Efficacy Testing of Biocidal Products Overview of Available Tests - Asw - Fbe - 080305 - Final PDF

October 2012
Efficacy testing of biocidal products

– overview of available tests
This report was produced by FB Engineering in 2008 at the request of the Swedish
Chemicals Agency.
Mall-id: MAG-0006, 2011-12-28
Kemikalieinspektionen
Postadress Besök Faktureringsadress Telefon & fax Internet Org nr

Box 2 Esplanaden 3A, FE 124 Telefon +46 8 519 41 100 www.kemi.se 202100-3880
172 13 Sundbyberg Sundbyberg 838 80 Hackås Fax +46 8 735 76 98 kemi@kemi.se
At the request of
SWEDISH CHEMICALS AGENCY (KEMIKALIEINSPEKTIONEN),
DEPARTMENT OF PESTICIDES AND BIOTECHNICAL PRODUCTS
EFFICACY TESTING
OF
BIOCIDAL PRODUCTS
- Overview of available tests
Göteborg 2008
Ann-Sofie Wernersson (asw@fbe.se)
FB Engineering AB
Skärgårdsgatan 1, Göteborg
Postal address: Box 12076, SE-402 41 Göteborg, Sweden
Phone: +46 31 775 1000
www.fbe.se
1 ACRONYMS AND ABBREVIATIONS
Acronym Full names Web page to organisations (if available)

AATCC American Association of Textile www.aatcc.org/
Chemists and Colors
AFNOR Association française de /ww.afnor.fr/
normalisation (NF standards)
AOAC Association of Official Analytical www.aoac.org/
Chemists
ASTM American Society of Testing and www.astm.org/)
Materials
ATCC American Type Culture Collection
BBA Federal Biological Research Centre www.bba.de
for Agriculture and Forestry
(Biologische Bundesanstalt für Land
- Und Forstwirtschaft
Bundesrepublik Deutschland)
BP Biocidal Product
BPD Biocidal Product Directive (referring
to 98/8/EG)
BSI British Standards Institute (BS www.bsi.org.uk/
standards)
CA Competent Authority
CEB Commission Des Essais www.afpp.net/commande/commissions/CEB.htm
Biologiques
CEFIC European Chemical Industry www.cefic.org
Council
CEN European Committee for www.cenorm.be
Standardisation
CEPE European council of paint, printing www.cepe.org
inks and artist’s colours industry
CSMA Chemical Specialties Manufactures www.csma.org
Association
CTBA Centre Technique du Bois et de www.ctba.fr
l’Ameublement, Bordeaux
EBPF European Biocidal Product Forum
EPA United States Environmental www.epa.gov
Protection Agency
EPPO European and Mediterranean Plant www.eppo.org
Protection Organization
ISO International Standards www.iso.org/iso/home.htm
Organisation
MAFF Ministry of Agriculture Fisheries and
Foods
MS Malaysian Standards http://msonline.sirim.my/msonline
OECD Organisation for Economic Co- www.oecd.org
operation and Development
OPPTS Office of Prevention, Pesticides and www.epa.gov/internet/oppts/
Toxic Substances, United States
Environmental Protection Agency
PT Product Type
SABS South African Bureau of Standards www.sabs.co.za
1
2 SUMMARY
Efficacy of a biocidal product can be defined as the power to produce an effect,

and is related to the intrinsic efficacy of the active substance/s of the product and
is influenced by the presence of non-active substances and the use pattern. In the
European Community, authorisation of biocidal products is regulated by the
Biocidal Products Directive (98/8/EC). Whereas active substances are assessed at
Community level, the actual products are subject to national authorisation.
Annex IIB of the directive requires likely information for the basis of a label claim
but there is at present no international consensus on specific data requirements
for different product types. For several product types there is no information in the
TNsG on Product Evaluation, on how the tests are performed and whether there
are standards available. In Sweden, information about efficacy was previously not
regarded as important for the approval decision-making in national product
applications. The aim of this project was therefore to provide the Swedish
Chemicals Agency with an overview and brief description of available efficacy test
methods (both standards and others) for all product types and information on how
the tests are evaluated and what efficacy criteria are applied.
Test references and, to some extent, test descriptions were found by searching for
available test procedures and contact persons on web pages, by e-mail and phone
correspondence with authorities, industry organisations, standardization
organizations, individual manufacturers and test/research institutes. The present
overview covers all product types except PT 16, 22 and 23 and includes 387 test
references, of which 341 are standard methods. For one third of the tests, the
scope, design, applicability and/or efficacy criteria is briefly summarised. In
addition, the test references of more than 250 standards have been checked to
make sure that the newest version available is included in the overview and to
exclude tests that have been withdrawn.
The numerous ways of testing and measuring efficacy makes comparisons

between products and the setting of performance standards difficult. Lack of
performance standards and specific data requirements could result in inconsistent
evaluations in different Member States. Therefore, authorities, standardisation
organisations and industry representatives need to develop pass/fail criteria, new
test standards, identify reference products and establish lists of recommended
tests for particular product applications. As a start, the lists of available tests in the
current version of the TNsG on Product Evaluation need to be updated.
2
3 SAMMANFATTNING
Effektiviteten hos biocidprodukter kan definieras som förmågan att åstadkomma en effekt
och beror av den inneboende effektiviteten hos den aktiva substansen i produkten. Den
påverkas dock även av andra substanser i produkten och användningsmönstret. Inom EG
regleras biocidprodukter av biociddirektivet (98/8/EC). Aktiva substanser bedöms på EG
nivå medan produkterna regleras nationellt.
I direktivets Annex IIB beskrivs de informationskrav som ligger till grund för den påstådda
effektiveteten hos produkten. Det råder dock för närvarande ingen internationell
samstämmighet när det gäller specifika datakrav för respektive produkttyp för att styrka
den påstådda effektiviteten. I ”TNsG on Product Evaluation” saknas information för flera
produkttyper när det gäller testdesign och huruvida det förekommer några standarder. I
Sverige ansågs tidigare att effektivitetsdata inte var viktiga för tillståndsprövningen av
biocidprodukter. Syftet med föreliggande rapport är därför att förse Kemikalieinspektionen
med en sammanställning och kort beskrivning (inklusive hur de utvärderas och vilka
effektivitetskriter som råder) av tillgängliga effektivitetstester (både standarder och övriga),
för samtliga produkttyper.
Test referenser och till viss del även testbeskrivningar erhölls genom att söka efter
testmetoder och kontaktpersoner via hemsidor, e-mail och telefonkontakt med
myndigheter, branschorganisationer, standardiseringsorganisationer, tillverkare,
kommersiella laboratorier och forskningsinstitut. Sammanställningen täcker in alla
produkttyper förutom PT 16, 22 och 23 och innehåller referenser till 387 effektivitetstester,
varav 341 är standarder. För en tredjedel av testerna summeras eller citeras syfte, design,
applicerbarhet och/eller effektivitetskriteria. Dessutom har mer än 250 testreferenser
kontrollerats för att säkerställa att den senaste versionen tas med i sammanställningen
och för att förhindra att tester som inte längre är giltiga är med.
Eftersom effektivitet kan undersökas och mätas på så många olika sätt blir det svårt att
jämföra effektiviteten hos olika produkter och att enas om krav på en viss prestanda hos
produkterna inom samma typ och användningsområde. I kombination med att det saknas
specifika testkrav finns det risk för en inkonsekvent bedömning i olika medlemsländer. Det
är därför nödvändigt att myndigheter, standardiseringsorganisationer och industrin
tillsammans sätter prestandakrav, utecklar nya standardtester, tar fram referensprodukter
samt arbetar fram listor på rekommenderade tester för specifika produkttyper och
användningsområden. En första början är att uppdatera de listor över tillgängliga tester
som finns i den nuvarande versionen av "TNsG on Product Evaluation".
3
TABLE OF CONTENTS
1 ACRONYMS AND ABBREVIATIONS ................................................................. 1
2 SUMMARY .......................................................................................................... 2
3 SAMMANFATTNING .......................................................................................... 3
4 BACKGROUND AND AIM OF THE PRESENT OVERVIEW ............................... 6
4.1 BACKGROUND .................................................................................................. 6
4.1.1 Biocidal Products Directive .............................................................................. 6
4.1.2 Data requirements related to efficacy .............................................................. 6
4.2 AIMS OF THIS OVERVIEW ................................................................................ 8
5 APPROACH ........................................................................................................ 9
6 INTRODUCTION ............................................................................................... 10
6.1 PERFORMANCE STANDARDS vs LABEL CLAIMS ....................................... 10
6.2 EXPERIMENTAL DESIGN OF EFFICACY TESTS ........................................... 10
6.3 STANDARDISED AND "IN HOUSE" PROCEDURES ...................................... 11
6.3.1 PT 1-5 ............................................................................................................... 11
6.3.2 PT 6: In-can preservatives .............................................................................. 12
6.3.3 PT 8: Wood preservatives ............................................................................... 13
6.3.4 PT 10: Masonry biocides ................................................................................. 14
6.3.5 PT 13: Metalworking fluid preservatives ........................................................ 14
6.3.6 PT 14: Rodenticides ........................................................................................ 14
6.3.7 PT 18: Insecticides, acaricides and products to control other

arthropods ....................................................................................................... 15
6.3.8 PT 21: Antifouling products ............................................................................ 15
7 RESULTS.......................................................................................................... 15
4
7.1 TEST REFERENCES FOUND IN THIS OVERVIEW ......................................... 15
7.1.1 International standardisation organisations – own searches ...................... 16
7.1.2 Tests used in other MSs – TM and national guidelines ................................ 17
7.1.3 Methods in use according to manufacturers and industry

organisations ................................................................................................... 18
7.1.4 Test endpoints ................................................................................................. 19
7.2 LIMITATIONS AND KNOWLEDGE GAPS ........................................................ 19
8 DISCUSSION .................................................................................................... 20
8.1 EVALUATING EFFICACY – COMPARING TEST RESULTS TO

LABEL CLAIMS AND PERFORMANCE STANDARDS ................................... 20
9 CONCLUSIONS AND SUGGESTIONS............................................................. 22
10 APPENDIX I. TEST PROCEDURES (STANDARDS AND OTHER

REFERENCES). ................................................................................................ 23
5
4 BACKGROUND AND AIM OF THE PRESENT OVERVIEW
4.1 BACKGROUND
4.1.1 Biocidal Products Directive
In the European Community, authorisation of Biocidal Products (BP) is regulated

by the Biocidal Products Directive (BPD) 1. In regulating the use of BPs, the
benefits are to be balanced against the risks. The authorisation of a BP should
only be granted if that product is shown to be sufficiently effective (article 5(1)(b) of
BPD) without posing an unacceptable risk to the environment. There should also
be no other unacceptable effects (such as development of resistance).
Efficacy can be defined as the power to produce an effect, and is described in

Annex IIB and VI of the BPD as the ability to fulfil label claims. The efficacy of a
BP determines the lower concentration or dose limit that should be used. Efficacy
is not only related to the intrinsic efficacy of the active substance/s of the product
but also affected by non-active substances (such as surfactants and solvents)
included in the product. The efficacy of a product is also related to the use pattern
of the product. Knowledge about how BPs are used is therefore essential in the
assessment of the product.
Active substances used in BPs placed on the market are assessed at Community
level and must first be listed on Annex I, IA or IB of the BPD. In order to include an
active substance on Annex I, IA or IB, the notifier needs to demonstrate that the
active substance is effective in at least one application.
Products are subject to national authorisation (BPD Annex VI, articles 51, 52, 92,
93). Label claims are a central issue in the assessment of efficacy. The applicant
needs to submit data on the product to substantiate the label claim and intended
uses, and efficacy claims must be presented on the product label and in product
information. Efficacy claims that could cause the product to be used in an
inappropriate way should be avoided, in order to minimize biocide use and
unwanted side effects.
4.1.2 Data requirements related to efficacy
Data requirements are listed in Annex IIA, IIB, IIIA, IIIB as well as IVA and IVB of
the BPD. Product related data is found in part B of these Annexes, whereas data
1
BPD: Directive 98/8/EC of the European parliament and of the council of 16 February 1998
concerning the placing of biocidal products on the market.
6
on the active substance is found in part A. The data and information required,
relevant to the effectiveness of the BP is found in Annex IIB and the corresponding
section 5 of the Technical Notes for Guidance (TNsG) on Data Requirements 2.
However, there is no international consensus on what data are required to prove

the efficacy of a BP. In the TNsG on Product Evaluation 3, it is stated that there are
no existing guidelines on how to test the efficacy of BPs and no international
agreements on label claims regarding supportive data or quality assurance
aspects of how such data are produced.
4
In an OECD survey from 1999, some questions on how authorities regarded
data on the efficacy of BPs were included. At the time of the survey, not all
countries (including Sweden) had an approval and/or notification process for all
5
Product Types (PTs) , which means that there were also no efficacy data
requirements for these PTs. Also, not all countries regarded efficacy data as
important for the approval process. Ireland did not require any efficacy data at
all, and Sweden did not consider them important for approval decision-making
since they believed that the market will remove products that are not effective
enough.
On the other hand, among the European countries in the survey, Belgium,
Denmark, France, Germany, Greece, Hungary, Netherlands, Portugal,
Switzerland and the UK considered ”insufficient efficacy” as a cause for not
authorising a specific biocide product, at least for some product types. In
Belgium, tests simulating practical use conditions were required for biocides for
non agricultural use. For use in agriculture, efficacy data were needed for
disinfectants for veterinary use and for products against ecto-parasites. In
Denmark, disinfectants were validated on data from a European suspension
test or similar. Nordic standards were used to assess wood preservatives and
paper industry was evaluating effectiveness of slimicides for pulp production. In
Germany, efficacy data were the most important data for approval of vector
control products (insecticides, acaricides and rodenticides in the field of
hygiene) and the actual testing was performed by the Robert-Koch Institute
(disinfectants), and by the Federal Environmental Agency. The testing included
laboratory and field tests, and was based on vector extermination. Although
Finland did not consider ”insufficient efficacy” as a cause for not authorising a
specific biocide product, wood impregnation chemicals were tested and
assessed by the Nordic Wood Preservation Council (non obligatory inspection)
and the Agricultural Research Centre tested the efficacy of all insecticides,
rodenticides and repellents.
2
Technical Guidance Document in support of the directive 98/8/ec concerning the placing of
biocidal products on the market - guidance on data requirements for active substances and biocidal
products
3
TNsG in support of Annex VI of directive 98/8/EC of the European Parliament and the Council
concerning the placing of biocidal products on the market. Common principles and practical
procedures for the authorisation and registration of products. Ver 10.0, July 2002.
4
OECD Environmental Health and Safety Publications, Series on Pesticides No 9. “Report of the
Survey of OECD Member Countries’ Approaches to the Regulation of Biocides. Paris 1999.
ENV/JM/MONO(99)11. This report (and its annexes) can be found at
http://www.oecd.org/document/18/0,3343,en_2649_32159259_32480722_1_1_1_1,00.html
5
The 23 different Product Types (PTs) are described in Annex V of BPD.
7
In chapter 7 of the TNsG on Product Evaluation, there is some guidance on how to
proceed. It focuses particularly on information that is needed for stating label
claims, study robustness and report details, Quality Assurance procedures,
evaluation of data and decision making. Any efficacy claims will depend on e.g.
the PT use patterns and desired effects (including target organisms). If the product
has a broad label claim, suitable principal organisms should be identified as the
target organisms.
Annex IIB of the BPD requires likely information for the basis of a label claim. This
includes PT, spectrum of biological activity [target organisms and their
development stage and function as well as mode of action (used also to evaluate
potential and existing pest resistance)], area of use/site of application,
geographical variability, duration of control/effect, directions for use, other relevant
information (including target dose rate, variability and application method), efficacy
or inefficacy of product under certain conditions (nature of infestation, density of
microorganisms, application temperature).
4.2 AIMS OF THIS OVERVIEW
The guidance provided in the TNsG on Product Evaluation is not in the form of a
checklist and there are no rigid criteria to evaluate efficacy data. Expert scientific
judgement is therefore necessary on a case by case basis. As stated in section
7.1.2. of the TNsG on Product Evaluation, the lack of harmonisation of efficacy
requirements can result in uncertainty, confusion, inconsistency and
misunderstandings regarding the extent of efficacy data required by the regulatory
authorities.
The TNsG on Product Evaluation includes references to available standard tests

for Product Type (PT) 1-6, 8, 10, 13-15, 18 and 21. Nevertheless, for several PTs
there is no information on how the tests are performed and whether there are
standards available (PT 7, 9, 11, 12, 16, 17, 19, 20, 22, 23).
In Sweden, information about efficacy was not regarded as important for the
approval decision-making in national BP applications (dossiers). Moreover, some
of the PTs that are now regulated by the BPD were previously exempted from
regulation (including food and feed area disinfectants, in-can preservatives and
metal working fluid preservatives) and some regulated PTs were not subject to an
authorisation process (human and veterinary hygiene biocide products,
preservatives for food and feedstocks, embalming and taxidermist fluids and
biocides for “other vertebrates”).
8
In order to be better prepared for evaluating the efficacy data of BPs according to
the BPD, the aims of this project were to provide the Swedish Chemicals Agency
(Kemikalieinspektionen, KemI), i.e. CA Sweden with
• an overview and brief description of available efficacy test methods
(standards and "in house") for BPs, including all PTs
• information on how the tests are evaluated and what efficacy criteria are
applied.
Knowledge gaps should also be identified.
5 APPROACH
The present overview includes references to both standards and non standard ("in
house") tests submitted by individual companies. Test references and, to some
extent, test descriptions were found by searching for available test procedures and
contact persons on web pages, e-mail and phone correspondence with authorities,
industry organisations, standardization organizations, individual manufacturers
and test institutes. Some material (including protocols from a Technical Meeting in
2005 at ECB) were also provided by KemI (CA Sweden).
Much effort was made to contact individual persons in companies and test
institutes, either by e-mail and/or phone, in order to include also test references to
"in house" tests and information on what standards that are actually used by
individual companies.
The web pages of the following standardisation organisations were searched for
efficacy tests currently available: ASTM, CEN, EPPO, ISO and OECD. Search
phrases included "efficacy", "antiseptics", "disinfectants", "antifouling",
"bactericides", "preservatives". It was also valuable to search standards through
the Technical Committees and Working Groups, such as E35 for Pesticides under
ASTM.
In order to obtain a starting point for available efficacy tests, the TNsG on Product
Evaluation and national guidelines (UK and NL) were also consulted.
The project was performed during the period of November-December in 2007.
9
6 INTRODUCTION
6.1 PERFORMANCE STANDARDS vs LABEL CLAIMS
Performance standards refer to a predetermined efficacy required by the

authorities and should, according to the TNsG on Product Evaluation, be the same
within all Member States for a particular use and situation. The performance
standard ("pass/fail criteria") can be expressed in quantitative or qualitative 6 terms.
Label claims are made by the manufacturer and written on the product label. It
makes a claim about the function of the product, such as pest organisms that it
controls, duration of control, type of control (kill, repel etc), and directions for use
(including method and application rate).
6.2 EXPERIMENTAL DESIGN OF EFFICACY TESTS
The experimental design of efficacy tests can be divided into three different types:
screening tests, simulation tests and field tests.
Screening tests are usually laboratory studies of either the active substance or
simple formulations and often performed during a relatively short period of time.
Studies may include dose-response tests and several replicates. Laboratory data
are usually not sufficient to predict actual treatment levels or effectiveness in
service under real situations, but can be used to estimate the innate efficacy of the
product. An untreated control should be included.
Simulation tests are performed in artificial environments that resemble the real
conditions during use and often the actual product is used in the test. An untreated
control should be included.
Field studies generate data based on actual use of the product as prescribed by
the product label. Even if the exposure conditions are more realistic than the
screening tests it should be kept in mind that they are still only representing a
particular situation and efficacy could be influenced if changing e.g. the level of
pest pressure or application technology. In addition, even if time scales are
generally longer, it is frequently not possible to perform the test during the full life
cycle of the product. PT 8 tests last e.g. 5 years but the service life might be 60
years.
6
including e.g. percentage kill, extent of remaining population, greatest dilution of product still
producing the desired effect etc
10
The product should preferably be tested at a variety of application rates, including
levels below those suggested in a real use situation. The last point ensures that
the biocidal product dose will be as low as possible.
As in any laboratory testing of chemicals, and also stated in the TNsG on Product
Evaluation, it is appropriate to include a negative (untreated) and/or positive (a
product/test material with the same use pattern and with proven efficacy, often
called a reference product) control. The negative control provides important
information to estimate the extent of the problem without any treatment. However,
for field tests, there are often limited possibilities to include replicates in the study
and to include untreated areas as a control. The latter aspect is sometimes
approached by including pre- and posttreatment assessments. Standard BPs, to
be used as positive controls in the efficacy tests, have been developed for e.g. PT
18 and 8 and can facilitate the ranking of products but also help to control the
reproducibility of a test and facilitates comparisons with other test facilities.
6.3 STANDARDISED AND "IN HOUSE" PROCEDURES
As stated in BPD Annex VI, efficacy testing should be carried out according to
Community guidelines if available and applicable, but other methods may also be
used, including other international standards (e g CEN, ISO), national standards,
industry standards, individual producer standards or data from product
development, as long as these are accepted by the Competent Authority (CA).
The availability of international standards varies between PTs. In the TNsG on

Product Evaluation, there are several standards listed for PT [1-5] 7, [8] 8, [14] 9 and
[18] 10; and a few for PT [6] 11, [10] 12, [13] 13, [15] 14 and [21] 15. For PT 1-5, 6, 8, 10,
13, 14, 18 and 21 some of the information and guidance provided in the TNsG on
Product Evaluation is summarised below. For further details, the reader should
consult the original reference.
6.3.1 PT 1-5
• Typical use patterns: health care area for hard surfaces (including medical
equipment), public area for reduction of nuisance, pathogens or algae to
acceptable level, for veterinary or other animal accommodation areas to
prevent/control outbreak of diseases, as bacteriostats against
7
from CEN, AOAC/EPA, ASTM and MAFF
8
all from CEN; in particular, EN599 describes pass/fail criteria
9
from EPA/OPP, BBA, EPPO and ASTM
10
from a number of sources, including BS, US CSMA Aerosol guide, AFNOR, WHO, South African
Bureau of standards, EPPO, ASTM and AATCC
11
from AFNOR, DIN, SABS, ASTM
12
from BS and ENV
13
from ASTM
14
from ASTM and EPA
15
from CEPE and ASTM
11
microorganisms causing aesthetic problems (odour) in presence of
moisture.
• The EN approach is to divide the tests into phase 1, phase 2 and phase 3
tests, corresponding to testing of innate activity, simulated use and "field"
testing (under practical conditions) respectively. Phase 2 tests are
subdivided into 2 steps (suspension tests and others):
o Phase 1: Suspension tests to test innate bactericidal, fungicidal
and/or sporicidal activity; defining minimum standards.
o Phase 2, step 1: Suspension tests, simulating the conditions during
practical use
o Phase 2, step 2: Other tests (handwash, handrub, surface test),
simulating the conditions during practical use
o Phase 3: Field tests (practical conditions)
• Broad label claims for PT 2, if intended to be used as a disinfectant in public
hygiene, should indicate efficacy against Gram negative and Gram positive
bacteria. Furthermore, if intended to destroy tuberculosis bacteria, claims
against Mycobacterium tuberculosis will need to be substantiated.
• Influencing factors to consider during efficacy testing of disinfectant
biocides include water hardness, interfering substances, contaminants,
temperature, contact time, pH and the roughness of the surface.
6.3.2 PT 6: In-can preservatives

• Spoilage of in-can and in-tank products may be the result of
microorganism’s (primarily bacteria 16 but also yeast 17 and moulds18)
feeding on substances present in the product, leading to reduced integrity.
By-products of microbial growth can also contribute to spoilage and
consequences include discoloration, gassing, viscocity loss etc.
• Laboratory tests are performed either as MIC (Minimal Inhibitory
Concentration), challenge testing or heat stability testing. The usual method
for evaluating in-can preservatives in paints or other aqueous products is
the challenge test.
o MIC determinations are conducted on a dilution series of the active
substance. The minimum amount of biocide that is required to inhibit
microbial growth is identified and efficacy is assessed against a
range of bacterial, fungal and yeast spoilage organisms. Results are
presented as the concentration required to inhibit the growth of a
particular test organism and are useful for determining the spectrum
of activity of an active substance.
o In the challenge test, microbial cells are added to the test sample
and the survival or death rate of these cells is monitored with respect
to time.
16
Common species are Alcagenes spp, Micrococcus luteus, E. coli, Proteus vulgaris.
17
Common species are Aspergillus spp, Geotrichium candidum, Penicillum spp.
18
Common species are Candida albicans, Rhodotorula rubra, Saccharomyces cerevisiae
12
o The level of active substance is usually measured by a suitable
chromatographic method at time zero and after incubation at an
elevated temperature to determine whether the biocide has
degraded. Incubation time and temperature varies between tests and
the results indicate the stability of the active substance in a particular
product formulation.
• US EPA assessment criteria require that the active substances should
show effectiveness against bacteria in at least two representative
formulations in which the biocide is intended for use and tested (simulated-
use) in at least three replicates of each of the two product formulations.
Actual bacterial isolates (identified at least to genus) from spoiled product
and/or ATCC (American Type Culture Collection) spoilage bacteria should
be employed as test inocula. Mixed bacterial and fungal inocula are not
acceptable in demonstrating bacterial deterioration. Bacteriological
sampling should be quantitative and product quality observed concurrently.
Test duration: 6 m to 1 yr. Conditions in the negative controls should cause
significant growth and deteriorative (physical and chemical) changes.
6.3.3 PT 8: Wood preservatives

• Durability of treated products will rely on the residue in wood which remains
active for the claimed service life. Efficacy will to a large extent depend on
climate (extremely variable situations exist within the European
Community), target organisms, wood species, application process19 and
exposure class (see below).
• Label claims should include a statement whether the product is aimed at
fungal, insect or attack from marine borers, and whether it is a preventive or
remedial (curative, eradicant)20 product. The test data required for remedial
claims may need to consider both preventive and eradicant action.
• The data required will depend on the Use/Hazard classes (see e.g. EN 335-
1, -2, -3), based on increasing level of wetting degree and exposure
severity: Hazard class 1: Above ground (Dry) exposure; class 2: Above
ground (wetting, protected from the weather); class 3: Above ground
(Exposed to weathering, but not in ground contact); class 4: Timbers in
contact with the ground or fresh water, or above ground if water trapping or
logging exist. Hazard class 5: Timbers in the marine environment 21.
19
There are three types of application methods: Penetrating treatments (including “double
vacuum”, “vacuum-pressure” and “diffusion”), surface treatments (including brush and spray
techniques and “dipping” processes) and other treatment methods.
20
The minimum performance requirements for eradicant BPs are given in prEN 14128:2001.
Remedial (in situ) treatment systems will be more varied than preventive treatments, depending on
e.g. a variety of likely treatment methods.
21
Data required to support claims for preventive efficacy, for Hazard class 1: Laboratory test on
wood boring insects (no fungal hazard); for class 2: Laboratory test on Wood rotting
basidiomycetes - brown rot; for class 3: Laboratory test on wood rotting basidiomycetes - brown rot
and in some cases white rot, and an optional field L-joint test (5 year); for class 4: Laboratory test
on Wood rotting basidiomycetes - white rot and brown rot, Laboratory test on soft rot microfungi,
and an optional field Stake test (5 year); for class 5: Laboratory test on wood rotting
basidiomycetes - white rot and brown rot, lab test on soft rot microfungi, lab test on Marine borers,
and a compulsory field Marine test (5 year).
13
6.3.4 PT 10: Masonry biocides
• Effectiveness (often described as kill or prevention of regrowth) depends on
substrate, target organisms (including dry rot fungus, fungus/yeasts,
lichens, mosses and liverworts), penetration, concentration used etc.
• In laboratory tests for general surface BP use, different concentrations of
the active substance is absorbed on assay discs applied to agar that is
seeded with fungal or algae. The zone of growth inhibition is determined
after incubation. There are also simulated use and field tests.
• To test products intended for the control of dry rot, simulated use tests are
based on employing mortar treated with the product and then exposed to
challenge by Serpula lacrymans. Growth over or through the treated
substrate is evaluated. Outbreaks of dry rot are unique and to control growth it is
neccary to implement a number of measures. It is therefore difficult to include
"untreated controls" and to design field tests.
6.3.5 PT 13: Metalworking fluid preservatives

• Simple laboratory tests generally determine MIC (Minimum Inhibitory
Concentrations). Many plant variables (such as machine characteristics,
metal, and the present microorganisms - both bacteria and fungi and yeast)
can influence overall preservative efficacy. Field use levels are therefore
often difficult to predict based on laboratory tests.
• In simulated use tests the fluid and preservative is generally placed in a
vessel and tested in a shaker, aeration or pump system, regulated in a
manner that simulates the plant conditions. Microbial inoculation is
performed repeatedly and the fluid is monitored regarding visual
appearance, pH, oxygen uptake and the presence/survival of
microorganisms.
• Field (in-use) tests will only be applicable to the particular conditions during
the test and it may be difficult to run concurrent negative controls.
6.3.6 PT 14: Rodenticides

• Rodent species can cause montory loss (through consumption or spoilage
of foodstuff or damage due to gnawing and burrowing) and spread disease.
Major target organisms include rats and house mice. Selected test species
should be relevant to the geographic region and specified on the label.
• Laboratory test methods for products include “no choice” tests (test
organisms are only offered contaminated food) and “choice” tests (both
contaminated and uncontaminated food is offered) as well as studies
related to specific product types (contact rodenticides or gassing agents) or
related to specific efficacy claims (for use in damp situations).
14
• In field trials /under seminatural conditions/ rodent activity on the site should
be determined before and after treatments and amount of bait consumed
should be monitored.
6.3.7 PT 18: Insecticides, acaricides and products to control other arthropods

• The standards can be used for testing efficacy against flying, crawling,
larval insects and also to test fumigants.
• Product applications include spray, textiles and fumigation. The effects
include knockdown, kill, residual22, flushing, ovicidal/larvicidal and the
control of resistant pests. Treatment methods include general surface
treatment, crack/crevice treatment, contact (direct) space spray, spot
treatment and baits.
• The tests are performed either as lab/screening tests, simulated use tests
or field tests.
6.3.8 PT 21: Antifouling products
• Although there are a few standards related to antifouling products, these

are usually related to leaching rate rather than efficacy. The two standards
included produce simulated field data (raft testing) during which panels
coated with the test formulation are immersed in water under static
conditions. Other test principles could be in vitro (lab test) and field (patch)
test.
7 RESULTS
7.1 TEST REFERENCES FOUND IN THIS OVERVIEW
National and international standards, in house tests and other test references are
included in the list presented in Appendix I, comprising in total 387 test references.
Of these, 341 are standards and 46 are "in house" and other non standardised
procedures developed either by the manufacturers or external test institutes and
research laboratories.
The standard test references already listed in TNsG of Product Evaluation are all
included in the list, except tests that are not considered "efficacy tests" (see below)
and tests that have since been withdrawn. In addition, web pages of some of the
standardisation organisations were searched for the latest versions and currently
22
Residual effects are due to active substances that remain active for a long time (weeks to
months) in order to control e.g. walking insects (that are unlikely to react to space sprays).
Treatments are applied at high dosages to surfaces or as palatable baits.
15
available tests (see next section). Manufacturers, industry organisations, other
MSs and test institutes and laboratories were also contacted regarding tests that
they use or know are being used.
About 90 manufacturers, test institutes and industry organisations were contacted.

