Development and implementation of evidence-based laboratory safety management tools for a public health laboratory.

We developed an evidence-based continuous quality improvement (CQI) cycle for laboratory safety as a method of utilizing survey data to improve safety in a public health laboratory setting. • Expert Opinion: The CQI cycle begins with the solicitation of laboratory staff input via an annual survey addressing potential chemical, physical and radiological hazards associated with multiple laboratory activities. The survey collects frequency, severity and exposure data related to these activities in the context of the most pathogenic organisms handled at least weekly. • Gap Analysis: Step 2 of the CQI cycle used survey data to identify areas needing improvement. Typically, the traditional two-dimensional risk assessment matrix is used to prioritize mitigations. However, we added an additional dimension - frequency of exposure - to create three-dimensional risk maps to better inform and communicate risk priorities. • Mitigation Measures: Step 3 of the CQI cycle was to use these results to develop mitigations. This included evaluating the identified risks to determine what risk control measures (elimination, substitution, engineering, administrative or PPE) were needed. In the 2016 iteration of the CQI cycle described here, all mitigations were based on administrative controls. • Evaluation and Feedback: The last step of the CQI cycle was to evaluate the inferred effects of interventions through subsequent surveys, allowing for qualitative assessment of intervention effectiveness while simultaneously restarting the cycle by identifying new hazards. Here we describe the tools used to drive this CQI cycle, including the survey tool, risk analysis method, design of interventions and inference of mitigation effectiveness.

Analgesia during Monkeypox Virus Experimental Challenge Studies in Prairie Dogs (Cynomys ludovicianus).

Because human patients with monkeypox virus (MPXV) infection report painful symptoms, it is reasonable to assume that animals infected with MPXV experience some degree of pain. Understanding whether and how analgesics affect MPXV disease progression is crucial when planning in vivo challenge experiments. In the current study, we challenged prairie dogs with a low dose (4 ×10³ pfu) of MPXV and treated with meloxicam (NSAID) or buprenorphine (opioid); control animals did not receive analgesia or received analgesia without MPXV challenge. Subsets of animals from each group were serially euthanized during the course of the study. Disease progression and viral kinetics were similar between groups, but MXPVinfected, meloxicam-treated animals showed increasing trends of morbidity and mortality compared with other groups. Differences between no-analgesia MPXV-infected control animals and MPXV-infected animals treated with buprenorphine were minimal. The findings in the current study allow more informed decisions concerning the use of analgesics during experimental MPXV challenge studies, thereby improving animal welfare. In light of these findings, we have modified our pain scale for this animal model to include the use of buprenorphine for pain relief when warranted after MPXV challenge.

Humoral immunity to smallpox vaccines and monkeypox virus challenge: proteomic assessment and clinical correlations.

Despite the eradication of smallpox, orthopoxviruses (OPV) remain public health concerns. Efforts to develop new therapeutics and vaccines for smallpox continue through their evaluation in animal models despite limited understanding of the specific correlates of protective immunity. Recent monkeypox virus challenge studies have established the black-tailed prairie dog (Cynomys ludovicianus) as a model of human systemic OPV infections. In this study, we assess the induction of humoral immunity in humans and prairie dogs receiving Dryvax, Acam2000, or Imvamune vaccine and characterize the proteomic profile of immune recognition using enzyme-linked immunosorbent assays (ELISA), neutralization assays, and protein microarrays. We confirm anticipated similarities of antigenic protein targets of smallpox vaccine-induced responses in humans and prairie dogs and identify several differences. Subsequent monkeypox virus intranasal infection of vaccinated prairie dogs resulted in a significant boost in humoral immunity characterized by a shift in reactivity of increased intensity to a broader range of OPV proteins. This work provides evidence of similarities between the vaccine responses in prairie dogs and humans that enhance the value of the prairie dog model system as an OPV vaccination model and offers novel findings that form a framework for examining the humoral immune response induced by systemic orthopoxvirus infection.

Establishment of the black-tailed prairie dog (Cynomys ludovicianus) as a novel animal model for comparing smallpox vaccines administered preexposure in both high- and low-dose monkeypox virus challenges.

The 2003 monkeypox virus (MPXV) outbreak and subsequent laboratory studies demonstrated that the black-tailed prairie dog is susceptible to MPXV infection and that the ensuing rash illness is similar to human systemic orthopoxvirus (OPXV) infection, including a 7- to 9-day incubation period and, likely, in some cases a respiratory route of infection; these features distinguish this model from others. The need for safe and efficacious vaccines for OPVX in areas where it is endemic or epidemic is important to protect an increasingly OPXV-naïve population. In this study, we tested current and investigational smallpox vaccines for safety, induction of anti-OPXV antibodies, and protection against mortality and morbidity in two MPXV challenges. None of the smallpox vaccines caused illness in this model, and all vaccinated animals showed anti-OPXV antibody responses and neutralizing antibody. We tested vaccine efficacy by challenging the animals with 10(5) or 10(6) PFU Congo Basin MPXV 30 days postvaccination and evaluating morbidity and mortality. Our results demonstrated that vaccination with either Dryvax or Acambis2000 protected the animals from death with no rash illness. Vaccination with IMVAMUNE also protected the animals from death, albeit with (modified) rash illness. Based on the results of this study, we believe prairie dogs offer a novel and potentially useful small animal model for the safety and efficacy testing of smallpox vaccines in pre- and postexposure vaccine testing, which is important for public health planning.