/

What is Biosafety?
All possible safety measures employed when
handling biohazardous materials to avoid infecting
oneself, others or the environment.
Biosafety ensures

BIOSAFETY HANDOUT

Biosafety means practicing

That biological research is conducted in a safe fashion

That biological research meets regulatory requirements
Support the Institutional Biological Safety Committee
(IBSC)

safe science
Risk Assessment

Ask Questions BEFORE you start

Plan safety into your experiment
Biosafety manuals - each lab should have these and
provide training on their contents
Good lab practices/microbiological techniques
Hazard communication - you must notify anyone who
enters your area of risks
Reduce risks to acceptable levels
Concern for others and the environment - we have an
obligation to protect the public and the environment

Risk assessments on lab procedures should be done before

initiating them. They should answer the following:
What are the hazards?
What might happen?
How likely is it to happen?
How serious are the consequences if it happens?
What are the possible exposures?
How can I mitigate exposure?
What is the WORST that can happen?

Biosafety learning objectives

1. Understand the role and responsibilities of the
experimenter with respect to Biosafety
2. Understand the underlying principles of biosafety as they
relate to conducting safe and effective science requiring the
use of biohazardous materials in laboratories
3. Be able to locate resources that will facilitate the
determination of the relative biosafety risk associated with
a planned experiment
4. Recognize the significance and need for conducting
experiments at different biosafety containment levels
5. Understand the differences among BSLI, 2 and 3

What is a Biohazard?

Biosafety learning objectives continued:

6. Recognize the role each piece of Personal Protective
Equipment plays in safeguarding the health and welfare of
the laboratorian and community
7. Be familiar with the proper use of Biological Safety
Cabinets (BSC)
8. Recognize the inberent risk with using centrifuges in the
laboratory with a special emphasis on risks associated
with infectious agents
9. Understand the significance of good laboratory
housekeeping
10. Be able to clean up a minor biological spill within a BSC
11. Understand the significance of working with
biohazardous materials specifically the role of the PI in
informing the experimenter of potential hazards in the
workplace as well as the personal responsibility required
of each employee and other workers

Universal Biohazard Symbol

Universal

Symbol

A potential hazard to humans, animals or the

environment caused by a biological organism, or
by material produced by such an organism

Communicates
exposure

Examples

present

Viruses, bacteria, fungi, and parasites and their toxins.

Blood and body fluids, as well as tissues from humans
and animals.
Transformed
acids.

cell lines and certain types of nucleic

Typically

potential

red or orange

Symbol should be defaced

when hazard is no longer
Use sparingly,

explicitly

Use for cultures

pathogens

of

Human blood, tissue

Equipment

Storage

used with above

Cages of infected

Entry door of a laboratory

working with pathogens

areas of above
animals

Safety Risk Groups

Introduction

\VHO Risk Group 1 (no or low individual and community risk)

A microorganism that is unlikely to cause human disease or animal disease
\VHO Risk Group 2 (moderate individual risk. low community risk)
A pathogen that can cause human or animal disease but is unlikely to be a
serious hazard to laboratory workers, the community, livestock or the
environment
Laboratory exposures may cause serious infection. but effective treatment and
preventative measures are available and the risk of spread of infection is
limited.
WHO Risk Group J (high individual risk. low community risk).
A pathogen that usually causes serious human or animal disease but does not
ordinarily spread from one infected individual to another.
Effective treatment and preventive measures are available.
WHO Risk Group 4 (high individual and community risks)
A pathogen that usually causes serious human or animal disease and that can
be readily transmitted from one individual to another, directly or indirectly.
Effective treatment and preventive measures are not usually available.

Chain of Infection

Biosafety Levels

Conventional Agents
Risk

Individual

Community

Group

Risk

Risk

Low

Low

Levell

'coli.

Lenl~

E.coli 0J57:H7,

eIn
01' 1InIm_

I
..,c_~

ModerateR .