Out of these, 37 responded. In some cases there was a high interest in the project
but due to the limited timeline of the project and/or other restrictions some
companies could not contribute with specific test references to particular products
to be included in the overview. In addition, some of the contacted companies did
not manufacture and/or test /the efficacy of/ BPs. Nevertheless, 14 companies
contributed with a total of 130 test references (including both standards and other
test methods) to the overview, and one manufacturer will send references
separately at a later stage.
7.1.1 International standardisation organisations – own searches
The web pages of the following standardisation organisations were also searched
for efficacy tests currently available: ASTM, CEN, EPPO, ISO and OECD. The
results are included in Appendix I.
Some additional CEN standards only indirectly related to actual efficacy testing
include (but is probably not limited to) EN 1014 part 1-4 (methods of sampling and
analysis of creosote), EN212:2003 (General guidance on sampling and
preparation for analysis of wood preservatives and treated timber), EN 335-1 and -
2 s well as EN 599-1 and -2 and EN 14128:2003 (related to use and hazard
classes, performance criteria and labelling regarding durability of wood and wood-
based products). These tests are not included in Appendix I.
EPPO is primarily focused on Plant Protection Products, although some standards

can also be applicable to some biocidal product types, such as rodenticides (PT
14) and fumigants (PT 18). Except for the standards included in Appendix I, they
have also published a “Good Plant Protection Practice” for rodent control for crop
protections and on farms 23.
ASTM has several standards applicable to microorganisms and wood

preservatives but also other PTs. Most ASTM standards have not been ring
tested, except some of the standards that correspond to USEPA methods.
Working groups searched are E35 (Pesticides, including subcommittees 35.12
Insect control agents, 35.22 Pesticide formulations and delivery systems, 35.15
Antimicrobial agents), E47.01 (Aquatic Assessment and Toxicology), D31.02
(Leather, subcommittee wet blue), D01.28 (Biodeterioration) and D01.45 (Marine
Coatings), both subcommittees under D01 (Paint and related coatings, materials
23
PP 2/5(1) Rodent control for crop protection and on farms. OEPP/EPPO Bulletin 25, 709-736. It
was edited as an EPPO standard in 1998 and can be found at
http://archives.eppo.org/EPPOStandards/gpp.htm
16
and applications), G03.04 (Weathering and durability, biological deterioration),
D14.30 (Wood adhesives, subcommittee under D13 Adhesives).
Some additional ASTM standards only indirectly related to actual efficacy testing
include (but is not limited to): ASTM E1326-06 (Standard guide for evalutating
nonconventional microbiological tests used for enumerating bacteria), ASTM
D5259-92(2006) (Standard Test Method for Isolation and Enumeration of
Enterococci from Water by the Membrane Filter Procedure), ASTM E1427-00e1
(Standard Guide for Selecting Test Methods to Determine the Effectiveness of
Antimicrobial Agents and Other Chemicals for the Prevention, Inactivation and
Removal of Biofilm), D283-84(1990)e1 and D284-88(1993)e1 (related to chemical
analysis of active substances in antifouling products) and D5108-90 (Standard
Test Method for Organitin Release Rates of Antifouling Coating Systems in Sea
Water). These are not included in the list.
ISO standards have been ring tested but there are only a few available published
efficacy standards, included in Appendix I.
As opposed to e.g. fate and ecotoxicity tests, efficacy testing is covered only to a
little extent by OECD guidelines. There are only a few applicable OECD guidelines
published, although there are some drafts, see Appendix I. Five new test methods
to evaluate efficacy of antimicrobials on hard surfaces will be investigated in ring
trial. For the particular case of testing piscicides (PT17), it is possible to use test
guidelines that were developed for aquatic toxicity testing.
The US EPA Office of Prevention, Pesticides and Toxic Substances (OPPTS) has
published updated test guidelines in ten series, of which 810 is related to product
performance 24. Nine guidelines related to Product Performance were finalized and
published on the web page at the time of the present project and included in the
overview.
7.1.2 Tests used in other MSs – TM and national guidelines
In the second Technical Meeting at ECB in 2005 (TMII05), the MSs were asked to
submit references to any efficacy tests that they knew were being used within their
territory, based on a list provided by COM. CA France provided a detailed
response and test references on their list have been included in Appendix I, if also
found on the homepages of the standardisation organisations. In the German list,
some standards that are not actually efficacy tests were indicated and therefore
erased from Appendix I.
In the UK, detailed national guidance documents can be found at the Health and
Safety Executive (HSE) website 25 for wood preservatives, rodenticides, anti-fouling
products, surface biocides, as well as insecticides and acaricides. These
24
http://www.epa.gov/opptsfrs/home/guidelin.htm
25 http://www.hse.gov.uk/pesticides/application/index.htm
17
guidelines outline the nature and extent of testing required to gain approval and all
but the antifouling guideline include lists of available standards. Upon contacting
the HSE, it was found that the most commonly used efficacy standards they have
encountered so far are EN standards, but ASTM standards are probably the next
most frequently submitted. They state that industry tests are also common,
especially for some PTs. Wood preservatives have generally been tested
according to CEN standards; insecticides according to in house tests (but where
guidelines are used, they are most frequently WHO tests); rodenticides according
to non standard tests or EPPO guidelines; anti-fouling paints were normally tested
in industry raft tests or patch tests and finally surface biocides were most
commonly tested according to non standard test methods. A list of references to
tests for some PTs that was so far included in the authorisation process was also
provided and these references have been included in Appendix I.
In Denmark, wood preservatives are efficacy tested according to CEN standards.
In the Netherlands, The Board for the Authorisation of Plant Protection Products
and Biocides (Ctgb) 26 is responsible for the authorisation of biocides. On its
website, efficacy concept guidance documents are available for disinfectants,
wood preservatives and rodenticides. There is also a guidance document coming
up on insecticides and a new version of the rodenticide guideline is presently
being negotioated in Technical Meetings. References found in the last version of
the rodenticide guidance document as well as the guidance for wood preservatives
were included in Appendix I.
7.1.3 Methods in use according to manufacturers and industry organisations
In the TNsG on Product Evaluation as as well as national guidelines (UK and NL)
there are several lists of available standard tests available for efficacy testing of
biocide products (see above). However, they do not make any comments on the
suitability of particular test methods and how frequently they are used. In order to
obtain an indication as to what tests are actually applied on the BPs by different
companies today (i.e. what test data the CA Sweden – and other Member States –
may expect to receive in applications), manufacturers were also contacted to ask
about what tests they use on their products. In addition, several tests available are
not in the form of standards but rather “in house” methods, sometimes developed
by the companies themselves and therefore not listed in the TNsG on Product
Evaluation.
Companies were initially contacted in Sweden but because suitable contact

persons were usually found to be located abroad, the companies were later mainly
contacted internationally. In addition, test institutes were also contacted, regarding
available efficacy tests on BPs and these references are also included in Appendix
I.
26
http://www.ctb.agro.nl
18
7.1.4 Test endpoints
In the TNsG on Product Evaluation it is pointed out that Good Laboratory Practice
(GLP) is not required for efficacy tests, although the spirit of these principles
should be applied at least during documentation. Due to limited access to full text
test descriptions and the short time period of the project, an in-depth evaluation of
any particular tests could not be performed. The individual design, applicability and
efficacy criteria, was therefore only briefly cited or summarised in Appendix I, if
information was available from either the home page of the standardisation
organisations or from full text references. However, it can be concluded that the
results of efficacy tests are expressed in numerous ways, depending on PT, test
design etc. Some examples of efficacy endpoints are "mean time to 50%
knockdown of flies", % kill, EC50 ("concentration causing the measured effect in
50% of the test population"), "elapsed time to first bite", “% ants crossing a tile
within 60 minutes”, “MIC (Minimum Inhibitory Concentration)”, ‘”% reduction in
CFU (Colony Forming Units)” etc, see Appendix I for particular tests.
7.2 LIMITATIONS AND KNOWLEDGE GAPS
In this project, a priority was to cover most or preferably all PTs rather than obtain
in depth information on tests for a particular PT. Although for some PTs only a few
test references were found, Appendix I covers all PTs except PT 16, 22 and 23.
One reason for the major lack of information on some PTs may be that there are
only a few products that will belong to this group (particularly true for e.g. PT 20,
because these products are generally covered by other regulations), but also that
not all product types have been previously regulated in all Member States.
Almost 90 % of the tests included in the overview are standards, although the
situation can vary between different PTs. There are most likely several additional
"in house" and non standardised tests, not available to the author during the
relatively short time period of the project. The coverage related to non standard
tests is related to the contributions obtained from different manufacturers. There
are also several additional standards available, but in this project in-depth
searches were focused on international standardisation organisations rather than
national standards.
19
8 DISCUSSION
BPD Annex VI, paragraph 92 and 93:
”92. Member States shall not authorise a biocidal product which does not
possess acceptable efficacy when used in accordance with the conditions
specified on the proposed label or with other conditions of authorisation.
93. The level, consistency and duration of protection, control or other intended
effects must, as a minimum, be similar to those resulting from suitable reference
products, where such products exist, or to other means of control. Where no
reference products exist, the biocidal product must give a defined level of
protection or control in the areas of proposed use. Conclusions as to the
performance of the biocidal product must be valid for all areas of proposed use
and for all areas in the Member State except where the proposed label
prescribes that the biocidal product is intended for use in specific
circumstances. Member States shall evaluate dose response data generated in
trials (which must include an untreated control) involving dose rates lower than
the recommended rate, in order to assess if the recommended dose is the
minimum necessary to achieve the desired effect.”
8.1 EVALUATING EFFICACY – COMPARING TEST RESULTS TO LABEL

CLAIMS AND PERFORMANCE STANDARDS
The purpose of testing BPs using efficacy tests, is to determine whether the
products posess “acceptable efficacy” (see BPD Annex VI, para 92). Statistically
significant results in a quantitative test including a negative control indicates
whether the product is efficient compared to no treatment at all. However, as with
any laboratory testing, whether a particular result is statistically significant is
influenced by aspects such as the number of replicates tested and other factors
related to test design (incubation period, choice of test organism etc). Therefore, a
product may be "efficient" according to one particular test, but maybe not if
applying other test conditions.
Furthermore, “statistically significant” may not be efficient enough from a user’s

point of view. A product causing 20% reduction in the number of cockroaches is
probably not regarded as efficient enough. BPs need to be efficient also in “real
life” and should fulfil minimum requirements for a certain use. Therefore, test
results need to be compared to performance standards (pass/fail criteria).
Individual efficacy test methods usually do not include pass/fail criteria, although
there exists CEN standards with pass-fail criteria for PT 8 (EN599) and several
applications under PT 1-5 (prEN14885:2006). Therefore, in most cases, the
efficacy of BPs has to be evaluated by comparing test results to the label claims
made by the manufacturer. The evaluating authorities need good knowledge about
20
how the BP is used and how it was tested, and the label claim need to be detailed
enough to allow this evaluation.
There are so far very few test recommendations or “data requirements” (specifying
recommended tests) for efficacy testing of BPs. The meaning of certain definitions
used in the labels, such as "disinfectant" and “germicidal effect” also need to be
defined on an international level (today different countries use somewhat different
definitions). In addition, standards are not available or there are very few for
several applications, which means that the authorities in these cases need to
evaluate data from manufacturers that have used internal test protocols. This may
cause the need to evaluate not only the test data but also the test protocols used
to generate the data by different manufacturers.
One problem with this case by case approach is that it will be difficult for
authorities to rank different products within a particular PT. Efficacy tests are
performed, and results expressed, in numerous ways, making comparisons
impossible. In addition, the evaluation can become inconsistent between different
CAs.
The lists currently available in the TNsG on Product Evaluation do not make any
statements about the suitability of particular tests, only availability. To fully
harmonise all testing of BPs for a particular PT would be very difficult, due to the
large variety of BPs and use areas, application rates, time scales of the protective
effects etc. However, it would probably be beneficial for both the manufacturers
and the CAs if recommended standard tests or data requirements are available for
all PTs and the most common areas of use. Individual deviations from test
standards can then be allowed if necessary due to the specific use (and label
claim) of the product. These minor deviations in the test design might influence the
results, but difficulties in comparing the results can be partly overcome by using
positive controls.
In general, ranking of different products (under specific conditions) would be

facilitated by the use of standard BPs as positive controls in the test. A postive
control also helps controlling the reproducibility of a test and facilitates
comparisons with other test facilities. When developing these standard BPs, not
only efficacy of the product should be evaluated, but also negative side effects on
non target species. It may not be necessary for the positive control to be “the most
efficient” product available.
Performance standards should be developed in parallel to lists of recommended

tests and reference products, so that it is indeed possible to determine whether a
particular product fulfils the criteria by testing it according to a certain test. The
quality of the particular test protocols (robustness) needs to be evaluated before
inclusion in the list of recommended/required tests. Authorities, standardisation
organisations, industry associations and manufacturers should all be involved
during the development of pass/fail criteria, new test standards, reference
products identification and establishing listings of recommended tests for particular
product applications.
21
Even with long term field tests it is not easy to predict real performance of the
product. The problems associated with efficacy testing are not unique though;
predicting ecosystem effects of chemicals based on a few toxicity tests performed
in the laboratory is also not a simple task. However, if harmonising performance
standards in so far as possible for particular PTs, use situations and label claims,
the authorisation process as well as the choice of test method and design will be
facilitated.
9 CONCLUSIONS AND SUGGESTIONS
• In total, 387 test references are included in this overview, covering all PTs
except 16, 22 and 23. The overview is based on references from the TNsG
on Product Evaluation and information from manufacturers, MSs and
national guidelines, test institutes and own searches of tests available on
the homepage of some international standardisation organisations. About
90% of the available tests included are standards.
• For one third of the test references included in this overview, the scope,
design, applicability and/or efficacy criteria is briefly summarised as far as
possible if information was available.
• The test references of more than 250 standards have been checked to
make sure that the newest version available is included in the overview.
• Efforts should be made to develop

o performance standards (pass/fail criteria) to minimize inconsistent
evaluation of efficacy
o additional standard methods in order to cover the most frequent
applications
o suitable reference substances, to facilitate ranking of different BPs
that were tested using different procedures and also the evaluation
of the reproducibility of test results.
o lists of recommended tests for particular products and use situations.
• Authorities, standardisation organisations and industry representatives

need to be involved during the development of pass/fail criteria, new test
standards, reference products identification and to establish lists of
recommended tests for particular product applications. As a start, the lists
of available standards for efficacy testing of BPs need to be updated in the
TNsG on Product Evaluation, since several tests have been withdrawn or
replaced by newer versions. It is the hope of the author that the present
overview may be of assistance in this process.
22
10 APPENDIX I. TEST PROCEDURES (STANDARDS AND OTHER
REFERENCES).
The following table includes relevant standards from TNsG on Product Evaluation,
list of standards used in France, national guidelines (UK, NL), results from own
searches on the web page of standardisation organisations, standards and non
standardized test refered to and/or supplied by manufacturers, industry
organizations and performing laboratories. The standards related to multiple
sources are updated to the most recent version if applicable. References in italics
have been checked by searching the web page of the standardisation organisation
(reference number, title) and, if applicable, been updated to the most recent
version of the test.
The suitability of particular test methods have not been evaluated, but for several
tests (if information was available to the author), the test procedures and
evaluation criteria have been briefly summarised, based or cited mainly from
information found in the description of the tests available on the web pages of the
standardisation organisations, full test description and/or from the TNsG on
Product Evaluation (TNsG PE).
For ASTM methods, the number after the hyphen is refering to the last revision
date of the test; the number in parenthesis is referring to the last reapproval of the
test.
The table includes only standards that are related to efficacy testing (as far as this
has been possible to read out from the title and available descriptions of the
standard), not e.g. use class standards (for PT8), leaching rate standards (for PT
21) etc. Standards that may have been present in the TNsG on Product Evaluation
but withdrawn since then (such as several ASTM standards related to avicides,
PT15) are also not included. Section 810.2000 of the OPPTS of US EPA is related
to antimicrobials in public health, but none of the guidelines were found on the
EPA web page at the time of the project. Although several EPA guidelines related
to PT 1-5 were included in the TNsG on Product Evaluation, these are not
included in table 1 due to uncertain reference details (titles are missing in the
TNsG PE).
The list is not exhaustive and the PT designation of a particular test reference may
not always reflect possible applications of the test method and in some cases it
was difficult to assign the test to a particular PT.
23
Table 1. Overview of efficacy test references, in order of product type.
REFERE TITLE PT SHORT TEST DESCRIPTION TYPE OF

NCE (IF TEST METHOD REFERENCE
AVAILABLE OR SOURCE
INFORMATION PROVIDED
FROM ELSEWHERE)
ASTM Standard Guide for ? Own searches
E1891- Determination of a
97(2002) Survival Curve for
Antimicrobial Agents
Against Selected
Microorganisms and
Calculation of a D-
Value and
Concentration
Coefficient
ASTM Test Method for ? This test method assesses the Own searches
WK15324 Determining the Time- microbicidal activity of
Kill Kinetics Of antimicrobial materials,
Antimicrobial whereby the survival of
Compounds organisms exposed to a water
miscible antimicrobial agent is
determined as a function of
time. The primary purpose of
this method is to provide a set
of standardized conditions and
test organisms to facilitate
comparative assessments of
antimicrobial materials
miscible in aqueous systems.
ASTM Standard Practice for 8- A wide variety of properties Own searches
G160-03 Evaluating Microbial 10? may be affected by microbial
Susceptibility of attack depending on material
Nonmetallic Materials or item characteristic.
by Laboratory Soil Standard methods (where
Burial available) should be used for
each different property to be
evaluated. This practice does
not attempt to enumerate all of
the possible properties of
interest nor specify the most
appropriate test for those
properties. Test methods must,
however, be appropriate to the
material being tested.
Evaluation of a nonmetallic
material's microbiological
susceptibility when in contact
with the natural environment of
the soil and is intended for use
on material test specimens
that are approximately 2 cm
(3/4 in.) thick and 100 cm2 (20
in2) or less. It is recommended
that this practice be combined
24
AVAILABLE OR SOURCE
FROM ELSEWHERE)
with appropriate environmental
exposures (for example,
sunlight simulating weathering
devices, the hydrolytic effects
of extended aqueous contact,
or extraneous nutrients) or
fabrication into articles (for
example, adhesive bonding of
seams) which may promote
microbiological susceptibility
during the service life of
material.
Microbiological susceptibility
may be reflected by a number
of changes including staining,
weight loss, or reduction in
tensile or flexural strength.
This practice may be applied
to articles that do not spend
the majority of their service life
in soil.
Alleman, Comparative 1-5 Microorganisms were exposed Manufacturer
J.E., Evaluation of to a mixture of biocide in
Etzel, Alternative Halogen- solution for 4 or 16 minutes.
J.E., based Disinfection The percentage survival was
Gendron, Strategies assessed under various
D., conditions such as high or low
Kirsch, nitrogen (as ammonium), high
E.J., pH or low temperatures.
Conley,
J.,
Fidelle,
T.,
Handy,
F., and
Hildebran
dt, M.
nd
42
Purdue
Industrial
Waste
Conferen
ce, May
1987,
519-524
American Standard Methods for 1-5 A swimming pool trial was Manufacturer
Public the Examination of performed over a three month
Health Water and Wastewater period to demonstrate the
Associati (SMEWW). disinfection efficacy of the
on, 16th product. Water samples were
Edition, collected for microbial
1985 analysis. Water and, air
25
AVAILABLE OR SOURCE
FROM ELSEWHERE)
temperature, number of
bathers (at the time when
samples were taken and total
bathers for the day), eye and
skin irritation and water clarity
were also measured.
Criteria for effective control
was determined as follows:
The state of California,
Department of Health Services
requirement for pool water is;
Bacteriological quality of water
in the swimming pool shall be
such that not more than two
consecutive samples, taken
when the pool is in use shall:
1. Contain more than 200
bacteria per ml, as determined
by the standard plate count; or
2. Contain a total coliform
organism count of 2.2 or
greater per 100 ml of sample
3. Chemical quality of water in
the pool shall not cause
irritation of eyes or skin of the
bathers, or have other
objectionable physiological
effects on bathers.
AOAC “Disinfectant (water) 1-5 Manufacturer
(Associati for Swimming Pools”
on of official method of
Official Analysis, 16th Edition,
Analytical 1995
Chemists
) official
method
965.13
ASTM Standard Test Method 1-5 Laboratory suspension test Manufacturer
E1052- for Efficacy of that determines the
96(2002) Antimicrobial Agents effectiveness of antimicrobial
Against Viruses in solutions against designated
Suspension prototype viruses. The
effective antimicrobial
concentration should be
determined using cell cultures
as the host system for specific
viruses. For special
applications of virucides, such
as inactivation of viruses in
contaminated liquid wastes,
and as a first stage in
determining virucidal potential
26
AVAILABLE OR SOURCE
FROM ELSEWHERE)
of liquid chemical germicides,
liquid hand soaps, OTC
topicals or other skin products.
ASTM Standard Test Method 1-5 Laboratory test method. Manufacturer

E1053- for Efficacy of Virucidal Evaluates the virucidal efficacy
97(2002) Agents Intended for of liquid, aerosol, or trigger
Inanimate spray antimicrobial solutions
Environmental on inanimate nonporous
Surfaces environmental surfaces,
against designated prototype
viruses. The effective
antimicrobial concentration
should be determined utilizing
cell cultures as the host
system for specific viruses.
Efficacy is measured by a
percentage reduction in titer.
ASTM Standard Test 1-5 Effectiveness of procedures Own searches
E1054-02 Methods for Evaluation and agents for inactivating
of Inactivators of (neutralizing, quenching) the
Antimicrobial Agents microbiocidal properties of
antimicrobial agents and to
ensure that no components of
the neutralizing procedures
and agents, themselves, exert
an inhibitory effect on
microorganisms targeted for
recovery.
ASTM Test Method for 1-5 Own searches
E1115-02 Evaluation of Surgical
Hand Scrub
Formulations
ASTM Standard Test Method 1-5 Own searches
E1153-03 for Efficacy of
Sanitizers
Recommended for
Inanimate Non-Food
Contact Surfaces
E1173- of a Evaluation of a
01e1 Preoperative,
Precatheterization, or
Preinjection Skin
Preparations
E1174-06 Evaluation of the
Effectiveness of Health
Care Personnel
Handwash
Formulations
27
AVAILABLE OR SOURCE
FROM ELSEWHERE)
E1327-07 for Evaluation of
Antimicrobial
Handwash
Formulations by
Utilizing Fingernail
Regions
E1482-04 for Neutralization of
Virucidal Agents in
Virucidal Efficacy
Evaluations
E1589-05 for Evaluation of First
Aid Antiseptic Drug
Products
E1766- for Determination of
95(2002) Effectiveness of
Sterilization Processes
for Reusable Medical
Devices
E1837- to Determine Efficacy
96(2002) of Disinfection
Processes for
Reusable Medical
Devices (Simulated
Use Test)
E1838-02 for Determining the
Virus-Eliminating
Effectiveness of Liquid
Hygienic Handwash
and Handrub Agents
Using the Fingerpads
of Adult Volunteers
ASTM Standard Test Method 1-5 This test method determines Own searches
E1882-05 for Evaluation of the antibacterial activity and
Antimicrobial persistence of test
Formulations by the formulations, as measured by
Agar Patch Technique the inhibition of a test
organism on an agar surface
exposed to test sites on
human skin treated with the
formulations. This procedure
can be used to evaluate
formulations containing
ingredients intended to inhibit
growth of bacteria on intact
skin and measures the
difference, post-product-
exposure, between numbers of
28
AVAILABLE OR SOURCE
FROM ELSEWHERE)
bacterial colonies on active
test formulation plates and
numbers on control plates,
expressed as percent
inhibition.
This procedure may also be
used to test for persistence of
activity, as a function of time
elapsed between application of
active test formulation and
application of active test
plates. Because no procedure
for neutralization of the
antimicrobial action of active
ingredients can be included in
the test, the agar patch
method is limited to the extent
that results expressed as
percent inhibition do not
differentiate between
bacteriostatic and bacteriocidal
effects and, hence, must not
be portrayed as “reductions.”
E1883-02 for Assessment of an
Antibacterial
Handwash Product by
Multiple Basin Wash
Technique
Handwashing
Formulations for Virus-
Eliminating Activity
Using the Entire Hand
ASTM Standard Quantitative 1-5 This test can be performed Own searches
E2111-05 Carrier Test Method to with or without a soil load to
Evaluate the determine the effect of such
Bactericidal, loading on microbicide
Fungicidal, performance. The soil load
Mycobactericidal, and developed for this test is a
Sporicidal Potencies of mixture of three types of
Liquid Chemical proteins (high molecular
Microbicides weight proteins, low molecular
weight peptides, and mucous
material) to represent the body
secretions, excretions, or other
extraneous substances that
chemical microbicides may
encounter under field
condititions.
This test method is designed
29
AVAILABLE OR SOURCE
FROM ELSEWHERE)
for use in product development
and for the generation of
product potency data. This test
method is fully quantitative and
it also avoids any loss of viable
organisms through wash off. It
permits the loading of each
carrier with a known volume of
the test organism. The
incorporation of controls can
also determine the initial load
of colony forming units (CFU)
of organisms on the test
carriers and any loss in CFU
after the mandatory drying of
the inoculum. This test method
is designed to have survivors
and also to be used with a
performance standard. The
surviving microorganisms on
each test carrier are compared
to the mean of no less than
three control carriers to
determine if the performance
standard has been met. To
allow proper statistical
evaluation of results, the size
of the test inoculum should be
sufficiently large to take into
account both the performance
standard and the experimental
variation in the results.
ASTM Standard Quantitative 1-5 The method is designed to Own searches
E2197-02 Disk Carrier Test evaluate the ability of liquid
Method for chemical germicides to
Determining the inactivate vegetative bacteria,
Bactericidal, Virucidal, viruses, fungi, mycobacteria
Fungicidal, and bacterial spores in the
Mycobactericidal and presence of a soil load on disk
Sporicidal Activities of carriers that represent
Liquid Chemical environmental surfaces and
Germicides medical devices. It is also
designed to have survivors
that can be compared to mean
of no less than three control
carriers to determine if the
performance standard has
been met. For proper statistical
evaluation of the results, the
size of the test inoculum
should be sufficiently large to
take into account both the
30
AVAILABLE OR SOURCE
FROM ELSEWHERE)
performance standard and the
experimental variation in the
results. The test protocol does
not include any wiping or
rubbing action. It is, therefore,
not designed for testing
germicide-soaked wipes.
Laundry Sanitizers and
Disinfectants
E2276- for Determining the
03e1 Bacteria-Eliminating
Effectiveness of
Hygienic Handwash
and Handrub Agents
of Adult Subjects
E2314-03 for Determination of
Effectiveness of
Cleaning Processes
Instruments Using a
Microbiologic Method
(Simulated Use Test)
ASTM Standard Guide for 1-5 This guide covers examples of Own searches
E2315-03 Assessment of a basic method to measure the
Antimicrobial Activity changes of a population of
Using a Time-Kill aerobic microorganisms within
Procedure a specified sampling time
when tested against
antimicrobial test materials in
vitro. Several options for
organism selection and
growth, inoculum preparation,
sampling times and
temperatures are provided.
Antimicrobial activity of specific
materials, as measured by this
technique, may vary
significantly on variables
selected.
ASTM Standard Guide for 1-5 This guide covers test Own searches
E2361-04 Testing Leave-On methods and sampling
Products Using In-Situ procedure options for leave-on
Methods products (such as alcohol
hand rubs and lotions
containing antimicrobial
ingredients) for consumer and
31
AVAILABLE OR SOURCE
FROM ELSEWHERE)
hospital personnel. These
products are distinguished
from conventional washing and
scrubbing preparations in that
they do not rely on the rinsing,
physical removal, and
antimicrobial action in
determining their
effectiveness. Although
agitation and friction may
serve to release organisms
from the skin and folds and
crevices, organisms are then
killed in situ and are not rinsed
from the skin surface before
sampling. Appropriate test
methods for the hands have
been published, while other
sampling methods will be
needed for testing body areas
other than the hands.
ASTM Standard Practice for 1-5 Own searches
E2362-04 Evaluation of Pre-
saturated or
Impregnated
Towelettes for Hard
Surface Disinfection

