MICROBIOLOGY

SAFETY ON THE CLINICAL

MICROBIOLOGY LABORATORY
NAME OF LECTURER
DATE OF LECTURE

● Clinical Laboratory and Standards Institute (CLSI)

OUTLINE ○ Universal precautions as set of preventive measures
I. Laboratory Safety B. Sterilization designed to reduce the risk of transferring HIV, hepa
A. Possible Routes for C. Disinfection B virus, and other bloodborne pathogens in the
Infection D. Antiseptics health care setting
II. Exposure Control Plan E. Biocides ○ Apply to human blood and other body fluids w visible
III. Universal and Standard F. Liquid Decontaminants blood but does not apply to feces, nasal secretions,
Precautions VIII. Fire Safety saliva (unless they contain visible blood)
A. Safety Guidelines for A. Type A ● Standard precautions
Laboratory B. Type B ○ Set of preventive measures applied to all patients
IV. Personal Protective C. Type C that are designed to reduce the risk of infection in
Equipment D. Type D the health care setting
V. Engineering Controls E. Class K ● Potentially infectious
A. Biosafety Levels IX. Chemical Safety ○ Blood, tissue, body fluids, secretions, excretions
B. Biosafety Cabinets A. Material Safety Data (except sweat)
VI. Post Exposure Plan Sheet
VII. Disposal of Hazardous B. Hazardous Chemicals A. POSSIBLE ROUTES FOR INFECTION
Waste 1. No food or drink
A. Autoclave 2. No smoking
3. No cosmetics
4. Protective eyewear must be worn
5. Wear face shields
I. LABORATORY SAFETY
6. Clothing: gowns, aprons lab coats, closed shoes
7. Hair should be tied up
A. POSSIBLE ROUTES FOR INFECTION 8. Wash hands frequently and thoroughly
● Airborne 9. Eyewash stations: 100 feet or 10 secs away from any area in
○ Aerosols from centrifugation of unstopped tubes or the lab
from heating cultures or specimens too rapidly, 10. Emergency stations: shower
removing stoppers from tubes, leakage from a 11. Respirators: available where there is air contamination
container that holds contaminated specimens 12. Mouth pipetting: ingestion of microorganisms; use pipette
● Ingestion bulbs/automated devices
○ Infection as a result of failure to wash hands or 13. Glassware and sharps: in puncture-resistant containers
eating, drinking, smoking, applying cosmetics, or 14. Centrifuges: minimize the production of aerosols
pipetting w the mouth
● Direct inoculation IV. UNIVERSAL AND STANDARD PRECAUTIONS
○ Infection from needlesticks, broken glass, animal ● Protective laboratory clothing disposable gloves, eye
bites, small scratches on the dingers protection, face masks
● Mucous membrane contact ● Barrier protection: to prevent skin and mucous membrane
○ Infection if the organism can directly enter through contamination
the mucous membranes (conjunctiva of the eye) ● Gloves: worn when there is potential for skin contact w
● Arthropod vectors potentially infectious materials; worn in phlebotomy and
○ Infectious sources include ticks, fleas, and capillary puncture
mosquitoes ● Facial protection and body covering: worn when splashes
w blood and other body fluids
II. EXPOSURE CONTROL PLAN ● High-efficiency particulate air respirators: should be fit
Exposure control plan tested for each person; those who encounter mycobacteria
● The risk of exposure to infectious agents for all job through contact w either patient or specimen
classifications and explains exposure-reduction methods must ● Lab employees who come in contact w body fluids should be
include: offered hepa B vaccinations for free
○ Safety education
○ Universal precautions and safety standard V. ENGINEERING CONTROLS
precautions ● To protect employees from the hazards that may occur during
○ Engineering controls the performance of lab procedures
○ PPE ● All labs must adhere to a minimum of Biosafety Level 2
○ Disposal of hazardous waste guidelines
○ Post Exposure procedures