Limited

HiCh

Low

Levell

'''yc.,.. ",lou.If

HI.h

Hi.h

I w"" "":::;~::'>."". I

several
&. salmonella
species. adenovirvses.
Hepatitis
A. B &. C InjIuOlZD. Measles

Biosafety Level 2

Biosafety Levell
:;'SuitJ.ble for work involving wellcharacterized agents not known to
cause disease in healthy adult
humans and of minimal potential
hazard to laboratory
personnel
and the environment

,Suitable
Ior work
agents or moderate
hazard to personnel
environment.

Involving
potential
and the

ssrepsococcus

huto\_OI'~"dis._.

Bacillus sub/iUs

c_

UnIIkefyto
hMlthy wortwr.

Examples

Containment

Level 4

Bacillus anthraW.
Mycobocle.rium
tuberculosis.
Hanus virus. HIV. Yellow fever
virvs
WS:iO virus. Ebola viruJ.
Marburg virus

).-Examples

<. 6adllus
<.

Canine

;"Examples
Mnsltsvlrus

sub,ilis
hepatitis

virus

.:. E. coli
-:. Variulla (Cnteken

.:- SIII",."d/ae

.:. Toxopllls",. species

POI)

Biosafety Level 3
,. Recommended for work with
infectious

agents

cause serious
lethal disease

which

Risk Group-l

Biosafety Level 4

may

or

potentially
as a result of

exposure.

.> Recommended for work with

dangerous and exotic agents that
pose high individual and
community risks.

.:. Mycobacurium tuberculosis

.:. Bacillus anthracis
.:. West Nile virus
.:. Encephalitis virus
':'SARS
.:. Yellow fever
.:. Influenza virus HINI
.:. Francisdla tularensis

is a Grarn+ve.

.,. B. subtilis

is not considered

(BSL-l): Bacillus subtilis

sporulating
a human

rod-shaped
pathogen:

commonly

it may contaminate

found

food but

rarely causes food poisoning.

Risk Group-2 (BSL-2)

Pathogenicity:
Host range:

acute disease,

fatality <

0.5% - 250;;,

Human

.:. Ebola virus

.:. Marburg virus

Transmission:

.:. Hanta virus

Vaccine

primarily

droplet

spread

available

Hepatitis Virus
-Parhogeniciry. asymptomatic

':'HlV

and symptomatic

infections

-Lcng-term fatality = 2.30/0.950/ofadu1t infections

.:. HistoplllSmll

bacterium

Measles Virus

,Example:

)- Example

>B. subtilis,
in soil.

self-limiting

cllpsulatum

-Host range:

Human (chimpanzees

are susceptible)

-Vaccine available

Risk group - 4: Ebola virus

Risk Group 3 (BSL-3)

Mycobacterium
,.1. tuberculosis
Mycobaclerium
Host Range:

tuberculosis

Ebala is responsible
for
the
disease
EOOla hemorrhagic rever (EHF). Electron
micrographs
show
long
filaments,

(MTB) is a pathogenic bacterial species in the genus

and the causative agent of most cases of tuberculosis.
Primarily

human,

cattle, primates

etc.

characteristic

of this viral family.

The virus interferes with the endothelial

cells
lining the interior surface of blood vessels and

'

platelet cells.
As the blood vessel walls become damaged
and the platelets
are unable to coagulate,
patients succumb to hypovolemic
shock.

Bacillus anthracis
-Bacillus

anthracis

causative

agent of Anthrax

is a Gram-positive

spore-forming,

rod-shaped

bacterium.

the

Ebcla is transmitted primarily through bodily

fluids and to a limited extent through
mucous membrane contact.

skin and

BSL-4 laboratory
recommended even for
clinical work

Elements that can modify risk

Levels of Biohazards

Are host factors that change the risk

Deficiencies

in host" immunity

- Pre-existing

medical

conditions

- for example:

eczema
Reproductive

hazards

- Pregnancy,
Rubella,

teratogens,

mutagens

Toxoplasma,

- for example

Behavioral

vaccine

constituents,

antimicrobial

therapies

Animal dander, egg proteins. latex

training,

experience,

motivation,

I
I

BSL2

'Ii

elements

- Education,

so does;

- the risk of the organism to humans, animals,

plants and/or the environment

BSL 3

Chlamydia

Allergies
- Foreign proteins,

As the level increases

BSL4

attentiveness

BSL 1

-the procedural and facility requirements

- the level of containment required
- the degree of protection for personnel, the
environment and the community,

Infection/Biohazard
Unconventional

Control

agents

TSE prion diseases; lethal transmissible

neurodegenerative conditions
Creutzfeld-Jakob disease, Variant C-J Disease, Mad
Cow Disease! Scrapie, Chronic Wasting Disease.

