Lecture 6 Laboratory Safety in Microbiology
Lecture 6 Laboratory Safety in Microbiology
Lecture 6 Laboratory Safety in Microbiology
INTRODUCTION
Microorganisms are normally present in the environment and many of which are carried on and in the human body.
The Microbiology laboratory poses the hazard of exposure to infectious agents. Laboratory personnel should observe safety precautions to avoid laboratoryacquired infections.
Laboratory acquired infections are often attributed to carelessness and poor technique.
2. Aseptic Technique
e.) Cuts, scratches, sores, and other lesions on the hands and exposed parts of the body should be covered with adhesive plasters.
g.) Decontaminate bench tops and other surfaces before and after leaving the laboratory or whenever there are spills (1% dilution of sodium hypochlorite/bleach, 70% isopropyl alcohol) h.) Laboratory work area should be kept clean and free of unnecessary clutter
Biosafety levels are determined by the WHO, NIH and CDC WHO recommends that the health authorities of each country should make lists of organisms and viruses in each Risk Group as appropriate to local circumstances so that suitable precautions may be applied.
Organisms involved
Food spoilage bacteria Common molds yeasts
Risk Group I
Risk Group II
Staphylococci Streptococci
Enterobacteria (except
Organisms involved
M. Tuberculosis Brucella Salmonella typhi
Risk Group IV
Classification of LABORATORIES
3 KINDS of LABORATORY
1. BASIC LABORATORY (Biosafety Level 1)
Simple; Adequate for work with organisms in Risk Group I and II Varies between rooms used at the Primary Health Care Level and the hospital diagnostic laboratory
Clean, comfortable, with enough space Adequate sanitary facilities, especially for handwashing
3 KINDS of LABORATORY
BASIC LABORATORY (Biosafety Level 2) same as Biosafety Level 1 Laboratory must be equipped with an autoclave and a Biological Safety Cabinet Doors are posted with a biohazard sign a separate bin for infectious and non infectious waste
3 KINDS of LABORATORY
2. CONTAINMENT LABORATORY (Biosafety Level 3)
More sophisticated; Used for the work with organisms in Risk Group III
A separate room with controlled access by authorized staff only Requires a microbiological safety cabinet Only highly skilled and experienced staff are allowed to work in this laboratory
3 KINDS of LABORATORY
3. MAXIMUM CONTAINMENT LABORATORY (Biosafety Level 4) Intended for work with viruses in Risk Group IV The most strict safety precautions are necessary Separate buildings with strictly controlled access through air locks and exit through decontaminant showers Laboratory staff is specially trained for the work
Class III cabinets are used for Risk Group IV viruses (i.e. smallpox, Ebola virus)
2. ASEPTIC TECHNIQUE
A method that prevents the introduction of unwanted organisms into an environment. In microbial cultures, it is used to prevent introducing additional organisms (contaminants) into the culture.
2. ASEPTIC TECHNIQUE
Purposes: To prevent microbial contamination of laboratory worker which may result in infection To prevent contamination of the environment they are working in (e.g. fomites) To prevent contamination of the specimen especially when a pure culture is desired
2. ASEPTIC TECHNIQUE
General Guidelines Always use the proper aseptic technique when transferring cultures from one medium to another.
Heat the entire piece of metal of the inoculation instrument in the flame: it should be RED HOT. Be sure to COOL your inoculation instrument before picking the inoculum. Use an inoculating needle for agar deeps and an inoculating loop for the agar plate and the broths. Label all test tubes and petri plates
2. ASEPTIC TECHNIQUE
General Guidelines Place test tubes in racks when working at your table Keep test tube caps and petri dish covers on media to reduce contamination ALWAYS check agar plates carefully to make sure that there are no mold or bacterial contaminants on the plate: if so, discard the plate. Do the same with any tubed media.
2. ASEPTIC TECHNIQUE
To practice ASEPTIC TECHNIQUES one must: Know how to prepare and clean a work area Know how to handle bacteria in different culture containers Know how to transfer organisms from one place to another without contamination
2. ASEPTIC TECHNIQUE
2.1 STREAK PLATE METHOD
The most common way of separating bacterial cells on the agar surface to obtain isolated colonies To isolate individual organisms into pure culture
2. ASEPTIC TECHNIQUE
2. ASEPTIC TECHNIQUE
2.2 BROTH TRANSFER METHOD the sterile technique of inoculating a bacterial organism into a liquid broth
3. DECONTAMINATION
A process of removing or neutralizing hazardous clinical and radioactive wastes by chemical or mechanical means TYPES OF DECONTAMINATION Decontamination by Chemical Disinfectants Decontamination by AUTOCLAVING Decontamination by INCINERATION
Alcohols
Chlorine Aldehydes
4 - 10 % formaldehyde
2% glutaraldehyde (e.g. Cidex) excellent laboratory disinfectant
B. Decontamination by AUTOCLAVING
one of the most dependable methods of sterilizing laboratory equipment and decontaminating biohazardous waste
C. Decontamination by INCINERATION
A treatment technology involving destruction of waste by controlled burning at high temperatures; to remove the water and reduce the remaining residues to a safe, non-burnable ash that can be disposed of safely on land, in some waters, or in underground locations.
CONCLUSION
Microbiology safety is largely in the hands of the laboratory worker, who even in the remote areas with few resources can achieve a considerable measure of protection by maintaining a high standard of personal hygiene and care, and by good laboratory practice.