SOP for Handling Spills of Biohazardous Material SOP Number: MB-13-0??


US Environmental Protection Agency
Office of Pesticide Programs

Office of Pesticide Programs

Microbiology Laboratory

Environmental Science Center, Ft. Meade, MD

SOP for Handling Spills of Biohazardous Material

SOP Number: MB-13-02
Date Revised: 01-24-11

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 1 of 14

EPA/OPP MICROBIOLOGY LABORATORY
ESC, Ft. Meade, MD

Standard Operating Procedure
for

Handling Spills of Biohazardous Material
SOP Number: MB-13-02
Date Revised: 01-24-11

Initiated By:		Date: / /

Print Name:	

Technical Review: 	Date: / /

Print Name:

Technical Staff

QA Review:		Date: / /

Print Name:	

QA Officer

Approved By:		Date: / /

Print Name:	

Branch Chief

SHEM Review: 	Date: / /

Print Name:

SHEM Manager

Effective Date:	/ /

Controlled Copy No.:	

Withdrawn By:	Date:	/	/

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 2 of 14

TABLE OF CONTENTS
Contents	Page Number

1.0 SCOPE AM) APPLICATION	4

2.0 DEFINITIONS	4

3.0 HEALTH AND SAFETY	5

4.0 CAUTIONS	6

5.0 INTERFERENCES	6

6.0 PERSONNEL QUALIFICATIONS	7

7.0 SPECIAL APPARATUS AND MATERIALS	7

8.0 INSTRUMENT OR METHOD CALIBRATION	8

9.0 SAMPLE HANDLING AND STORAGE	8

10.0 PROCEDURE AM) ANALYSIS	8

10.1. Reporting Instructions	8

10.2 Recommendations for Reducing Potential for Spills	8

10.3.	Spills of Biohazardous Organisms Requiring

Biosafety Levels 1 and 2 Containment	9

10.4.	Spills of Biohazardous Organisms Requiring

Biosafety Level 3 Containment (inside BSC)	10

10.5.	Spills with Potential for Exposure to Bacillus anthracis
(inside BSC)	 11

10.6	Spills of Biohazardous Organisms Requiring

Biosafety Level 3 Containment (outside BSC)	12

10.7	Decontamination of Clothing	13

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 3 of 14

TABLE OF CONTENTS (continued)

Contents	Page Number

11.0	DATA ANALYSIS/CALCULATIONS	14

12.0	DATA MANAGEMENT/RECORDS MANAGEMENT	14

13.0	QUALITY CONTROL	14

14.0	NONCONFORMANCE AND CORRECTIVE ACTION	 14

15.0	REFERENCES	 14

16.0	FORMS AND DATA SHEETS	 14

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 4 of 14

1.0 SCOPE AND APPLICATION:

1.1	The protocol presents guidelines for decontamination and clean up of
biohazardous spills. Periodically, the SOP distinguishes between large spills vs.
small spills, and spills inside vs. outside of the biological safety cabinet.
Procedures for responding to a spill may vary, depending upon the degree and
location of the spill of biohazardous material.

1.2	The OPP Microbiology Laboratory Branch (MLB) holds a select agent
registration to work with the select agent Bacillus anthracis in room B202. This
SOP also provides guidance on handling spills of B. anthracis.

2.0 DEFINITIONS:

2.1	Mycobacterium bovis (BCG) is a live attenuated vaccine strain used to vaccinate
humans against infection by Mycobacterium tuberculosis.

2.2	BSC = biological safety cabinet

2.3	PPE = personal protective equipment

2.4	CDC = Centers for Disease Control and Prevention

2.5	NIH = National Institutes of Health

2.6	"Appropriate" disinfectant = EPA-registered hospital disinfectant (efficacious
against S. aureus, P. aeruginosa, and S. choleraesuis), hospital disinfectant with
tuberculocidal claims (efficacious against S. aureus, P. aeruginosa, S.
choleraesuis, andM bovis (BCG)), or EPA-registered disinfectant towelette
efficacious against spore-former Clostridium difficile. All disinfectants must be
used according to the directions (e.g., use dilution, contact time, etc.) specified on
the labeling.