Disinfectants for Use
in High Efficiency
Washing Operations
ASTM Standard Test Method 1-5 This test method should be Own searches
E640-06 for Preservatives in used to determine if a
Water-Containing preservative or preservative
Cosmetics system has application for the
preservation of water-miscible
cosmetic products.
It sets minimal requirements
for preservative performance
in model formulations.
ASTM Standard Guide for 1-5 Own searches
WK12880 Evaluation of Clean
Room Disinfectants
WK4751 Selecting Test
Methods to Determine
the Efficacy of
32
AVAILABLE OR SOURCE
FROM ELSEWHERE)
and Other Chemicals
for Sanitization of
Produce
ASTM Guideline for 1-5 Own searches
WK9062 Evaluation of Residual
Effectiveness of
Antibacterial Personal
Cleansing Products
WK9378 for Determining the
Fungus-Eliminating
Effectiveness of
Hygienic Handwash
and Handrub Agents
of Adults
Bechert A new method for 1-5 The test is based on the Test institute
et al., screening anti-infective release of vital daughter cells
Nature biomaterials from the sample surface into
Medicine the surrounding. The
6, 1053- proliferation activity of these
1056 daughter cells, which are
(2000) responsible for infection
development, can be
monitored in a time course.
Antimicrobial activity is
monitored by the time needed
to reach a defined optical
density, which is dependent on
the number of released
daughter cells. All
measurements are performed
in comparison to an untreated
control (without antimicrobial
additive). The difference
between the reference (A) and
the test sample (B) to reach
the threshold OD gives the
degree of antimicrobial activity.
If all bacteria on the surface of
the material are prevented
from multiplying, no daughter
cells are produced and the test
object is considered
bactericidal. Materials can also
be antimicrobial, which means
that not all cells on the test
surface are prevented from
growing. Some cells are able
to divide and release daughter
cells into the surroundings,
which are then optically
33
AVAILABLE OR SOURCE
FROM ELSEWHERE)
registered in a so-called
growth curve. If surviving
daughter cells are grown under
controlled conditions over 48 h
(observation time), a higher
turbidity and therefore a bigger
signal is generated. At the
same time only vital and
proliferative cells are
accounted for.
In particular antimicrobial
samples will release daughter
cells into the surrounding.
Hence, microbial growth is first
observed noticeably later. This
right-shift towards longer times
is indicative for the
antimicrobial efficacy of the
tested samples. The so-called
onset OD serves as a
quantifiable parameter and is
equivalent to the required
number of hours required for
the surviving daughter cells to
grow to a predefined optical
density (OD = 0.2).
A material is defined to be
antimicrobial only if the
formation of at least 99.9%
(which is equivalent to a net
onset-OD of 6 hours and is
similar to a killing rate of 3
log10 steps) of the daughter
cells during the challenge time
is prevented in comparison
with the blank sample.
Marketed worldwide by Ciba

Specialty Chemicals under the
tradename NumetrikaTM.
BS General Purpose 1-5 Mycobacterium fortuitum Manufacturer
6734:200 Disinfection Test
4
CEN Washer-disinfectors - 1-5 Also ISO method. Acceptance Own searches
15883- Part 5: Test soils and criteria are included, based on
5:2005 methods for visual inspection and/or a
demonstrating microbiological end-point as
cleaning efficacy stated for each method. Where
chemical detection of residual
soiling is required/sought,
34
AVAILABLE OR SOURCE
FROM ELSEWHERE)
methods can be
complemented by the specific
determination of a residual
component of the applied test
soil.
EN Chemical disinfectants 1-5 Basic bacterial activity against Industry
1040:200 and antiseptics - of a test material against organizaton
5 Quantitative Staphylococcus aureus (ATCC
suspension test for the 6538) and Pseudomonas manufacturer
evaluation of basic aeruginosa (ATCC 15442). A Communication
bactericidal activity of test suspension of bacteria is with UK (HSE)
chemical disinfectants added to a prepared sample of
and antiseptics - Test the product under test. The TM II05 (Fr)
method and mixture is maintained at 20 oC.
requirements (phase At a specified contact time TNsG PE
1) chosen from one of the
following: 1 , 5, 15 , 30 , 45 or (updated)
60 minutes, an aliquot is taken.
The bactericidal action of this
aliquot is immediately
neutralised or suppressed by a
validated method. The method
of choice is dilution-
neutralisation. If a suitable
neutraliser cannot be found,
membrane filtration is used.
The number of surviving
bacteria in each sample is
determined and the reduction
in viable counts calculated.A
criterion for activity by this test
method is that the test material
should demonstrate at least a
5-log reduction in viable counts
of the test organisms in 60
minutes.
EN Chemical disinfectants 1-5 General guidance Industry
12353:20 and antiseptics - organizaton
06 Preservation of test
organisms used for the TM II05 (Fr)
determination of
bactericidal, TNsG PE
mycobactericidal,
sporicidal and
fungicidal activity
EN Chemical disinfectants 1-5 Basic fungicidal activity against Industry
1275:200 and antiseptics - Candida albicans (ATCC organizaton
5 Quantitative 10231) and Aspergillus niger
suspension test for the (ATCC 16404). A test Manufacturer
evaluation of basic suspension of yeast cells or
fungicidal or basic mould spores is added to a
yeasticidal activity of prepared sample of the
35
AVAILABLE OR SOURCE
FROM ELSEWHERE)
chemical disinfectants product under test. The
and antiseptics - Test mixture is maintained at 20 oC.
method and At a specified contact time
requirements (phase chosen from one of the
1) following 5, 15, 30 or 60 Communication
minutes, an aliquot is taken; with UK (HSE)
the fungicide action in this
portion is immediately TM II05 (Fr)
neutralised or suppressed by a
validated method. The method TNsG PE
of choice is dilution-
neutraliser cannot be found,
membrane filtration is used.
The number of surviving yeast
cells or mould spores in each
sample is determined and the
reduction in viable counts
calculated.
The criterion for activity by this
test is that the test material
minutes.
EN Chemical disinfectants 1-5 Dilution/Neutralisation . Industry
1276:199 and antiseptics - Efficacy determined by a organizaton
7 Quantitative reduction in CFU.
suspension test for the Manufacturer
evaluation of
bactericidal activity of
chemical disinfectants
and antiseptics used in
food, industrial,
domestic, and
institutional areas -
Test method and
requirements (phase
2, step 1)
TM II05 (Fr)
TNsG PE
EN Chemical disinfectants 1-5 Testing mainly bactericidal Industry

12791:20 and antiseptics - activity; in vivo test; phase 2 organizaton/
05 Surgical hand step 2 Manufacturer
disinfection - Test
method and TM II05 (Fr)
requirement (phase
2/step 2) TNsG PE
EN Chemical disinfectants 1-5 Industry
36
AVAILABLE OR SOURCE
FROM ELSEWHERE)
13610:20 - Quantitative organizaton
02 suspension test for the
evaluation of virucidal TM II05 (Fr)
activity against
bacteriophages of TNsG PE
used in food and
industrial areas - Test
method and
requirements (phase
2, step 1)
13624:20 and antiseptics - organizaton/ma
03 Quantitative nufacturer
suspension test for the
evaluation of fungicidal
activity of chemical
disinfectants for TM II05 (Fr)
instruments used in
the medical area - Test TNsG PE
method and
requirements (phase
2, step 1)
EN Chemical disinfectants 1-5 Dilution/Neutralisation Method. Industry
13697:20 and antiseptics - Efficacy measured by a organizaton
01 Quantitative non- reduction in CFU.
porous surface test for Manufacturers
the evaluation of
bactericidal and/or
fungicidal activity of
used in food,
industrial, domestic
and institutional areas
- Test method and
requirements without
mechanical action
(phase 2/step2)
TM II05 (Fr)
TNsG PE

13704:20 - Quantitative organizaton
02 suspension test for the
evaluation of sporicidal Manufacturer
disinfectants used in TM II05 (Fr)
food, industrial,
domestic and TNsG PE
Test method and
37
AVAILABLE OR SOURCE
FROM ELSEWHERE)
requirements (phase
2, step 1)
EN Chemical disinfectants 1-5 Efficacy determined by the Industry
13727:20 and antiseptics - reduction in CFU counts organizaton
03 Quantitative
evaluation of
for instruments used in
the medical area - Test
method and
requirements (phase
2, step 1)
TM II05 (Fr)
TNsG PE
EN Chemical disinfectants 1-5 Dilution/Neutralisation Method. Industry

14204:20 and antiseptics - Effiacy measured by reduction organisation
04 Quantitative in CFU.
evaluation of
mycobactericidal
activity of chemical TM II05 (Fr)
disinfectants and
antiseptics used in the
veterinary area - Test
method and TNsG PE
requirements (phase
2, step 1)
EN Chemical disinfectants 1-5 Basic sporicidal activity against Industry
14347:20 and antiseptics - Basic dormant spores of Bacillus organizaton
05 sporicidal activity - subtilis (ATCC 6633) and
Test method and Bacillus cereus (ATCC 12826). Manufactuer
requirements (phase A prepared sample of the
1) product under test is added to
a test suspension of bacterial
spores. The mixture is Communication
maintained at 20 oC or any with UK (HSE)
other temperature to be
defined. At a specified contact TM II05
time chosen from one of the
following: 30, 60 and 120 TNsG PE
minutes, an aliquot portion is
taken and the sporicidal as
well as sporistatic action in this
portion is neutralised. The
method of choice is dilution-
neutralisation. The number of
surviving bacterial spores is
determined in parallel and the
38
AVAILABLE OR SOURCE
FROM ELSEWHERE)
reduction in viable counts
calculated. The effectiveness
of neutralisation is controlled in
the test.
The criterion for activity by this
test is that the test material
minutes.Medical area
Veterinary area, Food,
industrial, domestic and
institutional hygiene
EN Chemical disinfectants 1-5 Efficacy measured by a Industry
05 Quantitative
evaluation of
mycobactericidal
disinfectants in the
medical area including
instrument
disinfectants - Test
methods and
requirements (phase
2, step 1)
EN Chemical disinfectants 1-5 Non-porous surface test, Industry
14349:20 and antiseptics - bactericidal, for disinfectants organizaton
07 Quantitative surface used in the veterinary
test for the evaluation applications
of bactericidal activity Dilution/Neutralisation Method. Manufacturer
of chemical By one manufacturer, the test
disinfectants and material is deemed to have
antiseptics used in passed the test and be
veterinary area on efficacious if it demonstrates a TM II05 (Fr)
non-porous surfaces log 4 or more reduction in
without mechanical viable counts under the
action - Test method conditions defined in the test. TNsG PE
and requirements Reduction in viable microbial
(phase 2, step 2) counts compared with water
controls.
05+A1:20 Virucidal quantitative
06 suspension test for Manufacturer
human medicine - Test TM II05 (Fr)
method and
requirements (phase
2, step 1)
39
AVAILABLE OR SOURCE
FROM ELSEWHERE)
06 Quantitative carrier
test for the evaluation Manufacturer
of bactericidal activity
for instruments used in TM II05 (Fr)
method and
requirements (phase
2, step 2)
06 Quantitative carrier
test for the evaluation Manufacturer
of fungicidal or
yeasticidal activity for
instruments used in TM II05 (Fr)
method and
requirements (phase
2, step 2)
06 Quantitative
evaluation of virucidal
disinfectants and TM II05 (Fr)
method and
requirements (phase
2, step 1)
EN Ophtalmic optics - 1-5 Also ISO method. Own searches
14730: contact lense care
2000 products -
antimicrobial
preservative efficacy
testing and guidance
on determining discard
rate
prEN Chemical disinfectants 1-5 Industry
06 Application of
European Standards Manufacturer
for chemical
disinfectants and
antiseptics
TM II05 (Fr)
EN Chemical disinfectants 1-5 Applied in medical area Industry

40
AVAILABLE OR SOURCE
FROM ELSEWHERE)
7 Hygienic handwash -
Test method and Manufacturer
requirements (phase
2/step 2)
TM II05 (Fr)
TNsG PE

7 Hygienic handrub -
Test method and Manufacturer
requirements (phase
2/step 2) TM II05 (Fr)
TNsG PE

7 Quantitative
evaluation of fungicidal
activity of chemical TM II05 (Fr)
disinfectants and
antiseptics used in
food, industrial, TNsG PE
domestic, and
Test method and
requirements (phase
2, step 1)
EN 1656 Chemical disinfectants 1-5 Dilution/Neutralisation Method. Manufacturers
: 2000 and antiseptics - Efficacy measured by a
Quantitative reduction in CFU.
evaluation of
veterinary field - Test
method and
requirements (phase
2/step 1)
Industry
organisation
TM II05 (Fr)
TNsG PE
EN Chemical disinfectants 1-5 By one manufacturer, the Manufacturers

1657:200 and antiseptics - product was to be deemed to
41
AVAILABLE OR SOURCE
FROM ELSEWHERE)
5/AC:200 Quantitative have passed the test if it
7 suspension test for the demonstrated a 10E5
evaluation of fungicidal logarithmic reduction in
or yeasticidal activity viability.
of chemical
disinfectants and By one manufacturer a
antiseptics used in the reduction in viable microbial Industry
veterinary area - Test counts is compared with water organizaton
method and controls.
requirements (phase The test material is deemed to TM II05 (Fr)
2, step 1) have passed the test and be
efficacious if it demonstrates a
log 4 or more reduction in TNsG PE
viable counts under the
conditions defined in the test
MAFF Disinfectants for use 1-5 TNsG PE
(1969) specifically against: a)
anthrax, brucellosis,
contagious bovine
pleuro-pneumonia and
glanders; b) For use
against tuberculosis; c)
For use against foot-
and-mouth disease; d)
For use against fowl
pest (Newcastle
disease fowl plague).
NF T72- Water-miscible, 1-5 Manufacturer
230 neutralizable
August antiseptics and
1988 disinfectants used in
liquid form.
Determination of
sporicidal activity.
Dilution-neutralization
method.
NF T72- Methods of airborne 1-5 TM II05 (Fr)
281 disinfection of
Septemb surfaces. Determiation
er 1986 of bactericidal,
fongicidal and
Not Assessment of the 1-5 Various concentrations of Manufacturer
available biocide efficacy using product was mixed with algal
three algal species. In- suspensions (Chlorella
house method vulgaris
Navicula pelliculosa
Anabaena flos-aquae) in glass
conical flasks All flasks were
incubated and shaken in an
orbital shaker. Samples were
taken at 96h and the
42
AVAILABLE OR SOURCE
FROM ELSEWHERE)
chlorophyll a concentration
determined. Efficacy was
measured by determining the
EC50 value obtained for algae.
Not Disinfectant for 1-5 Using a growth inhibition test Manufacturer
available swimming pools for method for recording MIC
control of bacteria, values to test the efficacy of
fungi and algae. the product at various diluted
Determination of concentrations. MIC value was
Minimum Inhibitory determined using microdilution
Concentration Against technique in microtiter plates
Bacteria, Fungi, Algae Tests were carried out in
and Cyanobacteria (in quadruplicate. The ppm level
house method) of the product in the last
microtiter well demonstrating
no growth was determined as
the MIC for the product against
that microorganism.
Not Disinfectant treatment 1-5 Several concentrations of the Manufacturer
available of waste water. A product were added to river
laboratory study water and allowed to react for
involving disinfection 30 mins. Total counts and
of untreated river coliforms were determined.
water, as a surrogate Efficacy was determined
for a full field trial. (in- based on reduction in CFU
house methods) counts.
Not Disinfectant treatment 1-5 A product was used to treat Manufacturer
available of waste water. Full waste water at 3
scale field trial concentrations under real field
conditions involving a waste
water treatment plant; variable
flow and bacterial load, and
operating temperatures.
Efficacy was measured based
on a log removal of bacteria
after 15 and 30 mins.
Not Disinfectant treatment 1-5 Experiments were conducted Manufacturer
available of waste water. to investigate in pilot scale the
Inactivation of enteric disinfection efficiency against
micro-organisms in enteric micro-organisms in
tertiary treated tertiary treatment waste waters
municipal waste and the effect of dose and
waters (in-house contact time on disinfection
methods) efficiency. After incubation
bacterial colonies were
counted and microbial
numbers calculated as cfu/
100 ml.
Efficacy was measured by
percentage reductions of
microorganisms after 8 and 18
minutes contact time.
43
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Not Disinfectant treatment 1-5 A purpose built disinfection, Manufacturer
available of waste water. Pilot continuous flow, Pilot Plant
studies to assess the was specially constructed and
disinfection installed at a functioning
performance of on a wastewater treatment plant.
physicochemical Prior to installation the
wastewater effluent. wastewater was treated only
by chemically assisted
sedimentation, making
disinfection particularly difficult.
As part of this pilot study,
modifications to the Plant and
process were explored and
adopted on the basis of
experience and experimental
data using standard
methodologies. Disinfection
efficiency under a number of
conditions and wastewater
loads were explored. Method
for recording faecal coliforms:
American Public Health
association; American Water
Works Association and Water
Environment federation (1998)
Standard Methods for the
Examination of Water and
Wastewater 20th Ed.;
Washington DC, USA; Method
9222D. Method for recording
Enterococcus, coliphage
Clostridium
according to laboratory
methods. Efficacy was
determined by measured Log
reductions (or removal) in
counts. Colonies were counted
before and after disinfection.
Not Swimming pool 1-5 A residential spa was used to Manufacturer
available disinfectant: test the efficacy of the product
Residential spa field in typical field conditions. The
test maintenance pattern used was
considered to be typical of the
“average spa owner”, i.e.
conforming to minimum and
not optimum use pattern and
maintenance procedures.
Bacteriological analysis was
conducted on 150 samples
were taken over the 90 day
test period. Efficacy was
determined by a reduction in
44
AVAILABLE OR SOURCE
FROM ELSEWHERE)
cfu/100ml.
OECD Proposed 1-5 Own searches
(ENV/JM/ development of an
BCID(200 OECD Guidance
7)2) Document for
establishing the
efficacy of biocides
used in swimming
pools and spas
Powers Biocidal Efficacy of a 1-5 Biocidal emergency water Manufacturer
et al. Flocculating purification tablets were tested
Applied Emergency Water for bactericidal, virucidal and
and Purification Tablet. cysticidal efficacy in water at
Environm temperatures ranging from 5 to
ental 25ºC to validate and verify the
Microbiol effectiveness of the tablets for
ogy, 60: the destruction and removal of
2316 – microorganisms. Counts of
2323 surviving organisms were
(1994) compared with the initial
counts to calculate the log-
reduction. Replicate results
were averaged
prEN Chemical disinfectants 1-5 Own searches
13623 and antiseptics -
Quantitative
evaluation of
bactericidal activity
against Legionella
pneumophila of
for aqueous systems -
Test method and
requirements (phase
2, step 1)
prEN Chemical disinfectants 1-5 Manufacturer
Quantitative carrier
test for the evaluation TM II05 (Fr)
of mycobactericidal or
tuberculocidal activity
of chemical
disinfectants used for
instruments in the
medical area - Test
method and
requirements (phase
2, step 2)
ASTM Standard Test Method 1-5 + This test method specifies the Own searches
E2562-07 for Quantification of 12 operational parameters
Pseudomonas required to grow a repeatable
45
AVAILABLE OR SOURCE
FROM ELSEWHERE)
aeruginosa Biofilm Pseudomonas aeruginosa
Grown with High biofilm under high shear (1).
Shear and Continuous The resulting biofilm is
Flow using CDC representative of generalized
Biofilm Reactor situations where biofilm exists
under high shear rather than
representative of one particular
environment. The biofilm
generated in the CDC biofilm
reactor is also suitable for
efficacy testing. 3 This test
method describes how to
sample and analyze biofilm for
viable cells. Biofilm population
density is recorded as log
colony forming units per
surface area. After the 48 h
growth phase is complete, the
user may add the treatment in
situ or harvest the coupons
and treat them individually.
This test method uses the
Centers for Disease Control
and Prevention (CDC) biofilm
reactor. The CDC biofilm
reactor is a continuously
stirred flow reactor with high
wall shear. Although it was
originally designed to model a
potable water system for the
evaluation of Legionella
pneumophila, the reactor is
versatile and may also be used
for growing and/or
characterizing biofilm of
varying species.
ASTM Standard Test Method 6 Own searches
D2574-06 for Resistance of
Emulsion Paints in the
Container to Attack by
Microorganisms
ASTM Standard Test 6 Determination of the TNsG on Prod
D4783- Methods for resistance of liquid adhesive Eval
01e1 Resistance of preparations to microbial
Adhesive Preparations attack in the container by
in Container to Attack challenging adhesive
by Bacteria, Yeast, specimens with cultures of
and Fungi bacteria, yeast, or fungi, and
checking for their ability to
return to sterility.
These test methods return
qualitative results
46
AVAILABLE OR SOURCE
FROM ELSEWHERE)
ASTM Standard Practice for 6 The procedure should be used Own searches
E1259-05 Evaluation of to evaluate the relative efficacy
Antimicrobials in Liquid of microbicides in liquid fuels
Fuels Boiling Below boiling below 390°C. The
390°C effect of environmental
conditions, such as a variety of
fuel additives, metal surfaces,
and climatology, are variables
that can be included in specific
tests using this protocol.
ASTM Guideline For 6 This method uses an agar- Own searches
WK5097 Evaluating The dilution procedure to determine
Potential For Decline the Minimum Inhibitory
In Planktonic Concentration (MIC) and
Microorganism subsequently to screen for the
Susceptability To development of decreased
Antimicrobial susceptibility for test
Compounds: formulations versus bacteria or
Healthcare yeast species. The inocula
Applications used on the second, third,
fourth, and fifth test cycles will
be prepared from the microbial
growth present at the previous
day’s end point;i.e., that agar-
dilution plate containing the
highest concentration of the
test formulation (lowest
dilution) that allows microbial
growth.
Not Other in can 6 The method was conducted by Manufacturer

available preservatives. In- inoculating mixed bacteria and
house method based mixed fungi into a interior paint
on ‘Antimicrobial formulation containing various
Preservative concentrations of the active
Effectiveness substance which was repeated
Challenge Test’ and on a weekly basis. After 0, 7
DAB 10 guidelines 14, 21 and 28 days samples of
the formulations were placed
in a preservative neutralizer
and then plated. After plate
incubation the number of
surviving organisms was
compared to the control. The
criteria for measuring good
preservation efficacy was a
microbial count of <10 cfu/ml
at day 7, 14 , 21 and 28 days
Not Preservative for 6 The method was conducted by Manufacturer
available detergents. In-house inoculating mixed bacteria and
method based on mixed fungi into a washing up
‘Antimicrobial liquid formulation containing
47
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Preservative various concentrations of the
Effectiveness active substance which was
Challenge Test’ and repeated on a weekly basis.
DAB 10 guidelines After 0, 7 14, 21 and 28 days
samples of the formulations
were placed in a preservative
neutralizer and then plated.
After plate incubation the
number of surviving organisms
was compared to the control .
The criteria for measuring
good preservation efficacy was
a microbial count of <10 cfu/ml
at day 7, 14 , 21 and 28 days.
SABS Bacterial efficacy of 6 Efficacy test for in can TNsG on Prod
1102 biocides used in water- preservatives in paints Eval
(1987) based emulsion paints (emulsion) against bacteria.
SM020 S&M Fako test 6 Preserving effect of chemical Manufacturer
(in preservatives in water
house) containing coatings, adhesives
and other water containing
technical emulsions or
dispersions (in can).
Test batch: unpreserved
samples with different
concentrations of test
preservatives, inoculated and
streaked on agar plates.
Test germ: G+, G-, Moulds,
Yeasts
"Well preserved" if after 6w no
microbial growth can be
observed.
Corresponds to 2 years of
microbial stability.
4 evaluation levels
SM021 S&M KOKO test 6 Preservatives in cosmetic Manufacturer
(in formulations, e g creams,
house) shampoos etc (in can)
Yeasts
"Well preserved" if after 6w no
microbial growth can be
observed.
Corresponds to 30 months of
4 evaluation levels
48
AVAILABLE OR SOURCE
FROM ELSEWHERE)
SM026 S&M Betoko test: 6 Preserving effect of chemical Manufacturer
(in Determination of the preservatives in concrete
house) preserving effect of admixtures (in can
chemical preservatives preservation).
in concrete admixtures Test batch: unpreserved
Yeasts
Free of growth during 4 weeks:
"Well preserved"
(4 evaluation levels)
Corresponds to 1 year of
microbiological stability
SM029 S&M FeuTuKo test 6 Preservatives in wet tissues in Manufacturer
(in their original packaging.
house) Inoculation of whole packaging
and after 3w of incubation,
pieces of tissue is placed on
agar plates.
Yeasts
3d bacteria, 7d moulds. 4
evaluation levels.
SM036 S&M WiWako test 6 Preservation of fountain Manufacturer
(in solutions for offset printing.
house) Test batch: unpreserved
Yeasts
12 cycles
4 evaluation levels.
SM037 S&M TaBaKo test 6 Preservation of paint baths. Manufacturer
(in Test batch: unpreserved
house) samples with different
Yeasts
12 cycles. 4 evaluation levels.
SM044 Determination of the 6 Preserving effect of chemical Manufacturer
(in preserving effect of preservatives in household
house) chemical preservatives formulations, e g softener,
in household washing up liquids, all purpose
formulations cleaner (in can preservation).
49
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Yeasts
Free of growth during 6 weeks:
"Well preserved". (4 evaluation
levels). Corresponds to 30
months of microbiological
stability
NF X41- Protection. Testing 6+ TNsG on Prod
520 method for resistance 7 Eval
March of paints to TMII05
1968 microorganisms and
their protective power.
ASTM Standard practice for 7 This practice covers Own searches
G21- determining resistance determination of the effect of
96(2002) of synthetic polymeric fungi on the properties of
materials to fungi synthetic polymeric materials
in the form of molded and
fabricated articles, tubes, rods,
sheets, and film materials.
Conditions favorable for attack,
namely, a temperature of 2 to
38°C (35 to 100°F) and a
relative humidity of 60 to 100
% need to be established.
Expected effects include
surface attack, discoloration,
loss of transmission (optical),
and removal of susceptible
plasticizers, modifiers, and
lubricants, resulting in
increased modulus (stiffness),
changes in weight,
dimensions, and other physical
properties, and deterioration of
electrical properties such as
insulation resistance, dielectric
constant, power factor, and
dielectric strength.
Since attack by organisms
involves a large element of
chance due to local
accelerations and inhibitions,
the order of reproducibility may
be rather low. To ensure that
estimates of behavior are not
too optimistic, the greatest
observed degree of
50
AVAILABLE OR SOURCE
FROM ELSEWHERE)
deterioration should be
reported.
Conditioning of the specimens,
such as exposure to leaching,
weathering, heat treatment,
etc., may have significant
effects on the resistance to
fungi. Determination of these
effects is not covered in this
practice.
ASTM Standard Test Method 7 This test method describes a Own searches
D3273- for Resistance to small environmental chamber
00(2005) Growth of Mold on the and the conditions of operation
Surface of Interior to evaluate reproducibly in a 4-
Coatings in an week period the relative
Environmental resistance of paint films to
Chamber surface mold fungi, mildew
growth in a severe interior
environment. The apparatus is
designed so it can be easily
built or obtained by any
interested party and will
duplicate results obtained in a
large tropical chamber.
An accelerated test for
determining the resistance of
interior coatings to mold
growth; useful in estimating
the performance of coatings
designed for use in interior
environments that promote
mold growth and in evaluating
compounds that may inhibit
such growth and the aggregate
levels for their use.
Used to evaluate the
comparative resistance of
interior coating to accelerated
mildew growth. Performance at
a certain rating (in accordance
with Test Method D3274) does
not imply any specific period of
time for a fungal free coating.
However, a better rated
coating nearly always performs
better in actual end use. This
test method is intended for the
accelerated evaluation of an
interior coatings' resistance to
fungal defacement. Use of this
test method for evaluating
exterior coatings' performance
51
AVAILABLE OR SOURCE
FROM ELSEWHERE)
has not been validated, nor
have the limitations for such
use been determined. Any
accelerated weathering
(leaching, weathering machine
exposure, etc.,) should be
reported and should also bear
reference to the fact that it is
beyond the current scope of
this test method.
Temperature and humidity
must be effectively controlled
within the relatively narrow
limits specified in order for the
chamber to function
reproducibly during the short
test period. Severity and rate
of mold growth on a film is a
function of the moisture
content of both the film and the
substrate. A relative humidity
of 95 to 98 % at a temperature
of 32.5+ 1oC (90+ 2oF ) is
necessary for test panels to
develop rapidly and maintain
an adequate moisture level to
support mold growth.
ASTM Standard Practice for 7 This practice provides Own searches
D3456- Determining by guidelines for determining the
86(2002) Exterior Exposure susceptibility of paint films to
Tests the Susceptibility microbiological attack on
of Paint Films to exterior exposure. The degree
Microbiological Attack to which microbiological
discoloration occurs is the
primary concern. This practice
covers the preparation of
coatings for testing, their
application on substrates, and
the arrangement of the coated
panels on exterior test fences
to determine the degree of
microbiological attack that may
occur on the surface of the
coatings over a period of time.
This practice is intended to
provide guidelines for, and a
discussion of, the various
factors critical in selection of
exterior coatings resistant to
discoloration or disfigurement
by algae and fungi.
ASTM Standard Test 7 Tests the ability of adhesive Own searches
52
AVAILABLE OR SOURCE
FROM ELSEWHERE)
D4300-01 Methods for Ability of films to inhibit or support the
Adhesive Films to growth of selected fungal
Support or Resist the species growing on agar plates
Growth of Fungi by providing means of testing
the films on two agar
substrates, one which
promotes microbial growth,
and one which does not.
constituents.
These test methods are not

appropriate for all adhesives.
The activity of certain biocides
may not be demonstrated by
these test methods as a result
of irreversible reaction with
some of the medium. As an
example, quaternary
ammonium compounds are
inactivated by agar. A test
method is included for use with
low-viscosity adhesives along
with an alternative method for
use with mastic-type
adhesives.
ASTM Standard Test Method 7 This test method covers an Own searches
D5589- for Determining the accelerated method for
97(2002) Resistance of Paint determining the relative
Films and Related resistance of a paint or coating
Coatings to Algal film to algal growth.
Defacement This test method should not be
used as a replacement for
exterior exposure since many
other factors, only a few of
which are listed will affect
those results.
Films and Related resistance of two or more
Coatings to Fungal paints or coating films to fungal
Defacement by growth.
Accelerated Four- This test method should not be
Week Agar Plate used as a replacement for
Assay exterior exposure (that is,
Practice D 3456) since many
other factors, only a few of
which are listed will affect
those results.
Comparative evaluation of
different coating formulations
for their relative performance
53
AVAILABLE OR SOURCE
FROM ELSEWHERE)
under a given set of
conditions. It does not imply
that a coating that resists
growth under these conditions
will necessarily resist growth in
the actual application.
Round-robin testing of this test

method versus exterior
exposure is planned.
ASTM Standard Test Method 7 This test method provides a Own searches
E1428- for Evaluating the technique for evaluating
99(2004) Performance of antimicrobials in or on
Antimicrobials in or on polymeric solids against
Polymeric Solids staining by Streptoverticillium
Against Staining by reticulum, and should assist in
Streptoverticillium the prediction of performance
reticulum (A Pink Stain of treated articles under actual
Organism) field conditions. Conditioning
of the specimens, such as
exposure to leaching,
weathering, and heat
treatment, may have
significant effects on
performance of antimicrobials
against staining. Determination
of these effects is not included
in this test method. This test
method is also not suitable for
evaluating dark-pigmented test
samples.
ASTM Standard Test Method 7 This test method is used for Own searches
E2196-07 for Quantification of a growing a repeatable
Pseudomonas Pseudomonas aeruginosa
aeruginosa Biofilm biofilm in a continuously stirred
Grown with Shear and flow reactor. In addition, the
Continuous Flow test method describes how to
Using a Rotating Disk sample and analyze biofilm for
Reactor viable cells. In this test
method, biofilm population
surface area.
ASTM Standard Practice for 7 Determination of the Own searches
G29- Determining Algal susceptibility of plastic films to
96(2002) Resistance of Plastic the attachment and
Films proliferation of surface-growing
algae, produced in bodies of
water, such as swimming
pools, artificial ponds, and
irrigation ditches that are lined
with plastic films.
54
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Evaluating the degree and
permanency of protection
against surface growth of
algae afforded by various
additives incorporated in the
film.
EN Paints and varnishes - 7 Manufacturer
15457:20 Laboratory method for
07 testing the efficacy of
film preservatives in a
coating against fungi
EN Paints and varnishes - 7 Own searches
coating against algae
ASTM Standard 8 Field test: aids in evaluating Own searches
D1006-93 Recommended the performance of house and
Practice for trim paints to new, previously
Conducting Exterior unpainted wood exposed to
Exposure Tests of the environment.
Paints on Wood Exposures in several locations
with different climates which
represent a broad range of
anticipated service conditions
are recommended and several
years of repeat exposures are
needed to get an “average”
test result for a given location.
Solar radiation varies
considerably as function of
time of year, and can cause
large differences in the
apparent rate of degradation in
many polymers. Comparing
results for materials exposed
for short periods (less than
onee year) is not
recommended unless
materials are exposed at the
same time in the same
location.
CEN/TR Wood preservatives - 8 Pretreatment procedure before Own searches
15046:20 Artificial weathering of efficacy testing
05 treated wood prior to
biological testing - UV-
radiation and water-
spraying procedure
CEN/TS Wood preservatives - 8 TM II05 (Fr)
12037:20 Field test method for
03 determining the
relative protective
55
AVAILABLE OR SOURCE
FROM ELSEWHERE)
effectiveness of a
wood preservative
exposed out of ground
contact - Horizontal
lap-joint method
CTBA- Field ageing test on 8 TM II05 (Fr)
BIO-E treated soils
001
BIO-E treated masonry
002
CTBA- Field test on bait 8 TM II05 (Fr)
BIO-E system for termite
003 control
CTBA- Laboratory tests for 8 TM II05 (Fr)
BIO-E bait system for termite
004 control
CTBA- Laboratory tests for 8 TM II05 (Fr)
BIO-E bait system for termite
005 control
CTBA- Action of U.V. light on 8 TM II05(Fr)
BIO-E physico-chemical
006 barrier for termite
CTBA- Action of alcalin 8 TM II05 (Fr)
BIO-E conditions on physico-
007 chemical barrier for
termite control
BIO-E physico-chemical
008 barrier for termite
control
CTBA- Forced test on termite 8 TM II05 (Fr)
BIO-E bait control
009
CTBA- Choice test on termite 8 TM II05 (Fr)
BIO-E bait control
010
EN Wood preservatives - 8 Laboratory test. Efficacy TNsG on Prod
113:1996 Test method for criteria based on weight loss of Eval
/A1:2004 determining the untreated specimen.
protective TM II05 (Fr)
effectiveness against
wood destroying NL guidance
basidiomycetes -
Determination of the UK guidelines
toxic values
Test institute
Manufacturer
EN Wood preservatives - 8 TNsG on Prod

