Laboratory Safety and Standards Precautions MODULE

Microbiology

6
Notes
LABORATORY SAFETY AND
STANDARDS PRECAUTIONS

6.1 INTRODUCTION
Working in a laboratory usually involves working with various chemical,
physical, and biological hazards. Because the hazards vary from laboratory to
laboratory, employers must address the hazards specific to their laboratories.
Standard precautions are meant to reduce the risk of transmission of blood borne
and other pathogens from both recognized and unrecognized sources. They are
the basic level of infection control precautions which are to be used, as a
minimum, in the health care settings.

OBJECTIVES
After reading this lesson, you will be able to
z describe preparing the laboratory
z explain about common symbols used in laboratories
z explain fire safety and prevent fire accidents in the laboratories
z describe and follow equipment safety
z explain and follow universal standard precautions

6.2 PREPARING FOR LABORATORY WORK

Before starting to work in a laboratory, you must familiarize with the following:
z The hazards of the materials in the lab, as well as appropriate safe handling,
storage and emergency protocols. Read labels and material safety data

MICROBIOLOGY 67
 MODULE Laboratory Safety and Standards Precautions

Microbiology sheets (MSDSs) before moving, handling or opening chemicals. Never use
a product from an unlabeled container, and report missing labels to your
supervisor.
z The agents, processes and equipment in the laboratory. If you are unsure
of any aspect of a procedure, check with your supervisor before proceeding.
z The location and operation of safety and emergency equipment such as fire
Notes extinguishers, eye wash and shower, first aid and spill response kits, fire
alarm pull stations, telephone and emergency exits
z Emergency spill response procedures for the materials you will handle

During laboratory work

z Restrict laboratory access to authorized persons only. Children are not
permitted in labs.
z Smoking; eating; drinking; storing food, beverages or tobacco; handling
contact lenses are not permitted in laboratories.
z Wear lab coats (knee length) and safety glasses in laboratories employing
chemicals, biohazards or radioisotopes. Open shoes, such as sandals, should
never be worn in the lab.
z Keep work places clean and free of unwanted chemicals, biological
specimens, Avoid leaving reagent bottles, empty or full, on the floor.
z Work only with materials once you know their safe handling and storage.
z Never pipette by mouth; use mechanical transfer devices.
z Keep exits and passageways clear at all times.
z Ensure that access to personal protective equipments are not blocked.
z Report accidents and dangerous incidents (“near-misses”) promptly to your
supervisor
z Wash your hands thoroughly before leaving the laboratory.
z Perform procedures that liberate infectious bio-aerosols in a biological
safety cabinet
z Handle all human blood and body fluids as if potentially infectious.

Cleaning up before leaving

Perform a safety check at the end of each experiment and before leaving the lab.
make sure to:
z Turn off gas, water, electricity, vacuum and compression lines and heating
apparatus

68 MICROBIOLOGY
Laboratory Safety and Standards Precautions MODULE
z Return unused materials, equipment and apparatus to their proper storage Microbiology
locations
z Dispose of all waste material.
z Remove defective or damaged equipment immediately, and arrange to have
it repaired or replaced
z Decontaminate any equipment or work areas that may have been in contact
with hazardous materials. Notes
z Leave behind protective clothing (lab coats, gloves, etc.) when leaving the
laboratory.

6.3 SYMBOLS TO BE IDENTIFIED BY ALL

LABORATORY TECHNICIANS

MICROBIOLOGY 69
 MODULE Laboratory Safety and Standards Precautions

Microbiology
6.4 FIRE SAFETY
Laboratory fires can by caused by bunsen burners, runaway chemical reactions,
electrical heating units, failure of unattended or defective equipment, or
overloaded electrical circuits. Familiarize yourself with the operation of the fire
extinguishers and the location of pull stations, emergency exits and evacuation
routes where you work. In the event that the general alarm is sounded use the
Notes evacuation routes established for your area and follow the instructions of the
Evacuation Monitors. Once outside of the building, move away from the doors
to enable others to exit.

The fire triangle

Fire cannot occur without an ignition source, fuel and an oxidizing atmosphere
(usually air), the three elements that comprise what is called the “fire triangle”:

Fire will not be initiated if any one of these elements is absent, and will not be
sustained if one of these elements is removed. This concept is useful in
understanding prevention and control of fires. For example, the coexistence of
flammable vapours and ignition sources should be avoided, but when flammable
vapours cannot be controlled elimination of ignition sources is essential.
Learn how to use the extinguisher in your lab, as there will be no time to read
instructions during an emergency. Attempt to fight small fires only, and only if
there is an escape route behind you. Remember to have the extinguisher
recharged after every use.
z P: Pull and twist the locking pin to break the seal.
z A: Aim low, and point the nozzle at the base of the fire.
z S: Squeeze the handle to release the extinguishing agent.
z S: Sweep from side to side until the fire is out.
z Be prepared to repeat the process if the fire breaks out again

70 MICROBIOLOGY
Laboratory Safety and Standards Precautions MODULE
Microbiology
6.5 GLASSWARE SAFETY
Use a dustpan and brush, not your hands, to pick up broken glass. Discard broken
glass in a rigid container separate from regular garbage and label it appropriately.