2.7	Spill = A spill is defined as a biohazardous material out of control. The quantity
of the biohazardous material spilled is not the sole determining factor in deciding
whether or not an event is classified as a spill. Rather, the essential issue is
whether the biological agent, the location, and the quantity collectively cause the
situation to be beyond the control of the laboratory worker. A large spill is one
that cannot be handled safely by laboratory employees in the immediate area. A
small spill is one which can be handled by the laboratory workers in the
immediate area without posing a serious threat to their health and safety, and that
can be cleaned up with available absorbents and disinfectants.

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 5 of 14

2.8	OEP = Occupant Emergency Plan

2.9	CHP = Chemical Hygiene Plan

2.10	Restrooms = Restrooms equipped with showers (e.g., D222 and D223)

2.11	References to water mean reagent-grade water

2.12	P100 HEPA = Oil proof High-Efficiency Particulate Air filter, 99.97% efficient in
removing particles 0.3 microns or larger

HEALTH AND SAFETY:

3.1	To protect the laboratory worker from possible infection by microorganisms, the
health and safety guidelines provided in SOP MB-01 (Biosafety in the
Laboratory) and in the Centers for Disease Control and Prevention/National
Institutes of Health (CDC/NIH) "Biosafety in Microbiological and Biomedical
Laboratories," (BMBL) 5th ed. (see ref. 15.1) manual must be followed. All
laboratory personnel are required to read and familiarize themselves with SOP
MB-01 and sections on Biosafety Levels 2 and 3 in the BMBL.

3.2	Spills and accidents are handled according to the practices outlined in this SOP, as
well as procedures referenced in the ESC OEP and CHP. See section 10.1 for
instructions for reporting spills to the Branch Chief and SHEM (Safety, Health,
and Environmental Management) manager.

3.3	Procedures for handling spills of biosafety level 3 microorganisms, including
Bacillus anthracis, are divided into two categories: 1) spills occurring inside of
the BSC, and 2) spills occurring outside of the BSC. Serious or lethal disease
may result from inhalation of a biosafety level 3 microorganism. If the BSC
remains operational following a spill, infectious aerosols resulting from a spill
inside of the BSC are contained, and the analysts do not need to evacuate the
laboratory to avoid exposure. Infectious aerosols resulting from spills outside of
the BSC are not contained; hence the need for different responses, depending
upon the location of the spill. In the event of a spill of a biosafety level 3
microorganism outside of the BSC, workers are to evacuate the laboratory.

3.4	In the event that analysts are unable to reach the SHEM manager, they are to call
security at extension 2800 and report the spill. Security will locate the SHEM
manager or his designee.

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 6 of 14

4.0 CAUTIONS:

4.1	Lack of use or understanding of this SOP may negatively impact the
decontamination efforts of laboratory staff and hence, cause unnecessary exposure
of employees to biohazardous microorganisms.

4.2	Room-specific procedures (see Table 4) for addressing clothing and skin
contaminated with a biosafety level 3 microorganism result from the difference in
the structure of the double-door access rooms in B202 and B207. The B202
double-door access room has a sink. The B207 double-door access room does not
have a sink.

4.3	Failure to clean the ultraviolet lamps in the BSCs will reduce the lamps'
effectiveness. Periodically clean the ultraviolet lamps in the BSCs with a lint-free
cloth dampened with alcohol.

4.4	Do not use a liquid bleach solution to decontaminate stainless steel surfaces (e.g.,
BSC) following a spill of biohazardous material, including spore-forming bacteria
(e.g., Bacillus subtilis), unless the microorganism is a select agent (i.e., Bacillus
anthracis). In the event that a spill occurs in the BSC involving Bacillus
anthracis, treat the stainless steel surfaces with a liquid bleach solution as
described in section 7.6.2. Following treatment, the surface must be rinsed with
water, 70% ethanol, or an EPA-registered liquid disinfectant to remove excess
sodium hypochlorite.

