florida, Tampa, Florida, USA.
J Am Assoc Lab Anim Sci. 2019 Mar 1;58(2):208-215. doi: 10.30802/AALAS-JAALAS-18-000112. Epub 2019 Feb 22.
Exposing immunodeficient mice to opportunistic microbes introduces risks of data variability, morbidity, mortality, and the invalidation of studies involving unique human reagents, including the loss of primary human hematopoietic cells, patient-derived xenografts, and experimental therapeutics. The prevalence of 15 opportunistic microbes in a murine research facility was determined by yearlong PCR-based murine and IVC equipment surveillance comprising 1738 specimens. Of the 8 microbes detected, 3 organisms- , , and biotype Heyl-were most prevalent in both murine and IVC exhaust plenum specimens. Overall, the 8 detectable microbes were more readily PCR-detectable in IVC exhaust airways than in murine specimens, supporting the utility of PCR testing of IVC exhaust airways as a component of immunodeficient murine health surveillance. Vaporized hydrogen peroxide (VHP) exposure of IVC equipment left unassembled (that is, in a 'static-open' configuration) did not eliminate PCR detectable evidence of microbes. In contrast, VHP exposure of IVC equipment assembled 'active-closed' eliminated PCR-detectable evidence of all microbes. Ensuring data integrity and maintaining a topographically complex immunodeficient murine research environment is facilitated by knowing the prevalent opportunistic microbes to be monitored and by implementing a PCR-validated method of facility decontamination that mitigates opportunistic microbes and the risk of invalidation of studies involving immunodeficient mice.
将免疫缺陷小鼠暴露于机会性微生物会带来数据变异性、发病率、死亡率以及涉及独特人类试剂的研究无效的风险,包括原代人类造血细胞、患者来源的异种移植物和实验性治疗药物的损失。通过为期一年的基于PCR的小鼠和独立通风笼具(IVC)设备监测(共1738个样本),确定了小鼠研究设施中15种机会性微生物的流行情况。在检测到的8种微生物中,3种微生物—— 、 和Heyl生物型——在小鼠和IVC排气室样本中最为普遍。总体而言,8种可检测到的微生物在IVC排气气道中比在小鼠样本中更容易通过PCR检测到,这支持了将IVC排气气道的PCR检测作为免疫缺陷小鼠健康监测组成部分的实用性。未组装的IVC设备(即“静态开放”配置)暴露于汽化过氧化氢(VHP)并不能消除微生物的PCR可检测证据。相比之下,组装为“主动封闭”的IVC设备暴露于VHP可消除所有微生物的PCR可检测证据。了解要监测的常见机会性微生物,并实施经过PCR验证的设施净化方法,以减轻机会性微生物以及涉及免疫缺陷小鼠的研究无效的风险,有助于确保数据完整性并维持地形复杂的免疫缺陷小鼠研究环境。