117:2005 Determination of toxic Eval
56
AVAILABLE OR SOURCE
FROM ELSEWHERE)
values against
Reticulitermes species TM II05 (Fr+De)
(European termites)
(Laboratory method) NL guidance
UK guidelines

118:2005 Determination of Eval
preventive action
against Reticulitermes TM II05 (Fr+De)
species (European
termites) (Laboratory NL guidance
method)
UK guidelines
EN Durability of wood and 8 TM II05 (Fr)

12490:19 wood-based products -
98 Preservative-treated
solid wood -
Determination of the
penetration and
retention of creosote in
treated wood
EN Wood preservatives - 8 TNsG Prod Eval
1390:200 Determination of the
6 eradicant action TM II05 (Fr)
against Hylotrupes
bajulus (Linnaeus) NL guidance
larvae - Laboratory
method UK guidelines
EN 152-1 Test methods for wood 8 TNsG on Prod

(1988) preservatives - Eval
Laboratory method for
determining the TM II05 (Fr)
preventive
effectiveness of a NL guidance
preservative treatment
against blue stain in UK guidelines
service - Part 1:
Brushing procedure
EN 152- Test methods for wood 8 TNsG on Prod
2:1988/A preservatives - Eval
C1:1989 Laboratory method for
protective
effectiveness of a NL guidance
against blue stain in UK guidelines
service - Part 2:
Application by
57
AVAILABLE OR SOURCE
FROM ELSEWHERE)
methods other than
brushing
EN 20- Wood preservatives - 8 TNsG on Prod
1:1992 Determination of the Eval
protective
effectiveness against TM II05 (Fr)
Lyctus Brunneus
(Stephens) - Part 1: NL guidance
Application by surface
treatment (laboratory UK guidelines
method)
protective
effectiveness against TM II05 (Fr)
Lyctus brunneus
(Stephens) - Part 2: NL guidance
Application by
impregnation UK guidelines
(Laboratory method)
EN Field test method for 8 Field test, in ground contact TNsG on Prod
252:1989 determining the Eval
/AC1:198 relative protective
9 effectiveness of a Test institute
wood preservative in
ground contact
NL guidance
UK guidelines
TM II05 (Fr)
EN Wood preservatives - 8 Marine field test, 5 year TNsG on Prod

275:1992 Determination of the minumum Eval
protective
effectiveness against Test institute
marine borers
TM II05 (Fr)
NL guidance
UK guidelines
58
AVAILABLE OR SOURCE
FROM ELSEWHERE)
330:1993 Field test method for Eval
determining the
relative protective Test institute
effectiveness of a
wood preservative for
use under a coating TM II05 (Fr)
and exposed out of
ground contact: L-joint NL guidance
method
UK guidelines
EN 350- Durability of wood and 8 TM II05 (Fr)

Natural durability of
solid wood - Part 1:
Guide to the principles
of testing and
classification of the
natural durability of
wood
Guide to natural
durability and
treatability of selected
wood species of
importance in Europe
Preservative-treated
Classification of
preservative
penetration and
retention
Guidance on sampling
for the analysis of
preservative-treated
wood
eradicant efficacy in
preventing emergence TM II05 (Fr)
of Anobium punctatum
59
AVAILABLE OR SOURCE
FROM ELSEWHERE)
(De Geer) NL guidance
UK guidelines

solid wood - Guide to
the durability
requirements for wood
to be used in hazard
classes
preventive action
against Hylotrupes TM II05 (Fr+De)
bajulus (Linnaeus) -
Part 1: Larvicidial NL guidance
effect (Laboratory
method) UK guidelines
EN 46- Wood preservatives - 8 TM II05 (De)

preventive action NL guidance
against Hylotrupes
Part 2: Ovicidal effect
(laboratory method)
47:2005/ Determination of the Eval
AC:2007 toxic values against
larvae of Hylotrupes TM II05 (Fr+De)
UK guidelines

eradicant action
against larvae of TM II05 (Fr+De)
Anobium punctatum
(De Geer) (laboratory NL guidance
method)
UK guidelines

protective
effectiveness against TM II05 (Fr+De)
Anobium punctatum
60
AVAILABLE OR SOURCE
FROM ELSEWHERE)
(De Geer) by egg- NL guidance
laying and larval
survival - Part 1: UK guidelines
treatment (Laboratory
method)
protective
effectiveness against TM II05 (Fr+De)
Anobium punctatum
(De Geer) by egg- NL guidance
laying and larval
survival - Part 2: UK guidelines
Application by
impregnation
(Laboratory method)
EN Wood preservatives - 8 Pretreatment procedure before TNsG on Prod
73:1988/ Accelerated ageing of efficacy testing Eval
AC:1992 treated wood prior to
biological testing - TM II05 (Fr)
Evaporative ageing
procedure NL guidance
UK guidelines
EN Wood preservatives - 8 Pretreatment procedure before TNsG on Prod

84:1997 Accelerated ageing of efficacy testing Eval
treated wood prior to
Leaching procedure
NL guidance
UK guidelines
EN/TR Durability of wood and 8 Own searches

04 Determination of
treatability of timber
species to be
impregnated with
wood preservatives -
Laboratory method
EN/TR Wood preservatives - 8 Own searches
04 preventive efficacy
against wood
destroying
basidiomycetes fungi
EN/TS Wood preservatives - 8 Field test, out of ground Test institute
12037:20 Field test method for contact
61
AVAILABLE OR SOURCE
FROM ELSEWHERE)
03 determining the
relative protective
effectiveness of a UK guidelines
wood preservative
contact - Horizontal
lap-joint method
EN/TS Wood preservatives - 8 Own searches
05 preventive
sapstain fungi and
mould fungi on freshly
sawn timber - Field
test
15397:20 Method for natural
06 preconditioning out of
ground contact of
treated wood
specimens prior to
biological laboratory
test
ENV Durability of wood and 8 TM II05 (Fr)
02 Wood-based panels - UK guidelines
Method of test for
determining the
resistance against
wood-destroying
basidiomycetes
ENV Wood preservatives - 8 Laboratory test TM II05 (Fr)
soft rotting micro-fungi
and other soil NL guidance
inhabiting micro-
organisms UK guidelines
ENV Wood preservatives - 8 TM II05 (Fr)

protective NL guidance
wood destroying UK guidelines
basidiomycetes -
treatment
EPPO Blue-Stain fungi of 8 Own searches
PP Softwood
1/194(2)
(1995)
JIS K Test methods for 8 Own searches
62
AVAILABLE OR SOURCE
FROM ELSEWHERE)
1571:200 determining the
4 effectiveness of wood
preservatives and their
performance
requirements
NF B50- Durability of wood and 8 TM II05 (Fr)
100-4 wood-based products -
October Definition of use
2007 classes - Part 4 :
national declaration on
the situation of
biological agents
105-3 wood based products -
February Preservative treated
2008 solid wood - Part 3 :
wood preservation
performance and
treatment certificate -
Adaptation to France
metropolitan territory
and DOM
NF FD Protection - Termites - 8 TM II05 (Fr)
X40-501 Buildings protection
Novembe against termite
r 2005 infestation
NF T72- Wood preservatives. 8 TM II05 (Fr)
050 Technical grade borax.
Septemb Specifications and
er 1986 tests.
052 Boric acid.
Septemb Specifications and
er 1986 tests.
054 Technical grade
Septemb pentachlorophenol.
er 1995 Specificaitons and
tests.
061 Technical grade
Septemb sodium
er 1995 pentachlorophenate.
Specifications and
tests.
065 Technical grade
Septemb sodium pentaborate.
er 1986 Specifications and
tests.
NF T72- Products for protecting 8 TM II05
083/A1 wood surfaces -
63
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Novembe Method of testing
r 1979 resistance to
microorganisms
086 Lasures. Natural
Septemb weathering test.
er 1991
NF X41- Protection of wood. 8 TM II05 (Fr)
521 July Testing methods for
1968 the corrosive action of
wood protection
products on metals.
NF X41- Wood preservatives. 8 TNsG on Prod
541 Determination of the Eval
Septemb protective
er 1994 effectiveness against TM II05
termites of
products designed for
walls, foundations and
masonry. Laboratory
method.
547 Determination of Eval
Decembe longicide efficacy of
r 1992 temporary wood TM II05
protectives for green
sawn timber.
Laboratory method.
NF X41- Wood preservatives. 8 TM II05 (Fr)
548 Determination of
Decembe fongicide efficacy of
r 1992 temporary wood
protectives for fresh
cut wood billets.
Laboratory method.
555 Determination of the
August toxic values against
1982 {chaetomium}
{globosum} kunze.
Soft rotting agent.
NF X41- Wood preservatives – 8 TM II05 (Fr)
580 part Physicochemical
1-10 testing
May 2006
NF XP Wood preservatives. 8 TM II05 (Fr)
X41-540 Termites.
Novembe Determination of anti-
r 1995 termites action for
products used in liquid
phase for ground
64
AVAILABLE OR SOURCE
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treatment (laboratory
method).
NF XP Wood preservatives. 8 TNsG on Prod
X41-540 Termites. Eval
Novembe Determination of anti-
r 1995 termites action for
products used in liquid
phase for ground
method).
X41-542 Anti-termite treatment
Septemb product for floors,
er 1995 walls, foundations and
masonry work.
Accelerated ageing
test of treated
materials prior to
biological testing.
Percolation test.
NF XP Wood preservatives - 8 TM II05 (Fr)
X41-549 Evaluation of fongicide
Decembe efficacy of temporary
r 1999 wood protectives for
green sawn timber -
Site method
NF XP Wood preservatives - 8 TNsG on Prod
X41-549 Evaluation of fongicide Eval
Decembe efficacy of temporary
r 1999 wood protectives for
green sawn timber -
Site method
NWPC Standard for 8 Test institute
testing of wood
preservatives.
Mycological test.
"Jordburk" method - A
NWPC soil block test with
1.4.1.1./7 wood-rotting
0 Basidiomycetes.
testing of wood
preservatives.
NWPC Mycological test.
Standard "Mullåde" method - A
1.4.1.2./7 soil block test in
0 unsterile soil.
Mycological testing of 8 Test institute
NWPC anti-stain
Standard preservatives for
1.4.1.3./7 freshly sawn timber.
9 The Miniboard
65
AVAILABLE OR SOURCE
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method. (Mykologisk
provning av
träskyddsmedel mot
blånad och mögel på
nysågat virke.
Minibrädmetoden.)
Standard for testing of 8 Test institute
NWPC wood preservatives
Standard Mycological test. Field
1.4.2.1./7 test - A field test with
1 stakes.
wood preservatives
NWPC Marine test - A test
Standard against marine
1.4.2.2./7 woodboring organisms
3 in sea water.
prEN 807 Wood preservatives - 8 Own searches
rev Determination of the
and other soil
inhabiting micro-
organisms
AATCC Antibacterial Finishes 9 Applied to textiles Manufacturer
100 - on Textile Materials:
1999 Assessment of
Antibacterial Activity 9 Applied to textiles Manufacturer
AATCC Assessment of Textile
147 - Materials: Parallel
1998 Streak Method
Antifungal Activity, 9 Applied to textiles Manufacturer
Assessment on Textile
Materials:
Mildew and Rot
AATCC Resistance of Textile
30 - 1999 Materials
ASTM Standard Test Method 9 Determination of mold growth
D4576- for Mold Growth resistance of wet blue subject
01(2006) Resistance of Wet to storage and shipping
Blue requirements and intended for
use in leather manufacturing,
assisting the prediction of
storage time before molding
occurs. This test method may
not be suitable to evaluate
fungicides that are inactivated
by proteins, including
alkyldimethylbenzyl
ammonium chlorides.
To allow use of this test
method by any laboratory,
66
AVAILABLE OR SOURCE
FROM ELSEWHERE)
flexibility has been permitted in
times, temperature, and
humidity of incubation,
inoculum, hide sampling area,
and choice of control. These
may be adjusted to fit local
conditions but must be
standardized.
Conclusions about mold
growth resistance are drawn
from the results by comparing
the test with a simultaneously
run control of known
resistance. Success or failure
is determined by the amount of
mold growth relative to the
control.
The degree of correlation

between this test and
commercial quantities of wet
blue in storage or shipment
situations, or both, has not
been fully determined.
ASTM Determining the 9 Evaluates the resistance of Manufacturer,
E2149-01 Antimicrobial Activity non-leaching antimicrobial applied to
of Immobilized treated specimens to the textiles
Antimicrobial Agents growth of microbes under
Under Dynamic dynamic contact conditions.
Contact Conditions This test determines the
antimicrobial activity of treated
specimen by shaking samples
of surface bound materials in a
concentrated bacterial
suspension for a one hour
contact time or other contact
times as specified by the
investigator. The suspension is
serially diluted both before and
after contact and cultured. This
dynamic shake flask test was
developed for routine quality
control and screening tests in
order to overcome difficulties
(including ensuring contact of
inoculum to treated surface) in
using classical antimicrobial
test methods to evaluate
substrate-bound
antimicrobials. This test also
allows for the versatility of
testing contamination due to
67
AVAILABLE OR SOURCE
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such things as hard water,
proteins, blood, serum, various
chemicals, and other
contaminates or
physical/chemical stresses or
manipulations of the
specimens of interest.
The number of viable
organisms in the suspension is
determined and the percent
reduction is calculated based
on initial counts or on retrievals
from appropriate untreated
controls. his method is
intended for those surfaces
having a percent reduction
activity of 50 % to 100 % for
the specified contact time.
Surface antimicrobial activity is
determined by comparing
results from the test sample to
simultaneously run controls.
The presence of a leaching
antimicrobial is both pre- and
post-determined by the
presence of a zone of
inhibition.
ASTM Standard Test Method 9 This test method is designed Own searches
E2180-07 for Determining the to evaluate (quantitatively) the
Activity of Incorporated antimicrobial effectiveness of
Antimicrobial Agent(s) agents incorporated or bound
In Polymeric or into or onto mainly flat (two
Hydrophobic Materials dimensional) hydrophobic or
polymeric surfaces. The
method focuses primarily on
assessing antibacterial activity;
however, other
microorganisms such as yeast
and fungal conidia may be
tested using this method. The
vehicle for the inoculum is an
agar slurry which reduces the
surface tension of the saline
inoculum carrier and allows
formation of a "pseudo-
biofilm," providing more even
contact of the inoculum with
the test surface. This test
method facilitates the testing of
hydrophobic surfaces by
utilizing cells held in an agar
68
AVAILABLE OR SOURCE
FROM ELSEWHERE)
slurry matrix.
This method can confirm the
presence of antimicrobial
activity in plastics or
hydrophobic surfaces and
allows determination of
quantitative differences in
antimicrobial activity between
untreated plastics or polymers
and those with bound or
incorporated low water-soluble
antimicrobial agents.
Comparisons between the
numbers of survivors on
preservative-treated and
control hydrophobic surfaces
may also be made. This test
method, as written, is
inappropriate to determine
efficacy against biofilm cells,
which are different both
genetically and metabolically
than planktonic cells used in
this test. The procedure also
permits determination of
"shelf-life" or long term
durability of an antimicrobial
treatment which may be
achieved through testing both
non-washed and washed
samples over a time span.
ASTM Standard Test Method 9 Rapid screening evaluating Own searches
E2471-05 for Using Seeded-Agar (qualitatively) the presence of
for the Screening antimicrobial (both
Assessment of antibacterial and antifungal)
Antimicrobial Activity activity in or on the carpet face
In Carpets fiber or incorporated into the
backing structure of the carpet
(or both). The method
simulates actual use
conditions that may occur on
carpets (for example, food and
beverage spills, soiling from
foot traffic, prolonged moisture
exposure) and provides a
means to screen for activity
and durability of an
antimicrobial treatment under
conditions of organic loading.
This test method provides for
the simultaneous assessment
of multiple carpet components
69
AVAILABLE OR SOURCE
FROM ELSEWHERE)
for antimicrobial activity.
Carpets may be cleaned prior
to testing in order to assess
the durability of the
antimicrobial effect. The
method can assess the
durability of the antimicrobial
treatments on new carpets,
and on those repeatedly
shampooed or exposed to in-
use conditions.
Half strength (nutrient and
agar) tryptic soy agar is used
as the inoculum vehicle for
bacteria and half strength
potato dextrose agar as the
inoculum vehicle for mold
conidia. Use of half strength
agars may reduce undue
neutralization of an
antimicrobial due to excessive
organic load.
This test method
simultaneously evaluates (both
visual and stereo-microscopic)
antimicrobial activity both at
the fiber layer and at the
primary backing layer of
carpet.
ASTM Revision of E2180-01 9 Evaluates (quantitatively) the Manufacturer

WK16397 Standard Test Method antimicrobial effectiveness of (updated),
for Determining the agents incorporated or bound applied to
Activity of Incorporated into or onto mainly flat (two textiles.
Antimicrobial Agent(s) dimensional) hydrophobic or
In Polymeric or polymeric surfaces. The
Hydrophobic Materials method focuses primarily on
assessing antibacterial activity;
however, other
microorganisms such as yeast
and fungal conidia may be
tested using this method.This
method can confirm the
presence of antimicrobial
activity in plastics or
hydrophobic surfaces. The
vehicle for the inoculum is an
agar slurry which reduces the
surface tension of the saline
biofilm," providing more even
70
AVAILABLE OR SOURCE
FROM ELSEWHERE)
contact of the inoculum with
the test surface.
The method allows
determination of quantitative
differences in antimicrobial
activity between untreated
plastics or polymers and those
with bound or incorporated low
water-soluble antimicrobial
agents. Comparisons between
the numbers of survivors on
preservative-treated and
control hydrophobic surfaces
may also be made. The
procedure also permits
determination of "shelf-life" or
long term stability of an
antimicrobial treatment which
may be achieved through
testing both non-washed and
washed samples over a time
span.
ASTM Standard Test Method 9 Designed to evaluate Own searches
WK4757 for the Assessment of (qualitatively) the presence
Antimicrobial Activity and effectiveness of
In Carpets; Seeded- antimicrobial preservatives in
Agar Overlay Screen or on carpets. This method can
be used to evaluate both the
antibacterial and antifungal
activity
This method is useful for
assessing the durability of the
antimicrobial treatments
because it can be performed
on both new carpets and those
that have been repeatedly
shampooed or exposed to in-
use conditions.
This method utilizes either
tryptic soy agar as the
inoculum vehicle for bacteria
or potato dextrose agar as the
inoculum vehicle for mold
conidia. . Full compliment
agars can be used to mimic
high soil or organic loads on
carpets or partial nutrient
complement agars can be
used to mimic lightly soiled
conditions on carpet. This
method allows for the
simultaneous evaluation (both
71
AVAILABLE OR SOURCE
FROM ELSEWHERE)
visual and stereo-microscopic)
of antimicrobial activity both at
carpet.
Determination of the 9 Manufacturer
BS resistance of textiles
6085:199 to microbiological
2 deterioration
EN ISO Textiles - 9 Manufacturer
11721- Determination of the
1:2001 resistance of cellulose-
containing textiles to
micro-organisms - Soil
burial test - Part 1:
Assessment of rot-
retardant finishing
(ISO 11721-1:2001)
EN ISO Textiles - 9 Own searches
11721- Determination of the
2:2003 resistance of cellulose-
containing textiles to
micro-organisms - Soil
burial test - Part 2:
Identification of long-
term resistance of a rot
retardant finish (ISO
11721:2003)
EN ISO Textile fabrics - 9 Manufacturer
20645:20 Determination of
04 antibacterial activity -
Agar diffusion plate
test (ISO 20645:2004)
EN ISO Textiles - 9 Applied to textiles Manufacturer
07 antibacterial activity of Reference material: non Laboratory
antibacterial finished treated polyester
products (ISO
20743:2007)
JIS L Testing for 9 Antimicrobial activity. Test institute
1902: antibacterial activity Reference material: e.g. non
2002 and efficacy treated polyester.
on textile products
Manufacturer
Antimicrobial Products 9 Applied to textiles Manufacturer
JIS Z – Test for antimicrobial
2801: activity Test institute
2000 and efficacy
JIS Z Methods of test for 9 Applied to textiles Manufacturer
72
AVAILABLE OR SOURCE
FROM ELSEWHERE)
2911: fungus resistance
1992 (PT
9)
NF X41- Protection of plastics. 9 TM II05
513 Part 1. Testing method
August for resistance of
1961 ingredients to
microorganisms.
NF X41- Protection of the 9 TM II05
515 plastic materials - Part
March 3: Test method of the
1962 resistance of materials
and apparatus to
microorganisms
NF XP Properties of textiles - 9 Manufacturer
G39-010 Textiles and polymeric
May 2000 surfaces having
antibacterial properties
- Characterisation and
measurement of
antibacterial activity
OECD Guidance document 9 Own searches
(ENV/JM/ on the evaluation of
BCID(200 the efficacy of
7)5) antimicrobial treated
articles with claims for
external effects
Determination of the 9 Applied to textiles Manufacturer
SN 195 Antibacterial Activity,
920 - Agar Diffusion Plate
1994 Test
SN 195 Antimycotic Activity
1994 Test
SN 195 Determination of the 9 Applied to textiles Manufacturer
924 - antibacterial activity,
1983 germ count method
ASTM Standard Test Method 10 This test method can be used Own searches
WK8681 for Resistance to Mold to evaluate the comparative
Growth on Interior resistance of coated building
Coated Building products to accelerated mold
Products in an growth. Performance at a
Environmental certain rating does not imply
Chamber any specific period of time for
a mold free surface. However,
a better rated panel would be
expected to perform better in
actual end use.
An environmental chamber
and the conditions of operation
to evaluate in a 4-week period
73
AVAILABLE OR SOURCE
FROM ELSEWHERE)
the relative resistance of
coated building products to
surface mold growth in a
controlled interior environment
is described. The apparatus is
designed so it can be easily
interested party.
ASTM Test Method for 10 This test method can be used Own searches
WK14960 Standard Test Method to evaluate the comparative
for the Resistance of resistance of uncoated building
Mold Growth on products to accelerated mold
Uncoated Interior growth. The apparatus
Building Products in an (environmental chamber) is
Environmental designed so it can be easily
Chamber. built or obtained by any
interested party.
4-week evaluation period.
Relative resistance of
uncoated interior building
products to surface mold
growth in a controlled interior
environment is evaluated, but
performance at a certain rating
does not imply any specific
period of time for a mold free
surface.
BS 3900 Methods of test for 10 Provides only a methodology TNsG Prod Eval
paints. Part G6. for production of a test surface
Assessment of for exposure by inoculation UK guidelines
resistance to fungal with mold growth. The test
growth. BSI London, therefore has to be modified to
UK be used as a test method for
assessing interior surface
biocides.
ENV Durability of wood and 10 Simulated use test TNsG Prod Eval
97 Assessment of the TM II05 (Fr)
effectiveness of a
masonry fungicide to UK guidelines
prevent growth into
wood of Dry Rot
Serpula lacrymans
(Schumacher ex Fries)
S.F. Gray - Laboratory
method
OECD Update on work to 10 Own searches
(ENV/JM/ validate efficacy
BCID(200 methods for
7)3) antimicrobials used on
hard surfaces
SM022 Determining the 10 Resistance of masonry Manufacturer
74
AVAILABLE OR SOURCE
FROM ELSEWHERE)
(in resistance to fungal coatings to fungal growth.
house) growth Test substrate: Masonry
coatings on paper
Test germ: Aspergillus niger
and Penicillum funiculosum
"Sufficiently finished against
fungal growth" if max 1%
growth.
SM022a Determining the 10 Resistance of masonry Manufacturer
(in resistance to fungal coatings to fungal growth.
house) growth Test substrate: Masonry
coatings on paper
Test germ: Aspergillus niger
and Penicillum funiculosum
"Sufficiently finished against
fungal growth" if max 1%
growth.
SM023 Determining the 10 Resistance of masonry Manufacturer
(in resistance to algal coatings to algal attack.
house) growth Test substrate: masonry
coatings on paper.
Test germ: Scenedesmus
vacuolatus.
No algal growth on the test
pieces after 2 weeks:
"Effectively protected against
algal growth"
ASTM Standard Test Method 11 Efficacy of microbicides Manufacturer
E645-07 for Efficacy of (algicides, bactericides, and
Microbicides Used in fungicides), evaluated using
Cooling Water simulated or real cooling tower
Systems water against (1) microbes
from cooling water, (2)
microbes in microbiological
deposits (biofilms) from
operating cooling systems, or
(3) microorganisms known to
contaminate cooling water
systems, or a combination
thereof. Choice of enumerating
method may vary, e g pour
plate, spread plate, MPN
techniques etc. Test
concentrations also vary but
are usually between 1 and 50
mg/l. Exposure time varies
with mode of action but
includes 3h contact time. For
bacteria 48 h incubation, fungi
75
AVAILABLE OR SOURCE
FROM ELSEWHERE)
48h-7d, for algae 5-14d.
Even if using cooling water
and deposits/biofilm obtained
from the field, laboratory
results may not be totally
predictive of microbicidal
effectiveness in the field (due
to e g variability in
environmental factors). If
solvents are added, a solvent
control is included. If initial
count in cooling water control
samples are <10E5 bact/ml (or
<10E3 CFU/ml fungi or algae),
the test is invalid. In addition,
the viable counts of fungi/algae
in the control must at least be
equal to the numbers at time 0.
The untreated control should
show a stable population with
no more than 0.5 log increase
or decrease in growth during
the test perod. Results are
expressed as “Log reduction in
number of microorganisms at
each biocide concentration”:
log (initial count of
microorganisms of the control
sample) – log (number of
microorganisms detected at a
given biocide concentration
after a specific contact time). If
1 log reduction: corresponds to
90% kill, if 2 log reduction
corresponds to 99% kill, 3 log
reduction to 99.9% kill.
Not Field trial in a Plant 11 The antimicrobial effectiveness Manufacturer
available recirculating cooling of biocide was determined by
system monitoring chemical and
physical cooling water
characteristics, microbiological
parameters such as viable
counts of bacteria, fungi and
algae, and operational and
engineering parameters of the
tested cooling system
Analysis methods used for
microbial counting were from:
Standard methods for the
Waste Water (APHA 1992),
and in-house Laboratory
76
AVAILABLE OR SOURCE
FROM ELSEWHERE)
SOPs. Efficacy was
determined by a reduction in
microbial counts
SM019 S&M Boko test 11 Preservation of water diluted Manufacturer
(in coolants.
Test germ: bacteria, moulds,
Yeasts
4 evaluation levels. 12 cycles.
ASTM Standard Test Method 11 or Laboratory test method is used Communication
E723-07 for Efficacy of 12? to determine the efficacy of an with UK (HSE)
Antimicrobials as antimicrobial for preventing
Preservatives for bacterial spoilage of in-process
Aqueous-Based pigment suspensions, dye
Products Used in the solutions, pulp slurries, starch
Paper Industry solutions, polymers, sizing
(Bacterial Spoilage) agents, latex emulsions, and
other aqueous-based materials
used in the paper industry from
bacterial spoilage. Test
organisms may vary but
should lie above 10E6 CFU/ml.
Bacterial numbers in the
sample are determined at
variable time periods and
compared to a control with no
bactericide. Material to be
preserved is used as
substrate, but for some
materials nutrients may be
added. Results are expressed
as % kill: [(control plate count-
test plate count)/ control plate
count]x100 at each sampling
time.
ASTM Standard Test Method 11 or Laboratory method that Communication
E875- for Efficacy of Fungal 12? determines if a fungal control with UK (HSE)
00(2005) Control Agents as agent is effective to preserve
Preservatives for pigment suspensions, dye
Aqueous-Based solutions, pulp slurries, starch
Products Used in the solutions, polymers, sizing
Paper Industry agents, latex emulsions, and
other specific aqueous-based
materials to prevent spoilage
of in-process aqueous-based
products used in the paper
industry. Will also prevent
spore germination. Plates are
77
AVAILABLE OR SOURCE
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incubated for 7d or until control
plates shos sufficient growth
for rating. The exposure time
should correspond to the real
time desired, often 6w.
Separate evaluations should
be made on a representative
type for each specific class of
product to be preserved.
Growth is determined by
visible signs of deterioration in
the sample and by obtaining
fungal numbers and comparing
them to a sample without
fungal control agents. A proper
level of fungal control agent
prevents deterioration and
reduces and keeps organisms
to an acceptable level in the
test material, which is
determined by the tester or
user. Often a scale from 0 to 4
(heavy growth).
ASTM Test Method for 12 A procedure for testing the Own searches
WK Evaluating the Efficacy efficacy of a liquid microbicide
17314 of a Liquid against biofilm grown on
Microbiocide against coupons removed from biofilm
Biofilm Bacteria reactors standardized to grow
a repeatable biofilm, such as
the CDC Biofilm Reactor
(ASTM Method E 2562) or
Rotating Disk Biofilm Reactor
(ASTM Method E 2196). The
preparation of the liquid
microbicide and exposure time
are completed according to
manufacturer’s instructions for
use. The liquid microbicide is
tested at room temperature
under static conditions. Biofilm
population density is recorded
as log10 colony forming units
per surface area. Efficacy is
determined by calculating the
log reduction in viable cells.
The microbicide is tested at its
use dilution for the
recommended time.
ASTM Standard Guide for 12 The purpose of this guide is to Own searches
E1427- Selecting Test inform the investigator of
00e1 Methods to Determine methods that can be used for
the Effectiveness of biofilm formation and
78
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Antimicrobial Agents measurement, allowing
and Other Chemicals development of test
for the Prevention, procedures for determining the
Inactivation and effectiveness of chemical
Removal of Biofilm treatments for prevention,
inactivation, and removal of
unwanted biofilm. This guide is
not an exhaustive survey of
biofilm methods.
ASTM Standard test method 12 A procedure to evaluate the Communication
E1839- for efficacy of efficacy of slimicides for the with UK (HSE)
07 slimicides for the control of bacterial and fungal
paper industry – slimes in paper mill systems Manufacturer
bacterial and fungal and their counterparts. This
slime test method is run in acid,
alkaline, or acid and alkaline
conditions to determine the
efficacy of the slime control
agent. The test conditions may
be modified to reflect intended
use patterns in typical paper
mill systems, including use of
actual paper mill furnish.
Efficacy based on a %
reduction on CFU.
ASTM Revision of E1839- 12 Efficacy of slimicides for the Own searches
WK14214 96(2002) Standard control of bacterial and fungal
Test Method for slimes in paper mill systems
Efficacy of Slimicides and their counterparts.
for the Paper Industry-
-Bacterial and Fungal
Slime
EEC: 12 Used in the process of oil Communication
1988 Algal inhibition test recovery with UK (HSE)
Report Biological Test 12 Efficacy test against Alga: Communication
EPS Method: Growth Selenastrum capriconutum with UK (HSE)
1/RM/25 Inhibition Test Using
Environm the Freshwater Alga Used in the process of oil
ent Selenastrum recovery
Canada. capriconutum
1992
ASTM Standard Practice for 13 Laboratory procedures for TNsG on Prod
E2275- Evaluating Water- rating the relative inherent Eval
03e1 Miscible Metalworking bioresistance of water-miscible
(replaces Fluid Bioresistance metalworking fluids, the
D3946 and Antimicrobial bioresistance attributable to
and Pesticide Performance augmentation with
E686) antimicrobial pesticides or
both, for determining the need
for microbicide addition prior to
or during fluid use in
metalworking systems and for
79
AVAILABLE OR SOURCE
FROM ELSEWHERE)
evaluating microbicide
performance.
Relative bioresistance is
determined by challenging
metalworking fluids with a
biological inoculum that may
either be characterized
(comprised of one or more
known biological cultures) or
uncharacterized (comprised of
biologically contaminated
metalworking fluid or one or
more unidentified isolates from
deteriorated metalworking
fluid). Challenged fluid
bioresistance is defined in
terms of resistance to biomass
increase, viable cell recovery
increase, chemical property
change, physical property
change or some combination
thereof.
This practice is applicable to
antimicrobial agents that are
incorporated into either the
metalworking fluid concentrate
or end-use dilution. It is also
applicable to metalworking
fluids that are formulated using
non-microbicidal, inherently
bioresistant components.
The results of tests completed
in accordance with this
practice should be used only to
compare the relative
performance of products or
microbicide treatments
included in a test series.
Results should not be
construed as predicting actual
field performance.
ASTM Standard Test Method 13 TNsG on Prod
E979- for Evaluation of Eval
91(2004) Antimicrobial Agents
as Preservatives for
Invert Emulsion and
Other Water
Containing Hydraulic
Fluids
ASTM New Standard Test 13 Determines the relative Own searches
WK8252 Method for bioresistance of aqueous
Determining metalworking fluids towards
80
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Resistance of non-tuberculous (NTM), rapidly
Aqueous Metalworking growing( RGM), environmental
Fluids towards Non- mycobacteria by challenging
Tuberculous, them with a mycobacterial
Environmental inoculum isolated from actual
Mycobacteria spoiled metalworking fluid field
samples from the user/s site.
In order to simulate field
conditions, another challenge
inoculum consisting of a
mixture of common
metalworking fluid spoilage
microorganisms originating
from actual MWF field samples
is also used.
IBRG A Standardized 13 TNsG on Prod
(draft Screening Method for Eval
MWF) Determining the
(1993) Bioresistance of and
Evaluating Biocides in
Aqueous Metal
Working Fluids
Not Challenge Testing in 13 The purpose of the method Manufacturer
available Metal Working was to determine the efficacy
Emulsion (in-house of preservatives against
laboratory study) bacterial and fungal
contamination in Metal
Working Fluids (MWF).
The method was conducted by
inoculating once a week during
10 weeks mixed bacteria and
optionally mixed fungi into
water based MWF emulsions
containing various amounts of
the test substance diluted in
3% synthetic oil. After each
week a sample of the emulsion
was plated. After plate
compared to the control The
criteria for measuring good
microbial count of <10 cfu/ml
after 10 inoculations for all
microorganisms.
Rawlinso A recirculating test rig 13 TNsG on Prod
n and for the investigation of Eval
Shennan, metal-working fluid
1987. spoilage. In Industrial
microbiological testing.
Edited by Hopton and
81
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Hill, Blackwell
Scientific Publications,
Oxford. ISBN 0 632
01793 7. pp 227-231
RENAUL Evaluation of the 13 TNsG on Prod
T Biostability of Aqueous Eval
D551721 Metal Working Fluids
(1987)
SABS South African standard 13 TNsG on Prod
1435- specification for Eval
1987 biocides for use in
emulsions of aqueous
metal working fluid and
aqueous hydraulic
fluid.
UK MOD Cutting fluid, soluble, 13 TNsG on Prod
91-70 biostable joint service Eval
issue designation ZX-9
(1990)
BBA 9 - Richtlinie für die 14 TNsG on Prod
3.1 Prüfung von Eval
Nagetierbekämpfungs
mitteln gegen UK guidelines
Hausmaüse
Manufacturer