Equipment Safety
Every effort should be made to prevent equipment from becoming contaminated.
To reduce the likelihood of equipment malfunction that could result in leakage, Notes
spill or unnecessary generation of aerosolized pathogens:
z Review the manufacturer’s documentation. Keep for future reference.
z Use and service equipment according to the manufacturer’s instructions.
z Ensure that anyone who uses a specific instrument or piece of equipment
is properly trained in setup, use and cleaning of the item.
Ensure that equipment leaving the laboratory for servicing or disposal is
appropriately decontaminated. Complete a Certificate of Equipment
Decontamination.

Centrifuges
z Check glass and plastic centrifuge tubes for stresslines, hairline cracks and
chipped rims before use. Use unbreakable tubes whenever possible.
z Avoid filling tubes to the rim.
z Use caps or stoppers on centrifuge tubes. Avoid using lightweight materials
such as aluminum foil as caps.
z Use sealed centrifuge buckets (safety cups) or rotors that can be loaded and
unloaded in a biological safety cabinet. Decontaminate the outside of the
cups or buckets before and after centrifugation. Inspect o-rings regularly and
replace if cracked or dry.
z Ensure that the centrifuge is properly balanced.
z Do not open the lid during or immediately after operation, attempt to stop
a spinning rotor by hand or with an object, or interfere with the interlock
safety device.
z Decant supernatants carefully and avoid vigorous shaking when re-
suspending.

Heating baths, water baths

z Never use laboratory ovens for preparation of food for human consumption
z Glassware that has been rinsed with an organic solvent should be rinsed
with distilled water before it is placed in a drying oven.

MICROBIOLOGY 71
 MODULE Laboratory Safety and Standards Precautions

Microbiology
6.6 UNIVERSAL / STANDARD PRECAUTIONS
Universal Precautions
These guidelines refer to the precautions, consistently used for all patients
regardless of their infection status and diagnosis. The main objective is to
prevent exposure of staff and patients to blood and body fluids.

Notes z Don’t eat, drink, smoke or apply cosmetics (including lip balm).
z Don’t insert or remove contact lenses.
z Don’t bite nails or chew on pens.
z Don’t mouth pipette.
z Limit access to the laboratory to trained personnel only.
z Assume all patients are infectious for HIV or other blood borne pathogens.
z Use appropriate barrier precautions to prevent skin an mucous membrane
exposure, including wearing gloves at all times and masks, goggles, gowns
or aprons if there is a risk of splashes or droplet formation.
z Wash hands thoroughly and other skin surfaces after gloves are removed
and immediately after any contamination.
z Avoid injuries to sharps such as needles and scalpels.

Standard Precautions
In 1996, CDC developed a new system of standard precaution synthesizing the
features of universal precautions and body substance isolation. Standard
precautions are used in the care of all patients and apply to blood, all body fluids,
secretion and excretion except sweat regardless of whether they contain visible
blood.
Standard precautions are guidelines and procedures designed to reduce the risk
of transmission of microorganisms from both recognized and unrecognized
sources of infection in healthcare settings.

Standard Precautions Include

z Hand washing
z Barrier protection
z Safe handling of sharp items
z Safe handling of specimen (blood etc)
z Safe handling of spillage of blood/body fluid
z Use of disposable/ sterile items

72 MICROBIOLOGY
Laboratory Safety and Standards Precautions MODULE
Hand Washing Microbiology

Single most important method to limit cross transmission of nosocomial

pathogens
Multiple opportunities exist for HCW hand contamination by Direct patient care
and Inanimate environment
This is an ideal safety precaution and gloves should not be regarded as a Notes
substitute for hand washing.

For General Patient Care (Hand Contamination)

z Wash hands thoroughly in running water with soap without missing any
area. For effective hand washing first wash palms with fingers followed by
back of hands, knuckles, thumbs, fingertips and wrists,. Rinse and dry
thoroughly.
z Wash hand immediately after accidental contamination with blood/body
fluid, before eating and drinking and after removing gowns/coats and
gloves.
z Leave soap bars in dry containers to prevent contamination with
microorganism.

The 6 stages of effective hand hygiene

MICROBIOLOGY 73
 MODULE Laboratory Safety and Standards Precautions

Microbiology For Hand Disinfection (Hygienic Hand Wash)

Required in high risk health care settings (ICU, neonatal units, nursery)
z Use 2-4% chlorhexidine gluconate/ detergent solution, povidone iodine/
detergent solution containing 0.75% available Iodine.
z Do not use alcoholic hand rubs as substitute for hand washing except for
rapid hand decontamination between patient contacts.
Notes
Barrier Protection

Gloves
z Wear while collecting/ handling blood specimens and blood soiled items.
z Wear while disposing waste.
z Remove before handling door knobs, telephone, pens performing office
work.
z Discard if cracked, discoloured or punctured.
z Discard if blood spills on them.
z Don’t reuse disposable gloves.
z Wash hands when gloves are removed or changed.

