Adopted March 2006

PoIicy on Choice of Fumigant for Fumigation of MicrobioIogicaI

Safety Cabinets & Containment LeveI 3 Suites

1 Introduction
Until recently, the only accepted method for effective fumigation of a microbiological
Safety Cabinet [MSC] and Containment Level 3 facilities was to use formaldehyde.
However, formaldehyde poses a considerable risk to health [see below] and there is
an increasing impetus for finding a safer alternative. For that reason, the Safety
Office has been investigating an alternative method which involves the use of vapour
pressure hydrogen peroxide [VPHP]. The evaluation considered the following
aspects:

Safety
Effectiveness
Speed and convenience
Cost

The recommendations which follow are based on information from Bioquell, who
have given a presentation and demonstration of their system to Biological Safety
Officers , independent investigators at CAMR Porton and the advice of HSE.

2 Hazards associated with formaIdehyde

2.1 Toxic properties
Formaldehyde is a very hazardous substance. Among its adverse effects are the
following:
Causes burns. Very toxic by inhalation, ingestion and through skin absorption.
Readily absorbed through skin.. Mutagen. May cause damage to kidneys. May
cause allergic reactions. May cause sensitisation. May cause heritable genetic
damage. Lachrymator at levels from less than 20 ppm upwards. Very destructive of
mucous membranes and upper respiratory tract, eyes and skin. Because of this high
level of hazard, it is presently assigned a Work Exposure Limit [WEL] in air of 2 ppm
[2.5mg.m-3], with the presumption that ambient levels must be maintained as far
below that limit as is reasonably practicable. mportantly, formaldehyde was recently
reclassified by the nternational Agency for Research into Cancer [ARC} as a Class
1 carcinogen, and a cause of nasopharyngeal cancer. This reclassification has been
accepted by WHO and the HSE has plans to review the WEL for this substance in
the near future with a strong probability that it will classify it as a class 1 carcinogen.

t should be noted that some individuals are not able to detect its presence, even at
levels around or above the WEL and therefore may not be aware that they are being
exposed, unless an appropriate monitoring device is used.

Adopted March 2006
2.2 PhysicaI Properties

Formaldehyde is explosive at 7.75% in dry air. Above this explosive air-vapour
mixtures can be formed, unless the atmosphere is humid. Because it penetrates
poorly in very dry conditions, for both safety and efficacy the conditions should be
humid and warm [above 65% relative humidity and above 200C].
n order to achieve this, a proprietary fumigation kettle must be used to produce the
required vapour.

2.3 ChemicaI Interaction
Under certain conditions formaldehyde can react with hypochlorite and other chlorine
containing chemicals such as Chloros to form bis-(chloromethyl)-ether which is a
known lung carcinogen. Chlorine containing compounds must therefore be removed
from rooms and cabinets before fumigation.

3 Hazards associated with hydrogen peroxide [H
2
O
2
]
These are significantly less. The vapour is an irritant of the eyes, mucous
membranes and skin and may cause lung irritation if inhaled. Skin contact with liquid
H
2
O
2
can cause temporary bleaching of the skin or redness and blisters if not
washed away.


4 Comparison of formaIdehyde and VPHP incIuding the risks associated
with the two processes

These are outlined in a document produced by HSE , 'Fumigation operations in
microbiological containment laboratories. Guidance on the available technologies
and their application' [Appendix 1]. This is a technical supplement to ACDP's
guidance 'The management, design and operation of microbiological containment
laboratories'


4.1 Effectiveness
There is an historical belief that formaldehyde fumigation is always highly effective,
though it is not easy to find data from confirmatory tests or comparison with VPHP in
this regard [but see below]. Also, its effectiveness is greatly reduced if used at the
wrong ambient temperature or humidity. Temperature and relative humidity are
rarely controlled during routine fumigations.

VPHP has been reported in a number of studies to be active against a wide range of
organisms. The attached document, 'Hydrogen peroxide vapour biological efficacy'
gives details of the efficacy testing carried out by Bioquell.
http://www.bioquell.com/resources/BDS-3-BOEFFCACY-V5.0.pdf

Adopted March 2006
4.2 Independent study
The results of a recent independent study at Porton Down indicated that both
formaldehyde, when used under appropriate conditions, and VPHP possessed
effective disinfection capability against a wide range of biological agents, though
both had poor activity against prions.

The action of both disinfectants was affected by ambient temperature, with efficacy
reducing at low temperatures.

The action of VPHP was adversely affected by the presence of blood in the original
solution, presumably because the presence of proteins, etc protected the organisms.
Formaldehyde was also affected, but to a lesser extent.

VPHP efficacy appeared to be reduced by the presence of organisms which
produced catalase. This does not occur with formaldehyde.

The overall conclusions of this study for both formaldehyde and VPHP were:

[1 ] Efficacy of either gaseous decontaminant is dependant on the following
factors
Composition of suspending fluid
Microbial concentration
Presence of protective agents

[2] Pre-cleaning before gaseous disinfection is essential to reduce point loading
and dilute microbial contamination

Taken together, the evidence from this study indicates that the efficacy of VPHP is
at least equivalent to that of formaldehyde for the organisms tested, but the potential
for reduction in effectiveness due to the presence of catalase inhibitors or protective
organic material must be taken into account when planning the fumigation process.

