The RBL is one of several infectious-disease laboratories that make up the Judson Harper Research Complex on the University’s Foothills Campus, located five miles west of the main Fort Collins campus. The Foothills Campus also includes a $60-million BSL-3 Vector-Borne Diseases Lab operated by the CDC, as well as the U.S. Department of Agriculture’s 23-acre National Wildlife Research Laboratory, where a BSL-3 animal lab to study zoonotic diseases is slated to be built.

The National Institute of Allergy and Infectious Diseases (NIAID) awarded almost $30 million to Colorado State University in 2003 to build the RBL. Nearly two years later, Colorado State University was designated the lead institution for a Regional Center for Excellence (RCE), and funded the RBL operation with a four-year, $41-million grant.

The RBL is attached to an existing 12,000-sf building, which recently underwent a $3.7-million expansion, which includes 4,100 sf of BSL-3 labs and 2,000 sf of BSL-2 media prep space. A further $8-million expansion will soon be under construction, which will comprise 6,500 sf of BSL-3 space, including three up-to-date insectories.

“We think this complex comprises a real national strength in this region,” says Ralph Smith, professor of microbiology in the College of Veterinary Medicine and Biomedical Sciences at Colorado State.

The RBL is located on an outlying campus partly because the real estate was available, and partly because of the nature of the work. By definition, BSL-3 laboratories handle potentially lethal infectious agents.  The RBL is charged with researching these specific agents, some of which have the potential to be used as biological weapons:

• Burkholderia mallei and pseudomallei, which cause glanders in horses and melidosis in humans. Melidosis is a chronic disease that can lie dormant for 30 years after exposure, then cause non-respiratory disease leading to organ failure. It is spread through the skin.
• Eastern and Venezuelan Equine Encephalitis viruses
• Japanese Encephalitis virus
• Francisella tularensis, which causes Tularemia, or “rabbit fever,” and is highly infectious
• Yersinia pestis, a bacteria carried by fleas that causes plague
• Coxiella burnetii, the causitive agent of Q fever

“When we finish this building complex, we will have 50,000 sf of BSL-2 support space and BSL-3 research space under one roof,” says Smith. “We are really delighted about that.”

Having both laboratory levels in one building is beneficial for management, training, maintenance, and security, he explains. For example, virologists now work with uninfected mosquitoes in the Arthropod-Borne Infectious Disease Lab, a BSL-2 lab across the quad from the RBL. When the mosquitoes are infected with a Select Agent, they are in the BSL-3 building attached to the complex that includes the RBL.

The reverse of that occurs when researchers in a BSL-3 laboratory need BSL-2 space to do a work-up of an agent. In the current set of facilities, they have to walk to the Arthropod-Borne Infectious Disease Lab or drive to the main campus, says Smith. Having all of these required facilities under one roof facilitates stronger research programs and allows for critical experimentation that could not previously be done because of safety concerns.

“It’s a lot better to simply go out of the BSL-3 barrier and down the hall without leaving the building,” he says.

From Labs to Antidotes

The RBL contains three identical suites that integrate BSL-3 laboratories and rooms for mice and rats. The NIAID asked that the suites be designed to minimize the researchers’ risk of contamination by minimizing their exposure to the mice.

Researchers access the suite through a single entrance that leads to women’s and men’s changing and showers rooms. The entrance also leads to an alcove containing a large autoclave capable of taking an entire rack of mouse cages. The changing rooms lead to a corridor with three adjacent BSL-3 laboratories. The researchers never have contact with the more than 4,500 mice housed in the suite unless they need to. If they need to enter the area of the suite where mice are housed, they pass through an ante room and into the “mouse corridor,” which leads to the tissue culture and necropsy rooms—complete with two laminar flow biosafety cabinets and a Class-III glove cabinet. The corridor also leads to a laboratory containing an inhalation chamber, and two mouse holding rooms, both of which are preceded by an ante room and have only one point of access.

In addition to mice being housed at the laboratory, thousands of mosquitoes infected with virus pathogens that cause encephalitis will inhabit small mesh boxes in three insectaries in the newly expanded wing of the building. They present a different challenge when it comes to biocontainment.

“Mosquitoes are obviously much smaller and more difficult to contain than mice,” says Smith. “This is a very tightly controlled environment, but simple solutions work, like negative air pressure. We also hang white sheets over the entrances so that if an insect were to escape, it would be brushed off a person leaving the insectary and kept in the containment room.”

Since work on the Select Agents is intended to produce vaccines, diagnostics, and therapeutics, the RBL also includes BSL-3 proteomics, genomics, and product development laboratories, as well as a BSL-2 manufacturing suite that employs Good Manufacturing Practices (GMP). The product development laboratories and manufacturing suite are located on the same corridor and each is accessible through only one entrance.

These laboratories are used for the large-scale growth of BSL-3 agents. They are designed to be as clean as possible, with rounded corners, minimal seams in the floors and walls, and minimal overhead passage of pipes. When a product development is completed, it is made biologically inactive and the process is transported down the hall for manufacturing.

The manufacturing suite is a pilot plant intended to produce only a few dozen vials of a product, says Smith. Under extreme conditions, it might have the capacity to go to Phase II clinical trials, he says. The manufacturing and final fill rooms are rated Class 10,000, while the rest of the suite is rated Class 100,000.

A Complicated Mission

Correct operation of the facility rests on the people who work there, so training is paramount.

“We are planning for there to be period of time when we have finished construction and yet not become Select-Agent certified where we will do intensive training in one of the BSL-3 suites,” says Smith. “The training will be done by our University biosafety officers and operation managers; the PIs are required to train their workers with cooperation from the biosafety officers.”

One of the greatest areas of training emphasis, says Smith, is of the mouse laboratory staff.

“We think there is a huge need nationally for well-trained laboratory workers,” he says. The following training modules have been established by the biosafety officers, and will last from one to four weeks:

• Select Agent Rules and Regulations
• Biosafety Cabinet Training
• Bench Training in the Training Lab
• Bench Training in the BSL-3 Labs
• Entry/Exit, Alarms, Safety, Occupational Health Program
• Packaging and Shipping

Once the facility is up and running, new researchers will receive several months of bench-side training, says Smith. Someone with more experience might receive a month of training.

The RBL serves many masters. Its region encompasses six states: Colorado, Wyoming, Utah, Montana, and North and South Dakota. It partners with more than a dozen institutions in those states. The RBL also is mandated to engage industry in product development, which further complicates the development of a long-term operation and management plan.

“I can tell you that we work on that plan almost every day,” says Smith. “The NIAID project manager will assist in the construction of the building through commissioning, and NIAID staff will share in the management of the facility once it is completed.”

The geographic size of the region also creates its own challenges.

“We serve a lot of square miles, and the institutions are scattered all around, some of which are very difficult to get to,” says Smith. “We communicate with them very easily with IT, but physical face-to-face meetings can take days.”

By Lisa Wesel