Construction and organization of a BSL-3 cryo-electron microscopy laboratory at UTMB.

A unique cryo-electron microscopy facility has been designed and constructed at the University of Texas Medical Branch (UTMB) to study the three-dimensional organization of viruses and bacteria classified as select agents at biological safety level (BSL)-3, and their interactions with host cells. A 200keV high-end cryo-electron microscope was installed inside a BSL-3 containment laboratory and standard operating procedures were developed and implemented to ensure its safe and efficient operation. We also developed a new microscope decontamination protocol based on chlorine dioxide gas with a continuous flow system, which allowed us to expand the facility capabilities to study bacterial agents including spore-forming species. The new unified protocol does not require agent-specific treatment in contrast to the previously used heat decontamination. To optimize the use of the cryo-electron microscope and to improve safety conditions, it can be remotely controlled from a room outside of containment, or through a computer network world-wide. Automated data collection is provided by using JADAS (single particle imaging) and SerialEM (tomography). The facility has successfully operated for more than a year without an incident and was certified as a select agent facility by the Centers for Disease Control.

Design, Implementation, and Evaluation of an N95 Respirator Decontamination and Reuse Program for Healthcare Workers During the COVID-19 Pandemic.

Early in the COVID-19 pandemic, substantial disruptions in personal protective equipment (PPE) supply chains forced healthcare systems to become resourceful to ensure PPE availability for healthcare workers. Most worrisome was the global shortage of N95 respirators. In response, a collaboration between the Department of Infection Control and Healthcare Epidemiology and the Department of Biosafety at the University of Texas Medical Branch developed a PPE recycling program guaranteeing an adequate supply of respirators for frontline staff. The team successfully developed and implemented a novel workflow that included validated decontamination procedures, education, and training programs as well as transportation, labeling, and storage logistics. In total, 15,995 respirators of various types and sizes were received for recycling. Of these, 12,752 (80%) were recycled. Following the program's implementation, we surveyed 134 frontline healthcare workers who overwhelmingly graded our institution's culture of safety positively. Overall impressions of the N95 respirator recycling program were mixed, although interpretation of those results was limited by a lower survey response rate. In an era of increasing health security threats, innovative recycling programs like this one may serve as a model for other health systems to respond to future PPE supply chain disruptions.

Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 1. Biosafety Level 4 Suit Laboratory Suite Entry and Exit Procedures.

Biosafety level 4 (BSL-4) suit laboratories are specifically designed to study high-consequence pathogens for which neither infection prophylaxes nor treatment options exist. The hallmarks of these laboratories are: custom-designed airtight doors, dedicated supply and exhaust airflow systems, a negative-pressure environment, and mandatory use of positive-pressure ("space") suits. The risk for laboratory specialists working with highly pathogenic agents is minimized through rigorous training and adherence to stringent safety protocols and standard operating procedures. Researchers perform the majority of their work in BSL-2 laboratories and switch to BSL-4 suit laboratories when work with a high-consequence pathogen is required. Collaborators and scientists considering BSL-4 projects should be aware of the challenges associated with BSL-4 research both in terms of experimental technical limitations in BSL-4 laboratory space and the increased duration of such experiments. Tasks such as entering and exiting the BSL-4 suit laboratories are considerably more complex and time-consuming compared to BSL-2 and BSL-3 laboratories. The focus of this particular article is to address basic biosafety concerns and describe the entrance and exit procedures for the BSL-4 laboratory at the NIH/NIAID Integrated Research Facility at Fort Detrick. Such procedures include checking external systems that support the BSL-4 laboratory, and inspecting and donning positive-pressure suits, entering the laboratory, moving through air pressure-resistant doors, and connecting to air-supply hoses. We will also discuss moving within and exiting the BSL-4 suit laboratories, including using the chemical shower and removing and storing positive-pressure suits.