4.3 HSE Attitude

HSE recognises the considerable safety issue with formaldehyde and now accept
that VPHP is an effective and much safer alternative for the fumigation of MSCs
providing it can be shown to be effective against the organisms in use.


4.4 Speed and convenience
There are currently two companies that supply equipment to perform this process.
Steris and Bioquell. Bioquell also offer an Equipment Bio-decontamination Service
[EBDS] for MSCs, incubators and other items that may require fumigation. This can
is carried out by their service engineers, and can be done as part of the routine
service of the MSC. n addition they also offer a Room Bio-decontamination Service
for the fumigation of CL 3 suites. This avoids the capital outlay to purchase the
equipment. Steris supply similar equipment to Bioquell but do not appear to offer any
decontamination service.

Adopted March 2006
The recommended contact time for formaldehyde is 4 t0 6 hours which has meant
that people tend to set this up to run overnight. The gas is then vented off the next
day until levels are well below the WEL this can take up to 2 hours. This is a high
risk operation and can only be carried out by highly trained technical staff. Because
of the danger of leaks areas laboratories often have to be closed during the
fumigation process.

Fumigation carried out by the Bioquell engineer will take less than 2 hours in total.
However given our policy of only fumigating cabinets when absolutely necessary, in
most circumstances, fumigation will not be a pre-requisite for service and testing.
Where the nature of the work in an area means that fumigation of several cabinets
is mandatory on cabinet can be being tested whilst another is fumigated. Staff need
not be excluded from the area so providing their movement does not affect the OPF
test the lab can stay in operation.

4.5 Cost
There are no capital costs or conversion costs for the MSCs if Schools use Bioquell
to carry out the fumigation and the subsequent service of the cabinet.
Recently one School obtained a quotation from Bioquell [ Appendix 2]. in taking out
a service contract with Bioquell which includes fumigation by VPHP. This might still
appear to be high compared to the very small cost of formaldehyde. However, the
far greater downtime and the need to involve University staff in the fumigation, rather
than it being done entirely by Bioquell mean that the relative costs may be even
higher for formaldehyde fumigation.


5 ConcIusions

(1) There is a substantially greater risk to safety from the use of formaldehyde
compared to that with VPHP.
(2) The effectiveness of VPHP is now established, although there are
considerations which need to be taken into account [see above]. Bioquell
have also offered to collaborate in testing efficacy for organisms where the
existing data is insufficient.
(3) The Bioquell method offers a much more rapid and convenient means of
fumigation
(4) Although an exact cost comparison is difficult to make, the higher unit cost
of each VPHP fumigation is at least substantially offset by the reduced
need for UoN staff time and down time of the equipment and laboratory.

t is clear that replacement of formaldehyde fumigation with VPHP is reasonably
practicable and offers a significant reduction in risk to Health.


6 Future PoIicy

VPHP fumigation of MSCs/ CL 3 suites at UoN should be introduced as the
method of choice as soon as possible. n the case of any organisms for which
there is not yet sufficient data on efficacy, validation tests will be required.

Adopted March 2006
The presumption will be against any future use of formaldehyde fumigation

Where Schools/Divisions choose not to adopt the use of VPHP as their
method of fumigation, this must be supported by an appropriate risk
assessment, signed by the HoS/HoD to show their endorsement of that
decision.












































Adopted March 2006
APPENDIX I
Fumigation operations in microbiological containment laboratories
Guidance on the available technologies and their application
[Guidance issued by HSE]

Introduction

This guidance is aimed at all those who have responsibility for assessing and
managing the risks from exposure to biological agents in a laboratory setting. ts
purpose is to provide information on the different decontamination technologies
available for those undertaking fumigation operations and focuses specifically on the
use of formaldehyde and hydrogen peroxide vapour although other gaseous
fumigation methods are briefly addressed.

n microbiological containment laboratories fumigation will often be performed to
decontaminate microbiological safety cabinets and less frequently entire
laboratories. The information provided in this guidance should be useful in
determining which technology is most suitable for the chosen application and
facilitate the production of a suitable and sufficient assessment of the risks. The
guidance is therefore, intended to help the employer comply with the Health and
Safety at Work etc Act (1974) and the Control of Substances Hazardous to Health
Regulations 2002 (as amended) (COSHH) and should be useful to safety advisors,
biological safety officers and other safety professionals who provide competent
advice to their employer.

Background

On occasions it will be necessary to decontaminate laboratories, animal containment
facilities and microbiological safety cabinets (MSCs) by fumigation when, for
example, there has been a spillage of infectious material or before servicing or
maintenance work is to be carried out.

Fumigation should always be a planned exercise with appropriate controls in place,
and with information and warnings provided for those that need to know. The
fumigation operation should only be carried out by named, trained personnel working
to an agreed plan and using a method that is known to be effective in the
circumstances of use.

Formaldehyde vapour has been known for many years as a highly effective biocidal
agent and is the fumigant most commonly used in laboratories. However,
methodology developed for use in pharmaceutical settings is now widely available
and is being marketed as a safe, environmentally friendly, alternative to
formaldehyde for decontamination of any enclosed area that may be contaminated
with microorganisms. n particular the use of automated vaporised hydrogen
peroxide systems have been actively promoted for use in hospital diagnostic
microbiology laboratories and research laboratories in both the academic and
commercial environment.