Nagetierbekämpfungs
mitteln gegen UK guidelines
Wanderratten
Manufacturer
CEB Laboratory test 14 Laboratory test method TMII Fr

(1981) method to evaluate the
efficacy of rodenticidal
products in rats
CEB Trial method to 14 TMII Fr

(1981) evaluate the efficacy of
rodenticidal products
against rats under
practical conditions
EPA/OPP Standard Norway Rat 14 Anticoagulant rodenticide test TNsG on Prod
Protocol and Roof Rat against Norway rat/Roof rat Eval
Number: Anticoagulant Liquid applied as liquid bait
1.201 Bait Laboratory Test NL guidance
Method
UK guidelines
EPA/OPP Standard House 14 Anticoagulant rodenticide test TNsG on Prod
82
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Protocol Mouse Anticoagulant against House mouse applied Eval
Number: Liquid Bait Laboratory as liquid bait
1.202 Test Method NL guidance
UK guidelines

Number: Anticoagulant Dry Bait applied as dry bait
1.203 Laboratory Test NL guidance
Method
UK guidelines

Number: Dry Bait Laboratory as Dry bait
UK guidelines
EPA/OPP Standard Norway 14 Anticoagulant rodenticide test TNsG on Prod

Protocol Rat/Roof Rat against Norway rat/Roof rat Eval
Number: Anticoagulant Tracking applied as Tracking powder
1.205 Powder Efficacy NL guidance
Laboratory Test
Method UK guidelines
EPA/OPP Standard Norway 14 Acute rodenticide test against TNsG on Prod

Protocol Rat/Roof Rat Acute Norway rat/Roof rat applied as Eval
Number: Liquid Bait Laboratory Liquid bait
UK guidelines
EPA/OPP Standard House 14 Acute rodenticide test against TNsG on Prod

Protocol Mouse Acute Liquid House mouse applied as Eval
Number: Bait Laboratory Test Liquid bait
1.208 Method NL guidance
UK guidelines

Number: Dry Bait Laboratory Dry bait
UK guidelines

Protocol Mouse Acute Dry Bait House mouse applied as Dry Eval
Number: Laboratory Test bait
83
AVAILABLE OR SOURCE
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UK guidelines

Number: Tracking Powder Tracking powder
1.211 Efficacy Laboratory NL guidance
Test Method
UK guidelines

Number: Tracking Powder as Tracking powder
1.212 Efficacy Laboratory NL guidance
Test Method
UK guidelines
EPA/OPP Standard Norway 14 Anticoagulant rodenticide test TNsG on Prod

Protocol Rat/Roof Rat against Norway rat/Roof rat Eval
Number: Anticoagulant Wax applied as Wax block and wax
1.213 Block and Wax Pellet pellet NL guidance
Laboratory Test

Number: Wax Block and Wax as Wax block and wax pellet
1.214 Pellet Laboratory Test NL guidance
Method
UK guidelines

Number: Anticoagulant applied as Placepark dry bait
1.217 Placepack Dry Bait NL guidance
Laboratory Test

Number: Placepack Penetration as Placepark penetration
Method
UK guidelines
EPA/OPP Standard Norway Rat 14 Acute rodenticide test against TNsG on Prod
Protocol and Roof Rat Acute Norway rat/Roof rat applied as Eval
Number: Placepack Penetration Placepark penetration
Method
UK guidelines
84
AVAILABLE OR SOURCE
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Protocol Mouse Acute House mouse applied as Eval
Number: Placepack Dry Bait Placepark dry bait
Method
UK guidelines
EPA/OPP Proposed Norway Rat 14 Anticoagulant rodenticide test TNsG on Prod

Protocol Anticoagulant against Norway rat applied as Eval
Number: Technical and Technical and concentrated
1.221 Concentrated Dry Bait dry bait NL guidance
Laboratory Test
EPA/OPP Proposed Norway Rat 14 Acute rodenticide test against TNsG on Prod
Protocol Acute Technical and Norway rat applied as Eval
Number: Concentrated Dry Bait Technical and concentrated
1.222 Laboratory Test dry bait NL guidance
Method
UK guidelines
EPA/OPP Proposed House 14 Anticoagulant rodenticide test TNsG on Prod

Protocol Mouse Anticoagulant against House mouse, applied Eval
Number: Technical and as Technical and
1.225 Concentrated Dry Bait concentrated dry bait NL guidance
Laboratory Test
EPA/OPP Proposed House 14 Acute rodenticide test against TNsG on Prod

Protocol Mouse Acute House mouse applied as Eval
Number: Technical and Technical and concentrated
1.226 Concentrated Dry Bait dry bait NL guidance
Laboratory Test
EPA/OPP Proposed House 14 Acute rodenticide test against TNsG on Prod

Protocol Mouse Acute Tracking House mouse applied as Eval
Number: Powder Efficacy Tracking powder
Method
UK guidelines
14 NL guidance
EPPO
PP Field rodents TM II05 (Fr)
1/169(2 (Microtus, Arvicola)
Field tests against 14 NL guidance
EPPO synanthropic rodents
PP (Mus musculus, Rattus TM II05 (Fr)
1/114(2) norvegicus, R. rattus)
EPPO Laboratory and field 14 NL guidance
PP tests for the evaluation
85
AVAILABLE OR SOURCE
FROM ELSEWHERE)
1/97(2) of rodenticidal dusts TM II05 (Fr)
Laboratory tests for 14 NL guidance
evaluation of the
toxicity and TM II05 (Fr)
acceptability of
EPPO rodenticides and Test institute
PP rodenticide
1/113(2) preparations
EPPO 14
PP Non-target effects of NL guidance
1/197(1) rodenticides
EPPO Rodent repellents 14 Own search
PP against debarking of
1/200(1) trees
EPPO 14 Own search
PP Rodent seed
1/199(1) repellents
Testing rodents for 14 NL guidance
EPPO resistance to
PP anticoagulant TM II05 (Fr)
EPPO Guidelines for the 14 TNsG on Prod
(1982) biological evaluation of Eval
rodenticides N°1.
Laboratory tests for UK guidelines
evaluation of the
toxicity and Manufacturer
acceptability of
rodenticides and
rodenticide
preparations
rodenticides. Field
tests against UK guidelines
synanthropic rodents
(Mus musculus, Rattus Manufacturer
norvegicus, rattus
rattus)
rodenticides.
Laboratory and field UK guidelines
tests for the evaluation
of rodenticidal dusts.
US EPA 15 TNsG on Prod
96-5 Avian Toxicants Eval
US EPA Avian Frightening 15 TNsG on Prod
96-7 Agents Eval
ASTM Standard practice for 17 Adaptable to both lotic and Communication
D4131- sampling fish with lentic situations in littoral and with UK (HSE)
84 Rotenone limnetic areas.
86
AVAILABLE OR SOURCE
FROM ELSEWHERE)
(2005)
Standard guide for 17 2 to 8-day exposures, Communication
conducting acute depending on the species. with UK (HSE)
toxicity tests on test Static, renewal, or flow-through
ASTM materials with fishes, procedures.
E729-96 macroinvertebrates Usually reported as LC50 or
(2007) and amphibians EC50.
ASTM Standard guide for 17 A flow through test, beginning Communication
WK6845 conducting early life- before hatch and ending after with UK (HSE)
[revision stage toxicity test with hatch. 28 to 120-day
of E fishes (depending on species)
1241- continuous exposure.
98 (2004)
]
EPA 72- 17 Communication
5 Life-cycle test of fish with UK (HSE)
Simulated or actual 17 Communication
EPA 72- field testing for aquatic with UK (HSE)
7 organisms
OECD 17 Communication
203 with UK (HSE)
(1992) Fish, Acute toxicity test
OECD Fish, Prolonged 17 Communication
204 toxicity test: 14-day with UK (HSE)
(1984) study
US EPA Acute toxicity test for 17 Communication
72-1 freshwater fish with UK (HSE)
US EPA 17 Communication
96-2 Fish control agents with UK (HSE)
810.3000 General 18 General guide Manufacturer
(1999) Considerations for
Efficacy of Invertebrate
Control Agents
UK guidelines
Assesment of the Anti- 18 Applied to textiles Manufacturer

House Dust Mite
Properties
AATCC of Textiles under Long-
194-2006 Term Test Conditions
ASTM Standard Test Method 18 Determines the effectiveness Own searches
E1517- for Determining the of ovicidal materials in liquid,
99(2006) Effectiveness of gel, cream, or shampoo form
Liquid, Gel, Cream, or against the ova (that is, eggs
Shampoo Insecticides or nits) of the human louse,
Against Human Louse Pediculus humanus.
OVA This test method consists of
five replicates for a statistical
87
AVAILABLE OR SOURCE
FROM ELSEWHERE)
comparison of formulations.
ASTM Standard Test Method 18 Determines the relative Own searches
E652- for Nonresidual Liquid efficiency of household and
91(2003) Household industrial-use, contact
Insecticides Against insecticides dissolved in base
Flying Insects oils and applied in spray
formulations. It is developed to
test insecticides against
house flies (Musca domestica,
L), but test data may also be
adequate to support label
claims for the use of the
products against mosquitoes,
gnats, flying moths, wasps,
and certain other small flying
insects. Not designed to
measure the residual action of
the spray formulation.
ASTM Standard Test Method 18 The test determines the UK guidelines
E653- for Effectiveness of relative efficacy of aerosol and
91(2003) Aerosol and pressurized space spray
Pressurized Space insecticide formulations
Spray Insecticides against house flies (Musca
Against Flying Insects domestica, L) strains and, with
modifications in dosage, other
flying insects. Test data
obtained by this test method
may also be adequate to
support label claims for the
use of the product against
mosquitoes, gnats, flying
moths, wasps, and certain
other small flying insects.
This test method is not
designed to measure the
residual activity.
The test may be conducted
using approximately 100
house flies per test (small
group) or 500 flies per test
(large group). Selected
reference standards are the
Official Test Aerosol II (OTA II)
for oil based aerosol products
and Tentative Official Aqueous
Pressurized Spray (TOAPS)
for water based aerosol
products. Aerosol test
knockdowns: % down of total
flies at 5, 10, 15 minutes after
application. Aerosol test knock
down mortality: dead knocked
88
AVAILABLE OR SOURCE
FROM ELSEWHERE)
down x100/total flies. These
numbers should on average be
equal to, greater than or no
more than 5% points below the
corresponding numbers of the
reference in order to meet the
standard. No statement on
precision or bias, only whether
conformance to criteria for
success specified in the
procedure.
ASTM Standard Test Method 18 Test of insecticides against UK guidelines
E654- for Effectiveness of crawling insects: cockroaches
96(2003) Aerosol and Determines the relative
Pressurized Spray efficiency of aerosol and
Insecticides Against pressurised spray formulations
Cockroaches against cockroaches, but test
data by this test method may
also be adequate to support
claims for use of the product to
control the exposed or
accessible stages of silverfish,
ants, centipedes, spiders, and
certain stored product pests.
Applied as direct sprays for 30
s. on last instar nymphs.
Observation period: 48h. The
test is not designed to
measure the residual action.
Ten groups with 20 organisms
in each. The test is run in
conjunction with the Official
Test Aerosol II (OTA II) (or
Tentative Official Aqueous
Pressurized Spray (TOAPS)
as the standard basis of
comparison. The mortality after
24h should be between 50 and
75% when testing with the
OTA. The test specimens meet
the standard if average %
dead and moribound is equal
to, above or within 10% points
less than average % dead of
the OTA series after 48h.
Precision or bias is not
specified, only states whether
conforms to efficacy criteria.
ASTM Standard Test Method 18 Test of insecticides against Own searches
E938-05 for Effectiveness of crawling insects: human louse.
Liquid, Gel, or Cream Only gels or creams that
Insecticides Against liquefy at 32°C (90°F) can be
89
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Adult Human Lice tested).
Five replicates, for a statistical
comparison of formulations.
BS 4172- Hand-held pressurized 18 TNsG on Prod
1:1999 aerosol dispensers Eval
against houseflies.
Specification for TM II05 (Fr)
insecticidal
performance UK guidelines

against houseflies.
Method for TM II05 (Fr)
determination of
insecticidal UK guidelines
performance
CEB 107 Trial method to 18
(1985) evaluate the efficacity
of insecticidal products
for the control of stable
flies in premises for
the rearing of domestic
animals under
CEB 135 Trial method to 18 Surface treatments of storage TM II05 (Fr)
(1987) evaluate the efficacy of premises of products
insecticidal or miticidal
products for
treatments of storage
premises of products
for animals or plants
CEB Laboratory test 18 Space treatments TM II05 (Fr)
135bis method to evaluate the
(1996) efficacy of insecticidal
products in premises
for the storage,
industrial processing
and sale of products
from animals or plants
CEB 159 Trial method to 18 TM II05 (Fr)
insecticidal products
for the control of
cockroaches in
buildings under
insecticidal bait
products against
common species
90
AVAILABLE OR SOURCE
FROM ELSEWHERE)
a fumigant for insect
control in premises for
the storage,
processing and
production of food
fumigants for insect
control in stored
products
EPPO Laboratory testing of 18 UK guidelines
PP plant protection
1/204(1) products against insect
and mite pests of
stored plant products
18 TM II05 (Fr)
EPPO Space and structural
PP treatments of store UK guidelines
1/202(1) rooms
EPPO The EPPO Conference 18 TNsG on Prod
Bulletin, on Fumigation, Paris, Eval
15 Pages 1983
1-119, UK guidelines
Paris
(1983)
EPPO, EPPO 18 TNsG on Prod
Paris Recommendations on Eval
(1982) fumigation standards
(2nd Edition) UK guidelines
ISO Textiles - 18 Applicable to all textiles TNsG on Prod
3998:197 Determination of containing animal fibres in any Eval
7 resistance to certain proportion. Conditioned
insect pests voracity control specimens and TM II05 (Fr)
test specimens of known mass
are placed in contact with UK guidelines
selected larvae for 14 days.
The loss in mass of all
specimens and the condition of
the test larvae are ascertained
to assess the resistance of
each test specimen.
MS 1004 Specification for 18 Manufacturer
part 1 mosquito vapourising
(2002) mat: part 1: physical
and chemical
requirements (first
revision)
(2002) mat: part 2: method for
91
AVAILABLE OR SOURCE
FROM ELSEWHERE)
evaluation of biological
efficacy - glass
chamber method (first
revision)
efficacy - glass
cylinder method
efficacy - peet grady
method
MS 1008 Method for 18 Manufacturer
(1986) determination of
delivery rate of aerosol
dispenser
(1992) household insecticidal
residual spray aerosol
space spray aerosol
brimful capacity of
aerosol cans
part 1 mosquito electric liquid
(1996) vaporizer: Part 1:
physical and chemical
requirement
(1996) vapourizer: part 2:
method for evaluation
of biological efficacy -
glass chamber method
method for evaluation
glass cylinder method
MS 1497 Methods of biological 18 Manufacturer
(2000) evaluation of the
efficacy of repellent -
bioassay method for
mosquito repellent on
92
AVAILABLE OR SOURCE
FROM ELSEWHERE)
human skin
part 1 mosquito coils: Part 1:
(1998) physical and chemical
requirements (third
revision)
(1996) method for evaluation
(first revision)
(1998) method for evaluation
peet grady method
NF G39- Properties of textiles - 18 Manufacturer
011 April Textiles and polymeric
2001 materials having TM II05 (Fr)
antiacarien properties -
Characterisation and
measurement of
antiacarien activity
NF T72- Insecticides for flying 18 TNsG on Prod
320 insects. Insecticide Eval
March distributed under
1977 pressure ("aerosol" TM II05 (Fr)
type). Determination of
the efficiency rating. UK guidelines
Insecticides for flying 18 TM II05 (Fr)

insects. Permanent
NF T72- insecticide distributor.
March efficiency rating and
1977 the regularity rating.
NF X41- Protection of textiles. 18 TM II05 (Fr)
516 Protection against
January certain insect pests.
1980 Methods of testing.
Not Field Evaluation of in a 18 Structures in this study were Manufacturer
available Termite Control Baiting supplied by cooperating pest
System control companies. A number
of the structures had been
identified as "problem houses"
by the pest control operator.
Stations were inspected at
approximately monthly (when
active), and bi-monthly (when
not active) intervals for termite
activity, with the condition of
93
AVAILABLE OR SOURCE
FROM ELSEWHERE)
the station (active, inactive and
baited) being noted on an
inspection form. When activity
was found in a station, bait
was placed in the station.
The thin wood strips of the
baiting station were fan
sprayed with product solution
using a compressed air
sprayer. Product was mixed in
the acetone such that when
the solution was applied to the
strips, various concentrations
of product were imparted to
the wood strips .A tumble
technique was employed to
prevent uneven concentrations
of active ingredient in the
sawdust matrix. Efficacy was
measured by elimination of
termite infestation
Not In house Laboratory 18 Formulation, applied as a Manufacturer
available Study of the Efficacy residual spray, was assessed
against Reticulitennes for efficacy against
fIavipes cockroaches (Blatta orientalis,
Blatella germanica and
Periplaneta americana) under
laboratory conditions. The
spray deposits were assessed
at intervals up to 24 weeks
post application. Efficacy of the
formulation at each interval, in
terms of knockdown and
mortality, was evaluated over a
period of 96 hours post
treatment application
Efficacy was measured when
all termites in a container had
died, the length of time of
consumption by the termites of
the treated sawdust was
recorded
Not Insecticidal activity of 18 10 German cockroaches Manufacturer
available an oil based aerosol (Blattella germanica) were
against German released into a plastic
cockroach, Blattella container within a glass
germanica (in-house cylinder and the aerosol was
laboratory study) sprayed into the cylinder.
Knocked down insects were
counted at intervals up to 20
minutes.
Not Insecticidal efficacy of 18 6 adult American cockroaches Manufacturer
94
AVAILABLE OR SOURCE
FROM ELSEWHERE)
available an oil based aerosol (Periplaneta Americana) were
against American released into plastic container
cockroach, Periplaneta which was placed in the centre
Americana at the bottom of the glass
cylinder and 1013mg of
aerosol was sprayed into the
cylinder and was then covered
by a glass lid. Knocked down
insects were counted at
intervals up to 20 minutes.
Not Insecticidal efficacy of 18 100 houseflies were released Manufacturer
available an oil-based aerosol into a Peet-Grady chamber
against Housefly, and aerosol was sprayed into
Musca domestica. the chamber. Knocked down
Not Insecticidal efficacy of 18 50 mosquitoes (Culex pipiens Manufacturer
available an oil-based aerosol pallens) were released into a
against mosquito, Peet-Grady chamber and
Culex pipiens pallens aerosol was sprayed into the
chamber. Knocked down
The chamber was then
ventilated by an exhaust fan
and all insects were
transferred to a clean recovery
container with diet and water
within 20 minutes
Not Insecticide primarily for 18 Various concentrations of Manufacturer
available the control of termites: product were impregnated in a
Laboratory Study sawdust matrix readily
against Reticulitennes consumed by termites. Product
fIavipes was incorporated into the
sawdust by placing each batch
of solution and one batch of
sawdust in the mixing bowl of
a Kitchenaid Model K5SS
mixer. One untreated batch of
sawdust served as a control.
Sterile sand and distilled water
was mixed to form a
moistened sand substrate to
sustain the termites as they
fed upon the samples.
Eight replications of each
concentration plus the control
were tested. Termite activity
was observed in each of the
containers for four days.
elimination of termite
95
AVAILABLE OR SOURCE
FROM ELSEWHERE)
infestation.
Not Laboratory studies to 18 Product was applied as a Manufacturer
available assess products direct spray, was assessed for
applied as direct efficacy against cockroaches
sprays for efficacy (Blatta orientalis, Blatella
against a range of germanica and Periplaneta
crawling insects americana) under laboratory
conditions. Efficacy criteria
based on mortality and
knocked down insects
OPPTS Soil treatments for 18 Own searches
810.3100 imported fire ants
OPPTS Livestock, poultry, fur- 18 Own searches
810.3200 and wool-bearing
animal treatments
OPPTS Treatments to control 18 UK guidelines
810.3300 pests of humans and
pets
OPPTS Mosquito, black fly, 18 Test of insecticides against UK guidelines

810.3400 and biting midge (sand flying insects: Mosquito
fly) treatments Black Fly
Biting Midge (Sand Fly)
OPPTS Premises treatments 18 General guideline Manufacturer
810.3500
UK guidelines
OPPTS Methods for efficacy 18 Own searches

810.3800 testing of termite baits
SABS Pesticides: Biological 18 Manufacturer
st
233 1 evaluation of mists and
rev fogs - first revision
SABS Pesticides – Rearing 18 Manufacturer
303 and handling of the
human body louse
(Pediculus humanus
humanus L.) - first
revision
common clothes moth
(Tineola bisseliella
Hummel) - second
revision
German cockroach
(Blatella germanica
(L.)) - second revision
SABS Pesticides – Biological 18 Manufacturer
576 evaluation of
insecticidal oil-based
96
AVAILABLE OR SOURCE
FROM ELSEWHERE)
space spray in low-
pressurized
dispensers - first
revision
583 evaluation of the
contact efficacy of
liquid residual
insecticides - first
revision
6136 evaluation of materials
(2003) that release an
insetticide upon
heating
rd
689 3 evaluation of knock-
ed (2002) down and killing
proprieties of liquid
and aerosol
formulation (al posto
di Standard methods
SABS Method 8689-
first revision)
SABS Pesticides: biological 18 Manufacturer
(DRAFT) proprerties of solid fly
baits - DRAFT
SABS Methods for testing 18 TNsG on Prod
807 insecticides against Eval
flying and crawling
insects. Manufacturer
UK guidelines
SABS Insecticidal space 18 Manufacturer

