Reemergence of the Murine Bacterial Pathogen Chlamydia muridarum in Laboratory Mouse Colonies

Abstract

Chlamydia muridarum (Cm) was detected in mice from 2 colonies with lymphoplasmacytic pulmonary infiltrates using PCR and immunohistochemistry. This discovery was unexpected as Cm infection had not been reported in laboratory mice since the 1940s. A Cm specific PCR assay was developed and testing implemented for resident colonies from 8 vivaria from 3 academic institutions, 58 incoming mouse shipments from 39 academic institutions, and mice received from 55 commercial breeding colonies (4 vendors). To estimate Cms global prevalence in laboratory colonies, a database containing 11,387 metagenomic fecal microbiota samples from 120 institutions and a cohort of 900 diagnostic samples from 96 institutions were examined. Results indicate significant prevalence amongst academic institutions with Cm detected in 62.9% of soiled bedding sentinels from 3 institutions; 32.7% of incoming mouse shipments from 39 academic institutions; 14.2% of 120 institutions submitting microbiota samples; and 16.2% of the diagnostic sample cohort. All samples from commercial breeding colonies were negative. Additionally, naive NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice exposed to Cm shedding mice and their soiled bedding developed clinical disease 21 to 28 days following exposure. These mice had a moderate-to-severe histiocytic and neutrophilic bronchointerstitial pneumonia with respiratory epithelium demonstrating inclusions, chlamydial major outer membrane protein immunostaining, and hybridization with a Cm reference sequence (GenBank accession no. U68436). Cm was isolated on HeLa 229 cells from lungs, cecum, and feces of a Cm infected NSG mouse. The considerable prevalence of Cm is likely attributed to widespread global interinstitutional distribution of unique mouse strains and failure to recognize that some of these mice were from enzootically infected colonies. Given that experimental Cm colonization of mice results in a robust immune response and, on occasion, pathology, natural infection may confound experimental results. Therefore, Cm should be excluded and eradicated from endemically infected laboratory mouse colonies.