Engineering Controls
Administrative Controls

Resistant to destruction by procedures that

normally inactivate viruses.

Practices and Procedures

Thus, containment, procedures, waste disposal

etc. are very different in this situation.

Personal Protective Equipment

Engineering controls (contd.)

Engineering Controls

Primary barriers- containment

Technology based, reduce or eliminate

exposure to hazards by changes at the source
of the hazard.
Containment:
Primary vs Secondary
Containment levels

~
)0

.,.

or agent at source

Biological safety cabinet

Animal caging
Specialized lab equipment (centrifuges, fermenters.etc.)

Secondary barriers- contain the agent within the room or facility in

case an agent escapes
barriers

).
~

from the primary

Building & room construction

HV AC Issues:
.:.
.:.

Directional air now

Exhaust filtration

};- Other Engineering Controls:

.:. Solid waste treatment
.:. Wastewater treatment

Administrative

Controls
Practices and Procedures

Program based, information

minimize risk of exposure.

and methods to

Risk assessment
Medical Surveillance
Training/Education
Resources
Inspections
Signs & Labeling

General Safety Guidelines

Good Microbiological Techniques
Handwashing
Specific Procedures
Centrifuges
Needles & Syringes and other sharps
Pipettes
Blenders, Grinders, Sonicators & Lyophilizers
Inoculation Loops
Cryostats

Biosafety:
All procedures
production

Standard

involving

are performed

Laboratory

live pathogens

Practices

and toxins with risk of aerosol

inside the biosafery

Identifying types of PPE to protect against exposure by

different routes

cabinet

No work with open vessels on open bench

Use of paper covering
Use of appropriate
Substitute

Ingestion

on work surface

4;;U

Inhalation

disinfectant

plastic for glass

Careful pipetting

techniques

Wash hands often

Contact

No mouth pipetting
No eating or drinking
Minimize

aerosol

Decontaminate

in lab

generation

Percutaneous /'

work surfaces

Safe sharps handling

Wear appropriate

PPE

Biosafety: Personal Protective Equipment

Types ofPPE
.,. Gloves

PPE: Policies and procedures

Staff must have appropriate

,. Respirator

For example,

membrane or

barrier
against
skin,
respiratory exposure to

must

be chosen

based

on the hazards

- How to use the PPE they have selected Donning

gloves

to be handled

Purpose
Provides

PPE training

- Which PPE to use when

., Appropriate footwear
:; Lab coats
). Eye and face protection

);P

or PPE

mucous

infectious

agents
;,..To prevent spread of contamination
Limitations
:;..Does not eliminate the haunts
,. Integrity wanes with use (change gloves
frequently)
,. Not all glo\'r5 created equal-select best glove for
the task

(putting

Aseptic removal

on) and Doffing

procedures

(taking

ofT) procedures

gloves last

- PPE cleaning
Personal protective equipment is NOT worn outside of the
lab or taken home to be laundered!
Reusable protective clothing should be auteclaved on-site

- How to perform PPE maintenance

Need policy on frequent)'

Respirators
testing

require

medical approval

and fit

Example of PPE for work with

Mycobacterium tuberculosis

PPE: Safety features

Equipment

Safety features

Hazard corrected

Laboratory coats,
gowns. coveralls

Contamination

Plastic Aprons

Contamination

Back opening

of clothing

Cover street clothing

Back closing lab coat

-

Wrap around gowns with tight cuffs

Gloves
-

Impact and Splash

Closed Toe

Goggles and
Safety Spectacles

Impact and Splash

Impact-resistant
lenses,
Side shields

Face shields

Impact and splash

Single pair for entry

Double gloving for work in BSes.
and spill clean-up

transport.

