Maximum Containment Laboratories Research Articles

Potential Impact of a 2-Person Security Rule on BioSafety Level 4 Laboratory Workers

Directors of all major BioSafety Level 4 (BSL-4) laboratories in the United States met in 2008 to review the current status of biocontainment laboratory operations and to discuss the potential impact of a proposed 2-person security rule on maximum-containment laboratory operations. Special attention was paid to the value and risks that would result from a requirement that 2 persons be physically present in the laboratory at all times. A consensus emerged indicating that a video monitoring system represents a more efficient, economical standard; provides greater assurance that pathogens are properly manipulated; and offers an increased margin of employee safety and institutional security. The 2-person security rule (1 to work and 1 to observe) may decrease compliance with dual responsibilities of safety and security by placing undue pressure on the person being observed to quickly finish the work, and by placing the observer in the containment environment unnecessarily.

Defense against filoviruses used as biological weapons

The filoviruses, Marburg and Ebola, are classified as Category A biowarfare agents by the Centers for Disease Control. Most known human infections with these viruses have been fatal, and no vaccines or effective therapies are currently available. Filoviruses are highly infectious by the airborne route in the laboratory, but investigations of African outbreaks have shown that person-to-person spread requires direct contact with virus-containing material. In consequence, filovirus epidemics can be halted by isolating patients and instituting standard infection control and barrier nursing procedures. The filovirus disease syndrome resembles that caused by other hemorrhagic fever viruses, necessitating studies in a biocontainment laboratory to confirm the diagnosis. Some progress has been made in developing vaccines and antiviral drugs, but efforts are hindered by the limited number of maximum containment laboratories. Terrorists might have great difficulty acquiring a filovirus for use as a weapon, but my attempt to do so because of the agents’ ability to inspire fear. Accurate information is the best tool to prevent panic in the event of an attack.

Multiplex analysis of cytokines in the blood of cynomolgus macaques naturally infected with Ebola virus (reston serotype)**

Ebola virus (EBO) causes the most severe form of viral hemorrhagic fever in humans and nonhuman primates with up to 90% of infections culminating in death. The requirement of maximum containment laboratories for Ebola virus research has limited opportunities to study the pathogenesis of EBO infections. While tissue damage does occur, often it would appear not to be sufficient to explain death, indicating that soluble mediators play an important role in disease progression. In previous studies, fatal human infections with the Zaire subtype of Ebola (EBO-Z) were associated with an increase in the levels of inflammatory cytokines. In this investigation, a new multiplex assay was developed and used to measure circulating levels of cytokines and chemokines in cynomolgus macaques infected with the Reston subtype of EBO (EBO-R). Increased levels of IL-6, TNF-alpha, IFN-gamma, IL-2, IL-4, IL-8, IL-10, and GM-CSF were detected in infected animals, and the increase in circulating cytokines correlated with an increase in circulating viral antigen. Blood samples from animals showing high levels of cytokines were also tested for the chemokines: MCP-1, IL-1beta, MIP-1alpha, MIP-1beta, IP-10, and RANTES. High levels of MCP-1 and MIP-1beta, and RANTES were found in infected primates and, while levels were more variable, IL-1beta was detected only in infected animals.

SMALLPOX: AN ANNIVERSARY AND A DILEMMA.

This autumn marks the tenth anniversary of the last endemic case of smallpox in the world. But what should be done with the remaining smallpox virus now held in archival stocks in two maximum-containment laboratories, one in …

Inactivation of Lassa, Marburg, and Ebola viruses by gamma irradiation.

Because of the cumbersome conditions experienced in a maximum containment laboratory, methods for inactivating highly pathogenic viruses were investigated. The infectivity of Lassa, Marburg, and Ebola viruses was inactivated without altering the immunological activity after radiation with Co60 gamma rays. At 4 degrees C, Lassa virus was the most difficult to inactivate with a rate of 5.3 X 10(-6) log 50% tissue culture infective dose per rad of CO60 radiation, as compared with 6.8 X 10(-6) log 50% tissue culture infective dose per rad for Ebola virus and 8.4 X 10(-6) log 50% tissue culture infective dose per rad for Marburg virus. Experimental inactivation curves, as well as curves giving the total radiation needed to inactivate a given concentration of any of the three viruses, are presented. We found this method of inactivation to be superior to UV light or beta-propiolactone inactivation and now routinely use it for preparation of material for protein-chemistry studies or for preparation of immunological reagents.

Marburg-Like Virus: Ultrastructural Observations of an Isolate from Kenya, 1980

In February of this year specimens were sent from the Medical Research Center, Nairobi, Kenya to the Center for Disease Control (CDC), Atlanta, Georgia for isolation and identification of an agent responsible for a particularly severe disease clinically suggestive of a viral-induced hemorrhagic fever. Formalin-fixed liver biopsy from an acutely ill patient was sent for microscopic examination. Serum from this patient had been inoculated into Vero (continuous cell line derived from Ceropithecus aethiops) cell culture at the Medical Research Center. This cell culture material was further passaged in Vero cells upon receipt at the Maximum Containment Laboratory (MCL) of the CDC. For negative contrast examination, virus was purified from the cell supernatant fluids of the 3rd passage, 3-day postinfected Vero cells. For thin-section examination virus-infected Vero cells were prepared by gently scraping cells from a 3rd passage, 4-day postinfected cell culture, washing the cells in phosphate buffered saline, and pelleting the cells at 500Xg for 5 minutes in 2.5% buffered glutaraldhyde. The cell pellet was fixed In 2.5% buffered glutaraldehyde for 72 hours before it was processed and embedded for electron microscopy (EM).