4.5	Recently, the Agency has registered sodium hypochlorite towelettes which claim
effectiveness against the spore-forming microorganism, Clostridium difficile. In
the event that a spill of C. difficile occurs inside the BSC (i.e., steel surface), these
towelette products may be used to disinfect the surface. However, following the
duration of the towelette contact time, the surface must be rinsed with water, 70%
ethanol, or an EPA-registered liquid disinfectant to remove excess sodium
hypochlorite.

5.0 INTERFERENCES:

5.1	Failure to become familiar with and to put into practice the procedures set forth in
this SOP will result in analysts who are a danger to themselves, others, and the
environment.

5.2	Improper maintenance and/or expired certification may result in failure of the
BSCs to operate properly. Refer to proper use and maintenance procedures in
SOP QC-06, Use and Maintenance of Biological Safety Cabinets.

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 7 of 14

6.0 PERSONNEL QUALIFICATIONS:

6.1 Personnel are required to be knowledgeable of the procedures in this SOP.

Documentation of training and familiarization with this SOP can be found in the
training file for each employee.

7.0 SPECIAL APPARATUS AND MATERIALS:

7.1	Autoclave

7.2	Biohazard bags (clear in color, autoclavable) or containers inside and outside of
the biological safety cabinets for collection and storage of biohazardous waste.

7.3	When specified, personal protective equipment (PPE) such as gloves, safety
glasses, lab coats, disposable laboratory garments, shoe covers, respiratory
protection (respirators with P100 HEPA filter cartridges), and temporary clothing
(i.e., scrubs).

7.4	Biosafety Spill Kit containing items such as autoclave bag, latex gloves, gloves
for handling broken glass, dustpan/brush, shoe covers, disposable lab coat, paper
towels, disinfectant (with hospital disinfectant and tuberculocidal efficacy), and
safety glasses.

7.5	Appropriate signs to identify biohazardous materials and to limit access to
laboratories.

7.6	Bleach solutions made fresh as needed. Discard solution at the end of the day.
When a new container of bleach is opened, the analyst will note the date opened
on the container. The container of bleach will be discarded six months from the
date of receipt.

7.6.1	1:10 diluted bleach solution for decontamination purposes (vegetative
cells). Using an EPA-registered sodium hypochlorite product, dilute as
follows: 1 part bleach, 9 parts water.

7.6.2	1:10 diluted bleach solution at neutral pH for decontamination
purposes (spore-forming microorganisms). Using an EPA-registered
sodium hypochlorite product containing approximately 6% sodium
hypochlorite, dilute as follows: 1 part bleach, 8.4 parts water, and 0.6
parts 5% white vinegar or 5% lab grade acetic acid.

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 8 of 14

7.7	Appropriate disinfectants for microorganisms specified in Attachment A of SOP

MB-01, Biosafety in the Laboratory.

7.7.1	For microorganisms in vegetative form, use a 1:10 diluted bleach
solution as described in section 7.6.1 or EPA- registered hospital
disinfectant/tuberculocide.

7.7.2	For spore-forming microorganisms, prepare the adjusted bleach
solution described in section 7.6.2.

7.8	Key card readers are used to limit access to testing laboratories. Only authorized

personnel are permitted to enter.

8.0 INSTRUMENT OR METHOD CALIBRATION: Not applicable
9.0 SAMPLE HANDLING AND STORAGE: Not applicable
10.0 PROCEDURE AND ANALYSIS:

10.1	Guidance for Spills of Biohazardous Material - Reporting Instructions

10.1.1	Accidents are handled according to the practices outlined in this
subpart, as well as procedures referenced in the OEP and the ESC
CHP.

10.1.2	All spills and accidents involving a Biosafety Level 2 or 3
microorganism, regardless of how minor a spill, are reported to the
Branch Chief and the SHEM manager (or call security desk at
extension 2800).