The ability to safely decontaminate microbiological containment laboratories and
MSCs is essential for their safe operation and the introduction of any new technology
Adopted March 2006
should be carefully controlled and monitored to ensure both the health and safety of
staff working in laboratories and others who may be affected by the work.

Given the relative novelty of these new technologies the majority of users will have
little experience of the methodology involved and may be unfamiliar with the specific
issues that relate to its application.

LegisIative controIs

COSHH requires that every employer shall ensure that the exposure of his
employees to substances hazardous to health, which includes disinfectants and
fumigants, is either prevented or where this is not reasonably practicable, adequately
controlled. The first requirement of COSHH is to prevent the exposure to substances
hazardous to health where it is reasonably practicable (i.e. the costs in reducing
exposure would not be grossly disproportionate to the benefits).

This can be achieved by:

1. Changing the process so that the substance is no longer used
2. Replacing it with a safer alternative; or
3. Completely enclosing the process

Changing the process is not feasible here and fumigation remains the simplest way
of dealing with accidental spillages of infectious material in laboratories where the
spill has occurred outside of the confines of a microbiological safety cabinet.

A safer alternative should only be considered following a suitable and sufficient
assessment of the risks. Where low hazard alternatives are available they should
always be considered as part of the risk assessment process. For example, whilst
glutaraldehyde is an effective disinfectant it has long been recognised as a cause of
ill health, with dermatitis and respiratory problems being the most significant effects.
Consequently the use of glutaraldehyde in the laboratory is no longer recommended
and lower hazard disinfectants are used in its place. Successful decontamination is
then reliant on the efficacy and effectiveness of the alternative substance chosen
and HSE inspectors as part of planned inspection visits may ask to see validation
documents for any agent used.

All fumigation operations are performed in totally enclosed spaces and COSHH
requires that at Containment Level 3 (CL3) the workplace is sealable to permit
disinfection.

Where exposure cannot be prevented it must be adequately controlled and to assist
in this process the Health and Safety Commission (HSC) has established
"Workplace Exposure Levels (WELs) for a number of substances hazardous to
health. These are intended to prevent excessive exposure to specified hazardous
substances by containing exposure below a set limit. WELs are concentrations of
hazardous substances in the air, averaged over a specified period of time referred to
as a time weighted average (TWA). Two time periods are used: long term (8 hours)
and short term (15 minutes). Short-term exposure limits (STELs) are set to prevent
Adopted March 2006
acute effects, such as eye irritation, which may occur after only a few minutes
exposure.

A WEL is therefore the maximum concentration of an airborne substance averaged
over a reference period, to which employees may be exposed by inhalation. t is
important to recognise that WELs should not be considered a hard and fast line
between safe and unsafe.

EH40/2005 Workplace exposure limits includes the list of substances assigned
WELs. t also provides more detailed guidance on the use of WELs and describes
eight principles of good practice for the control of exposure to substances hazardous
to health. The principles require the degree to which exposure is reduced below the
WEL to be proportionate to the health risk. f employers apply the principles of good
practice for the control of substances hazardous to health correctly, exposure should
be below any relevant WEL.

Both formaldehyde and hydrogen peroxide have been assigned WELs.
Formaldehyde currently has a workplace exposure limit of 2ppm (both 8 hour TWA
and short term), however, HSC/E plan to review the limit values for this substance.
Formaldehyde is toxic by inhalation or contact with skin. t can cause sensitisation by
skin contact and presents the risk of possible irreversible effects. Hydrogen peroxide
is set a long-term exposure level of 1ppm (8-hour TWA) and a short-term exposure
limit of 2ppm. Hydrogen peroxide like formaldehyde may cause burns but because
the hydrogen peroxide vapour readily breaks down into water and oxygen, the
process has none of the environmental concerns associated with formaldehyde.

n addition, all biocides are subject to the Biocidal Products Regulations 2001 (BPR).
The BPR aim to ensure all biocidal products, which include disinfectants, are safe
when used properly. Further information can be found in the HSE guidance: A guide
to the Biocidal Products Regulations for users of biocidal products (HSG 215).

Fumigation Options

1. Hydrogen Peroxide

Aqueous hydrogen peroxide has a long history of use as a disinfectant, which has
been facilitated by its rapid antimicrobial efficacy and its ability to decompose on
reaction to generate environmentally innocuous residues (water vapour and oxygen).

2H
2
O
2
2H
2
O + O
2
hydrogen peroxide

water + oxygen

Aqueous hydrogen peroxide is active against a wide range of organisms and owes
its broad spectrum activity to its powerful oxidising capacity which is known to
damage cellular proteins, lipids and nucleic acids.

However manual application of liquid products for sanitisation can be time
consuming, difficult to control and difficult to validate, especially for large surface
applications. Fogging applications are also difficult to control and reproduce and so
Adopted March 2006
over the past ten years there has been considerable development of vapourised
hydrogen peroxide and hydrogen peroxide gas generator systems.

These systems have been used widely for sterilisation of pharmaceutical
applications, including production filling lines, sterility testing environments, sealable
enclosures, production rooms and lyophilisers and more recently for the
decontamination of laboratory animal, research and biosafety laboratory facilities and
of note, for decontamination of buildings following deliberate contamination with
Bacillus anthracis.