899 spray in pressurized
(1987) dispensers
US Test method for 18 Efficacy test against larvae TNsG on Prod
AATCC textiles to determine Eval
Technical resistance to insects
Manual (e.g. moths, carpet UK guidelines
Method beetles)
24 (1992)
US Test method for 18 Test of insecticides against TNsG on Prod
CSMA aerosol space sprays flying insects: Eval
Aerosol against flying insects
Guide UK guidelines
7 th
Edition, Manufacturer
pages
129-134
97
AVAILABLE OR SOURCE
FROM ELSEWHERE)
(1981)
US Test method for 18 Test of insecticides against TNsG on Prod
CSMA pressurised spray crawling insects: cockroaches Eval
Aerosol products against
Guide cockroaches UK guidelines
7 th
Edition,
pages
135-139
(1991)
Verwey & Liquid Electric test 18 For testing pyrethroids (draft Manufacturer
Sosa, method method) and natural actives
2007 (Pyrethrum extract) on
mosquitoues (knockdown).
Efficacy criteria: "effective
against mosquitoes for X
hours". Knockdown is
measured repeatedly for 2h
and mortality after 24h.
Control (no treatment)
knockdown: maximum 10%.
2-4 chamber replicates, 50
organisms in each. Mean and
Standard Deviations for each
time calculated as well as
KT50 and KT80 (Mean time to
50% and 80% knockdown
respectively).
WHO/CD Space spray 18 Brief description of the main TM II05 (Fr)
S/WHOP application of types of space spray
ES/GCD insecticides for vector equipment as well as the
PP/2003. and public health pest operational guidelines for
5 control – a space spray application of
practitioner’s guide insecticides.
WHO/VB Instructions for 18 TNsG on Prod
C/75.593 determining the Eval
(1981) susceptibility or
resistance of UK guidelines
cockroaches to
insecticides
resistance of mosquito
larvae to insect
development inhibitors
susceptibility or
resistance of adult UK guidelines
mosquitoes to
organochlorine,
98
AVAILABLE OR SOURCE
FROM ELSEWHERE)
organophosphate and
carbamate
insecticides, -
establishment of the
baseline.
susceptibility or
mosquitoes to
organochlorine,
organophosphate and
carbamate insecticides
- diagnostic test
resistance of mosquito UK guidelines
larvae to insecticides
resistance of body or
headlice to
insecticides
bed-bugs to
insecticides
blackflies, sandflies
and biting midges to
insecticides
resistance of blackfly UK guidelines
WHO/VB Instructions for 18 UK guidelines
C/81.812 determining the
larvae to insect
99
AVAILABLE OR SOURCE
FROM ELSEWHERE)
houseflies, tsetse flies,
stableflies, blowflies
etc. to insecticides
ticks to insecticides
resistance of fleas to UK guidelines
insecticides
Admixture of plant 18 + TM II05 (Fr)
protection products to 20
EPPO stored plant products
PP to control insects and
1/203(1) mites
Fumigants to control 18 + TM II05 (Fr)
EPPO insect and mite pests 20
PP of stored plant UK guidelines
1/201(1) products
ASTM Standard Test Method 19 Evaluates the repellency of Own search
E939- of Field Testing promising compounds that
94(2006) Topical Applications of have undergone primary
Compounds as laboratory studies and
Repellents for approved for skin application
Medically Important for secondary testing.
and Pest Arthropods The method is designed for the
(Including Insects, study of mosquito repellents,
Ticks, and Mites): but can be modified to
Mosquitoes determine the repellency of
candidate compounds for other
flying insects that attack
humans.
ASTM Standard Test 19 Can be used to test the Own search
E951- Methods for efficacy of repellent
94(2006) Laboratory Testing of compounds that can be diluted
Non-Commercial with ethanol, acetone etc. Both
Mosquito Repellent biological effectiveness and
Formulations On the persistence of the repellent
Skin can be assessed.
ED50 and ED95 are
determined for comparative
and practical purposes
respectively. Precision of the
test can be evaluated (confid
intervals).
Dautel H, Verwendung von 19 Dossier
Hilker M, Dodecansäure als
Kahl O, Zeckenrepellent
100
AVAILABLE OR SOURCE
FROM ELSEWHERE)
Siems K, (Patentschrift).
2001.
Fradin & Comparative efficacy 19 Human subjects: Arm in cage Dossier
Day, July of insect repellents studies (15 volunteers, 10
2002 against mosquito bites mosquitoues (Aedes aegypti)
N Engl J in each cage. Endpoint:
Med vol elapsed time to first bite.
347 vol Category of protection A-H
13-18 (significantly different mean
complete protection time;
ANOVA & Tukey's). No need
to recalculate the results to
"real condition" (simulate real
condition)
Hummel, Effect of the neem 19 Dossier
E., extract formulation
Kleeberg, neemazal-t/s on the
H. 1997. green pea aphid
in: acyrthosiphon pisum in
Practice the laboratory (1995),
orientate in: Practice orientated
d results results on use and
on use production of Neem-
and Ingredients and
productio Pheromones V
n of
Neem-
Ingredien
ts and
Pheromo
nes V.
Proceedi
ngs of the
5th
workshop
, Wetzlar,
Germany,
January
22-25,
1996
Not Repellency of Two 19 Vinyl floor tiles were sprayed Manufacturer
available Formulations against with the repellent solution and
Ants (in-house placed in cages. A food
laboratory study) attractant was placed in the
centre of each tile. The
number of ants crossing each
tile was counted at 30 and 60
minutes after tiles were placed
into cages. Data for 30 and 60
min was combined and
averaged. Repellency was
also measured the day after
101
AVAILABLE OR SOURCE
FROM ELSEWHERE)
the fourth consecutive
application. Sufficient
repellency was defined as
>65%.
Not Repellency of Two 19 The cockroaches had a choice Manufacturer
available product formulations of two shelters, one treated
against the German with the product and one
Cockroach (in-house untreated. Repellency after 1,
laboratory study) 3, 7 and 14 days was
recorded. Percent repellency
was calculated. Sufficient
>65%, excellent repellency
was defined as >85%.
OPPTS Insect Repellents For 19 Own search
810.3700 Human Skin and
“public Outdoor Premises
draft”
efficacy of mosquito
repellents - first
revision
US EPA Insect repellents for 19 UK guidelines
Guideline human skin and
OPPTS outdoor premises
810.3700
(1999)
AS Durability and 21 Simulated field test TM II05
1580.481 resistance to fouling -
.5 Marine underwater
(1994) paint systems
ASTM Standard Method for 21 Simulated field raft test, TNsG PE
D3623- Testing Antifouling screening test in shallow
78a(2004 Panels in Shallow marine environments TM II 05
) Submergence A standard antifouling panel of
known performance serves as UK guidelines
a control. Subcommittee
D01.45 has a revised rating
procedure now being
evaluated by round robin.
ASTM Standard Test Method 21 Simulated field test. TM II05
D4939-89 for Subjecting Marine Determination of antifouling
(2007) Antifouling Coating to performance and reduction of
Biofouling and Fluid thickness of marine antifouling
Shear Forces in coatings by erosion or under
Natural Seawater specified conditions of
hydrodynamic shear stress in
seawater alternated with static
exposure in seawater.
An antifouling coating system
of known performance is
102
AVAILABLE OR SOURCE
FROM ELSEWHERE)
included to serve as a control
in antifouling studies.
ASTM Standard Practice for 21 Testing in-situ partial TM II05
D5479- Testing Biofouling immersion exposure.
94(2007) Resistance of Marine A negative control (non toxic
Coatings Partially surface) is applied and should
Immersed show heavy fouling accretion
for the test to be valid.
ASTM Standard Test Method 21 Screening test, measuring TM II05
D5618- for Measurement of barnacle adhesion in shear to
94(2005) Barnacle Adhesion surfaces exposed in the
Strength in Shear marine environment. Surfaces
with known barnacle adhesion
strengths are included to serve
as controls.
ASTM Standard Practice for 21 Guidance to a panel inspector TM II05
D6990-05 Evaluating Biofouling for quantitative and consistent
Resistance and evaluation of coating
Physical Performance performance from test panels
of Marine Coating coated with marine antifouling
Systems coating systems.
CEPE Antifouling coatings - 21 Simulated field raft test TNsG PE
Antifoulin methods for the
g generation of efficacy TM II05
Working data
Group UK guidelines
1993
EN ISO Water quality - Marine 21 Growth inhibition test TM II05
10253:20 algal growth inhibition
06 test with Skeletonema
costatum and
Phaeodactylum
tricornutum (ISO
10253:2006)
103
Table 2. Overview of efficacy test references, in order of reference.
REFERE TITLE PT SHORT TEST TYPE OF

NCE DESCRIPTION (IF TEST REFERENCE
METHOD AVAILABLE OR SOURCE
INFORMATION
PROVIDED FROM
ELSEWHERE)
AATCC Antibacterial Finishes 9 Applied to textiles Manufacturer
100 - on Textile Materials:
1999 Assessment of
Antibacterial Activity 9 Applied to textiles Manufacturer
AATCC Assessment of Textile
147 - Materials: Parallel
1998 Streak Method
Assesment of the Anti- 18 Applied to textiles Manufacturer
House Dust Mite
Properties
AATCC of Textiles under Long-
194-2006 Term Test Conditions
Antifungal Activity, 9 Applied to textiles Manufacturer
Assessment on Textile
Materials:
Mildew and Rot
AATCC Resistance of Textile
30 - 1999 Materials
Alleman, Comparative Evaluation 1-5 Microorganisms were Manufacturer
J.E., of Alternative Halogen- exposed to a mixture of
Etzel, based Disinfection biocide in solution for 4 or
J.E., Strategies 16 minutes. The
Gendron, percentage survival was
D., Kirsch, assessed under various
E.J., conditions such as high or
Conley, low nitrogen (as
J., Fidelle, ammonium), high pH or low
T., Handy, temperatures.
F., and
Hildebran
nd
dt, M. 42
Purdue
Industrial
Waste
Conferenc
e, May
1987,
519-524
American Standard Methods for 1-5 A swimming pool trial was Manufacturer
Public the Examination of performed over a three
Health Water and Wastewater month period to
Associatio (SMEWW). demonstrate the
n, 16th disinfection efficacy of the
Edition, product. Water samples
1985 were collected for microbial
analysis. Water and, air
temperature, number of
bathers (at the time when
samples were taken and
104
INFORMATION
PROVIDED FROM
ELSEWHERE)
total bathers for the day),
eye and skin irritation and
water clarity were also
measured.
Criteria for effective control
was determined as follows:
The state of California,
Department of Health
Services requirement for
pool water is;
Bacteriological quality of
water in the swimming pool
shall be such that not more
than two consecutive
samples, taken when the
pool is in use shall:
1. Contain more than 200
bacteria per ml, as
determined by the standard
plate count; or
2. Contain a total coliform
organism count of 2.2 or
greater per 100 ml of
sample
3. Chemical quality of water
in the pool shall not cause
irritation of eyes or skin of
the bathers, or have other
objectionable physiological
effects on bathers.
AOAC “Disinfectant (water) for 1-5 Manufacturer
(Associati Swimming Pools”
on of official method of
Official Analysis, 16th Edition,
Analytical 1995
Chemists)
official
method
965.13
AS Durability and 21 Simulated field test TM II05
1580.481. resistance to fouling -
5 Marine underwater paint
(1994) systems
ASTM Standard practice for 7 This practice covers Own searches
G21- determining resistance determination of the effect
96(2002) of synthetic polymeric of fungi on the properties of
materials to fungi synthetic polymeric
materials in the form of
molded and fabricated
articles, tubes, rods,
sheets, and film materials.
105
INFORMATION
PROVIDED FROM
ELSEWHERE)
Conditions favorable for
attack, namely, a
temperature of 2 to 38°C
(35 to 100°F) and a relative
humidity of 60 to 100 %
need to be established.
Expected effects include
surface attack,
discoloration, loss of
transmission (optical), and
removal of susceptible
plasticizers, modifiers, and
lubricants, resulting in
increased modulus
(stiffness), changes in
weight, dimensions, and
other physical properties,
and deterioration of
electrical properties such
as insulation resistance,
dielectric constant, power
factor, and dielectric
strength.
Since attack by organisms
involves a large element of
chance due to local
accelerations and
inhibitions, the order of
reproducibility may be
rather low. To ensure that
estimates of behavior are
not too optimistic, the
greatest observed degree
of deterioration should be
reported.
Conditioning of the
specimens, such as
weathering, heat treatment,
etc., may have significant
effects on the resistance to
fungi. Determination of
these effects is not covered
in this practice.
ASTM Standard Test Method 11 or Laboratory test method is Communication
E723-07 for Efficacy of 12? used to determine the with UK (HSE)
Antimicrobials as efficacy of an antimicrobial
Preservatives for for preventing bacterial
Aqueous-Based spoilage of in-process
Products Used in the pigment suspensions, dye
Paper Industry solutions, pulp slurries,
106
INFORMATION
PROVIDED FROM
ELSEWHERE)
(Bacterial Spoilage) starch solutions, polymers,
sizing agents, latex
emulsions, and other
aqueous-based materials
used in the paper industry
from bacterial spoilage.
Test organisms may vary
but should lie above 10E6
CFU/ml.
Bacterial numbers in the
sample are determined at
variable time periods and
compared to a control with
no bactericide. Material to
be preserved is used as
substrate, but for some
materials nutrients may be
added. Results are
expressed as % kill:
[(control plate count-test
plate count)/ control plate
count]x100 at each
sampling time.
ASTM Standard Test Method 11 or Laboratory method that Communication
E875- for Efficacy of Fungal 12? determines if a fungal with UK (HSE)
00(2005) Control Agents as control agent is effective to
Preservatives for preserve pigment
Aqueous-Based suspensions, dye solutions,
Products Used in the pulp slurries, starch
Paper Industry solutions, polymers, sizing
agents, latex emulsions,
and other specific aqueous-
based materials to prevent
spoilage of in-process
aqueous-based products
used in the paper industry.
Will also prevent spore
germination. Plates are
incubated for 7d or until
control plates shos
sufficient growth for rating.
The exposure time should
correspond to the real time
desired, often 6w.
Separate evaluations
should be made on a
representative type for
each specific class of
product to be preserved.
Growth is determined by
visible signs of
107
INFORMATION
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deterioration in the sample
and by obtaining fungal
numbers and comparing
them to a sample without
fungal control agents. A
proper level of fungal
control agent prevents
deterioration and reduces
and keeps organisms to an
acceptable level in the test
material, which is
determined by the tester or
user. Often a scale from 0
to 4 (heavy growth).
ASTM Standard Test Method 11 Efficacy of microbicides Manufacturer
E645-07 for Efficacy of (algicides, bactericides,
Microbicides Used in and fungicides), evaluated
Cooling Water Systems using simulated or real
cooling tower water against
(1) microbes from cooling
water, (2) microbes in
microbiological deposits
(biofilms) from operating
cooling systems, or (3)
microorganisms known to
contaminate cooling water
systems, or a combination
thereof. Choice of
enumerating method may
vary, e g pour plate, spread
plate, MPN techniques etc.
Test concentrations also
vary but are usually
between 1 and 50 mg/l.
Exposure time varies with
mode of action but includes
3h contact time. For
bacteria 48 h incubation,
fungi 48h-7d, for algae 5-
14d.
Even if using cooling water
and deposits/biofilm
obtained from the field,
laboratory results may not
be totally predictive of
microbicidal effectiveness
in the field (due to e g
variability in environmental
factors). If solvents are
added, a solvent control is
included. If initial count in
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INFORMATION
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cooling water control
samples are <10E5 bact/ml
(or <10E3 CFU/ml fungi or
algae), the test is invalid. In
addition, the viable counts
of fungi/algae in the control
must at least be equal to
the numbers at time 0. The
untreated control should
show a stable population
with no more than 0.5 log
increase or decrease in
growth during the test
perod. Results are
expressed as “Log
reduction in number of
microorganisms at each
biocide concentration”: log
(initial count of
microorganisms of the
control sample) – log
(number of microorganisms
detected at a given biocide
concentration after a
specific contact time). If 1
log reduction: corresponds
to 90% kill, if 2 log
reduction corresponds to
99% kill, 3 log reduction to
99.9% kill.
ASTM Standard Test Method 6 Own searches
D2574-06 for Resistance of
Emulsion Paints in the
Container to Attack by
Microorganisms
ASTM Standard Test Method 10 This test method can be Own searches
WK8681 for Resistance to Mold used to evaluate the
Growth on Interior comparative resistance of
Coated Building coated building products to
Products in an accelerated mold growth.
Environmental Chamber Performance at a certain
rating does not imply any
specific period of time for a
mold free surface.
However, a better rated
panel would be expected to
perform better in actual end
use.
An environmental chamber
and the conditions of
operation to evaluate in a
109
INFORMATION
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4-week period the relative
resistance of coated
building products to surface
mold growth in a controlled
interior environment is
described. The apparatus
is designed so it can be
easily built or obtained by
any interested party.
ASTM Test Method for 12 A procedure for testing the Own searches
WK 17314 Evaluating the Efficacy efficacy of a liquid
of a Liquid Microbiocide microbicide against biofilm
against Biofilm Bacteria grown on coupons
removed from biofilm
reactors standardized to
grow a repeatable biofilm,
such as the CDC Biofilm
Reactor (ASTM Method E
2562) or Rotating Disk
Biofilm Reactor (ASTM
Method E 2196). The
preparation of the liquid
microbicide and exposure
time are completed
according to
manufacturer’s instructions
for use. The liquid
microbicide is tested at
room temperature under
static conditions. Biofilm
population density is
recorded as log10 colony
forming units per surface
area. Efficacy is
determined by calculating
the log reduction in viable
cells.
The microbicide is tested at
its use dilution for the
recommended time.
ASTM Test Method for 10 This test method can be Own searches
WK14960 Standard Test Method used to evaluate the
for the Resistance of comparative resistance of
Mold Growth on uncoated building products
Uncoated Interior to accelerated mold growth.
Building Products in an The apparatus
Environmental (environmental chamber) is
Chamber. designed so it can be easily
interested party.
4-week evaluation period.
110
INFORMATION
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Relative resistance of
uncoated interior building
products to surface mold
growth in a controlled
interior environment is
evaluated, but performance
at a certain rating does not
imply any specific period of
time for a mold free
surface.
ASTM Standard Test Method 18 Determines the Own searches
E1517- for Determining the effectiveness of ovicidal
99(2006) Effectiveness of Liquid, materials in liquid, gel,
Gel, Cream, or cream, or shampoo form
Shampoo Insecticides against the ova (that is,
Against Human Louse eggs or nits) of the human
OVA louse, Pediculus humanus.
This test method consists
of five replicates for a
statistical comparison of
formulations.
ASTM Standard 8 Field test: aids in Own searches
D1006-93 Recommended Practice evaluating the performance
for Conducting Exterior of house and trim paints to
Exposure Tests of new, previously unpainted
Paints on Wood wood exposed to the
environment.
Exposures in several
locations with different
climates which represent a
broad range of anticipated
service conditions are
recommended and several
years of repeat exposures
are needed to get an
“average” test result for a
given location. Solar
radiation varies
considerably as function of
time of year, and can cause
large differences in the
apparent rate of
degradation in many
polymers. Comparing
results for materials
exposed for short periods
(less than onee year) is not
recommended unless
materials are exposed at
the same time in the same
location.
111
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ASTM Standard Test Method 7 This test method describes Own searches
D3273- for Resistance to a small environmental
00(2005) Growth of Mold on the chamber and the conditions
Surface of Interior of operation to evaluate
Coatings in an reproducibly in a 4-week
Environmental Chamber period the relative
resistance of paint films to
surface mold fungi, mildew
growth in a severe interior
environment. The
apparatus is designed so it
can be easily built or
obtained by any interested
party and will duplicate
results obtained in a large
tropical chamber.
An accelerated test for
determining the resistance
of interior coatings to mold
growth; useful in
estimating the performance
of coatings designed for
use in interior environments
that promote mold growth
and in evaluating
compounds that may inhibit
such growth and the
aggregate levels for their
use.
Used to evaluate the
comparative resistance of
interior coating to
accelerated mildew growth.
Performance at a certain
rating (in accordance with
Test Method D3274) does
not imply any specific
period of time for a fungal
free coating. However, a
better rated coating nearly
always performs better in
actual end use. This test
method is intended for the
accelerated evaluation of
an interior coatings'
resistance to fungal
defacement. Use of this
test method for evaluating
exterior coatings'
performance has not been
validated, nor have the
112
INFORMATION
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limitations for such use
been determined. Any
accelerated weathering
(leaching, weathering
machine exposure, etc.,)
should be reported and
should also bear reference
to the fact that it is beyond
the current scope of this
test method.
Temperature and humidity
must be effectively
controlled within the
relatively narrow limits
specified in order for the
chamber to function
reproducibly during the
short test period. Severity
and rate of mold growth on
a film is a function of the
moisture content of both
the film and the substrate.
A relative humidity of 95 to
98 % at a temperature of
32.5+ 1oC (90+ 2oF ) is
necessary for test panels to
develop rapidly and
maintain an adequate
moisture level to support
mold growth.
ASTM Standard Practice for 7 This practice provides Own searches
D3456- Determining by Exterior guidelines for determining
86(2002) Exposure Tests the the susceptibility of paint
Susceptibility of Paint films to microbiological
Films to Microbiological attack on exterior
Attack exposure. The degree to
which microbiological
discoloration occurs is the
primary concern. This
practice covers the
preparation of coatings for
testing, their application on
substrates, and the
arrangement of the coated
panels on exterior test
fences to determine the
degree of microbiological
attack that may occur on
the surface of the coatings
over a period of time.
This practice is intended to
113
INFORMATION
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provide guidelines for, and
a discussion of, the various
factors critical in selection
of exterior coatings
resistant to discoloration or
disfigurement by algae and
fungi.
ASTM Standard Method for 21 Simulated field raft test, TNsG PE
D3623- Testing Antifouling screening test in shallow
78a(2004) Panels in Shallow marine environments TM II 05
Submergence A standard antifouling
panel of known UK guidelines
performance serves as a
control. Subcommittee
D01.45 has a revised rating
procedure now being
evaluated by round robin.
ASTM Standard practice for 17 Adaptable to both lotic and Communication
D4131-84 sampling fish with lentic situations in littoral with UK (HSE)
(2005) Rotenone and limnetic areas.
ASTM Standard Test Methods 7 Tests the ability of Own searches
D4300-01 for Ability of Adhesive adhesive films to inhibit or
Films to Support or support the growth of
Resist the Growth of selected fungal species
Fungi growing on agar plates by
providing means of testing
the films on two agar
substrates, one which
promotes microbial growth,
and one which does not.
constituents.
These test methods are not

appropriate for all
adhesives. The activity of
certain biocides may not be
demonstrated by these test
methods as a result of
irreversible reaction with
some of the medium. As an
example, quaternary
ammonium compounds are
inactivated by agar. A test
method is included for use
with low-viscosity
adhesives along with an
alternative method for use
with mastic-type adhesives.
ASTM Standard Test Method 9 Determination of mold
D4576- for Mold Growth growth resistance of wet
01(2006) Resistance of Wet Blue blue subject to storage and
114
INFORMATION
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shipping requirements and
intended for use in leather
manufacturing, assisting
the prediction of storage
time before molding occurs.
This test method may not
be suitable to evaluate
fungicides that are
inactivated by proteins,
including
alkyldimethylbenzyl
ammonium chlorides.
To allow use of this test
method by any laboratory,
flexibility has been
permitted in times,
temperature, and humidity
of incubation, inoculum,
hide sampling area, and
choice of control. These
may be adjusted to fit local
conditions but must be
standardized.
Conclusions about mold
growth resistance are
drawn from the results by
comparing the test with a
simultaneously run control
of known resistance.
Success or failure is
determined by the amount
of mold growth relative to
the control.
The degree of correlation

between this test and
commercial quantities of
wet blue in storage or
shipment situations, or
both, has not been fully
determined.
ASTM Standard Test Methods 6 Determination of the TNsG on Prod
D4783- for Resistance of resistance of liquid Eval
01e1 Adhesive Preparations adhesive preparations to
in Container to Attack microbial attack in the
by Bacteria, Yeast, and container by challenging
Fungi adhesive specimens with
cultures of bacteria, yeast,
or fungi, and checking for
their ability to return to
sterility.
115
INFORMATION
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These test methods return
qualitative results
ASTM Standard Test Method 21 Simulated field test. TM II05
D4939-89 for Subjecting Marine Determination of antifouling
(2007) Antifouling Coating to performance and reduction
Biofouling and Fluid of thickness of marine
Shear Forces in Natural antifouling coatings by
Seawater erosion or under specified
conditions of hydrodynamic
shear stress in seawater
alternated with static
exposure in seawater.
An antifouling coating
system of known
performance is included to
serve as a control in
antifouling studies.
ASTM Standard Practice for 21 Testing in-situ partial TM II05
D5479- Testing Biofouling immersion exposure.
94(2007) Resistance of Marine A negative control (non
Coatings Partially toxic surface) is applied
Immersed and should show heavy
fouling accretion for the test
to be valid.
Films and Related resistance of a paint or
Coatings to Algal coating film to algal growth.
Defacement This test method should not
be used as a replacement
for exterior exposure since
many other factors, only a
few of which are listed will
affect those results.
Films and Related resistance of two or more
Coatings to Fungal paints or coating films to
Defacement by fungal growth.
Accelerated Four-Week This test method should not
Agar Plate Assay be used as a replacement
for exterior exposure (that
is, Practice D 3456) since
many other factors, only a
few of which are listed will
affect those results.
Comparative evaluation of
different coating
formulations for their
116
INFORMATION
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relative performance under
a given set of conditions. It
does not imply that a
coating that resists growth
under these conditions will
necessarily resist growth in
the actual application.
Round-robin testing of this

test method versus exterior
exposure is planned.
ASTM Standard Test Method 21 Screening test, measuring TM II05
D5618- for Measurement of barnacle adhesion in shear
94(2005) Barnacle Adhesion to surfaces exposed in the
Strength in Shear marine environment.
Surfaces with known
barnacle adhesion
strengths are included to
serve as controls.
ASTM Standard Practice for 21 Guidance to a panel TM II05
D6990-05 Evaluating Biofouling inspector for quantitative
Resistance and and consistent evaluation
Physical Performance of coating performance
of Marine Coating from test panels coated
Systems with marine antifouling
coating systems.
ASTM Standard Test Method 1-5 Laboratory suspension test Manufacturer
E1052- for Efficacy of that determines the
96(2002) Antimicrobial Agents effectiveness of
Against Viruses in antimicrobial solutions
Suspension against designated
prototype viruses. The
determined using cell
cultures as the host system
for specific viruses. For
special applications of
virucides, such as
inactivation of viruses in
contaminated liquid wastes,
and as a first stage in
determining virucidal
potential of liquid chemical
germicides, liquid hand
soaps, OTC topicals or
other skin products.
ASTM Standard Test Method 1-5 Laboratory test method. Manufacturer

E1053- for Efficacy of Virucidal Evaluates the virucidal
97(2002) Agents Intended for efficacy of liquid, aerosol,
117
INFORMATION
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Inanimate or trigger spray
Environmental Surfaces antimicrobial solutions on
inanimate nonporous
environmental surfaces,
against designated
prototype viruses. The
determined utilizing cell
cultures as the host system
for specific viruses. Efficacy
is measured by a
percentage reduction in
titer.
ASTM Standard Test Methods 1-5 Effectiveness of Own searches
E1054-02 for Evaluation of procedures and agents for
Inactivators of inactivating (neutralizing,
Antimicrobial Agents quenching) the
microbiocidal properties of
antimicrobial agents and to
ensure that no components
of the neutralizing
procedures and agents,
themselves, exert an
inhibitory effect on
microorganisms targeted
for recovery.
E1115-02 Evaluation of Surgical
Hand Scrub
Formulations
E1153-03 for Efficacy of Sanitizers
Recommended for
Inanimate Non-Food
Contact Surfaces
E1173- of a Evaluation of a
01e1 Preoperative,
Precatheterization, or
Preinjection Skin
Preparations
E1174-06 Evaluation of the
Effectiveness of Health
Care Personnel
Handwash Formulations
ASTM Standard Practice for 6 The procedure should be Own searches