10.1.3	The Branch Chief is responsible for documenting spills and accidents.

10.1.4	The Branch Chief and SHEM manager will determine if written
documentation or follow-up is warranted.

10.2	Recommendations for Reducing Potential for Spills of Biohazardous

Material

10.2.1 Use secondary containment (e.g., autoclave bin) when transporting
live cultures in liquid media or on solid media.

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 9 of 14

10.2.2 Use secondary containment to store biohazardous waste (plates and
tubes) that is generated during the course of an assay.

10.2.3 Prepare the least amount of culture necessary for an assay.

10.2.4 Maintain a clean, well-organized work environment.

10.3 Guidance for Spills of Biohazardous Organisms Requiring Biosafety Level 1
and 2 Containment, (see Attachment A) - Spills Outside and Inside the BSC.

See the summary table (Table 1) below:

Table 1: Biosafety Level 1 and 2 Spill Control Procedures

Spills Outside & Inside of the BSC

Notify Workers/SHEM
Manager/Laboratory Director (or
Security)

Alert workers in the laboratory that a spill has occurred. Contact the SHEM Manager (or
call security desk at extension 2800) and Branch Chief.

Maintain BSC air flow

Keep the BSC operational. Do not turn off the blower for anv reason.

Treat Human Injury

Treat injury or call 911, depending upon the severity of the injury.

Control the Source

Stand the spilled bottle upright, and unplug any small contaminated equipment (i.e., vortex,
timer).

Control the Spread

Cover the spill with a paper towel (not plastic-backed material) or other appropriate
absorbent material (e.g., Isosorb 13000) to prevent the spill area from growing in size.

Remove Contaminated Clothing

Remove any contaminated gloves, lab coat, and street clothing and place in an autoclavable
container or bag. Wash contaminated skin as described below. Replace any contaminated
street clothing with temporary clothing such as scrubs (located in B202 and B207), or
jumpsuit from the shower area of the restrooms D222 and D223.

Wash Contaminated Skin

For small spills on the skin, wash skin thoroughly with soap and water and don temporary
clothing. For large spills to a significant portion of the body and clothing, disrobe, put on
temporary clothing and shoe covers (to protect against shedding of infectious
microoganisms in the corridor), wash hands with soap and water, exit the laboratory and
proceed to the restroom (D222 or D223) to fully shower for 10 minutes. Discard
temporary clothing and shoe covers in autoclavable container or bag. Don new set of
temporary clothing (e.g., jumpsuit in restroom).

Decontaminate Area

Vegetative cells: Saturate the paper towel or other absorbent material, starting with the
edges of the spill and working towards the center, with liquid disinfectant (e.g., 1:10 bleach
solution, section 7.6.1; EPA registered hospital disinfectant, section 7.7.1) and let stand for
20 to 30 minutes. Do not use a bleach solution on stainless steel surfaces of the BSC.

Spore-forming bacteria: For spills of spore-forming bacteria on non-stainless steel surfaces
outside of the BSC, use a 1:10 diluted bleach solution at neutral pEl (section 7.6.2) for a 60-
minute contact time. For spills of C. difficile in the BSC (stainless steel), wipe surface
with a sodium hypochlorite-containing towelette registered by the Agency for effectiveness
against the microorganism. After the duration of the label-specified contact time, wash the
surface with water, 70% ethanol, or an EPA-registered disinfectant to remove excess

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 10 of 14

sodium hypochlorite. For spills of other BSL-1 and -2 spore-forming bacteria in the BSC
(stainless steel), wipe or spray surface with a non-bleach liquid disinfectant and wipe up
spill with paper towels to physically remove spores. Turn on UV light overnight.

Clean Up

Pick up absorbent material and place in autoclavable container or bag. Use thick gloves,
scoop, and brush found in biosafety spill kit to clean up broken glass. Place in
autoclavable container.

Inspect the Area

Carefully check the entire affected area and beyond for remaining spill or cleanup residue.