Wet versus Dry

There are two schools of thought about gaseous hydrogen peroxide surface
decontamination. The traditional thinking is that it is a dry gas process and any
condensation should be avoided as it causes the process to become uncontrollable.
The other is that condensation is unavoidable given the usual operating parameters
and is the primary method of causing the decontamination.

As a consequence, hydrogen peroxide vapour systems may be classified as "wet or
"dry processes. Hydrogen peroxide vapour can be introduced into a given area up
to a certain concentration, dependant on the isolator temperature and humidity, to a
saturation level or dew point. f the concentration of hydrogen peroxide increases
above this level it will condense onto the surfaces of the isolator (condensation or
micro-condensation). n the case where micro-condensation is formed and
maintained during the cycle, this is considered a "wet process. f the vapour
concentration is maintained below the dew point during the cycle this is essentially a
"dry process. Although this description is useful in distinguishing between the two
different processes, the term "wet refers only to the fact that micro-condensation
has occurred and this may not be visible to the naked eye.

t is claimed that the antimicrobial efficacy, cycle characteristics, material
compatibility and safety aspects for both processes are distinct and should be
considered separate.

AvaiIabIe systems

Dry process:

Vapourised hydrogen peroxide (VHP) may only be used in sealed enclosures to
avoid uncontrolled leakage of hydrogen peroxide as well as the inflow of
unconditioned air. The gas should be uniformly distributed within the space in a
constant, minimum bactericidal and sporicidal concentration. A range of delivery and
control systems are available for decontamination of small, medium and large
enclosed areas. The VHP concentration is maintained below the condensation point
to prevent condensation of liquid peroxide on surfaces.


VHP is a dry process as the concentration in the enclosure is maintained below the
condensation point. The process can be carried out at temperatures ranging from 4
to 80
o
C.
Adopted March 2006

The cycle consists of four phases:

1. Dehumidification
2. Conditioning
3. Sterilisation
4. Aeration

The process parameters of the dehumidifying, conditioning, sterilisation and aeration
phases are determined on the basis of measurements of the air temperature and
humidity in the enclosure. During the dehumidification phase, the relative humidity
and temperature are adjusted to a range of values in which no condensation of the
VHP can occur.

During aeration VHP is no longer introduced and the residual vapour is catalytically
decomposed into water vapour and oxygen. The aeration phase is completed when
the content of hydrogen peroxide has fallen to < 1ppm.

A microprocessor control automatically monitors, controls and records the process
parameters during each cycle. Because application conditions vary, a cycle
development guide is used to develop biodecontamination cycles for different
applications. Spores of Bacillus stearothermophilus are generally used to verify and
validate these decontamination cycles and times will depend on factors such as VHP
concentration, and enclosure temperature and volume.

Wet Process:

Commercially available hydrogen peroxide gas generators are designed to provide
biological decontamination within enclosures and rooms, providing a layer of
hydrogen peroxide micro-condensation, deactivating microorganisms on all exposed
surfaces. The decontamination cycle consists of three phases:

1. Pre-conditioning
2. Gassing
3. Aeration

During the gassing phase liquid hydrogen peroxide is pumped onto a hot plate in an
air stream and is flash evaporated turning all the liquid into hydrogen peroxide
vapour. The hydrogen peroxide vapour is delivered to the chamber or enclosure at
an elevated temperature. As the flash evaporation process continues the
concentration of the vapour in the enclosure increases until the vapour dew point is
reached. At this point the vapour will start to condense onto all cooler surfaces and it
is the deposition of hydrogen peroxide condensate that facilitates the
decontamination.


When an appropriate contact time between all surfaces of the enclosure and the
vapour mixture has been reached, hydrogen peroxide is passed through a catalyst
and decomposed to water and oxygen. A drier removes water vapour produced in
Adopted March 2006
this process. This aeration phase continues until all of the peroxide has been
removed to a safe level.

The cycle is optimised to achieve the most rapid conditions for biodecontamination
whilst minimising the time required for vapour removal or catalytic conversion
(aeration) at the end of the cycle.

The suppliers claim that the microcondensation process does not damage materials
or equipment and that extensive test data is available to support this.

Efficacy and VaIidation

t is claimed the mode of action of vapourised hydrogen peroxide is distinct from
liquid hydrogen peroxide but regardless of the exact mode, there is increasing
evidence that hydrogen peroxide is a broad spectrum, rapid antimicrobial and this
broad spectrum efficacy has been shown against a wide range of microorganisms
over the last ten years.

However, as for any biocide, the efficacy of hydrogen peroxide can be affected by
the presence of both organic (e.g. proteins, lipids) and inorganic materials, which
may reduce the penetration and activity of the agent. This is certainly the case with
Mycobacterium species, which are considered to be more resistant than other
vegetative bacteria due to their unique lipophilic cell wall structure.

Bacterial spores, in addition, are generally considered to be the most resistant
organisms to disinfection and sterilisation processes and are used as an indicator
that successful decontamination has been achieved. However, there is evidence to
suggest that in certain circumstances organisms that produce catalase (a hydrogen
peroxide degrading enzyme) may be more resistant and this should be considered in
the risk assessment.

n the majority of studies, this technology has been used for fumigation of clean
environments for product protection purposes and results may be significantly
different in the presence of contaminating material. t is therefore very important that
the use of hydrogen peroxide should be evaluated relative to the type, scope and
source of contamination and should reflect worst-case conditions as defined by the
presence of the most resistant organism on the most resistant material.