E1259-05 Evaluation of used to evaluate the
Antimicrobials in Liquid relative efficacy of
118
INFORMATION
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Fuels Boiling Below microbicides in liquid fuels
390°C boiling below 390°C. The
effect of environmental
conditions, such as a
variety of fuel additives,
metal surfaces, and
climatology, are variables
that can be included in
specific tests using this
protocol.
Antimicrobial Handwash
Formulations by
Utilizing Fingernail
Regions
ASTM Standard Guide for 12 The purpose of this guide is Own searches
E1427- Selecting Test Methods to inform the investigator of
00e1 to Determine the methods that can be used
Effectiveness of for biofilm formation and
Antimicrobial Agents measurement, allowing
and Other Chemicals development of test
for the Prevention, procedures for determining
Inactivation and the effectiveness of
Removal of Biofilm chemical treatments for
prevention, inactivation,
and removal of unwanted
biofilm. This guide is not an
exhaustive survey of biofilm
methods.
ASTM Standard Test Method 7 This test method provides a Own searches
E1428- for Evaluating the technique for evaluating
99(2004) Performance of antimicrobials in or on
Antimicrobials in or on polymeric solids against
Polymeric Solids staining by
Against Staining by Streptoverticillium
Streptoverticillium reticulum, and should
reticulum (A Pink Stain assist in the prediction of
Organism) performance of treated
articles under actual field
conditions. Conditioning of
the specimens, such as
weathering, and heat
treatment, may have
significant effects on
performance of
antimicrobials against
staining. Determination of
these effects is not
included in this test
119
INFORMATION
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method. This test method is
also not suitable for
evaluating dark-pigmented
test samples.
E1482-04 for Neutralization of
Virucidal Agents in
Virucidal Efficacy
Evaluations
E1589-05 for Evaluation of First
Aid Antiseptic Drug
Products
E1766- for Determination of
95(2002) Effectiveness of
Sterilization Processes
Devices
E1837- to Determine Efficacy of
96(2002) Disinfection Processes
Devices (Simulated Use
Test)
E1838-02 for Determining the
Virus-Eliminating
Effectiveness of Liquid
Hygienic Handwash and
Handrub Agents Using
the Fingerpads of Adult
Volunteers
ASTM Standard test method 12 A procedure to evaluate the Communication
E1839-07 for efficacy of slimicides efficacy of slimicides for the with UK (HSE)
for the paper industry – control of bacterial and
bacterial and fungal fungal slimes in paper mill Manufacturer
slime systems and their
counterparts. This test
method is run in acid,
alkaline, or acid and
alkaline conditions to
determine the efficacy of
the slime control agent.
The test conditions may be
modified to reflect intended
use patterns in typical
paper mill systems,
including use of actual
paper mill furnish. Efficacy
based on a % reduction on
120
INFORMATION
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CFU.
ASTM Standard Test Method 1-5 This test method Own searches
E1882-05 for Evaluation of determines the antibacterial
Antimicrobial activity and persistence of
Formulations by the test formulations, as
Agar Patch Technique measured by the inhibition
of a test organism on an
agar surface exposed to
test sites on human skin
treated with the
formulations. This
procedure can be used to
evaluate formulations
containing ingredients
intended to inhibit growth of
bacteria on intact skin and
measures the difference,
post-product-exposure,
between numbers of
bacterial colonies on active
test formulation plates and
numbers on control plates,
expressed as percent
inhibition.
This procedure may also
be used to test for
persistence of activity, as a
function of time elapsed
between application of
active test formulation and
application of active test
plates. Because no
procedure for neutralization
of the antimicrobial action
of active ingredients can be
included in the test, the
agar patch method is
limited to the extent that
results expressed as
percent inhibition do not
differentiate between
bacteriostatic and
bacteriocidal effects and,
hence, must not be
portrayed as “reductions.”
E1883-02 for Assessment of an
Antibacterial Handwash
Product by Multiple
Basin Wash Technique
ASTM Standard Guide for ? Own searches
121
INFORMATION
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E1891- Determination of a
97(2002) Survival Curve for
Against Selected
Microorganisms and
Calculation of a D-Value
and Concentration
Coefficient
Handwashing
Formulations for Virus-
Eliminating Activity
Using the Entire Hand
ASTM Standard Quantitative 1-5 This test can be performed Own searches
E2111-05 Carrier Test Method to with or without a soil load to
Evaluate the determine the effect of
Bactericidal, Fungicidal, such loading on
Mycobactericidal, and microbicide performance.
Sporicidal Potencies of The soil load developed for
Liquid Chemical this test is a mixture of
Microbicides three types of proteins
(high molecular weight
proteins, low molecular
weight peptides, and
mucous material) to
represent the body
secretions, excretions, or
other extraneous
substances that chemical
microbicides may
encounter under field
condititions.
This test method is
designed for use in product
development and for the
generation of product
potency data. This test
method is fully quantitative
and it also avoids any loss
of viable organisms through
wash off. It permits the
loading of each carrier with
a known volume of the test
organism. The
incorporation of controls
can also determine the
initial load of colony
forming units (CFU) of
organisms on the test
carriers and any loss in
122
INFORMATION
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CFU after the mandatory
drying of the inoculum. This
test method is designed to
have survivors and also to
be used with a
performance standard. The
surviving microorganisms
on each test carrier are
compared to the mean of
no less than three control
carriers to determine if the
been met. To allow proper
statistical evaluation of
results, the size of the test
inoculum should be
sufficiently large to take
into account both the
performance standard and
the experimental variation
in the results.
ASTM Determining the 9 Evaluates the resistance of Manufacturer,
E2149-01 Antimicrobial Activity of non-leaching antimicrobial applied to textiles
Immobilized treated specimens to the
Antimicrobial Agents growth of microbes under
Under Dynamic Contact dynamic contact conditions.
Conditions This test determines the
antimicrobial activity of
treated specimen by
shaking samples of surface
bound materials in a
concentrated bacterial
suspension for a one hour
contact time or other
contact times as specified
by the investigator. The
suspension is serially
diluted both before and
after contact and cultured.
This dynamic shake flask
test was developed for
routine quality control and
screening tests in order to
overcome difficulties
(including ensuring contact
of inoculum to treated
surface) in using classical
antimicrobial test methods
to evaluate substrate-
bound antimicrobials. This
test also allows for the
123
INFORMATION
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versatility of testing
contamination due to such
things as hard water,
proteins, blood, serum,
various chemicals, and
other contaminates or
physical/chemical stresses
or manipulations of the
specimens of interest.
The number of viable
organisms in the
suspension is determined
and the percent reduction
is calculated based on
initial counts or on
retrievals from appropriate
untreated controls. his
method is intended for
those surfaces having a
percent reduction activity of
50 % to 100 % for the
specified contact time.
Surface antimicrobial
activity is determined by
comparing results from the
test sample to
simultaneously run
controls. The presence of a
leaching antimicrobial is
both pre- and post-
determined by the
presence of a zone of
inhibition.
ASTM Standard Test Method 9 This test method is Own searches

E2180-07 for Determining the designed to evaluate
Activity of Incorporated (quantitatively) the
Antimicrobial Agent(s) antimicrobial effectiveness
In Polymeric or of agents incorporated or
Hydrophobic Materials bound into or onto mainly
flat (two dimensional)
hydrophobic or polymeric
surfaces. The method
focuses primarily on
assessing antibacterial
activity; however, other
microorganisms such as
yeast and fungal conidia
may be tested using this
method. The vehicle for the
inoculum is an agar slurry
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which reduces the surface
tension of the saline
biofilm," providing more
even contact of the
inoculum with the test
surface. This test method
facilitates the testing of
hydrophobic surfaces by
utilizing cells held in an
agar slurry matrix.
This method can confirm
the presence of
antimicrobial activity in
plastics or hydrophobic
surfaces and allows
determination of
antimicrobial activity
between untreated plastics
or polymers and those with
bound or incorporated low
agents. Comparisons
between the numbers of
survivors on preservative-
treated and control
hydrophobic surfaces may
also be made. This test
method, as written, is
inappropriate to determine
efficacy against biofilm
cells, which are different
both genetically and
metabolically than
planktonic cells used in this
test. The procedure also
permits determination of
"shelf-life" or long term
durability of an
antimicrobial treatment
which may be achieved
through testing both non-
washed and washed
ASTM Standard Test Method 7 This test method is used for Own searches
E2196-07 for Quantification of a growing a repeatable
Pseudomonas Pseudomonas aeruginosa
aeruginosa Biofilm biofilm in a continuously
Grown with Shear and stirred flow reactor. In
125
INFORMATION
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Continuous Flow Using addition, the test method
a Rotating Disk Reactor describes how to sample
and analyze biofilm for
viable cells. In this test
method, biofilm population
surface area.
ASTM Standard Quantitative 1-5 The method is designed to Own searches
E2197-02 Disk Carrier Test evaluate the ability of liquid
Method for Determining chemical germicides to
the Bactericidal, inactivate vegetative
Virucidal, Fungicidal, bacteria, viruses, fungi,
Mycobactericidal and mycobacteria and bacterial
Sporicidal Activities of spores in the presence of a
Liquid Chemical soil load on disk carriers
Germicides that represent
environmental surfaces and
medical devices. It is also
designed to have survivors
that can be compared to
mean of no less than three
control carriers to
determine if the
been met. For proper
statistical evaluation of the
results, the size of the test
inoculum should be
sufficiently large to take
into account both the
performance standard and
the experimental variation
in the results. The test
protocol does not include
any wiping or rubbing
action. It is, therefore, not
designed for testing
germicide-soaked wipes.
Disinfectants
ASTM Standard Practice for 13 Laboratory procedures for TNsG on Prod
E2275- Evaluating Water- rating the relative inherent Eval
03e1 Miscible Metalworking bioresistance of water-
(replaces Fluid Bioresistance and miscible metalworking
D3946 Antimicrobial Pesticide fluids, the bioresistance
and E686) Performance attributable to
augmentation with
antimicrobial pesticides or
126
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both, for determining the
need for microbicide
addition prior to or during
fluid use in metalworking
systems and for evaluating
microbicide performance.
Relative bioresistance is
determined by challenging
metalworking fluids with a
biological inoculum that
may either be
characterized (comprised
of one or more known
biological cultures) or
uncharacterized
(comprised of biologically
contaminated metalworking
fluid or one or more
unidentified isolates from
deteriorated metalworking
fluid). Challenged fluid
bioresistance is defined in
terms of resistance to
biomass increase, viable
cell recovery increase,
chemical property change,
physical property change or
some combination thereof.
This practice is applicable
to antimicrobial agents that
are incorporated into either
the metalworking fluid
concentrate or end-use
dilution. It is also applicable
to metalworking fluids that
are formulated using non-
microbicidal, inherently
bioresistant components.
The results of tests
completed in accordance
with this practice should be
used only to compare the
relative performance of
products or microbicide
treatments included in a
test series. Results should
not be construed as
predicting actual field
performance.
E2276- for Determining the
127
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03e1 Bacteria-Eliminating
Effectiveness of
the Fingerpads of Adult
Subjects
E2314-03 for Determination of
Effectiveness of
Cleaning Processes for
Reusable Medical
Instruments Using a
Microbiologic Method
(Simulated Use Test)
ASTM Standard Guide for 1-5 This guide covers Own searches
E2315-03 Assessment of examples of a basic
Antimicrobial Activity method to measure the
Using a Time-Kill changes of a population of
Procedure aerobic microorganisms
within a specified sampling
time when tested against
antimicrobial test materials
in vitro. Several options for
organism selection and
growth, inoculum
preparation, sampling times
and temperatures are
provided. Antimicrobial
activity of specific
materials, as measured by
this technique, may vary
significantly on variables
selected.
ASTM Standard Guide for 1-5 This guide covers test Own searches
E2361-04 Testing Leave-On methods and sampling
Products Using In-Situ procedure options for
Methods leave-on products (such as
alcohol hand rubs and
lotions containing
antimicrobial ingredients)
for consumer and hospital
personnel. These products
are distinguished from
conventional washing and
scrubbing preparations in
that they do not rely on the
rinsing, physical removal,
and antimicrobial action in
determining their
effectiveness. Although
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agitation and friction may
serve to release organisms
from the skin and folds and
crevices, organisms are
then killed in situ and are
not rinsed from the skin
surface before sampling.
Appropriate test methods
for the hands have been
published, while other
sampling methods will be
needed for testing body
areas other than the hands.
ASTM Standard Practice for 1-5 Own searches
E2362-04 Evaluation of Pre-
saturated or
Impregnated Towelettes
for Hard Surface
Disinfection
Disinfectants for Use in
High Efficiency Washing
Operations
ASTM Standard Test Method 9 Rapid screening evaluating Own searches
E2471-05 for Using Seeded-Agar (qualitatively) the presence
for the Screening of antimicrobial (both
Assessment of antibacterial and antifungal)
Antimicrobial Activity In activity in or on the carpet
Carpets face fiber or incorporated
into the backing structure of
the carpet (or both). The
method simulates actual
use conditions that may
occur on carpets (for
example, food and
beverage spills, soiling
from foot traffic, prolonged
moisture exposure) and
provides a means to screen
for activity and durability of
an antimicrobial treatment
under conditions of organic
loading. This test method
provides for the
simultaneous assessment
of multiple carpet
components for
antimicrobial activity.
Carpets may be cleaned
129
INFORMATION
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prior to testing in order to
assess the durability of the
antimicrobial effect. The
method can assess the
durability of the
antimicrobial treatments on
new carpets, and on those
repeatedly shampooed or
exposed to in-use
conditions.
Half strength (nutrient and
agar) tryptic soy agar is
used as the inoculum
vehicle for bacteria and half
strength potato dextrose
agar as the inoculum
vehicle for mold conidia.
Use of half strength agars
may reduce undue
neutralization of an
antimicrobial due to
excessive organic load.
This test method
simultaneously evaluates
(both visual and stereo-
microscopic) antimicrobial
activity both at the fiber
layer and at the primary
backing layer of carpet.
ASTM Standard Test Method 1-5 + This test method specifies Own searches
E2562-07 for Quantification of 12 the operational parameters
Pseudomonas required to grow a
aeruginosa Biofilm repeatable Pseudomonas
Grown with High Shear aeruginosa biofilm under
and Continuous Flow high shear (1). The
using CDC Biofilm resulting biofilm is
Reactor representative of
generalized situations
where biofilm exists under
high shear rather than
representative of one
particular environment. The
biofilm generated in the
CDC biofilm reactor is also
suitable for efficacy testing.
3 This test method
describes how to sample
and analyze biofilm for
viable cells. Biofilm
population density is
130
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recorded as log colony
forming units per surface
area. After the 48 h growth
phase is complete, the user
may add the treatment in
situ or harvest the coupons
and treat them individually.
This test method uses the
Centers for Disease
Control and Prevention
(CDC) biofilm reactor. The
CDC biofilm reactor is a
continuously stirred flow
reactor with high wall
shear. Although it was
originally designed to
model a potable water
system for the evaluation of
Legionella pneumophila,
the reactor is versatile and
may also be used for
growing and/or
characterizing biofilm of
varying species.
ASTM Standard Test Method 1-5 This test method should be Own searches
E640-06 for Preservatives in used to determine if a
Water-Containing preservative or
Cosmetics preservative system has
application for the
preservation of water-
miscible cosmetic products.
It sets minimal
requirements for
preservative performance
in model formulations.
ASTM Standard Test Method 18 Determines the relative Own searches
E652- for Nonresidual Liquid efficiency of household and
91(2003) Household Insecticides industrial-use, contact
Against Flying Insects insecticides dissolved in
base oils and applied in
spray formulations. It is
developed to test
insecticides against house
flies (Musca domestica, L),
but test data may also be
products against
131
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Not designed to measure
the residual action of the
spray formulation.
ASTM Standard Test Method 18 The test determines the UK guidelines
E653- for Effectiveness of relative efficacy of aerosol
91(2003) Aerosol and and pressurized space
Pressurized Space spray insecticide
Spray Insecticides formulations against house
Against Flying Insects flies (Musca domestica, L)
strains and, with
modifications in dosage,
other flying insects. Test
data obtained by this test
method may also be
product against
This test method is not
residual activity.
The test may be conducted
using approximately 100
house flies per test (small
group) or 500 flies per test
(large group). Selected
reference standards are the
Official Test Aerosol II
(OTA II) for oil based
aerosol products and
Tentative Official Aqueous
Pressurized Spray
(TOAPS) for water based
aerosol products. Aerosol
test knockdowns: % down
of total flies at 5, 10, 15
minutes after application.
Aerosol test knock down
mortality: dead knocked
down x100/total flies.
These numbers should on
average be equal to,
greater than or no more
than 5% points below the
corresponding numbers of
the reference in order to
meet the standard. No
statement on precision or
bias, only whether
132
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conformance to criteria for
success specified in the
procedure.
ASTM Standard Test Method 18 Test of insecticides against UK guidelines
E654- for Effectiveness of crawling insects:
96(2003) Aerosol and cockroaches Determines
Pressurized Spray the relative efficiency of
Insecticides Against aerosol and pressurised
Cockroaches spray formulations against
cockroaches, but test data
by this test method may
also be adequate to
support claims for use of
the product to control the
exposed or accessible
stages of silverfish, ants,
centipedes, spiders, and
certain stored product
pests. Applied as direct
sprays for 30 s. on last
instar nymphs. Observation
period: 48h. The test is not
residual action.
Ten groups with 20
organisms in each. The test
is run in conjunction with
the Official Test Aerosol II
(OTA II) (or Tentative
Official Aqueous
Pressurized Spray
(TOAPS) as the standard
basis of comparison. The
mortality after 24h should
be between 50 and 75%
when testing with the OTA.
The test specimens meet
the standard if average %
dead and moribound is
equal to, above or within
10% points less than
average % dead of the
OTA series after 48h.
Precision or bias is not
specified, only states
whether conforms to
efficacy criteria.
Standard guide for 17 2 to 8-day exposures, Communication
ASTM conducting acute depending on the species. with UK (HSE)
E729-96 toxicity tests on test Static, renewal, or flow-
(2007) materials with fishes, through procedures.
133
INFORMATION
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macroinvertebrates and Usually reported as LC50
amphibians or EC50.
ASTM Standard Test Method 18 Test of insecticides against Own searches
E938-05 for Effectiveness of crawling insects: human
Liquid, Gel, or Cream louse. Only gels or creams
Insecticides Against that liquefy at 32°C (90°F)
Adult Human Lice can be tested).
Five replicates, for a
statistical comparison of
formulations.
ASTM Standard Test Method 19 Evaluates the repellency of Own search
E939- of Field Testing Topical promising compounds that
94(2006) Applications of have undergone primary
Compounds as laboratory studies and
Repellents for Medically approved for skin
Important and Pest application for secondary
Arthropods (Including testing.
Insects, Ticks, and The method is designed for
Mites): Mosquitoes the study of mosquito
repellents, but can be
modified to determine the
repellency of candidate
compounds for other flying
insects that attack humans.
ASTM Standard Test Methods 19 Can be used to test the Own search
E951- for Laboratory Testing efficacy of repellent
94(2006) of Non-Commercial compounds that can be
Mosquito Repellent diluted with ethanol,
Formulations On the acetone etc. Both biological
Skin effectiveness and
persistence of the repellent
can be assessed.
ED50 and ED95 are
determined for comparative
and practical purposes
respectively. Precision of
the test can be evaluated
(confid intervals).
ASTM Standard Test Method 13 TNsG on Prod
E979- for Evaluation of Eval
91(2004) Antimicrobial Agents as
Preservatives for Invert
Emulsion and Other
Water Containing
Hydraulic Fluids
ASTM Standard Practice for 8- A wide variety of properties Own searches
G160-03 Evaluating Microbial 10? may be affected by
Susceptibility of microbial attack depending
Nonmetallic Materials on material or item
by Laboratory Soil characteristic. Standard
Burial methods (where available)
134
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should be used for each
different property to be
evaluated. This practice
does not attempt to
enumerate all of the
possible properties of
interest nor specify the
most appropriate test for
those properties. Test
methods must, however, be
appropriate to the material
being tested. Evaluation of
a nonmetallic material's
microbiological
susceptibility when in
contact with the natural
environment of the soil and
is intended for use on
material test specimens
that are approximately 2
cm (3/4 in.) thick and 100
cm2 (20 in2) or less. It is
recommended that this
practice be combined with
appropriate environmental
exposures (for example,
sunlight simulating
weathering devices, the
hydrolytic effects of
extended aqueous contact,
or extraneous nutrients) or
fabrication into articles (for
example, adhesive bonding
of seams) which may
promote microbiological
susceptibility during the
service life of material.
Microbiological
susceptibility may be
reflected by a number of
changes including staining,
weight loss, or reduction in
tensile or flexural strength.
This practice may be
applied to articles that do
not spend the majority of
their service life in soil.
ASTM Standard Practice for 7 Determination of the Own searches
G29- Determining Algal susceptibility of plastic films
96(2002) Resistance of Plastic to the attachment and
Films proliferation of surface-
135
INFORMATION
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growing algae, produced in
bodies of water, such as
swimming pools, artificial
ponds, and irrigation
ditches that are lined with
plastic films.
Evaluating the degree and
permanency of protection
against surface growth of
algae afforded by various
additives incorporated in
the film.
WK12880 Evaluation of Clean
Room Disinfectants
ASTM Revision of E1839- 12 Efficacy of slimicides for Own searches
WK14214 96(2002) Standard Test the control of bacterial and
Method for Efficacy of fungal slimes in paper mill
Slimicides for the Paper systems and their
Industry--Bacterial and counterparts.
Fungal Slime
ASTM Test Method for ? This test method assesses Own searches
WK15324 Determining the Time- the microbicidal activity of
Kill Kinetics Of antimicrobial materials,
Antimicrobial whereby the survival of
Compounds organisms exposed to a
water miscible antimicrobial
agent is determined as a
function of time. The
primary purpose of this
method is to provide a set
of standardized conditions
and test organisms to
facilitate comparative
assessments of
antimicrobial materials
miscible in aqueous
systems.
ASTM Revision of E2180-01 9 Evaluates (quantitatively) Manufacturer
WK16397 Standard Test Method the antimicrobial (updated),
for Determining the effectiveness of agents applied to
Activity of Incorporated incorporated or bound into textiles.
Antimicrobial Agent(s) or onto mainly flat (two
In Polymeric or dimensional) hydrophobic
Hydrophobic Materials or polymeric surfaces. The
method focuses primarily
on assessing antibacterial
activity; however, other
microorganisms such as
yeast and fungal conidia
may be tested using this
136
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method.This method can
confirm the presence of
antimicrobial activity in
plastics or hydrophobic
surfaces. The vehicle for
the inoculum is an agar
slurry which reduces the
surface tension of the
saline inoculum carrier and
allows formation of a
"pseudo-biofilm," providing
more even contact of the
inoculum with the test
surface.
The method allows
determination of
antimicrobial activity
between untreated plastics
or polymers and those with
bound or incorporated low
agents. Comparisons
between the numbers of
survivors on preservative-
treated and control
hydrophobic surfaces may
also be made. The
procedure also permits
determination of "shelf-life"
or long term stability of an
antimicrobial treatment
which may be achieved
through testing both non-
washed and washed
WK4751 Selecting Test Methods
to Determine the
Efficacy of Antimicrobial
Agents and Other
Chemicals for
Sanitization of Produce
ASTM Standard Test Method 9 Designed to evaluate Own searches

WK4757 for the Assessment of (qualitatively) the presence
Antimicrobial Activity In and effectiveness of
Carpets; Seeded-Agar antimicrobial preservatives
Overlay Screen in or on carpets. This
method can be used to
evaluate both the
137
INFORMATION
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antibacterial and antifungal
activity
This method is useful for
assessing the durability of
the antimicrobial treatments
because it can be
performed on both new
carpets and those that
have been repeatedly
shampooed or exposed to
in-use conditions.
This method utilizes either
tryptic soy agar as the
inoculum vehicle for
bacteria or potato dextrose
agar as the inoculum
vehicle for mold conidia. .
Full compliment agars can
be used to mimic high soil
or organic loads on carpets
or partial nutrient
complement agars can be
used to mimic lightly soiled
conditions on carpet. This
method allows for the
simultaneous evaluation
(both visual and stereo-
microscopic) of
antimicrobial activity both at
carpet.
ASTM Guideline For 6 This method uses an agar- Own searches
WK5097 Evaluating The dilution procedure to
Potential For Decline In determine the Minimum
Planktonic Inhibitory Concentration
Microorganism (MIC) and subsequently to
Susceptability To screen for the development
Antimicrobial of decreased susceptibility
Compounds: Healthcare for test formulations versus
Applications bacteria or yeast species.
The inocula used on the
second, third, fourth, and
fifth test cycles will be
prepared from the microbial
growth present at the
previous day’s end
point;i.e., that agar-dilution
plate containing the highest
concentration of the test
formulation (lowest dilution)
138
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that allows microbial
growth.
ASTM Standard guide for 17 A flow through test, Communication

WK6845 conducting early life- beginning before hatch and with UK (HSE)
[revision stage toxicity test with ending after hatch. 28 to
of E 1241- fishes 120-day (depending on
98 (2004)] species) continuous
exposure.
ASTM New Standard Test 13 Determines the relative Own searches
WK8252 Method for Determining bioresistance of aqueous
Resistance of Aqueous metalworking fluids towards
Metalworking Fluids non-tuberculous (NTM),
towards Non- rapidly growing( RGM),
Tuberculous, environmental
Environmental mycobacteria by
Mycobacteria challenging them with a
mycobacterial inoculum
isolated from actual spoiled
metalworking fluid field
samples from the user/s
site.
In order to simulate field
conditions, another
challenge inoculum
consisting of a mixture of
common metalworking fluid
spoilage microorganisms
originating from actual
MWF field samples is also
used.
ASTM Guideline for Evaluation 1-5 Own searches
WK9062 of Residual
Effectiveness of
Antibacterial Personal
Cleansing Products
WK9378 for Determining the
Fungus-Eliminating
Effectiveness of
the Fingerpads of
Adults
Nagetierbekämpfungsm
itteln gegen Hausmaüse UK guidelines
Manufacturer
139
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Nagetierbekämpfungsm
itteln gegen UK guidelines
Wanderratten
Manufacturer
Bechert et A new method for 1-5 The test is based on the Test institute
al., Nature screening anti-infective release of vital daughter
Medicine biomaterials cells from the sample
6, 1053- surface into the
1056 surrounding. The
(2000) proliferation activity of
these daughter cells, which
are responsible for infection
development, can be
monitored in a time course.
Antimicrobial activity is
monitored by the time
needed to reach a defined
optical density, which is
dependent on the number
of released daughter cells.
All measurements are
performed in comparison to
an untreated control
(without antimicrobial
additive). The difference
between the reference (A)
and the test sample (B) to
reach the threshold OD
gives the degree of
antimicrobial activity. If all
bacteria on the surface of
the material are prevented
from multiplying, no
daughter cells are
produced and the test
object is considered
bactericidal. Materials can
also be antimicrobial, which
means that not all cells on
the test surface are
prevented from growing.
Some cells are able to
divide and release
daughter cells into the
surroundings, which are
then optically registered in
a so-called growth curve. If
surviving daughter cells are
140
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grown under controlled
conditions over 48 h
(observation time), a higher
turbidity and therefore a
bigger signal is generated.
At the same time only vital
and proliferative cells are
accounted for.
In particular antimicrobial
samples will release
daughter cells into the
surrounding. Hence,
microbial growth is first
observed noticeably later.
This right-shift towards
longer times is indicative
for the antimicrobial
efficacy of the tested
samples. The so-called
onset OD serves as a
quantifiable parameter and
is equivalent to the required
number of hours required
for the surviving daughter
cells to grow to a
predefined optical density
(OD = 0.2).
A material is defined to be
antimicrobial only if the
formation of at least 99.9%
(which is equivalent to a
net onset-OD of 6 hours
and is similar to a killing
rate of 3 log10 steps) of the
daughter cells during the
challenge time is prevented
in comparison with the
blank sample.
Marketed worldwide by
Ciba Specialty Chemicals
under the tradename
NumetrikaTM.
BS General Purpose 1-5 Mycobacterium fortuitum Manufacturer
6734:200 Disinfection Test
4
BS 3900 Methods of test for 10 Provides only a TNsG Prod Eval
paints. Part G6. methodology for production
Assessment of of a test surface for UK guidelines
141
INFORMATION
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resistance to fungal exposure by inoculation
growth. BSI London, UK with mold growth. The test
therefore has to be
modified to be used as a
test method for assessing
interior surface biocides.
against houseflies.
Specification for TM II05 (Fr)
insecticidal performance
UK guidelines

against houseflies.
Method for TM II05 (Fr)
determination of
insecticidal performance UK guidelines
Determination of the 9 Manufacturer

BS resistance of textiles
6085:199 to microbiological
2 deterioration
CEB Laboratory test method 14 Laboratory test method TMII Fr
(1981) to evaluate the efficacy
of rodenticidal products
in rats
CEB Trial method to evaluate 14 TMII Fr

(1981) the efficacy of
rodenticidal products
against rats under
CEB 107 Trial method to evaluate 18
(1985) the efficacity of
insecticidal products for
the control of stable flies
in premises for the
rearing of domestic
animals under practical
conditions
CEB 135 Trial method to evaluate 18 Surface treatments of TM II05 (Fr)
(1987) the efficacy of storage premises of
insecticidal or miticidal products
products for treatments
of storage premises of
products for animals or
plants
CEB Laboratory test method 18 Space treatments TM II05 (Fr)
135bis to evaluate the efficacy
142
INFORMATION
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(1996) of insecticidal products
in premises for the
storage, industrial
processing and sale of
products from animals
or plants
CEB 159 Trial method to evaluate 18 TM II05 (Fr)
insecticidal products for
the control of
cockroaches in
buildings under practical
conditions
insecticidal bait
products against
common species
(1999) the efficacy of a
fumigant for insect
control in premises for
the storage, processing
and production of food
(2001) the efficacy of fumigants
for insect control in
stored products
CEN Washer-disinfectors - 1-5 Also ISO method. Own searches
15883- Part 5: Test soils and Acceptance criteria are
5:2005 methods for included, based on visual
demonstrating cleaning inspection and/or a
efficacy microbiological end-point
as stated for each method.
Where chemical detection
of residual soiling is
required/sought, methods
can be complemented by
the specific determination
of a residual component of
the applied test soil.
CEN/TR Wood preservatives - 8 Pretreatment procedure Own searches
15046:20 Artificial weathering of before efficacy testing
05 treated wood prior to
biological testing - UV-
radiation and water-
spraying procedure
CEN/TS Wood preservatives - 8 TM II05 (Fr)
12037:20 Field test method for
03 determining the relative
protective effectiveness
143
INFORMATION
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of a wood preservative
contact - Horizontal lap-
joint method
CEPE Antifouling coatings - 21 Simulated field raft test TNsG PE
Antifouling methods for the
Working generation of efficacy TM II05
Group data
1993 UK guidelines

BIO-E 001 treated soils
BIO-E 002 treated masonry
CTBA- Field test on bait system 8 TM II05 (Fr)
BIO-E 003 for termite control
CTBA- Laboratory tests for bait 8 TM II05 (Fr)
BIO-E 004 system for termite
control
CTBA- Laboratory tests for bait 8 TM II05 (Fr)
BIO-E 005 system for termite
control
CTBA- Action of U.V. light on 8 TM II05(Fr)
BIO-E 006 physico-chemical
barrier for termite
CTBA- Action of alcalin 8 TM II05 (Fr)
BIO-E 007 conditions on physico-
chemical barrier for
termite control
BIO-E 008 physico-chemical
barrier for termite
control
CTBA- Forced test on termite 8 TM II05 (Fr)
BIO-E 009 bait control
CTBA- Choice test on termite 8 TM II05 (Fr)
BIO-E 010 bait control
Dautel H, Verwendung von 19 Dossier
Hilker M, Dodecansäure als
Kahl O, Zeckenrepellent
Siems K, (Patentschrift).
2001.
EEC: 12 Used in the process of oil Communication
1988 Algal inhibition test recovery with UK (HSE)
EN Chemical disinfectants 1-5 Basic bacterial activity Industry
1040:200 and antiseptics - against of a test material organizaton
5 Quantitative suspension against Staphylococcus
test for the evaluation of aureus (ATCC 6538) and manufacturer
basic bactericidal Pseudomonas aeruginosa Communication
activity of chemical (ATCC 15442). A test with UK (HSE)
disinfectants and suspension of bacteria is
144
INFORMATION
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antiseptics - Test added to a prepared TM II05 (Fr)
method and sample of the product
requirements (phase 1) under test. The mixture is TNsG PE
maintained at 20 oC. At a
specified contact time (updated)
chosen from one of the
following: 1 , 5, 15 , 30 , 45
or 60 minutes, an aliquot is
taken. The bactericidal
action of this aliquot is
immediately neutralised or
suppressed by a validated
method. The method of
choice is dilution-
neutraliser cannot be
found, membrane filtration
is used. The number of
surviving bacteria in each
sample is determined and
the reduction in viable
counts calculated.A
criterion for activity by this
test method is that the test
material should
demonstrate at least a 5-
log reduction in viable
counts of the test
organisms in 60 minutes.
EN Wood preservatives - 8 Laboratory test. Efficacy TNsG on Prod
113:1996/ Test method for criteria based on weight Eval
A1:2004 determining the loss of untreated specimen.
protective effectiveness TM II05 (Fr)
against wood destroying
basidiomycetes - NL guidance
toxic values UK guidelines
Test institute
Manufacturer