Shoe coverings
Face protection
-

Waterproof

of clothing

Footwear

Safety glasses

Respiratory
appropriate
SARS)

'Shield entire face

-Eastly removable

in case of accident

or goggles

protection may be
(e.g., M. tuberculosis,

Respirators

"Fun or half

race

air

purifying

-Full (ace or hooded powered air

purifying
Gloves

Biosafety Levell Lab Design

-Single-use disposable

Inhalation of aerosols

Disposable microbiologically
approved latex. vinyl or nitrile

Direct contact with

microorganism

Biosafety Levell

lab design

Requirements:
Labcratertes have doors
'Sink for hand W3shinc
-Work surfaces easuy cleaned
Bench tops are impervious to
water
-Sturdy furniture
-Windows fitted

wuh fly

screens

-Other design and

construction issues:
-Locatten - not separated
-Strueture - normal
construction
Ventilation - no special
requirements

Standard

Standard Microbiological

Practices

Restrict or limit access

when conducting research
Prohibit eating, drinking
and smoking
Prohibit mouth pipeting

Microbiological Practices

-Needles & sharps precautions

-

Use sharps containers

DON'T break, bend, resheath or reuse syringes

or needles

- Use alternatives to
needles when available
Minimize splashes and aerosols
Decontaminate work surfaces daily
Decontaminate infectious waste
Maintain an insect & rodent control programs

Biosafety Level 2

Biosafety Level 2
. BSLI

Requirements

plus:

-Laboratories
have
lockable doors
-Biological safety
cabinets installed as

needed
-Adequate

illumination

-Eyewash

readily

available
Location - separated
from public areas
-Ventilation

directional
Air flows into lab
without re-circulation
to non-lab areas

.1

Biosafety Level 3

Biosafety Level 3
BSL2 requirements plus:
-Enclosures
'Room

fur aerosol

penetrations

'Walls,

noors

generanng

equipment

sealed

and

cetungs

art

water

resistant

for

easy

dtaninz:
'BSe

d:U5 II or 111 to manipulate

-Separste

buildln:

-Directlonal
'Anteroom

-ncubte

Inward
doors

'Sin:ir-p3SS

or isolated

infectious

zone ",ithin

material
11

build in:

airflow

may be stlf-closing:

air; can be recirculated

and interlocking
if HEPA

filtered

door entry

-Additiena! requirements depending on agents:

'Effluent
-Persennet

decentamtnanon
snowers

Biosafetv Level Sumrnarv Table

Biosafety Level 4
BSL3 enhanced requirements plus:

"'"u'*<t"i...

.''''' ~ ". ~ 10141h" ' ~ ~r;1 ~IM .'''ob~k~J

._.u'. 1h."",I" ~.~It~)
I"r",,;;i .,

Either:

__~I.!'..I!:

Class III biosaftly

cabinet

Personnel
shower
room with cabinet

required

----:------------

+-

I . \:~.l' "D("".""jl'

to uit

- _-:

rl",:

i lI~l'll'f",":ko

- -

-l"nlf"d

E'" .,'

i.:r<-<1iooft,..,,,a:t<

[" ..n.I'''~<ltd''''(lIOI",

, ",1, ". ~tp<>."".

Or

\1'" ...

Suit deoontaminatlon
Personnel

Double

shower

HEPA-filtered

HEPA-filtered

supply

Decontamination
Sterilization
Redundant

shower
10 exn

of waste

~., "lnI"

u'lOll"iO)'

'" l.b'l ,,,,,,..,,~

.,

and materials
systems

lll.\.l-~ ,unit.

p"~"

",:.I,," .