Decontaminate all clean-up material
and PPE

Autoclave contaminated paper towel or other absorbent material and any contaminated
broken glass and writing utensils according to SOP MB-01, Biosafety in the Laboratory.
Treat contaminated gloves (for handling broken glass), scoop, and brush with disinfectant
or place under the UV light overnight. Treat contaminated PPE and any contaminated
street clothing with disinfectant or autoclave as described in section 10.7. Discard
autoclaved broken glass in a broken glass container.

10.4 Guidance for Spills of Biohazardous Organisms Requiring Biosafety Level 3
Containment (Excluding Bacillus anthracis) - Spills contained within the BSC
(bioaerosols will be contained by the BSC's laminar air flow). See the

summary table (Table 2) below:

Table 2: Biosafety Level 3 Spill Control Procedures (Excluding Bacillus anthracis)

Spills Inside of the BSC

Notify Workers/SHEM
Manager/Laboratory Director

Alert workers in the laboratory that a spill has occurred. Contact the SHEM Manager (or
call security desk at extension 2800) and the Branch Chief.

Maintain BSC Air Flow

Keep the BSC operational. Do not turn off the blower for anv reason.

Treat Human Injury

Treat injury or call 911, depending upon the severity of the injury.

Control the Source

Stand the spilled bottle upright, and unplug any small contaminated equipment (i.e., vortex,
timer).

Control the Spread

Cover the spill with a paper towel (not plastic-backed material) or other appropriate
absorbent material (e.g., Isosorb 13000) to prevent the spill area from growing in size.

Remove Contaminated Clothing

Remove contaminated gloves, lab coat, and street clothing and place in an autoclavable
container or bag. Wash contaminated skin as described below. Replace contaminated
street clothing with temporary clothing such as scrubs (located in B202 and B207 double-
door access rooms).

Wash Contaminated Skin

For small spills on the skin of the hands or forearms, wash skin thoroughly with soap and
water.

Decontaminate Area

Saturate the paper towel or other absorbent material, starting with the edges of the spill and
working towards the center, with liquid disinfectant (e.g., EPA registered hospital
disinfectant, section 7.7.1) and let stand for 20 to 30 minutes. Do not use a bleach solution
on stainless steel surfaces of the BSC.

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 11 of 14

Clean Up

Pick up absorbent material and place in autoclavable container or bag. Use thick gloves,
scoop, and brush found in biosafety spill kit to clean up broken glass. Place glass in
autoclavable container.

Inspect the Area

Carefully check the entire affected area and beyond for remaining spill or cleanup residue.

Decontaminate all clean-up material
and PPE

10.5 Guidance for Spills of Materials with Potential Risk of Exposure to Bacillus
anthracis - Spills contained within the BSC (bioaerosols will be contained by
the BSC's laminar air flow). See the summary table (Table 3) below:

Table 3: Spill Control Procedures for Bacillus anthracis
Spills Inside of the BSC

Notify Workers

Alert workers in the laboratory that a spill has occurred in the BSC. Notify the SHEM
Manager and Branch Chief.

Maintain BSC Air Flow

Keep the BSC operational. Do not turn off the blower for anv reason.

Keep Hands Inside the BSC

Analyst must keep his or her hands inside of the BSC until the spill is cleaned up and
the outer layer of gloves has been removed. Other analysts (uncontaminated) in the
laboratory will assist by supplying the necessary cleanup materials.

Spill onto Absorbent Padding

If the spill occurred onto absorbent padding, return the overturned flask, tube, etc., to
an upright position. Place the contaminated absorbent padding and other items into the
autoclavable waste container in the BSC. Wipe down the BSC with 70% ethanol or
EPA registered hospital disinfectant (section 7.7.1), replace the absorbent padding and
resume work. Upon completion of work in the BSC, turn on the UV light for a
minimum of 15 hours to decontaminate the area of the spill.

Spill directly on BSC Surface (no
absorbent padding)

If the spill occurred on the surface of the BSC, proceed through the steps below.