Masks
z Wear masks and protective glasses if splashing or spraying of blood/body
fluids is expected.
z Mask of cotton wool, gauze, or paper masks are ineffective. Paper mask
with synthetic material for filtration are an effective barrier against
microorganism.

Caps
z Cover hair completely in aseptic units, operating rooms, or performing
selected invasive procedure.

Gown and Aprons

z Wear clean clothes made up of a material easy to clean.
z Change after exposure to blood and body fluids.
z Wear gown or apron of plastic or water resistant paper when splashes of
blood or other body fluids are likely to occur e.g. during surgery, obstetric
procedure, invasive procedure, post mortem and embalming.

74 MICROBIOLOGY
Laboratory Safety and Standards Precautions MODULE
Occlusive Bandage Microbiology

z Cover all skin defects e.g. cuts, scratches or other breaks with waterproof
dressing before patient care.

Safe Handling of Sharps

z Take extreme care to avoid autoinoculation.
Notes
z Discard all chipped or cracked glassware in appropriate containers.
z Don’t manipulate disposable needles. Never bend, break, recap or remove
needle from syringe.
z Dispose your own sharps. Don’t pass used sharps directly from one person
to another.
z Discard needles in puncture proof rigid containers (Plastic or cardboard
boxes) after disinfection in 0.5-1% sodium hypochlorite solution. Use
needle shredder if available for needles or needles along with syringe
nozzle.
z Send sharp disposable containers for disposal when three-fourth full.

Safe Handling of Specimen

z Collect specimens, specially blood and body fluids in pre sterilized
containers properly sealed to prevent leakage or spillage.
z Use autoclaved/ pre-sterilized disposable syringes and needles for vene-
punture and lancets/ cutting needles foe finger pricks.
z Cover cuts in hands properly with waterproof adhesive bandages.
z Wear disposable gloves while collecting blood/body fluids and maintain
proper asepsis.
z Wash hands thoroughly with soap and water, particularly after handling
specimens.

Safe Handling of Blood/Body Fluid Spills

z Cover spills of infected or potentially infected material on the floor with
paper towel/ blotting paper/ newspaper.
z Pour 1% sodium hypochlorite solution on and around the spill area and cove
with paper for at least 30 minutes.
z After 30 minutes, remove paper with gloved hands and discard.

MICROBIOLOGY 75
 MODULE Laboratory Safety and Standards Precautions

Microbiology Use of Disposable Sterile Items

z Ensure proper handling of disposable/ sterile item before use.
z There should be no recirculation of disposable items.

WHAT HAVE YOU LEARNT

Notes
z The laboratory needs to be prepared for work both before and during the
work
z Laboratory fires can by caused by bunsen burners, runaway chemical
reactions, electrical heating units, failure of unattended or defective
equipment, or overloaded electrical circuits
z Fire cannot occur without an ignition source, fuel and an oxidizing
atmosphere (usually air), the three elements that comprise what is called
the “fire triangle”:
z Every effort should be made to prevent equipment from becoming
contaminated.
z Take appropriate measures to reduce the likelihood of equipment malfunction
that could result in leakage, spill or unnecessary generation of aerosolized
pathogens
z Glassware that has been rinsed with an organic solvent should be rinsed
with distilled water before it is placed in a drying oven.
z Standard precautions include Hand washing, Barrier protection, Safe
handling of sharp items, Safe handling of specimen (blood etc), Safe
handling of spillage of blood/body fluid, Use of disposable/ sterile items
z Hand Washing is an ideal safety precaution and gloves should not be
regarded as a substitute for hand washing.
z Wash hands thoroughly in running water with soap without missing any
area. For effective hand washing first wash palms with fingers followed by
back of hands, knuckles, thumbs, fingertips and wrists,. Rinse and dry
thoroughly.
z Wash hand immediately after accidental contamination with blood/body
fluid, before eating and drinking and after removing gowns/coats and
gloves.
z Use barrier protection like Gloves, Masks, Caps, Gown and Aprons,
Occlusive bandage
z Sharps must be handled safely and injury must be prevented

76 MICROBIOLOGY
Laboratory Safety and Standards Precautions MODULE
z Safety precautions are to be followed while handling specimens and Blood/ Microbiology
body fluid spills

TERMINAL QUESTIONS
1. What are standard precautions
Notes
2. How would you handle blood/body fluid spills
3. Write measures to prevent fire accidents
4. Explain steps of hand washing

MICROBIOLOGY 77