As indicated, the activity of any biocide or physical decontamination process will also
vary depending upon the type of surface being decontaminated. Porous materials, in
particular, demonstrate poor penetration of hydrogen peroxide in comparison with
other materials and may require longer cycle times.

Risk assessment

t is important that this technology should only be adopted following a suitable and
sufficient assessment of the risks. More information on making a suitable and
sufficient risk assessment is available in the approved code of practice attached to
COSHH. However, clearly the risk assessment should address how this technology
will be applied.
Adopted March 2006

t is extremely important that these systems are tailored to the local environment and
the needs of the purchaser, taking into account the type, scope and source of the
contamination as well as the geographical layout and topography of any large areas
that will need decontaminating. ssues concerning the biological agents in use,
material compatibility and the nature of biological challenge testing will need to be
considered.

Access to expert advice is essential to enable the definition of the process
requirements and ensure that the equipment purchased achieves the prescribed
objectives.

Information, instruction and training

The provision of suitable and sufficient information, instruction and training is
essential if the technology is to be used effectively and a requirement under COSHH.
This provision should include basic information on cycle development and validation
to reinforce the technical and service support that is available from the manufacturer,
but include for users, operational information and instruction that will facilitate the
development of risk assessments and standard operating procedures.

Issues to consider

MicrobioIogicaI safety cabinets (MSCs)

MSCs are used to work with biological agents of various hazard groups and a risk
assessment is needed to determine how often the cabinets should be
decontaminated. The exposed internal surfaces of the cabinet working space may be
decontaminated with a liquid disinfectant appropriate to the agent being handled but
for gaseous fumigation, depending upon the system adopted, alterations to the MSC
may need to be undertaken to allow delivery of the fumigant. This will have a cost
implication dependant on the number of MSCs is use.



Containment IeveI 3 and room fumigation

CL3 laboratories are designed for work with Hazard Group 3 biological agents and
the requirements for safe working with these agents are stringent. Following a
significant spillage in a CL3 laboratory, outside of primary containment e.g. the
microbiological safety cabinet, it is recommended that, regardless of whether the
agent being handled presents a risk of aerosol transmission, the room should first be
cleared of infectious aerosol and then fumigated. Depending on the nature of the
spillage, contamination may be extensive and fumigation is recommended as it
facilitates safe cleaning of the laboratory. Further information on the action to take in
the event of a spillage can be found in appendix 3 of "The management, design and
operation of microbiological containment laboratories(HSE books).

n order to achieve controlled, reproducible area decontamination, use of VHP
requires the gas to be circulated to ensure uniform contact with all exposed surfaces.
Adopted March 2006
Depending upon the size, shape and geometry of the room, fans are often used to
facilitate this circulation. This is not a problem as far as routine fumigations are
concerned, e.g. prior to routine maintenance or servicing, because the room can be
entered prior to the fumigation operation to set up any fans indicated in validation
exercises. However, in an emergency situation following an accidental spillage of
infectious material, current guidance issued by the Advisory Committee on
Dangerous Pathogens (ACDP) indicates that the room should be evacuated and not
re-entered until fumigation has been completed.

t is very important that this situation is considered as part of the initial consultancy
with the supplier of the technology to ensure that the decontamination process in
these circumstances is properly validated. There is also value in considering these
issues whenever the opportunity of a new build is considered.

2. FormaIdehyde

Formaldehyde acts an an alkylating agent, inactivating microorganisms by reacting
with carboxyl, amino, hydroxyl and sulphydral groups of proteins as well as amino
groups of nucleic acid bases. For formaldehyde to act as a disinfectant it must
dissolve at adequate concentration, in a film of moisture in the immediate vicinity of
the organisms which are to be killed. The success of this process is well
demonstrated by the effectiveness of formaldehyde against serious human
pathogens such as Mycobacterium tuberculosis.

Formaldehyde gas has been used for many years as a bactericidal agent and has
long been used for fumigation purposes despite early evidence of undesirable effects
on the eyes and respiratory system. n recent years there have been concerns about
its general toxicity and potential as a carcinogen and this has led to reviews of its
use, particularly in procedures such as the disinfection of laboratory equipment and
rooms in which there is potential for exposure to high concentrations. However,
because formaldehyde fumigation is cheap and simple to use it remains the most
common routine fumigant used in UK microbiological containment laboratories both
for decontamination of microbiological safety cabinets and for gaseous
decontamination of CL3 laboratories.

Effective fumigation is typically achieved by heat-initiated vaporisation of a 40%
solution of formaldehyde vapour in water. The water is necessary to maintain a
humidity of about 70%, at which the gas has its maximum antimicrobial effect.
Fumigation is usually performed at 70-80
o
C to avoid the production of various
unwanted solid polymers, such as paraformaldehyde.

Formaldehyde fumigation has the advantage of being cheap; easy to handle; not
harmful to fabrics, paints or metals; and of being rapidly neutralised with ammonia.
ts pungent odour also gives adequate warning of its presence if any leakage should
occur.

