Identification and Control of an Ornithonyssus Bacoti Infestation in a Rodent Vivarium by Using Molecular Diagnostic Techniques.

Ornithonyssus bacoti, commonly known as the tropical rat mite, is a zoonotic ectoparasite that occasionally infests research rodent colonies. Most infestations have been attributed to wild rodents that harbor the mite and spread it to research animals, often during building construction or other activity that disrupts wild rodent populations. Although infestation may be clinically silent, severe outbreaks have been reported to cause pruritis, dermatitis, decreased reproductive performance, and anemia in rodents. In mid-2020, our institution experienced increased activity of wild mice, which were found to be infested with O. bacoti, diagnosed by microscopic exam and confirmed by fur swab PCR analysis. We elected to add O. bacoti to our quarterly health monitoring exhaust air dust (EAD) testing PCR panel, increase wild mouse control measures, and treat the environment with a sustained-release synthetic pyrethroid spray in an attempt to prevent colony animal infestation. Initial quarterly EAD health monitoring results in September of 2020 were negative for O. bacoti. However, in early 2021, multiple IVC racks tested positive for O. bacoti at quarterly testing. Treatment consisted of providing permethrin-soaked nesting material and surface spray treatment of the room and hallway with a sustained-release synthetic pyrethroid. Historically in the literature, O. bacoti outbreaks of research mice were not identified until mite burden was high enough to cause dermatitis on animal care workers. Due to modern molecular diagnostics and proactive PCR-based health monitoring surveillance, we were able to identify the outbreak earlier than would have otherwise been possible. To the best of our knowledge, this is the first report to successfully identify O. bacoti using environmental health monitoring PCR techniques. This outbreak demonstrates the importance of screening for O. bacoti in facilities with the potential for wild rodent infestation and highlights unique considerations when managing O. bacoti infestations. In addition, a novel permethrin-soaked enrichment item was developed for cage-level treatment.

Evaluation of Pain and Distress and Therapeutic Interventions for Rectal Prolapse in Mice to Reduce Early Study Removal.

Rectal prolapse (RP) is a common clinical condition in mice, that does not have a recognized or documented standard of care. At our institution, an average of 240 mice develop RP each year. Our practice has been to recommend euthanasia upon identifying a RP based on its appearance as a painful or distressful condition. This study aimed to assess treatment options that would maintain the RP mucosa and allow mice to reach their study endpoint, and to evaluate the perception of this condition as a painful or distressful event. This study used 120 mice with spontaneous RP, concurrently assigned to ongoing research protocols. Mice were randomly assigned to 1 of 3 treatment groups: petroleum jelly, lidocaine jelly, or no treatment. Fecal samples were collected for pathogen testing, and all mice received an initial base score, followed by weekly blind scores. Upon euthanasia, RP tissue was collected for histopathology. Of the 120 mice identified with RP, 47 mice were breeders; 28% successfully produced 22 additional litters after developing RP. Seventy-three were nonbreeders, with 92% reaching their research study endpoint. No statistically significant differences were detected between the 3 treatment groups based on gross mucosal health, pain and distress, or histopathology. In this study, none of the mice in any group were euthanized based on the RP endpoint scoring criteria. These findings demonstrate that treatment is unnecessary for RP, and mice with RP did not show signs of pain or distress. In adherence to the 3Rs, this study supports animal number reduction and clinical refinement, allowing mice with RPs to reach their intended research study endpoints or produce additional litters.

Diagnosis, Surveillance and Management of Streptococcus equi subspecies zooepidemicus Infections in Chinchillas (Chinchilla lanigera).

During a 6-mo period, two 5-6 mo old female chinchillas (Chinchilla lanigera) were examined at the University of Colorado Anschutz Medical Campus after the discovery of firm, nonmobile masses in the left ventral cervical and left axillary region. Other than these findings and mild weight loss, both chinchillas' physical exams were normal. Bloodwork revealed an inflammatory leukogram characterized by leukocytosis, toxic neutrophils, lymphopenia, and monocytosis with mild, nonregenerative anemia. At necropsy, both masses were identified as abscesses. Streptococcus equi, subspecies zooepidemicus (S. zooepidemicus) was isolated in pure culture. Histology of the lungs, liver, spleen, and kidneys showed a marked increase in the numbers of both polymorphonuclear leukocytes and lymphocytes. Both animals were deemed unsuitable for research and were euthanized under isoflurane anesthesia by an intracardiac injection of pentobarbital sodium solution. S. zooepidemicus is an opportunistic, commensal organism found in the upper respiratory tract of horses. This organism has been documented to cause disease in other species and is zoonotic. Infections in humans have been reported, resulting in glomerulonephritis, endocarditis, septic arthritis, osteomyelitis, meningitis, and death. To aid in diagnosis and prospective surveillance of this bacteria, oral and nasal swabs were collected from the remaining cohort of chinchillas, and a qPCR screening assay was implemented. Within 12 mo, 4 of 41 additional females tested positive by culture or qPCR, resulting in a disease prevalence of 14% (6 of 43). However, only 2 of the additional 4 S. zooepidemicus positive animals developed clinical signs. The potential for the spread of infection, zoonosis, and adverse effects on research demonstrate that surveillance for S. zooepidemicus should be considered in a biomedical research environment.

Low-cost, Small-scale Decontamination of Laboratory Equipment by Using Chlorine Dioxide Gas.

A significant concern in laboratory animal medicine is contamination due to pathogen outbreaks and how to adequately decontaminate small equipment. Many factors play a role in the selection of the decontamination method including cost, efficacy, personnel time and safety. Chlorine dioxide (ClO2) gas is an effective method, but decontamination often requires a ClO2 gas generator with a specialized air-tight exposure chamber. Although this method works well for large-scale decon- tamination, the use of a gas generator may be impractical and too costly for smaller-scale decontamination. The goal of this study was to create and validate an effective, small-scale decontamination method that uses ClO2 gas and which is an affordable, efficient, safe, and reproducible. First, we identified a product that generates ClO2 gas after the combination of 2 dry reagents. To find an affordable exposure chamber, we evaluated the ability of 4 household totes with gasket-seal lid systems to retain ClO2 gas and relative humidity (RH). The efficacy of decontamination was validated by concurrently using 2 different biologic indicators (BI), Bacillus atrophaeus (B.a.) and Geobacillus stearothermophilus (G.s.). All household totes evaluated held sufficient gas and RH for a 15-h cycle, providing adequate contact time to inactivate both BI evaluated. Our results suggest that a total exposure dose of 71 ± 42 ppm-h of ClO2 gas over 15 h at 90% or greater RH is adequate to inactivate both B.a. and G.s. There was no statistical significance between the 2 BI as indicators for decontamination; 65 of 230 (28.3%) B.a. and 75 of 230 (32.6%) G.s spore strips were positive for growth (P = 0.36). In conclusion, we successfully combined a variety of low-cost materials to establish an effective, small-scale method to decontaminate laboratory equipment. Depending on the size of the tote and whether BI are used, the cost of our method is roughly 1% that of large-scale ClO2 gas generators used with specialized air-tight exposure chambers.