MICROBIOLOGY

SAFETY ON THE CLINICAL

MICROBIOLOGY LABORATORY
NAME OF LECTURER
DATE OF LECTURE

OUTLINE o Apply to human blood and other body fluids w visible

I. Laboratory Safety B. Sterilization blood but does not apply to feces, nasal secretions,
A. Possible Routes for C. Disinfection saliva (unless they contain visible blood)
Infection D. Antiseptics  Standard precautions
II. Exposure Control Plan E. Biocides o Set of preventive measures applied to all patients
III. Universal and Standard F. Liquid Decontaminants that are designed to reduce the risk of infection in
Precautions VIII. Fire Safety the health care setting
A. Safety Guidelines for A. Type A  Potentially infectious
the Laboratory B. Type B o Blood, tissue, body fluids, secretions, excretions
IV. Personal Protective C. Type C (except sweat)
Equipment D. Type D
V. Engineering Controls E. Class K A. SAFETY GUIDELINES FOR THE LABORATORY
A. Biosafety Levels IX. Chemical Safety
1. No food or drink
B. Biosafety Cabinets A. Material Safety Data
2. No smoking
VI. Postexposure Plan Sheet
3. No cosmetics
VII. Disposal of Hazardous B. Hazardous Chemicals
4. Protective eyewear must be worn
Waste
5. Wear face shields
A. Autoclave
6. Clothing: gowns, aprons lab coats, closed shoes
7. Hair should be tied up
A. LABORATORY SAFETY 8. Wash hands frequently and thoroughly
9. Eyewash stations: 100 feet or 10 secs away from any area in
B. POSSIBLE ROUTES FOR INFECTION the lab
 Airborne 10. Emergency stations: shower
o Aerosols from centrifugation of unstopped tubes or from 11. Respirators: available where there is air contamination
heating cultures or specimens too rapidly, removing 12. Mouth pipetting: ingestion of microorganisms; use pipette
stoppers from tubes, leakage from a container that holds bulbs/automated devices
contaminated specimens 13. Glassware and sharps: in puncture-resistant containers
 Ingestion 14. Centrifuges: minimize the production of aerosols
o Infection as a result of failure to wash hands or eating,
drinking, smoking, applying cosmetics, or pipetting w the B. PERSONAL PROTECTIVE EQUIPMENT
mouth  Protective laboratory clothing disposable gloves, eye
 Direct inoculation protection, face masks
o Infection from needlesticks, broken glass, animal bites,  Barrier protection: to prevent skin and mucous membrane
small scratches on the dingers contamination
 Mucous membrane contact  Gloves: worn when there is potential for skin contact w
o Infection if the organism can directly enter through the potentially infectious materials; worn in phlebotomy and
mucous membranes (conjunctiva of the eye) capillary puncture
 Arthropod vectors  Facial protection and body covering: worn when splashes
o Infectious sources include ticks, fleas, and mosquitoes w blood and other body fluids
 High-efficiency particulate air respirators: should be fit
C. EXPOSURE CONTROL PLAN tested for each person; those who encounter mycobacteria
through contact w either patient or specimen
Exposure control plan
 Lab employees who come in contact w body fluids should be
 The risk of exposure to infectious agents for all job
offered hepa B vaccinations for free
classifications and explains exposure-reduction methods must
include:
o Safety education C. ENGINEERING CONTROLS
o Universal precautions and safety standard precautions  To protect employees from the hazards that may occur during
o Engineering controls the performance of lab procedures
o PPE  All labs must adhere to a minimum of Biosafety Level 2
o Disposal of hazardous waste guidelines
o Postexposure procedures
A. BIOSAFETY LEVELS FOR INFECTIOUS AGENTS
D. UNIVERSAL AND STANDARD PRECAUTIONS Table 1. Biosafety Levels
BSL Function
 Centers for Disease Control (CDC)
1 No known pathogenic potential for immunocompetent
o Universal precautions as recommendations that
individuals
describe handling of clinical specimens by health
Bacillus subtilis
care personnel; 1987
Most undergraduate lab courses operate at this level
 Clinical Laboratory and Standards Institute (CLSI) Adherence to std lab techniques
o Universal precautions as set of preventive measures
2 Level 1 + lab coats, protective gloves, limited access,
designed to reduce the risk of transferring HIV, hepa decontamination of all infectious waste, biohazard signs
B virus, and other bloodborne pathogens in the Apparatus includes partial containment equipment
health care setting
1
 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
 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
B. 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)
C. BIOSAFETY CABINETS
i. 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
B. 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
o Self-contained w 70% of the air recirculated
o Not required to be vented; acceptable for low- to
moderate-risk agents
o Most hospital microbio labs use this type
 Type II-B
o Must be vented w 30% of the air exhausted from the
cabinet and 70% recirculated back into the room
C. 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
D. 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
E. 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
 Hexachlorophene: bactericidal and broad spectrum but may
be toxic
E. BIOCIDES
 Chemical agents that inactivate microorganisms; either -static
and inhibit growth of the microbe or -cidal and kill the target
organism
2
  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 C. REVIEW QUESTIONS
 Question 1
G. FIRE SAFETY  Question 2
 Ignition sources: open flames; heating elements; spark  Question 3
gaps, from light switches or static electricity; electrical
instrumentation Answer 1
A. TYPE A Answer 2
Answer 3
 Water Fire Extinguisher
 Used on combustible materials for fires in paper, wood,
D. REFERENCES
rubber, sloth, plastics
 Extinguish fire w cooling effect  APA Format

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

F. 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

B. HAZARDOUS CHEMICALS
 Corrosive: causes visible destruction or irreversible damage
to human skin on contact
 Toxic: serious biological effects after inhalation, ingestion, or
skin contact w relatively small amts
 Carcinogenic: ability of chemical to induce a malignant tumor
 Ignitable: any chemical that can burn and includes both
combustible and flammable liquids