The disadvantages of using formaldehyde, apart from the health hazards, outlined
later, are poor penetration, which restricts its use to surfaces and the persistence of
the polymer, which may lead to later release of formaldehyde gas over a period of
time. Formaldehyde vapour, like any other gas, penetrates only slowly into air
Adopted March 2006
spaces of porous materials so the desired effect is only achievable with long time
exposure to surfaces, and 12 hours is typically recommended. Formaldehyde may
also form a potentially explosive mixture with air at concentrations of more than 7%
but such mixtures are not encountered during fumigation.

n addition, contact of formaldehyde with hydrochloric acid and other chlorinated
disinfectants forms bis (chloromethyl) ether, which is carcinogenic. Since these
chemicals and disinfectants are widely used in laboratories they should be removed
before any planned fumigation operation.

Efficacy

n terms of overall efficacy and suitability for purpose, formaldehyde is 2-15 times
more effective in the killing of bacterial vegetative cells than it is for killing of bacterial
spores but this kill ratio is actually less marked than for other biocidal products.
Formaldehyde is therefore broadly effective, but for ideal fumigation, precise process
temperature and relative humidity controls must be maintained.

HeaIth Effects

The first signs or symptoms noticed on exposure to formaldehyde at concentrations
ranging from 0.1 to 5ppm are burning of the eyes, tearing (lacremation) and general
irritation of the upper respiratory passages. Higher exposures (10-20ppm) may
produce coughing, tightening in the chest, a sense of pressure in the head and
palpitation of the heart. Exposure at 50-100ppm and above can cause serious injury
such as collection of fluid on the lungs (pulmonary oedema), inflammation of the
lungs (pneumonitis) or death. Dermatitis due to formaldehyde solutions or
formaldehyde containing resins is also a well-recognised problem.

Emerging Issues

Carcinogenicity

Airborne formaldehyde has long been recognised as an irritant towards the eyes,
nose and respiratory tract. Human experience has shown that a single exposure to
formaldehyde can cause sensory irritation and at sufficiently high levels cause tissue
damage in these regions. Repeated inhalation exposure can produce marked,
chronic inflammation of the upper respiratory tract in experimental animals.
Formaldehyde is also a direct-acting mutagen in vitro and has produced genetic
damage at the initial site-of-contact in experimental animals.

These two properties of irritancy and genotoxicity raise concerns for carcinogenic
potential. A number of new epidemiology studies have been reported since 2000.
Some of these provide further evidence in relation to formaldehyde exposure and
cancer of the upper respiratory tract. n 2004, the nternational Agency for Research
on Cancer (ARC) reappraised its position on the carcinogenic potential of
formaldehyde, taking these new studies into consideration, and reached the
following conclusion in relation to nasopharyngeal cancer:

Adopted March 2006
"Overall, the Working Group concluded that the results of the study of
industrial workers in the USA, supported by the largely positive findings
from other studies, provided sufficient epidemiological evidence that
formaldehyde causes nasopharyngeal cancer in humans."

Given the importance of formaldehyde as a chemical and the significance and
consequences of pronouncing a substance as a "human carcinogen, the advice of
HSC`s Advisory committee on toxic substances, WATCH sub-committee (Working
Group on Action to Control Chemicals) was sought on this matter in January 2005.

After careful consideration of the available evidence WATCH published its
findings:

1) Formaldehyde has probably caused nasopharyngeal cancer in
humans;
2) n relation to the apparent association seen in some studies between
formaldehyde exposure and leukaemia, based on recent reviews of
evidence, and also considering biological plausibility, there is no basis
for any significant concern for this cancer;
3) t is probable that formaldehyde exposure has caused nasopharyngeal
cancer in humans, via a mechanism to which it can be predicted that
both chronic inflammation (provoked by irritancy) and genotoxicity
contributed.

Based on these conclusions further advice and guidance will be produced and
distributed by the HSE, however, during the meantime, it makes sense for
formaldehyde to be handled in the workplace as a potential occupational
carcinogen. Safe levels of exposure to carcinogens have not been
demonstrated, but the development of cancer will be reduced by decreasing
exposure. As far as fumigation operations are concerned therefore,
engineering controls and stringent work practices should be employed to
reduce potential exposure to the lowest feasible limit. All laboratories should
have an up-to-date protocol for dealing with microbiological accidents and
emergencies, and this should include precise information about the use and
hazards of formaldehyde. All members of the laboratory staff should be made
aware that formaldehyde presents a serious hazard to health.

Issues to consider

MechanicaI evacuation of fumigant

Where an appropriately sealed area with a dedicated ventilation system is to be
treated, e.g. a CL3 laboratory, fumigation with formaldehyde vapour is preceded by
sealing off the area and shutting off the ventilation system. The fumigant is then
immediately released and dispersed, usually by hot plate or purpose-designed
vaporiser. Typically, this is done using formalin (40% formaldehyde); with equivalent
to 100ml of formalin plus 900ml of water for every 28.3m3 of space to be treated
(Bruce, 1995). n some facilities this is achieved with manual ventilation controls,
usually switched off overnight while fumigation takes place. With more modern
ventilation systems a pre-set timer may be used to first evacuate the fumigant at a
Adopted March 2006
pre-determined time. The full ventilation system is then re-started automatically.
The principle of such procedures is to fumigate the area in the absence of any direct
worker contact with the fumigant, so minimising occupational exposure risk to as low
as reasonably practicable.