D'"f".~I'Urir
~t<.t!
..-b'("PO'tW1~rhkot
H1.th "':dI' I.o
\.-r~'
I_" 11I,<4
1,,1., Ii
'''', ....,I.~,

1.1.0<,"~I'

~:~:~~~:,~

,.Jd

I;a~". mk

!__

I.d

p'Ufk.,plu,,
(10<~11(~UCtNltor.'II'
,<;h -.run;'
I d.< I~ '
i 1:><lIill
'

""U~II""';
<I..~< .'~d
f, ~II.~ .I~"''''''''W1I'''
IlI.UI!p!'[, . '1""''''11''''
lat ,.'~-n <lo.O,,~~: P"'_,

..,~\pir'f~',

lB'L.~
I

,~' '!;tk.'t"!;"(

"mu~ balrlfn: Um I
r 8~' ,' "' ..!>~\-'k~1

( .11
. "u,
, ().>'"'~ffllu'i.~ .1 ~IIn~I.
l1>.<
~~a".AlI.1I~t, al'."
' r"'lbj.~.
I.nu
! {b ...'.......

exhaust
air

of all effluents

back-up

1.1tir ~fI nOli<-

, 0, IlI"- k.,, """'"

'.,1u

pr I<-.,iu n

i.f

r,~,.

'AU
I.,,,,,tdwn<ndl<,.dil>
I"'h"IIIR<,(',,,, fl,I",

~,.I""'''''.

II ll'(, ill'
i'~ h,1I t.<>d~..
",,iljo. p ,.,,,."",

\.,rI...-.!.
.

I !I'SL.! pl.-: - P\J"tI(~

.p."._ ft_ ~
ronW',r,~rif-.-I,";i1I!

d~I''''-oJ._ 1""
[)bJ",t.';r .
,1<''''.,.'1:<.-:.''
1~1I1I<'"
~.tJI;o)u~.,,,

! 1"It.'I')b~lr","':

w"' '.1
.,,,

\;"~i-;-.

~,,,bl'~

1'r.'._I
! n

1 It.tJC~.11~ ",,(f.'tl~1 I~I

'1 I"u 1., C'L t 1')011

tlul< <t'l'!a< ..

lllbon

Posltln: pressure
suit ",Uh serrcontained
breathing
apparatus

tM,Ik'":~~::~I~~~:.;'.:.'
"1

iul;').

,10:-;01""-, 1"',,,.

_ __

:~~~:i:~::~:~';:,~"
L"'I01'

, ~:7~~~::E::":E:~""IE:;~~~.~?;":;':~~:::
M

bu,~ .",1

0, .
<iook,'.q""""
- -

a~t..-jp"" 'iif.p ~
blikh.,
orl'oOlatofd
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."d .rb
!.,.""'~I.
u.1
a

.~"

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<Ioo."'.'~"''''.lu .
.<:~

."''t'ilr'II'I''~ ~<':Ihll
1
I III ,Ii. ~.u

10

Biological Safety Cabinets (BSCs)

Primary Barriers-BSe

based protection

Primary means of containment

Designed to provide protection for

Personnel
Protection

Product
Protection

Environment
Protection

Fume Hoods

Yes

No

No

Flow Clean

No

Yes

No

Yes

No

Yes

Yes

Yes

Yes

Yes

Yes

Yes

~Personal
Chemical

~Product
~Environment

Laminar

Three design types

Benches

~Class I, Class II, and Class III

~Class II BSCs are further classified into
Class II A I, Class II A2, Class II B I and Class II B2

Class I Biological
Safety Cabinets

Class II Biological
Safety Cabinets

Class 1IIBiological
Safety Cabinets

HEPA Filter
HEP A = !!igh Efficiency ~articulate ~ir
Minimum efficiency of99.97% removal of 0.3 micron particles
HEPA filters do not filter out gases, vapors or volatile
chemicals, they only filter out particulates (bacteria and
viruses)

Biosafety Cabinet - Do's and Don't

The Cabinet must not be used unless it is working properly.
The glass viewing panel must not be opened when the cabinet is in
use.
Apparatus and materials in the cabinet must be kept to a minimum.
Bunsen burners must not be used in the cabinet..
All work must be carried out in the middle or rear part of the
working surface.
Traffic behind the operator should be minimized.
Operator should not disturb the airflow by repeated removal and
reintroduction of his or her arm.
The surface of the biological safety cabinet should be wiped using an
appropriate disinfectant.
The cabinet fan should be run for at least 5 min before beginning
work and after completion of work.