A. POSSIBLE ROUTES FOR INFECTION

III. UNIVERSAL AND STANDARD PRECAUTIONS
Table 1. Biosafety levels.
● Centers for Disease Control (CDC)
○ Universal precautions as recommendations that
BSL Function
describe handling of clinical specimens by health
care personnel; 1987 1 No known pathogenic potential for immunocompetent

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 individuals
Bacillus subtilis
Most undergraduate lab courses operate at this level
Adherence to std lab techniques
2 Level 1 + lab coats, protective gloves, limited access,
decontamination of all infectious waste, biohazard signs
Apparatus includes partial containment equipment
Lab acquired infections: HBV, HIV, Staphylococcus, enteric
pathogens such as Salmonella and Shigella
3 Level 2 + special lab clothing, controlled access
Mycobacterium tuberculosis, Brucella, Coccidiodes
immitis, Rickettsia, arbovirus
Air movement must be carefully contained
4 Level 3 + entrance through a separate room, street clothing à
lab clothing
Maximum containment
Filovirus, arenavirus
Biohazard label
● Used to identify those areas of the lab where infectious
specimens or cultures are stored/present
Figure 1. Biohazard label.
Sterilization
● Done by heat, UV light, passage of air through HEPA filter
BIOSAFETY LEVEL 2 CRITERIA FOR MICROBIOLOGY PRACTICES
● Limit access to lab when working w infectious agents
● Decontaminate work surfaces at least once a day and after
any spill
● Use mechanical pipetting devices
● Do not eat, drink smoke, or use cosmetics. Do not store food
in biological refrigerator.
● Wash hands thoroughly
● Minimize the creation of aerosols
● Wear lab coats, gowns, smocks and remove before leaving
the lab
● Avoid skin contamination by covering cuts w occlusive
bandages and using gloves
● Do not use needles that are bent, cut, or recapped
● Handle M. tuberculosis at BSL 3 in Class II BSC while
wearing a solid-front gown and personal respirator (or face-
folding mask); use sealed safety cups for centrifugation
● Handle mild-lime fungi in BSC; plated media for myco should
be sealed w a cellulose band or cellophane or labelling tape
(prevent dispersion of spores)
B. BIOSAFETY CABINETS
CLASS I BSC
● Provide partial containment for procedures involving
moderate- and high-risk microorganisms (BSL 2 and 3)
● Open-fronted, negative-pressure, ventilated cabinets
● Unsterilized air enters and circulates w/in the cabinet
● Exhaust air from cabinet is filter by HEPA filter
CLASS II BSC
● Provide partial containment for procedures involving
moderate- and high-risk microorganisms (BSL 2 and 3)
● Sterilize bot air entering and circulating w/in the cabinet and
the exhaust air
● Vertical laminar-flow biological cabinets
● Open-fronted; have HEPA filter, recirculated airflow w/in the
workspace
● Exhaust air is filtered, too, by the HEPA filter
● Type II-A
○ Self-contained w 70% of the air recirculated
○ Not required to be vented; acceptable for low- to
moderate-risk agents
○ Most hospital microbio labs use this type
● Type II-B
○ Must be vented w 30% of the air exhausted from the
cabinet and 70% recirculated back into the room
CLASS III BSC
● Provide the highest level of safety
● All air entering and leaving the cabinet is sterilized w a HEPA
filter
● Supply air: drawn through the HEPA filter
● Exhaust air: filtered through 2 HEPA filters
● System is entirely enclosed
● Infectious materials: handled w rubber gloves
VI. POST EXPOSURE PLAN
● All lab accidents or safety incidents: reported to the lab safety
officer or supervisor
● Immediate and appropriate medical treatment
● Administration of immunizations, hepa B immune
immunoglobulin, hepa B vaccinations
● Collection of serum samples at 6 weeks, 3 mos, and 6 mos
for antibodies to HIV or abraded skin exposure to HBV or HIV
● Documentation of the accident w a report
● Corrective actions
VII. DISPOSAL OF HAZARDOUS WASTE
● Microbiological waste must be decontaminated before
disposal
● Needles and sharps should be placed in puncture-resistant
containers
● Contaminated materials should be placed into two leak-proof
plastic bags
● Contaminated pipettes, swabs, and glass should be placed
into puncture-resistant burn boxes
A. AUTOCLAVE
● Decontaminates that uses saturated steam at 121oC and 15
psi of pressure for 1 hr to sterilize
● Infectious medical waste: 132oC for 1/2 -1 hr
● Monitored periodically
● Bacillus stearothermophilus: spore indicator; weekly
monitoring
B. STERILIZATION
● Physical or chemical process that kills all microorganisms,
including spores
● Moist heat or steam under pressure: autoclave
● Dry heat: 171oC for 1 hr, 160oC for 2 hrs, 121oC for 6 hrs
● Liquid: glutaraldehyde, 6-30% hydrogen peroxide, 6-8%
formaldehyde, chlorine dioxide
C. DISINFECTION
● Destroys most microbes but does not kill spores
● Moist heat: 75-100oC
● Liquid: glutaraldehyde, 3-6% hydrogen peroxide, 1-8%
formaldehyde, chlorine compounds, 70% isopropyl alcohol
● Liquid household bleach
● Quaternary ammonium compounds: benzalkonium chloride
D. ANTISEPTICS
● For skin or tissue; not to be substituted for a disinfectant
● Alcohols: 70% ethyl or isopropyl alcohol; not against spores
● Iodophors: iodine combined w an organic carrier molecule;
povidone-iodine and poloxamer-iodine
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 ● Hexachlorophene: bactericidal and broad spectrum but may B. HAZARDOUS CHEMICALS
be toxic ● Corrosive: causes visible destruction or irreversible damage
to human skin on contact
● Toxic: serious biological effects after inhalation, ingestion, or
E. BIOCIDES skin contact w relatively small amts
● Chemical agents that inactivate microorganisms; either -static ● Carcinogenic: ability of chemical to induce a malignant tumor
and inhibit growth of the microbe or -cidal and kill the target ● Ignitable: any chemical that can burn and includes both
organism combustible and flammable liquids
● Explosive: reactive and unstable substances that readily
F. LIQUID DECONTAMINANTS undergo violent chemical change
● 70% EtOH or 10% bleach (NaClO)
● For 10 mins to increase ability to decontaminate X. REVIEW QUESTIONS
● Question 1
VIII. FIRE SAFETY ● Question 2
● Ignition sources: open flames; heating elements; spark ● Question 3
gaps, from light switches or static electricity; electrical Answer 1
instrumentation Answer 2
Answer 3
A. TYPE A
● Water Fire Extinguisher XI. REFERENCES
● Used on combustible materials for fires in paper, wood, ● APA format
rubber, sloth, plastics
● Extinguish fire w cooling effect