ChemicaI neutraIisation

Formaldehyde vapour can be neutralised with either liquid ammonia in an open
vessel, or with carefully controlled ammonia vapour generation following the
fumigation step. The use of liquid ammonia is likely to be most appropriate for low
volume cabinet-based fumigation, where 300ml of ammonia is used in an open
vessel for every 28.3m
3
of treated space or equivalent.

Combined mechanicaI / chemicaI fumigant removaI

For some facilities the room ventilation design may prevent the isolated release of
fumigant and its controlled removal as described above. Also, any Class
microbiological safety cabinets without external venting require sealing prior to
gaseous fumigation, and fumigant must then be removed before the cabinet is re-
opened. f an externally ducted cabinet, e.g. a Class cabinet, is installed close by,
then this can be used to evacuate fumigant from a non-vented Class cabinet via a
flexible duct link. Alternatively, an independent formaldehyde removal unit may be
appropriate for both the above situations, and this can either be placed within a room
to remove fumigant in large volumes, or can be linked via ducting to a
microbiological safety cabinet to remove contained fumigant. Typically, such
equipment has a high volume (vacuum) flow rate that draws contaminated air over a
carbon filter. Chemical residues become bound to the filter and are therefore
immobilised for safe removal later. Carbon filters are re-usable and may be used for
a number of decontamination procedures.

EnvironmentaI concerns

Formaldehyde does decompose in air to form a variety of by-products, including
formic acid, carbon dioxide, carbon monoxide and water. However, it is also a stable
compound and such decomposition is more likely to occur in the outdoor
environment, where UV activity levels are higher and other reactive molecules are
present, e.g. ozone. Within the indoor environment formaldehyde is likely to persist
for longer unless it is evacuated in some way, or else neutralised by chemical
means.

The Environment Agency aims to ensure that environmental exposures to
formaldehyde are too low to harm human health. mpacts of release to air are
minimised by formaldehyde degredation as it is rapidly removed from the
atmosphere by reaction with free radicals. However, formaldehyde is categorised by
the Environment Agency as a hazardous volatile compound (VOC). As a VOC it can
be involved in reactions with other air pollutants that form ground-level ozone, which
can cause damage to crops and materials as well as having potential effects on
human health. Further information can be obtained from the Environment Agency.

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3. Other gaseous fumigation methods

The following information provides a general summary of methods available
for gaseous chemical sterilization:

EthyIene oxide is an alky epoxide agent and in the US is the predominant
gaseous chemical sterilant because its key properties approach what could be
considered almost ideal. Chemical agents within this category are relatively
stable and tend to combine with other chemistries to form active third
compounds that are typically able to penetrate most polymeric materials; a
quality that is useful in the context of fumigation, where packaging may be
present. Biocidal activities are often exhibited when such compounds are
dissolved in water, and concentrations required for biocidal effect tend to be
quite high, in the order of 400-2,000 mg/L. Ethylene oxide is often the gas
used for comparison with other methods, and although early methods for its
use were regarded as slow (though effective), modification to its use, such as
treatment and segregation of more defined loads, have speeded up its use.
Many of the materials that become the target of fumigation, such as metal and
glass, do not adsorb ethylene oxide. As a result, little or no residue removal is
required when using this fumigant. Ethylene oxide can be used where
lumens are present or where permeable polymers are present. t is ideal for
thermo-labile devices as its application is performed at low temperatures.
EH40/2005 Workplace exposure limits describes ethylene oxide as capable of
causing cancer and/or heritable genetic damage. t is given an 8 hour, time
weighted average (TWA), workplace exposure limit of 5ppm. Ethylene oxide
has a flammability range from 3-100% by volume in air. t therefore has to be
supplied with inerting chemicals, or else in low volume cassettes for use in
dissemination systems that are designed to contain any explosive event.

PropyIene oxide (PO) possesses many of the sterilant properties described
for ethylene oxide, but requires a longer processing time. Gaseous PO is
used within the food industry and it has a narrower flammability range of
between 2-37%. t does not explosively decompose compared with ethylene
oxide. PO hydrolyses in to non-toxic by-products (unlike ethylene oxide), to
form biodegradable propylene glycols that are used for food additives and in
cosmetics. t is capable of causing cancer and/or heritable genetic damage
and is given an 8 hour, time weighted average (TWA), workplace exposure
limit of 5ppm.

Peracetic acid (PA) is an oxidising agent used mainly in liquid form, and is
also sporicidal in the vapour phase, with optimal activity at 80% rel. humidity
(RH). t has a slower decay rate in an open chamber than hydrogen peroxide
and should be expected to survive longer in a process, allowing deeper
penetration in to materials. Decomposition by-products include acetic acid,
hydrogen peroxide, water and oxygen, and it is not absorbed as easily as
hydrogen peroxide by cellulose-based materials. However, when treating
packaged items, specialist packaging can be used, but choice is still wider
than that routinely available for hydrogen peroxide vapour treatments. Low
pressure, low concentration cycles are typically used for PA applications, and
vapours must be generated at low temperature to avoid potentially violent
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decomposition of the high-energy molecule. There is some concern regarding
salts formed by the reaction of PA with hydrogen peroxide, as toxic metal
acetates or oxides may be generated. There is no mechanism to remove
such by-products during the gaseous processes, and if present they have to
be removed by water rinsing, which may compromise the microbiological
integrity of treated equipment.