11

Sterilization
Sterilization refers to any process that effectively kills or eliminates
transmissible agents (such as fungi. bacteria, viruses, spore forms, etc.)
from a surface, equipment, article of food or medication, or bio1ogical
culture medium.
There are different types of sterilization:
Dry heat sterilization
Moist heat sterilization
Chemical sterilization etc.

Dry Heat Sterilization

Moist heat sterilization

I.

Below 100C:
Water bath - 56C for 60 min
Vaccine bath-. 60

Pasteurization

for 60 min

of milk

Holder method> peaks at 63C for 30 min

Flash method> peaks at 72C for 20 seconds
Ultra high temperature
for a few seconds
Fractional
2.

stertlizatlcn-.

(UUT) method

peaks at 11Soe

Serum inspissator

AlIOOC:
Bolling for 10 to 30 min

Dry heat sterilization of an article is one of the earliest forms of

sterilization practiced. Dry heat, as the name indicates, utilizes hot air that
is either free from water vapour. or has very little of it. and where this
moisture plays a minimal or no role in the process of sterilization.

Tyndallizer ; steaming for 3 successive days at l000e to kin all

organism in their vegetative forms by a.Howing the
spores time to hatch in between the heating periods.
Steam
J.

Above

sternuer , Steam at atmospheric

pressure

for 90 minutes

100C:

Autoclavlng

Autoclave (contd.)

A widely-used
method
for heat
sterilization is the autoclave. sometimes
called a converter.
Autoclaves commonly use steam
heated to 121C.
To achieve sterility, a holding time
of at least 15 minutes at 121C.
Following sterilization, liquids in a
pressurized autoclave must be cooled
slowly to avoid boiling over when the
pressure is released.

Chemical Sterilization
Chemicals are also used for sterilization. Although heating
provides the most reliable way to rid objects of all
transmissible agents, it is not always appropriate, because
it will damage heat-sensitive materials such as biological
materials, fibre optics, electronics, and many plastics. Low
temperature gas sterilizers function by exposing the articles
to be sterilized to high concentrations (typically 5-10%
v/v) of very reactive gases (alkylating agents such as
ethylene oxide, and oxidizing agents such as hydrogen
peroxide and ozone). Liquid sterilants and high
disinfectants typically include oxidizing agents such as
hydrogen peroxide and peracetic acid and aldehydes
such as glutaraldehyde.

12

Definitions
Sterilization - act or process, physical or chemical, that
destroys or eliminates all forms of life, especially
microorganisms

Decontamination
is part of
Biosafety requirements

Disinfectant - an agent, usually chemical, that inactivates

viruses or kills vegetative microbes but not necessarily
resistant forms such as spores
Antiseptic - a substance that prevents or arrests the growth
or action of microbes, either by inhibiting their activity or
by destroying them (living tissue use)
Decontamination - disinfection or sterilization of
contaminated articles to make them suitable for use
Sanitizer - an agent that reduces the numbers of vegetative
bacteria only

The Basics: Hand washing

In 1847, Dr. Ignaz Semmelweis, a physician
in a Vienna hospital, insisted that students
and physicians clean their hands with
chlorine prior to patient contact, especially
after leaving the autopsy floor.
Childbirth fatalities dropped dramatically
due to the new practice.

Surface decontamination
Typically
Factors

",

wipe or spray surface

to consider

Surface topography

Organic

(wooden

surfaces.

cracks, etc)

load

Concentration

Contact

Shelflife

time

Remember,
-

with liquid disinfectant

include

these are hazardous

chemicals,

so

Training

- PPE
-

Ventilation

- Regulatory issues
-

Hazardous

waste disposal

13

Choosing disinfectants

Disinfection: What to use for my organism?

