laboratory biosafety 2007

(CAREC)PAHO/WHO

Serving21 Member Countriesin the English & Dutch -Speaking Caribbean

An overview of Laboratory Safety

Biosafety

Dr. Ashok Rattan,Medical Microbiologist &

Laboratory Director,CAREC

Caribbean Epidemiology Centre (CAREC) PAHO/WHO

Before entering the water to swim you must learn all you can about the sharks that may lurk there

SAFETY IN THEMYCOBACTERIOLOGYLABORATORY


Protection:– workers– “products”– co-workers– lab support personnel– environment

IntroductionWhy Biosafety Practices?

2.1


Safety practices should be applied throughout the testing process:

Pre- analytical– Specimen collection

– Specimen preparation

– Specimen transport

Analytical– Testing

Post-analytical– Disposal


Examples of Laboratory Hazards

Growth of microorganisms

Water baths

Aerosols, splashing, tube breakage

Centrifuge

Accidental inoculation, aerosol, spillage

Needles

HazardOperation / Equipment


Safety Resources


IntroductionChain of Infection

Reservoir of pathogenReservoir of pathogen

Portal of escapePortal of escape

TransmissionTransmission

Route of entry/infectious doseRoute of entry/infectious dose

Susceptible hostSusceptible host

Incubation periodIncubation period

Risk A

ssessment

PPE

Immunization

Surveillance

Practices/

Equipment


BSL1 - agents not known to cause disease. BSL2 - agents associated with human disease. BSL3 - indigenous/exotic agents with potential for aerosol

transmission; disease may have serious or lethal consequences. BSL4 - dangerous/exotic agents which pose high risk of life-

threatening disease.

Principles Biosafety Levels of agents

2.1


Designing for Safety

General requirements– Facility design

– Water supply/sinks for hand washing

– Ventilation

– Standard lab practices

Safety equipment– Personal protective equipment (PPE)

– Biosafety cabinets


Biosafety Level 1 Standard Microbiological Practices 1/3

Restrict or limit access when working

Prohibit eating, drinking and smoking in the laboratory

Pipetting by mouth strictly forbidden

2.3


Biosafety Level 1 Standard Microbiological Practices 2/3

2.3


Standard practices also include: 3/3

Keep work areas uncluttered and clean No food in lab refrigerator Minimize splashes and aerosols Decontaminate work surfaces daily Maintain insect & rodent control program


Biosafety Level 2 Facility Design (Secondary Barriers) 1/8

Requirements: Laboratories have lockable doors Sink for hand washing Work surfaces easily cleaned Bench tops are impervious to water Sturdy furniture

2.4


Biosafety Level 2Laboratory Facilities (Secondary Barriers) 2/8

BSL-1 Facilities PLUS:– Autoclave available– Eyewash station

available

2.4


Requirements:– Location - separated from public areas

– Structure - normal construction

– Ventilation - directional

Biosafety Level 2 Facility Construction (Secondary Barrier) 3/8

2.4


Biosafety Level 2Safety Equipment (Primary Barriers) 4/8

In addition to BSL-1: Use biosafety cabinets (class II) for work with

infectious agents involving:– Aerosols and splashes– Large volumes

– High concentrations

2.4


Biosafety Level 2Special Practices 5/8

Needles & Sharps Precautions – Use sharps containers– DON’T break, bend, re-sheath or reuse

syringes or needles

2.4



Needles & Sharps Precautions DON’T place needles or sharps in office waste

containers

2.4



Needles and Sharps Precautions – DON’T touch broken glass with hands

2.4


Identify “clean” and “contaminated” areas – Use appropriate warning signs

Decontaminate work surfaces Report spills and accidents Remove gloves, lab coats before leaving

work area No animals in laboratories


2.4



BSL-1 and 2 Facilities PLUS:

– Separate building or isolated zone

– Double door entry

– Directional inward airflow

– Single-pass air; 10-12 air changes/hour

2.5



BSL-1 and 2 Facilities PLUS (cont.):

– Enclosures for aerosol generating equipment

– Room penetrations sealed

– Walls, floors and ceilings are water resistant for easy cleaning

2.5



BSL-1 and 2 Safety Equipment PLUS:– BSC class

II or III to manipulate infectious material

2.5



BSL-1 and 2 Safety Equipment PLUS:– Respiratory protection may be indicated

2.5


BSL-2 Special Practices PLUS:– Work in certified BSC– Use bioaerosol-

containing equipment– Decontaminate spills

promptly


2.5


Biosafety Level 4 – Maximum Containment BSL -3 practices plus:

– Clothing change before entering laboratory– Shower on exit– All materials decontaminated on exit from facility

Safety Equipment:– Class III Biosafety cabinet– Class I or II biosafety cabinet– WITH full-body, air supplied,

• positive personnel suit


Biological Waste

Types– cultures, stocks, isolates– materials containing or contaminated with blood– sharps– pipettes, wrappers, tips– All materials used in the lab


Specimen Disposal


DecontaminationDecontamination Sterilization Disinfection


DecontaminationDefinition

SterilizationThe use of a physical or chemical procedure to

destroy all microbial life, including large numbers of highly resistant bacterial spores.


DisinfectionThe use of a physical or chemical procedure to

virtually eliminate all recognized pathogenic microorganisms but not all microbial forms (bacterial endospores) on inanimate objects.

DecontaminationDefinition


DecontaminationMethods

Heat Chemical Radiation


Types– Moist – steam– Dry– Incineration

*The most effective method of sterilization

DecontaminationHeat


Types– Liquids, i.e. chlorox,

hydrogen peroxide– Gases, i.e. ethylene oxide

DecontaminationChemical


General Lab Use - Hypochlorite Solutions– Large Spills/Large Organic Load

• undiluted from bottle– Small Spills/Virus Inactivation

• 10% - 1:9– General Surface Disinfection

• 1% - 1:99

DecontaminationChemical


In case of a spill

Wear disposable gloves Cover large blood spill with paper towels and

soak with 1% (10000 ppm) of household bleach and allow to stand for at least 5 minutes

Small spill - wipe with paper towel soaked in 1% bleach

Discard contaminated towels in infective waste containers

Wipe down the area with clean towels soaked in a same dilution of household bleach


Safety Documentation & Records

Laboratory Safety Manual - Policies and Procedures Sample Contents:

– Housekeeping– Personal protection– Safe decontamination of equipment– Decontamination & Waste Disposal– Emergency procedures

In-lab first aid Accidental injury Post exposure prophylaxis Contacts

– Personnel responsibilities Hygiene


Action Plan for Implementing Safety Practices Identify hazards Assess level of risks

– Prioritize risk Establish and implement safety polices and

procedures Conduct safety specific training

– Must be a priority– Communication is key

Perform regular audits and assessments


In Case of Exposure

Be ready for the emergency before hand- Familiar with exposure specific

policies- Conduct drills - Keep post exposure medicines

available- Check periodically for stock and

expiry of medicines Report immediately Go to the nearest, first available doctor


Post Exposure

Write a report and reasons for accident Actions taken to avoid future accidents Training


SAFETY IN THEMYCOBACTERIOLOGYLABORATORY

laboratory biosafety 2007

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