ChIorine dioxide (ClO
2
) is another oxidising agent and, since it is known to
be sporicidal at low temperatures it has been investigated since the 1980s. t
is typically used at 25-30
o
C and at a concentration of 10-50mg/L at 80% RH.
Although better penetration and a generally good performance were expected
of this molecule compared with other oxidising agents, reports on its use have
not supported this. Like ethylene oxide, the greater degree of permeation of
the molecule into target materials requires an aeration period at treatment
completion, in order that residues are removed. Chlorine dioxide has 8 hour,
time weighted average (TWA), workplace exposure limit of 0.1ppm and a
short term exposure limit of 0.3 ppm (15-minute reference period. Applications
are limited because the molecule has a damaging effect on some materials,
e.g. uncoated aluminium foil, copper, carbon steel and polycarbonate. n view
of the above limitations, indoor applications within laboratory and hospital
working environments are restricted.

Ozone (O
3
) was recognised as an oxidising sterilant as early as 1899, though
further work found that its benefits were greatly limited by the presence of any
organic residues (soiling) on contact surfaces. Microorganisms can be
inhibited in broth, on agar and other solid surfaces when exposed to ozone,
and sporicidal activity has been reported. Effective applications have been
reported by use at between 2 and 10% by weight, and ozone has received
US-FDA approval for a limited number of food related applications. n order to
be effective, levels of ozone and RH must be closely monitored and controlled
at a pre-determined concentration for different temperatures. Because it can
corrode surfaces, the use of ozone is limited to those materials that will resist
such action, such as titanium, steel, ceramics and some polymers. Ozone is
known to be particularly effective against Listeria, Giardia, Escherichia coli
and Salmonella species.

4. Hydrogen peroxide pIasma appIications

The technology for plasma generation and its known inactivation of microorganisms
has been known since the 1950s, but commercial applications were not developed
until the early 1990s. Plasma is defined as a fourth state of matter that is
energetically distinguishable from solids, liquids and gases. t can be produced by
the action of very high temperatures or electric or magnetic fields, and is normally
composed of clouds of ions, electrons and neutral species.

The exact mechanism of HP plasma antimicrobial action is not known. However, an
electrical discharge can generate free radicals and other potentially biologically
active species from hydrogen peroxide. This is akin to the activity previously
described for hydrogen peroxide vapour. An added advantage of hydrogen peroxide
plasma technology over other chemical fumigants is that it leaves no residue and
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because it has a limited penetration of the target material it causes minimal material
damage with repeated applications.

Comparison of fumigation methods

Many of the available comparison studies have been conducted with gluteraldehyde,
rather than formaldehyde, and have addressed liquid surface disinfection rather than
gaseous fumigation. However, there is a clear overlap with issues of concern for
both gluteraldehyde and formaldehyde products, although delivery systems will differ
for fumigation, and gluteraldehyde is regarded as less toxic when used in some of its
newer product forms. Areas such as toxicity, residues, efficacy of treatment and
damage to equipment are commonly considered within these reports and are usually
relevant within the context of fumigation and liquid disinfection.


FormaIdehyde
Advantages Disadvantages
nexpensive Slow-acting, long exposure times
Easy-to-use Harmful residues
Claimed broad spectrum efficacy Extremely toxic, carcinogenic
Requires high humidity for efficacy
Not automated, difficult to validate
Surfaces need to be post-cleaned


VHP decontamination
Advantages Disadvantages
Broad-spectrum efficacy, including
sporicidal
Limitations with absorptive and
proteinaceous material
Automated Expensive
Validated, Reproducible Requires fans to circulate gas
Short cycle time Resistant organisms and catalase
producers may extend cycle times
Good safety profile Material compatibility
Non-toxic residues Surfaces ideally need to be
precleaned


Summary and key findings

A number of properties should be considered when undertaking gaseous fumigation,
with emphasis on the safety of the chemical product being used. The most desirable
of products would be effective in its application but non-toxic to the user, though in
reality most gaseous fumigants require some degree of containment in order to
remain effective and safe in use. An ideal fumigant should leave no residues, or else
should be capable of reduction to safe levels immediately after fumigation.

Compared to formaldehyde, hydrogen peroxide is a broad spectrum antimicrobial
with bactericidal, viricidal, fungicidal and sporicidal activity. n addition, because the
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vapour readily breaks down into water and oxygen there are none of the
environmental concerns associated with the use of formaldehyde.

Recent research has demonstrated that the efficacy of gaseous disinfection depends
upon: the resistance of the microorganism; the suspending fluid and loading; the
material the surface is made of and the presence of other inhibitors e.g. catalase for
VHP fumigation.

For vapourised hydrogen peroxide applications it is generally recommended that
precleaning is essential for effective fumigation but this is not always possible in
microbiological containment laboratories and this hazard needs to considered as part
of a formal risk assessment to protect workers or others who may be affected by the
work.

The key message in adopting any technology in this environment is to ensure that a
suitable and sufficient assessment of the risks has been completed and to ensure
the efficacy and effectiveness of the fumigant has been appropriately validated in
situ.

Further information and advice can be obtained from the HSE`s Biological Agents
Unit, Magdalen House, Stanley Precinct, Bootle, Merseyside L20 3QZ
Telephone: 0151 951 4000