The disinfectant needs to be appropriate for the

biological agent of interest

Bacteria

Viruses

Yegdatitt! bacteria (Ecal; S. uureus)

2"_ demesne

Prions
Bacterial spores
Coccidia (Cryptosporidium)
Mycobacterium
Nonlipid viruses
(e.g. Hep A, Polio)
Fungi
Rickettsiae, Chlamydiae
Vegetative bacteria
Lipid-containing viruses
(e.g. HIV)

Area Decontamination

Quaternary

6% Inrmulated

to".

domestic

peroxide

2% domestic:

Spore forming

Hydro:en

bllc:tu;1I

10% demesne

on risk assessment.
include:

to decontaminate

Formaldehyde

Vapor phase hydrogen

Chlorine

dioxide

C1uttnldehyde
Formaldehyde
6% fonnulalfd

Hydrogen

peroxide"

-Laboratory

Ethylene

oxide

and Decontamination

wastes

-Sclid, non-sharp

that may need decontamination

labware

-Tissue

or carcass

(nasks.lubu,

wasre

platts,

boWes.

vials)

(patholo:ical)

such as

Aspirates,

of areas

include

-aste. such as

'l!!!!!!

Sharps.

peroxide

peroxide

Formaldeh)'de

{Bacillus spp }

Sera.

Hydrogen

Gluteraldehyde

peroxide"

Liquids.

gas decontamination

peroxide"

blt:lch

equipment

After large spill

Most common
Include

Hydro:en

bleac:b

6-;. formulated

compounds

6% rormul:ated

anunonia

Non enveloped {Hepatitis.Adenovirus)

1S% Ethanol
Phmollc:

blt:lch

6*/* formulated

bleach

before removing from lab

-

Quatunary

Hydrogen

MycobaCleriQ and fungi

- Before maintenance
In airlock,

10% domestic:
75% Ethanol

ammonia

Waste Handling

\Vhen to do depends
Common applications
-

Enveloped (HI"'. II~I

bl~(h

75"_ Ethanol

culture

nuids,

rtoses, washes, ere.

body fluids

such as

(VHP)
Anything
with a poinl
human skin
Glass

or fdge capable

lab wa~e (sometimes

of piercing

also broken

or CUllin:

plastic:)

Large variation in price and equipment

required
-

Formaldehyde
generated by heating
paraforrnaldehyde
flakes to generate
formaldehyde
gas while VHP requires
specialized equipment

Some are human carcinogens

-F. etlity needs:

waste

designation

process

'Waste

collection.

packaging.

'Treatment
'Disposal

capabilities
options

and transport

(intermediate

and/or

procedures
final)

(backups)

-Trainine

14

All Sharps

Common Errors In Waste Handling

Improper labeling of waste
Improper storage of waste
Failure to cap waste bottles
Accumulation of excessive waste
Improper segregation of waste

Must be Autoclaved and NEVER placed in

general waste
Includes disposable syringes, needles, scalpels, etc. and
may include pipette tips, glass slides, glass vials, etc.
Place "sharps" and disposable syringes in approved red Sharps
Container.
2) Add water (-20 ml), "Building and Room Number" label and
autoclave tape.
3) Place "Sharps Container" in metal pan and autoclave for
decontamination and pick-up.
1)

Neither syringes nor needles may be sent to the landfill. They must
be discarded in "Sharps" Containers only, not in regular trash or
autoc1aved discard.

Plan - Do - Check - Act

Plan
- Planning, including identification of hazards and risks and
estabUshing program goals

Do
- Implementing, including training and operational issues

Check
- Checking, including monitoring and corrective action

Act
- Reviewing, including process innovation and acting to make
needed changes to the management system

Conclusion
It is up to you to protect yourself, your coworkers, the community, and the environment!
Good lab practices/good microbiological
techniques are fundamental
Always communicate about hazardous materials
Always PLAN, PLAN and PLAN
When in doubt, don't forget to ASK the competent
authority'! !
With proper knowledge, planning and care, a
biohazardous exposure is completely avoidable.

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