B. TYPE B
● Carbon dioxide fire extinguisher
● Used on extremely flammable liquids or electrical fires
including fires of oil, kerosene, gasoline, paints, fats, grease,
solvents
● Extinguish fire by eliminating oxygen

C. TYPE C
● Dry chemical extinguishers
● Used in electrical fires in wiring and other electrical sources or
equipment
● Extinguish fire bc chemicals do not conduct electricity

D. TYPE D
● Combustible metals
● Used for fires involving combustible metals: Na, K, Mg, Na-K
alloys
● NaCl salt wc forms a metal layer over the fire wc occludes the
oxygen
● Graphite, sodium carbonate based, copper based

E. CLASS K
● Dry and wet chemical extinguishers
● Used for kitchen fires involving combustible materials such as
oil or fat
● Extinguish fire by using various wet or dry chemical agents

IX. CHEMICAL SAFETY

● Every compound or reagent must have precautionary labels

A. MATERIAL SAFETY DATA SHEET

● Characteristics of hazardous compound chemicals

Table 2. Sections of MSDS

Section Information
I Manufacturer’s name and address, emergency tel. no., tel
no. for info about preparation
II Hazardous ingredient’s identity info, components, specific
chemical identity common name, PEL
III Physical/chemical characteristics
IV Fire and explosion hazard data
V Reactivity data
VI Health hazard data: routes of entry
VII Precautions of safe handling and use
VIII Control measures