Biosafety & Security
Biosafety is concerned with the principles, practices, and procedures implemented to ensure the safe handling, storage, and disposal of biological agents, such as bacteria, viruses, and toxins, to prevent accidental exposure or release into the environment.
Biosecurity, on the other hand, deals with the protection, control, and accountability of biological materials within laboratories and facilities to prevent their unauthorized access, theft, misuse, or intentional release.
Recombinant and synthetic nucleic acids are crucial components in biotechnology, defined by their ability to replicate in living cells or be chemically synthesized, and are governed by strict safety guidelines.
- Recombinant Nucleic Acids: These are molecules constructed by joining nucleic acid segments (DNA or RNA) that can replicate within a living cell. They are often created through techniques such as cloning, where DNA from different sources is combined to produce new genetic combinations.
- Synthetic Nucleic Acids: These are nucleic acid molecules that are chemically synthesized or amplified. They can be designed to base pair with naturally occurring nucleic acids and may include modifications that enhance their stability or functionality.
Biosafety in Microbiological and Biomedical Laboratories
NIH Guidelines for Research Involving Recombinant DNA Molecules
Lentiviruses comprise a genus of the Retroviridae family and include bovine lentiviruses (e.g., Bovine immunodeficiency virus, Jembrana disease virus); equine lentiviruses (e.g., Equine infectious anemia virus); feline lentiviruses (e.g., Feline immunodeficiency virus); Ovine/caprine lentivirus (e.g., Caprine arthritis-encephalitis virus, Ovine lentivirus, Visna virus); and Primate lentiviruses (e.g., Human immunodeficiency virus (HIV) types 1 – 3, Simian AIDS retrovirus SRV-1, Human T-cell lymphotropic virus type 4, and Simian immunodeficiency virus). Research with lentiviral vectors must be conducted in accordance with the guidance document and containment precautions issued by the Institutional Biosafety Committee.
Researchers should be familiar with the properties of the lentiviral vectors used in their labs prior to handling and before an exposure occurs. Minimally, staff should understand the function of the transgene, number of plasmids used to generate the vector, host range and percentage of genome deleted.Exposure to a lentiviral vector in the lab must be reported and treated without delay.
NIH Biosafety Considerations for Research with Lentiviral Vectors
Autoclaves provide a physical method for disinfection and sterilization. They work with a combination of steam, pressure and time. Autoclaves operate at high temperature and pressure in order to kill microorganisms and spores.
They are used to decontaminate certain biological waste and sterilize media, instruments and lab ware.
Wear Personal Protective Equipment
- Lab coat
- Eye protection
- Closed-toe shoes
- Heat-resistant gloves to remove items, especially hot glassware
Packaging and Loading
- Only designated individuals should be allowed to set and/or change parameters for the autoclaves.
- Before using the autoclave, check inside for any items left by the previous user that could pose a hazard.
- Clean the drain strainer before loading the autoclave.
- Always place items in a secondary container.
- Do not overload or package bags too tightly. Leave sufficient room for steam circulation. If necessary, place container on its side to maximize steam penetration and avoid entrapment of air.
- Use only autoclavable bags to package waste.
- Do not allow bags to touch the interior walls of the autoclave to avoid melting of plastic.
- Ensure sufficient liquid is packed with contents of autoclave bags if dry.
- Place soiled glassware and lab ware in secondary containers and autoclave them in the solids cycle. Do not fill containers more than 2/3 full with liquids. Loosen caps or use vented closures.
- In case of clean glassware and wrapped instruments, lay them in a secondary container before autoclaving in wrapped goods cycle.
- For secondary containment, use autoclave trays made out of polypropylene, polycarbonate or stainless steel. The trays should have a solid bottom and sides to contain the contents and catch spills.
- Choose appropriate cycle for the material. Incorrect selection of cycle may damage the autoclave, cause liquid to boil over or bottles to break.
- Start your cycle and fill out the autoclave user log. A completed cycle usually takes between 1 to 1.5 hours.
- Check chamber/jacket pressure gauge for minimum pressure of 20 pounds per square inch (psi).
- Close and lock door.
- Check temperature for 250⁰F (121⁰C) every load.
- Do not attempt to open the door while autoclave is operating.
Unloading
- Ensure cycle has completed and both temperature and pressure have returned to a safe range.
- Wear PPE described above, plus an apron and face shield if removing liquids. Stand back from the door as a precaution and carefully open door no more than 1 inch. This will release residual steam and allow pressure within liquids and containers to normalize.
- Allow the autoclaved load to stand for 10 minutes in the chamber. This will allow steam to clear and trapped air to escape from hot liquids, reducing risk to operator.
- Do not agitate containers of super-heated liquids or remove caps before unloading.
- Place liquids in an area which clearly indicates the items are “hot” until the items cool to room temp.
- Allow autoclaved materials to cool to room temperature before transporting. Never transport superheated materials.
- Place cooled autoclaved biohazard bag into regulated medical waste box. Autoclaved infectious liquids may be disposed of into the sanitary sewer.
Tape Indicators
Autoclave tape indicators are adhesive-backed paper tape containing indicator ink that changes color after exposure to the correct temperature, pressure, and time during an autoclave cycle, confirming the entire sterilization process was completed. Tape indicators change color or display diagonal stripes, the words “sterile” or “autoclaved” when exposed to temperatures of 121°C.
Autoclave tape indicators are typically placed on the exterior of the waste load. If the temperature sensitive tape does not indicate that a temperature of at least 121°C was reached during the sterilization process, the load is not considered decontaminated.
The Office of Safety requires that you DO NOT use autoclave tape as the only indicator of decontamination or sterilization.
If tape indicators fail on two consecutive loads, notify the Office of Safety.
NOTE: Autoclave tape is a chemical indicator for successful sterilization, while temperature tape is a general term for a heat-sensitive tape indicating only temperature achieved. Temperature tape only shows a specific temperature has been reached but doesn't confirm if steam penetration or cycle length was sufficient for sterilization. Only use autoclave tape for this process.
Integrated Chemical Indicator Strips
Integrated chemical indicator strips provide a limited validation of temperature and time by displaying a color change after exposure to normal autoclave operating temperatures of 121ºC for several minutes. Chemical color change indicators can be placed within the waste load. If the chemical indicators fail on two consecutive loads, notify the Office of Safety.
Biological Indicators
Biological indicator vials contain spores from Geobacillus stearothermophilus, a microorganism that is inactivated when exposed to 121.1oC saturated steam for a minimum of 20 minutes. Autoclaves used to treat biological waste will be evaluated with a biological indicator monthly.