Laboratory Biosafety Prototocols-Heat Disinfection & Sterilization

LABORATORY BIOSAFETY PROTOCOLS:
The protocols emphasize the use of good microbiological work practices, appropriate containment equipment, proper facility design, operation and maintenance, and administrative considerations to minimize the risk of worker injury or illness. “Laboratory biosafety” is the term used to describe the containment principles, technologies and practices that are implemented to prevent unintentional exposure to pathogens and toxins, or their accidental release. “Laboratory biosafety” refers to institutional and personal security measures designed to prevent the loss, theft, misuse, diversion or intentional release of pathogens and toxins.
MICROBIOLOGICAL RISKS:

1. Pathogenicity of the agent and infectious dose.
   
2. Potential outcome of exposure.
   
3. Natural route of infection.
   
4. Other routes of infection, resulting from laboratory manipulations (parenteral, airborne, ingestion).
   
5. Stability of the agent in the environment.
   
6. Concentration of the agent and volume of concentrated material to be manipulated.
   
7. Presence of a suitable host (human or animal).
   
8. Laboratory activity planned (sanitation, aerosolization, centrifugation, etc.).
   
9. Any genetic manipulation of the organism that may extend the host range of the agent or alter the agent’s sensitivity to known, effective treatment.
   

Plant Care

PERSONAL PROTECTION:

1. Laboratory overalls, gowns or uniforms must be worn at all times for work in the laboratory.
   
2. Appropriate gloves must be worn for all procedures that may involve direct or accidental contact with blood, body fluids and other potentially infectious materials or infected animals. After use, gloves should be removed aseptically and hands must then be washed.
   
3. Personnel must wash their hands after handling infectious materials and animals, and before they leave the laboratory working areas. 
4. Safety glasses, face shields (visors) or other protective devices must be worn when it is necessary to protect the eyes and face from splashes, impacting objects and sources of artificial ultraviolet radiation. 
5. It is prohibited to wear protective laboratory clothing outside the laboratory, e.g. in canteens, coffee rooms, offices, libraries, staff rooms and toilets. Open-toed footwear must not be worn in laboratories.
6. Eating, drinking, smoking, applying cosmetics and handling contact lenses is prohibited in the laboratory working areas.
7. Storing human foods or drinks anywhere in the laboratory working areas is prohibited. Protective laboratory clothing that has been used in the laboratory must not be stored in the same lockers or cupboards as street clothing.

Plant Care

LABORATORY WORKING AREAS

1. The laboratory should be kept neat, clean and free of materials that are not pertinent to the work.
   
2. Work surfaces must be decontaminated after any spill of potentially dangerous material and at the end of the working day.
   
3. All contaminated materials, specimens and cultures must be decontaminated before disposal or cleaning for reuse.
   
4. Packing and transportation must follow applicable national and/or international regulations.
   
5. When windows can be opened, they should be fitted with arthropod-proof screens.
   

CONTINGENCY PLAN
The contingency plan should provide operational procedures for:

1. Precautions against natural disasters, e.g. fire, flood, earthquake and explosion
   
2. Biohazard risk assessment
   
3. Incident-exposure management and decontamination
   
4. Emergency evacuation of people and animals from the premises
   
5. Emergency medical treatment of exposed and injured persons
   
6. Medical surveillance of exposed persons
   
7. Clinical management of exposed persons
   
8. Epidemiological investigation
   
9. Post-incident continuation of operations development of this plan the following items should be considered for inclusion: Identification of high-risk organisms
10. Location of high-risk areas, e.g. laboratories, storage areas, animal facilities Identification of at-risk personnel and populations
11. Identification of responsible personnel and their duties, e.g. biosafety officer, safety personnel, local health authority, clinicians, microbiologists, veterinarians, epidemiologists, and fire and police services.
12. Lists of treatment and isolation facilities that can receive exposed or infected persons.
13. Transport of exposed or infected persons.
14. Lists of sources of immune serum, vaccines, drugs, special equipment and supplies.
15. Provision of emergency equipment, e.g. protective clothing, disinfectants, chemical and biological spill kits, decontamination equipment and supplies.

EMERGENCY EQUIPMENT
The following emergency equipment must be available:

1. First-aid kit, including universal and special antidotes.
   
2. Appropriate fire extinguishers, fire blankets.
   

The following are also suggested but may be varied according to local circumstances:

1. Full protective clothing (one-piece overalls, gloves and head covering for incidents involving microorganisms in risk).
   
2. Full-face respirators with appropriate chemical and particulate filter canisters.
   
3. Room disinfection apparatus, e.g. sprays and formaldehyde vaporizers.
   
4. Stretcher.
   
5. Tools, e.g. hammers, axes, spanners, screwdrivers, ladders, ropes.
   
6. Hazard area demarcation equipment and notices.
   

DISINFECTION AND STERILIZATION
The basic knowledge of disinfection and sterilization is important for biosafety in the laboratory to handle the contaminated materials.
DEFINITIONS:
The following are the commonly used terms for disinfection and sterilization.
ANTIMICROBIAL:
An agent that kills microorganisms or suppresses their growth and multiplication.

ANTISEPTIC:
A substance that inhibits the growth and development of microorganisms without necessarily killing them. Antiseptics are usually applied to body surfaces. 
BIOCIDE:
A general term for any agent that kills organisms.
CHEMICAL GERMICIDE:
A chemical or a mixture of chemicals used to kill microorganisms.
DECONTAMINATION:
Any process for removing and/or killing microorganisms. The same term is also used for removing or neutralizing hazardous chemicals and radioactive materials.
DISINFECTANT:
A chemical or mixture of chemicals used to kill microorganisms, but not necessarily spores. Disinfectants are usually applied to inanimate surfaces or objects. 
DISINFECTION:
A physical or chemical means of killing microorganisms, but not necessarily spores.
MICROBICIDE:
A chemical or mixture of chemicals that kills microorganisms. The term is often used in place of “biocide”, “chemical germicide” or “antimicrobial”.
SPOROCIDE:
A chemical or mixture of chemicals used to kill microorganisms and spores.
STERILIZATION:
A process that kills and/or removes all classes of microorganisms and spores.
CHEMICAL DISINFECTION
The following chemicals or germicides are used to disinfect the materials.

1. Chlorine (Sodium hypochlorite)
   
2. Chloramines
   
3. Chlorine dioxide (ClO2)
   
4. Formaldehyde
   
5. Alcohols
   
6. Iodine and Iodophors
   
7. Hydrogen peroxide
   
8. Peracid

HEAT DISINFECTION AND STERILIZATION
Saturated steam under pressure (autoclaving) is the most effective way of sterilizing laboratory materials.
The following cycle will surely sterilize the correctly loaded materials.

1. 3 minutes holding time at 134oC
   
2. 10 minutes holding time at 126oC
   
3. 15 minutes holding time at 121oC
   
4. 25 minutes holding time at 115oC

Molecular Lab-Department of Plant Pathology-IUB

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Written By:

Dr.Qaiser Shakeel

Tahir Mahmood

Department of Plant Pathology-Agriculture

The Islamia University of Bahwalpur-Pakistan.