Control the Spread

Return overturned flask, tube, etc. to an upright position. Cover the spill with paper
towels (not plastic-backed paper) or other absorbent material to prevent the spill area
from growing in size. Unplug any small contaminated equipment (i.e., vortex, timer).

Pre-c lean/Physical Removal of Spores

Saturate paper towel or other absorbent material, starting with the edges and moving
towards the center, with a 1:10 diluted bleach solution at neutral pH (section 7.6.2).
Let stand for a minimum of ten minutes.

Clean Up

Use thick gloves, scoop, and brush found in biosafety spill kit to clean up broken glass.
Place glass in autoclavable container inside the BSC. Wipe up liquid with paper
towels. Flush the BSC with water, 70% ethanol, or EPA registered hospital
disinfectant (section 7.7.1) to remove excess sodium hypochlorite. Wipe up the liquid
with paper towels. Place all absorbent material/paper towels in the autoclave bin
inside the BSC.

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 12 of 14

Inspect the Area

Carefully check the entire affected area and beyond for remaining spill or cleanup
residue.

Turn on UV Light in BSC

Once the spill has been pre-cleaned and the liquid wiped up, turn on the UV light for a
minimum of 15 hours.

Decontaminate all clean-up material and
PPE

Autoclave biohazardous waste, including gloves/brush/scoop used to pick up broken
glass, using a 3 hour liquid kill cycle for liquid and/or solid waste at 121°C. Discard
autoclaved broken glass in a broken glass container.

10.6 Guidance for Spills of Biohazardous Organisms Requiring Biosafety Level 3
Containment - Spills Outside of the BSC. See the summary table (Table 4)
below:

Table 4: Biosafety Level 3 Spill Control Procedures

Spills Outside of the BSC

Notify Workers/SHEM
Manager/Laboratory Director (or
Security)

Alert workers in the laboratory that a spill has occurred. Contact SHEM Manager (or call
security desk at extension 2800) and Branch Chief.

Maintain BSC Air Flow

Keep the BSC operational. Do not turn off the blower for anv reason.

Evacuate Laboratory - non-select
agent microorganism

Remove labcoat, gloves, and shoe covers, discarding them in the laboratory biohazard bin.
Exit the laboratory to the anteroom.

Evacuate Laboratory - Bacillus
anthracis-specific

Remove labcoat, shoe covers, and outer layer of gloves and place in laboratory biohazard
bin. Keep on respirator and inner layer of gloves. Exit laboratory to anteoom. Remove
respirator and then inner gloves. Place in bin in anteroom.

Treat Human Injury

Treat injury or call 911, depending upon the severity of the injury.

Remove Contaminated Clothing

Once in the double-door access room, remove contaminated street clothing and place in an
autoclavable container or bag. Wash contaminated skin as described below. Replace
contaminated street clothing with temporary clothing such as scrubs (located in B202 and
B207 double-door access rooms). Wash hands in the sink in the double-door access room
(B202) or if the spill occurred in B207, exit the double-door access room and proceed to
another laboratory to wash hands.

Wash Contaminated Skin

For small spills on the skin in B202, wash skin thoroughly with soap and water and don
temporary clothing. For small spills on the skin in B207, put on a disposable lab
coat/tyvek sleeve and gloves (to protect against shedding of infectious microorganisms in
the corridor) and proceed to another laboratory to wash skin thoroughly with soap and
water. For large spills (in either laboratory) to a significant portion of the body and
clothing, disrobe in the double-door access room, put on temporary clothing, gloves, and
shoe covers, exit the laboratory and proceed to the restroom (D222 or D223) to fully
shower for 10 minutes. Discard temporary clothing, gloves, and shoe covers in an
autoclavable container or bag. Don new set of temporary clothing (e.g., jumpsuit in
restroom).

Direct Workers to Stay Out

Direct (via telephone, note on door, etc.) analysts not present in the laboratory to stay out

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 13 of 14

until spill clean up is complete.

Allow Facility'"s Ventilation to
Remove Aerosols

Wait at least 30 minutes before re-entering the laboratory to allow the LSC's exhaust air
ventilation system to remove bioaerosols.

Re-I 'liter the 1 .aboratory
(Sill ¦ M/Responders)

Re-enter the laboratory with appropriate level of PPL as determined by SI IFM and
internal/external responders. Survey the spill site for location and amount of spill.

Cover the Spill Area

Cover the spill w itli a paper towel (not plastic-backed material) or other appropriate
absorbent material (e.g., Isosorb 13000) as appropriate to site and location of spill. 1 Jnplug
any small contaminated equipment (i.e.. vortex, timer).

Decontaminate Area

Saturate the paper towel or other absorbent material, starting w ith the edges of the spill and
working towards the center, with liquid disinfectant (e.g.. 1:10 bleach solution, section
7.6.1: LPA registered hospital disinfectant, section 7.7.1) and let stand for 20 to 30
minutes. Do not use bleach solution on stainless steel surfaces.

For spills of Bacillus anlhracis (outside of 1 JSC/non-slainless steel surface): Use a 1:10
diluted bleach solution at neutral pi 1 (section 7.6.2) for a 60-minute contact time.

Clean Up

Wearing appropriate PPL, pick up absorbent material and place in autoclavable container
or bag. 1 Jse thick gloves, scoop, and brush found in biosalety spill kit to clean up broken
glass. Place glass in autoclavable container. Wash (with water, 70% ethanol. or LPA
registered hospital disinfectant) and dry all areas where sodium hypochlorite was applied.

Inspect the Area

Carefully check the entire affected area and beyond for remaining spill or cleanup residue.

Decontaminate all clean-up material
and PPL

Autoclave contaminated paper towel or other absorbent material and any contaminated
broken glass and writing utensils according to SOP MB-01, Biosalety in the Laboratory.
Treat contaminated gloves (for handling broken glass), scoop, and brush with disinfectant
or place under the IJV light overnight. Treat contaminated PPIand any contaminated
street clothing with disinfectant or autoclave as described in section 10.7. Discard
autoclaved broken glass in a broken glass container.

10.7 Decontamination of Cloth Lab Coats, Street Clothing, and Footwear

10.7.1 Decontaminate clothing with an appropriate disinfectant or by
autoclaving.

10.7.2 If using disinfection as a means of decontamination, treat area of
contamination and surrounding area with disinfectant for label-
specified dilution and contact time (section 7.7.1).

10.7.3 Clothing potentially contaminated with microorganisms in spore form
must be autoclaved according to MB-01.

10.7.4 It is less harmful to clothing to autoclave it in a tray than it is to bag it.
Do not put water in the tray with the lab coat. Rather, put a second
tray into the autoclave and add water to this tray.

-------
SOP No. MB-13-02
Date Revised 01-24-11
Page 14 of 14

10.7.5	After clothing is decontaminated (by disinfection or autoclaving),
immerse it in water containing detergent to aid physical removal of
decontaminated biohazardous material.

10.7.6	Rinse lab coat and set aside to be sent out with the weekly lab coat
laundry service.

10.7.7	Take street clothing and footwear home and launder.

11.0 DATA ANALYSIS/CALCULATIONS: None

12.0 DATA MANAGEMENT/RECORDS MANAGEMENT:

12.1 The Branch Chief is responsible for documenting spills and accidents.

13.0 QUALITY CONTROL:

13.1 Appropriate quality control measures are integrated into each SOP.

14.0 NONCONFORMANCE AND CORRECTIVE ACTION:

14.1 Strict adherence to the biosafety practices is required. Nonconformance will

result in notification, retraining, or disciplinary action of laboratory employees.

15.0 REFERENCES:

15.1 Centers for Disease Control and Prevention and National Institutes of Health,

2007. Biosafety in Microbiological and Biomedical Laboratories, 5th edition. U.S.
Department of Health and Human Services. U.S. Government Printing Office,
Washington, D.C.

16.0 FORMS AND DATA SHEETS: None

-------