117:2005 Determination of toxic Eval
values against
Reticulitermes species TM II05 (Fr+De)
(European termites)
UK guidelines
145
INFORMATION
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preventive action
against Reticulitermes TM II05 (Fr+De)
species (European
termites) (Laboratory NL guidance
method)
UK guidelines
EN Chemical disinfectants 1-5 General guidance Industry

06 Preservation of test
organisms used for the TM II05 (Fr)
determination of
bactericidal, TNsG PE
mycobactericidal,
sporicidal and fungicidal
activity
98 Preservative-treated
solid wood -
penetration and
retention of creosote in
treated wood
EN Chemical disinfectants 1-5 Basic fungicidal activity Industry
1275:200 and antiseptics - against Candida albicans organizaton
5 Quantitative suspension (ATCC 10231) and
test for the evaluation of Aspergillus niger (ATCC Manufacturer
basic fungicidal or basic 16404). A test suspension
yeasticidal activity of of yeast cells or mould
chemical disinfectants spores is added to a
and antiseptics - Test prepared sample of the
method and product under test. The
requirements (phase 1) mixture is maintained at 20
oC. At a specified contact
time chosen from one of Communication
the following 5, 15, 30 or 60 with UK (HSE)
minutes, an aliquot is
taken; the fungicide action TM II05 (Fr)
in this portion is
immediately neutralised or TNsG PE
suppressed by a validated
method. The method of
choice is dilution-
neutraliser cannot be
found, membrane filtration
is used. The number of
surviving yeast cells or
146
INFORMATION
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mould spores in each
sample is determined and
counts calculated.
The criterion for activity by
this test is that the test
material should
counts of the test
organisms in 60 minutes.
EN Chemical disinfectants 1-5 Dilution/Neutralisation . Industry
1276:199 and antiseptics - Efficacy determined by a organizaton
7 Quantitative suspension reduction in CFU.
test for the evaluation of Manufacturer
food, industrial,
domestic, and
institutional areas - Test
method and
requirements (phase 2,
step 1)
TM II05 (Fr)
TNsG PE
EN Chemical disinfectants 1-5 Testing mainly bactericidal Industry

12791:20 and antiseptics - activity; in vivo test; phase organizaton/
05 Surgical hand 2 step 2 Manufacturer
disinfection - Test
method and TM II05 (Fr)
requirement (phase
2/step 2) TNsG PE
EN Chemical disinfectants - 1-5 Industry
13610:20 Quantitative suspension organizaton
02 test for the evaluation of
virucidal activity against TM II05 (Fr)
bacteriophages of
chemical disinfectants TNsG PE
used in food and
industrial areas - Test
method and
step 1)
13624:20 and antiseptics - organizaton/man
03 Quantitative suspension ufacturer
147
INFORMATION
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test for the evaluation of
for instruments used in TM II05 (Fr)
method and TNsG PE
step 1)
EN Chemical disinfectants 1-5 Dilution/Neutralisation Industry
13697:20 and antiseptics - Method. Efficacy measured organizaton
01 Quantitative non-porous by a reduction in CFU.
surface test for the Manufacturers
evaluation of
bactericidal and/or
used in food, industrial,
domestic and
method and
requirements without
mechanical action
(phase 2/step2)
TM II05 (Fr)
TNsG PE
EN Chemical disinfectants - 1-5 Industry

13704:20 Quantitative suspension organizaton
02 test for the evaluation of
sporicidal activity of Manufacturer
used in food, industrial, TM II05 (Fr)
domestic and
institutional areas - Test TNsG PE
method and
step 1)
EN Chemical disinfectants 1-5 Efficacy determined by the Industry
03 Quantitative suspension
for instruments used in
method and
step 1)
TM II05 (Fr)
148
INFORMATION
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TNsG PE
EN Wood preservatives - 8 TNsG Prod Eval

6 eradicant action against TM II05 (Fr)
Hylotrupes bajulus
(Linnaeus) larvae - NL guidance
Laboratory method
UK guidelines
EN Chemical disinfectants 1-5 Dilution/Neutralisation Industry

14204:20 and antiseptics - Method. Effiacy measured organisation
04 Quantitative suspension by reduction in CFU.
mycobactericidal activity
of chemical
disinfectants and TM II05 (Fr)
method and
requirements (phase 2, TNsG PE
step 1)
EN Chemical disinfectants 1-5 Basic sporicidal activity Industry
14347:20 and antiseptics - Basic against dormant spores of organizaton
05 sporicidal activity - Test Bacillus subtilis (ATCC
method and 6633) and Bacillus cereus Manufactuer
requirements (phase 1) (ATCC 12826). A prepared
sample of the product
under test is added to a
test suspension of bacterial Communication
spores. The mixture is with UK (HSE)
maintained at 20 oC or any
other temperature to be TM II05
defined. At a specified
contact time chosen from TNsG PE
one of the following: 30, 60
and 120 minutes, an aliquot
portion is taken and the
sporicidal as well as
sporistatic action in this
portion is neutralised. The
method of choice is
dilution-neutralisation. The
number of surviving
bacterial spores is
determined in parallel and
counts calculated. The
effectiveness of
neutralisation is controlled
149
INFORMATION
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in the test.
The criterion for activity by
this test is that the test
material should
counts of the test
organisms in 120
minutes.Medical area
Veterinary area, Food,
industrial, domestic and
institutional hygiene
EN Chemical disinfectants 1-5 Efficacy measured by a Industry
mycobactericidal activity
of chemical
disinfectants in the
medical area including
instrument disinfectants
- Test methods and
step 1)
EN Chemical disinfectants 1-5 Non-porous surface test, Industry
14349:20 and antiseptics - bactericidal, for organizaton
07 Quantitative surface test disinfectants used in the
for the evaluation of veterinary applications
bactericidal activity of Dilution/Neutralisation Manufacturer
chemical disinfectants Method. By one
and antiseptics used in manufacturer, the test
veterinary area on non- material is deemed to have
porous surfaces without passed the test and be TM II05 (Fr)
mechanical action - efficacious if it
Test method and demonstrates a log 4 or
requirements (phase 2, more reduction in viable TNsG PE
step 2) counts under the conditions
defined in the test.
Reduction in viable
microbial counts compared
with water controls.
05+A1:20 Virucidal quantitative
06 suspension test for Manufacturer
human medicine - Test TM II05 (Fr)
method and
step 1)
150
INFORMATION
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06 Quantitative carrier test
for the evaluation of Manufacturer
bactericidal activity for
instruments used in the TM II05 (Fr)
medical area - Test
method and
step 2)
06 Quantitative carrier test
for the evaluation of Manufacturer
fungicidal or yeasticidal
activity for instruments
used in the medical TM II05 (Fr)
area - Test method and
step 2)
virucidal activity of
and antiseptics used in TM II05 (Fr)
the veterinary area -
Test method and
step 1)
EN Ophtalmic optics - 1-5 Also ISO method. Own searches
14730: contact lense care
2000 products - antimicrobial
preservative efficacy
testing and guidance on
determining discard rate
EN 1-5 Applied in medical area Industry
1499:199 organizaton
7
Manufacturer
Chemical disinfectants
and antiseptics -
Hygienic handwash - TM II05 (Fr)
Test method and
requirements (phase TNsG PE
2/step 2)
7 Hygienic handrub - Test
method and Manufacturer
151
INFORMATION
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requirements (phase
2/step 2) TM II05 (Fr)
TNsG PE
EN 152-1 Test methods for wood 8 TNsG on Prod

(1988) preservatives - Eval
Laboratory method for
preventive effectiveness
of a preservative NL guidance
treatment against blue
stain in service - Part 1: UK guidelines
Brushing procedure
EN 152- Test methods for wood 8 TNsG on Prod

2:1988/A preservatives - Eval
C1:1989 Laboratory method for
of a preservative NL guidance
treatment against blue
stain in service - Part 2: UK guidelines
Application by methods
other than brushing
EN Paints and varnishes - 7 Manufacturer
coating against fungi
EN Paints and varnishes - 7 Own searches
coating against algae
chemical disinfectants TM II05 (Fr)
food, industrial,
domestic, and TNsG PE
method and
step 1)
EN 1656 : Chemical disinfectants 1-5 Dilution/Neutralisation Manufacturers
2000 and antiseptics - Method. Efficacy measured
Quantitative suspension by a reduction in CFU.
152
INFORMATION
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veterinary field - Test
method and
requirements (phase
2/step 1)
Industry
organisation
TM II05 (Fr)
TNsG PE
EN Chemical disinfectants 1-5 By one manufacturer, the Manufacturers

1657:200 and antiseptics - product was to be deemed
5/AC:200 Quantitative suspension to have passed the test if it
7 test for the evaluation of demonstrated a 10E5
fungicidal or yeasticidal logarithmic reduction in
activity of chemical viability.
disinfectants and
antiseptics used in the By one manufacturer a
veterinary area - Test reduction in viable Industry
method and microbial counts is organizaton
requirements (phase 2, compared with water
step 1) controls. TM II05 (Fr)
The test material is
deemed to have passed
the test and be efficacious TNsG PE
if it demonstrates a log 4 or
more reduction in viable
counts under the conditions
defined in the test
against Lyctus TM II05 (Fr)
Brunneus (Stephens) -
Part 1: Application by NL guidance
surface treatment
(laboratory method) UK guidelines

against Lyctus TM II05 (Fr)
brunneus (Stephens) -
Part 2: Application by NL guidance
impregnation
153
INFORMATION
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(Laboratory method) UK guidelines
EN Field test method for 8 Field test, in ground contact TNsG on Prod
252:1989/ determining the relative Eval
AC1:1989 protective effectiveness
of a wood preservative Test institute
in ground contact
NL guidance
UK guidelines
TM II05 (Fr)
EN Wood preservatives - 8 Marine field test, 5 year TNsG on Prod

275:1992 Determination of the minumum Eval
against marine borers Test institute
TM II05 (Fr)
NL guidance
UK guidelines

330:1993 Field test method for Eval
determining the relative
protective effectiveness Test institute
of a wood preservative
for use under a coating
and exposed out of TM II05 (Fr)
ground contact: L-joint
method NL guidance
UK guidelines

154
INFORMATION
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Guide to the principles
of testing and
classification of the
natural durability of
wood
Guide to natural
durability and treatability
of selected wood
species of importance in
Europe
Classification of
preservative penetration
and retention
Guidance on sampling
for the analysis of
preservative-treated
wood
eradicant efficacy in
preventing emergence TM II05 (Fr)
of Anobium punctatum
(De Geer) NL guidance
UK guidelines

solid wood - Guide to
the durability
requirements for wood
to be used in hazard
classes
preventive action
against Hylotrupes TM II05 (Fr+De)
155
INFORMATION
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Part 1: Larvicidial effect NL guidance
(Laboratory method)
UK guidelines
EN 46- Wood preservatives - 8 TM II05 (De)

preventive action NL guidance
against Hylotrupes
Part 2: Ovicidal effect
(laboratory method)
47:2005/A Determination of the Eval
C:2007 toxic values against
larvae of Hylotrupes TM II05 (Fr+De)
UK guidelines

eradicant action against
larvae of Anobium TM II05 (Fr+De)
punctatum (De Geer)
(laboratory method) NL guidance
UK guidelines

against Anobium TM II05 (Fr+De)
punctatum (De Geer) by
egg-laying and larval NL guidance
survival - Part 1:
Application by surface UK guidelines
treatment (Laboratory
method)
against Anobium TM II05 (Fr+De)
punctatum (De Geer) by
egg-laying and larval NL guidance
survival - Part 2:
Application by UK guidelines
impregnation
(Laboratory method)
EN Wood preservatives - 8 Pretreatment procedure TNsG on Prod
156
INFORMATION
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73:1988/A Accelerated ageing of before efficacy testing Eval
C:1992 treated wood prior to
Evaporative ageing
procedure NL guidance
UK guidelines
EN Wood preservatives - 8 Pretreatment procedure TNsG on Prod

84:1997 Accelerated ageing of before efficacy testing Eval
treated wood prior to
Leaching procedure
NL guidance
UK guidelines
EN ISO Water quality - Marine 21 Growth inhibition test TM II05

10253:20 algal growth inhibition
06 test with Skeletonema
costatum and
Phaeodactylum
tricornutum (ISO
10253:2006)
EN ISO Textiles - Determination 9 Manufacturer
11721- of the resistance of
1:2001 cellulose-containing
textiles to micro-
organisms - Soil burial
test - Part 1:
Assessment of rot-
retardant finishing (ISO
11721-1:2001)
EN ISO Textiles - Determination 9 Own searches
11721- of the resistance of
2:2003 cellulose-containing
textiles to micro-
organisms - Soil burial
test - Part 2:
Identification of long-
term resistance of a rot
retardant finish (ISO
11721:2003)
EN ISO Textile fabrics - 9 Manufacturer
04 antibacterial activity -
Agar diffusion plate test
(ISO 20645:2004)
EN ISO Textiles - Determination 9 Applied to textiles Manufacturer
20743:20 of antibacterial activity
157
INFORMATION
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07 of antibacterial finished Reference material: non Laboratory
products (ISO treated polyester
20743:2007)
EN/TR Durability of wood and 8 Own searches
04 Determination of
treatability of timber
species to be
impregnated with wood
preservatives -
Laboratory method
EN/TR Wood preservatives - 8 Own searches
04 preventive efficacy
basidiomycetes fungi
EN/TS Wood preservatives - 8 Field test, out of ground Test institute
12037:20 Field test method for contact
03 determining the relative
of a wood preservative UK guidelines
contact - Horizontal lap-
joint method
05 preventive effectiveness
against sapstain fungi
and mould fungi on
freshly sawn timber -
Field test
15397:20 Method for natural
06 preconditioning out of
ground contact of
treated wood
specimens prior to
biological laboratory test
ENV Durability of wood and 8 TM II05 (Fr)
02 Wood-based panels - UK guidelines
Method of test for
determining the
resistance against
wood-destroying
basidiomycetes
ENV Durability of wood and 10 Simulated use test TNsG Prod Eval
97 Assessment of the TM II05 (Fr)
effectiveness of a
masonry fungicide to UK guidelines
158
INFORMATION
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prevent growth into
wood of Dry Rot
Serpula lacrymans
(Schumacher ex Fries)
S.F. Gray - Laboratory
method
ENV Wood preservatives - 8 Laboratory test TM II05 (Fr)
and other soil inhabiting NL guidance
micro-organisms
UK guidelines
ENV Wood preservatives - 8 TM II05 (Fr)

protective effectiveness NL guidance
basidiomycetes - UK guidelines
treatment
17 Communication
EPA 72-5 Life-cycle test of fish with UK (HSE)
Simulated or actual field 17 Communication
testing for aquatic with UK (HSE)
EPA 72-7 organisms
EPA/OPP Standard Norway Rat 14 Anticoagulant rodenticide TNsG on Prod
Protocol and Roof Rat test against Norway Eval
Number: Anticoagulant Liquid rat/Roof rat applied as
1.201 Bait Laboratory Test liquid bait NL guidance
Method
UK guidelines
EPA/OPP Standard House Mouse 14 Anticoagulant rodenticide TNsG on Prod

Protocol Anticoagulant Liquid test against House mouse Eval
Number: Bait Laboratory Test applied as liquid bait
UK guidelines

Number: Anticoagulant Dry Bait rat/Roof rat applied as dry
1.203 Laboratory Test Method bait NL guidance
UK guidelines

Protocol Anticoagulant Dry Bait test against House mouse Eval
Number: Laboratory Test Method applied as Dry bait
1.204 NL guidance
159
INFORMATION
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UK guidelines
EPA/OPP Standard Norway 14 Anticoagulant rodenticide TNsG on Prod

Protocol Rat/Roof Rat test against Norway Eval
Number: Anticoagulant Tracking rat/Roof rat applied as
1.205 Powder Efficacy Tracking powder NL guidance
Laboratory Test Method
UK guidelines
EPA/OPP Standard Norway 14 Acute rodenticide test TNsG on Prod

Protocol Rat/Roof Rat Acute against Norway rat/Roof rat Eval
Number: Liquid Bait Laboratory applied as Liquid bait
UK guidelines
EPA/OPP Standard House Mouse 14 Acute rodenticide test TNsG on Prod

Protocol Acute Liquid Bait against House mouse Eval
Number: Laboratory Test Method applied as Liquid bait
1.208 NL guidance
UK guidelines

Protocol Rat/Roof Rat Acute Dry against Norway rat/Roof rat Eval
Number: Bait Laboratory Test applied as Dry bait
UK guidelines

Protocol Acute Dry Bait against House mouse Eval
Number: Laboratory Test Method applied as Dry bait
1.210 NL guidance
UK guidelines

Protocol Rat/Roof Rat Acute against Norway rat/Roof rat Eval
Number: Tracking Powder applied as Tracking powder
1.211 Efficacy Laboratory Test NL guidance
Method
UK guidelines

Protocol Anticoagulant Tracking test against House mouse Eval
Number: Powder Efficacy applied as Tracking powder
1.212 Laboratory Test Method NL guidance
UK guidelines
160
INFORMATION
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EPA/OPP Standard Norway 14 Anticoagulant rodenticide TNsG on Prod

Protocol Rat/Roof Rat test against Norway Eval
Number: Anticoagulant Wax rat/Roof rat applied as Wax
1.213 Block and Wax Pellet block and wax pellet NL guidance
UK guidelines

Protocol Anticoagulant Wax test against House mouse Eval
Number: Block and Wax Pellet applied as Wax block and
1.214 Laboratory Test Method wax pellet NL guidance
UK guidelines

Number: Anticoagulant rat/Roof rat applied as
1.217 Placepack Dry Bait Placepark dry bait NL guidance
UK guidelines

Protocol Anticoagulant test against House mouse Eval
Number: Placepack Penetration applied as Placepark
1.218 Laboratory Test Method penetration NL guidance
UK guidelines
EPA/OPP Standard Norway Rat 14 Acute rodenticide test TNsG on Prod

Protocol and Roof Rat Acute against Norway rat/Roof rat Eval
Number: Placepack Penetration applied as Placepark
1.219 Laboratory Test Method penetration NL guidance
UK guidelines

Protocol Acute Placepack Dry against House mouse Eval
Number: Bait Laboratory Test applied as Placepark dry
1.220 Method bait NL guidance
UK guidelines
EPA/OPP Proposed Norway Rat 14 Anticoagulant rodenticide TNsG on Prod

Protocol Anticoagulant Technical test against Norway rat Eval
Number: and Concentrated Dry applied as Technical and
1.221 Bait Laboratory Test concentrated dry bait NL guidance
Method
UK guidelines
EPA/OPP Proposed Norway Rat 14 Acute rodenticide test TNsG on Prod
161
INFORMATION
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Protocol Acute Technical and against Norway rat applied Eval
Number: Concentrated Dry Bait as Technical and
1.222 Laboratory Test Method concentrated dry bait NL guidance
UK guidelines
EPA/OPP Proposed House Mouse 14 Anticoagulant rodenticide TNsG on Prod

Protocol Anticoagulant Technical test against House mouse, Eval
Number: and Concentrated Dry applied as Technical and
1.225 Bait Laboratory Test concentrated dry bait NL guidance
Method
UK guidelines
EPA/OPP Proposed House Mouse 14 Acute rodenticide test TNsG on Prod

Protocol Acute Technical and against House mouse Eval
Number: Concentrated Dry Bait applied as Technical and
1.226 Laboratory Test Method concentrated dry bait NL guidance
UK guidelines
EPA/OPP Proposed House Mouse 14 Acute rodenticide test TNsG on Prod

Protocol Acute Tracking Powder against House mouse Eval
Number: Efficacy Laboratory Test applied as Tracking powder
UK guidelines
Admixture of plant 18 + TM II05 (Fr)

protection products to 20
EPPO stored plant products to
PP control insects and
1/203(1) mites
EPPO Blue-Stain fungi of 8 Own searches
PP Softwood
1/194(2)
(1995)
14 NL guidance
EPPO
PP Field rodents (Microtus, TM II05 (Fr)
1/169(2 Arvicola)
Field tests against 14 NL guidance
EPPO synanthropic rodents
PP (Mus musculus, Rattus TM II05 (Fr)
1/114(2) norvegicus, R. rattus)
EPPO Fumigants to control 18 + TM II05 (Fr)
PP insect and mite pests of 20
1/201(1) stored plant products UK guidelines
EPPO Laboratory and field 14 NL guidance
PP tests for the evaluation
1/97(2) of rodenticidal dusts TM II05 (Fr)
EPPO Laboratory testing of 18 UK guidelines
162
INFORMATION
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PP plant protection
1/204(1) products against insect
and mite pests of stored
plant products
Laboratory tests for 14 NL guidance
evaluation of the toxicity
EPPO and acceptability of TM II05 (Fr)
PP rodenticides and
1/113(2) rodenticide preparations Test institute
EPPO 14
PP Non-target effects of NL guidance
EPPO Rodent repellents 14 Own search
PP against debarking of
1/200(1) trees
EPPO 14 Own search
PP
1/199(1) Rodent seed repellents
18 TM II05 (Fr)
EPPO Space and structural
PP treatments of store UK guidelines
1/202(1) rooms
Testing rodents for 14 NL guidance
EPPO resistance to
PP anticoagulant TM II05 (Fr)
rodenticides N°1.
Laboratory tests for UK guidelines
evaluation of the toxicity
and acceptability of Manufacturer
rodenticides and
rodenticide preparations
rodenticides. Field tests
against synanthropic UK guidelines
rodents (Mus musculus,
Rattus norvegicus, Manufacturer
rattus rattus)
rodenticides. Laboratory
and field tests for the UK guidelines
evaluation of
rodenticidal dusts.
EPPO The EPPO Conference 18 TNsG on Prod
Bulletin, on Fumigation, Paris, Eval
15 Pages 1983
1-119, UK guidelines
163
INFORMATION
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Paris
(1983)
EPPO, EPPO 18 TNsG on Prod
Paris Recommendations on Eval
(1982) fumigation standards
(2nd Edition) UK guidelines
Fradin & Comparative efficacy of 19 Human subjects: Arm in Dossier
Day, July insect repellents against cage studies (15
2002 mosquito bites volunteers, 10 mosquitoues
N Engl J (Aedes aegypti) in each
Med vol cage. Endpoint: elapsed
347 vol time to first bite. Category
13-18 of protection A-H
(significantly different mean
complete protection time;
ANOVA & Tukey's). No
need to recalculate the
results to "real condition"
(simulate real condition)
Hummel, Effect of the neem 19 Dossier
E., extract formulation
Kleeberg, neemazal-t/s on the
H. 1997. green pea aphid
in: acyrthosiphon pisum in
Practice the laboratory (1995),
orientated in: Practice orientated
results on results on use and
use and production of Neem-
production Ingredients and
of Neem- Pheromones V
Ingredient
s and
Pheromon
es V.
Proceedin
gs of the
5th
workshop,
Wetzlar,
Germany,
January
22-25,
1996
IBRG A Standardized 13 TNsG on Prod
(draft Screening Method for Eval
MWF) Determining the
(1993) Bioresistance of and
Evaluating Biocides in
Aqueous Metal Working
Fluids
ISO Textiles - Determination 18 Applicable to all textiles TNsG on Prod
164
INFORMATION
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3998:197 of resistance to certain containing animal fibres in Eval
7 insect pests any proportion. Conditioned
voracity control specimens TM II05 (Fr)
and test specimens of
known mass are placed in UK guidelines
contact with selected larvae
for 14 days. The loss in
mass of all specimens and
the condition of the test
larvae are ascertained to
assess the resistance of
each test specimen.
JIS K Test methods for 8 Own searches
1571:200 determining the
4 effectiveness of wood
preservatives and their
performance
requirements
JIS L Testing for antibacterial 9 Antimicrobial activity. Test institute
1902: activity and efficacy Reference material: e.g.
2002 on textile products non treated polyester.
Manufacturer
Antimicrobial Products – 9 Applied to textiles Manufacturer
JIS Z Test for antimicrobial
2801: activity Test institute
2000 and efficacy
JIS Z 9 Applied to textiles Manufacturer
2911:
1992 (PT Methods of test for
9) fungus resistance
MAFF Disinfectants for use 1-5 TNsG PE
(1969) specifically against: a)
anthrax, brucellosis,
contagious bovine
pleuro-pneumonia and
glanders; b) For use
against tuberculosis; c)
For use against foot-
and-mouth disease; d)
For use against fowl
pest (Newcastle
disease fowl plague).
(2002) mat: part 1: physical
and chemical
requirements (first
165
INFORMATION
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revision)
efficacy - glass chamber
method (first revision)
efficacy - glass cylinder
method
efficacy - peet grady
method
delivery rate of aerosol
dispenser
residual spray aerosol
space spray aerosol
(1994) determination of brimful
capacity of aerosol cans
(1996) vaporizer: Part 1:
physical and chemical
requirement
method for evaluation of
biological efficacy -
method for evaluation of
glass cylinder method
MS 1497 Methods of biological 18 Manufacturer
(2000) evaluation of the
166
INFORMATION
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ELSEWHERE)
efficacy of repellent -
bioassay method for
mosquito repellent on
human skin
(1998) physical and chemical
requirements (third
revision)
(1996) method for evaluation of
(first revision)
(1998) method for evaluation of
biological efficacy - peet
grady method
100-4 wood-based products -
October Definition of use classes
2007 - Part 4 : national
declaration on the
situation of biological
agents
105-3 wood based products -
February Preservative treated
2008 solid wood - Part 3 :
wood preservation
performance and
treatment certificate -
Adaptation to France
metropolitan territory
and DOM
NF FD Protection - Termites - 8 TM II05 (Fr)
X40-501 Buildings protection
November against termite
2005 infestation
NF G39- Properties of textiles - 18 Manufacturer
011 April Textiles and polymeric
2001 materials having TM II05 (Fr)
antiacarien properties -
measurement of
antiacarien activity
050 Technical grade borax.
Septembe Specifications and tests.
167
INFORMATION
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r 1986
052 Boric acid.
Septembe Specifications and tests.
r 1986
054 Technical grade
Septembe pentachlorophenol.
r 1995 Specificaitons and tests.
061 Technical grade sodium
Septembe pentachlorophenate.
r 1995 Specifications and tests.
065 Technical grade sodium
Septembe pentaborate.
r 1986 Specifications and tests.
NF T72- Products for protecting 8 TM II05
083/A1 wood surfaces - Method
November of testing resistance to
1979 microorganisms
086 Lasures. Natural
Septembe weathering test.
r 1991
NF T72- Water-miscible, 1-5 Manufacturer
230 neutralizable antiseptics
August and disinfectants used
1988 in liquid form.
Determination of
Dilution-neutralization
method.
NF T72- Methods of airborne 1-5 TM II05 (Fr)
281 disinfection of surfaces.
Septembe Determiation of
r 1986 bactericidal, fongicidal
and sporicidal activity.
NF T72- Insecticides for flying 18 TNsG on Prod
320 insects. Insecticide Eval
March distributed under
1977 pressure ("aerosol" TM II05 (Fr)
type). Determination of
the efficiency rating. UK guidelines
Insecticides for flying 18 TM II05 (Fr)

insects. Permanent
NF T72- insecticide distributor.
March efficiency rating and the
1977 regularity rating.
168
INFORMATION
PROVIDED FROM
ELSEWHERE)
NF X41- Protection of plastics. 9 TM II05
513 Part 1. Testing method
August for resistance of
1961 ingredients to
microorganisms.
NF X41- Protection of the plastic 9 TM II05
515 materials - Part 3: Test
March method of the
1962 resistance of materials
and apparatus to
microorganisms
NF X41- Protection of textiles. 18 TM II05 (Fr)
516 Protection against
January certain insect pests.
1980 Methods of testing.
NF X41- Protection. Testing 6+ TNsG on Prod
520 method for resistance of 7 Eval
March paints to TMII05
1968 microorganisms and
their protective power.
NF X41- Protection of wood. 8 TM II05 (Fr)
521 July Testing methods for the
1968 corrosive action of wood
protection products on
metals.
541 Determination of the Eval
Septembe protective effectiveness
r 1994 against termites of TM II05
products designed for
walls, foundations and
masonry. Laboratory
method.
547 Determination of Eval
December longicide efficacy of
1992 temporary wood TM II05
protectives for green
sawn timber. Laboratory
method.
548 Determination of
December fongicide efficacy of
1992 temporary wood
protectives for fresh cut
wood billets. Laboratory
method.
August toxic values against
169
INFORMATION
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1982 {chaetomium}
{globosum} kunze. Soft
rotting agent.
NF X41- Wood preservatives – 8 TM II05 (Fr)
580 part Physicochemical testing
1-10
May 2006
NF XP Properties of textiles - 9 Manufacturer
G39-010 Textiles and polymeric
May 2000 surfaces having
antibacterial properties -
measurement of
antibacterial activity
X41-540 Termites. Determination
November of anti-termites action
1995 for products used in
liquid phase for ground
method).
NF XP Wood preservatives. 8 TNsG on Prod
X41-540 Termites. Determination Eval
November of anti-termites action
1995 for products used in
liquid phase for ground
method).
X41-542 Anti-termite treatment
Septembe product for floors, walls,
r 1995 foundations and
masonry work.
Accelerated ageing test
of treated materials
prior to biological
testing. Percolation test.
NF XP Wood preservatives - 8 TM II05 (Fr)
X41-549 Evaluation of fongicide
December efficacy of temporary
1999 wood protectives for
green sawn timber -
Site method
NF XP Wood preservatives - 8 TNsG on Prod
X41-549 Evaluation of fongicide Eval
December efficacy of temporary
1999 wood protectives for
green sawn timber -
Site method
Not Assessment of the 1-5 Various concentrations of Manufacturer
available biocide efficacy using product was mixed with
170
INFORMATION
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three algal species. In- algal suspensions
house method (Chlorella vulgaris
Navicula pelliculosa
Anabaena flos-aquae) in
glass conical flasks All
flasks were incubated and
shaken in an orbital shaker.
Samples were taken at 96h
and the chlorophyll a
concentration determined.
determining the EC50
value obtained for algae.
Not Challenge Testing in 13 The purpose of the method Manufacturer
available Metal Working Emulsion was to determine the
(in-house laboratory efficacy of preservatives
study) against bacterial and fungal
contamination in Metal
Working Fluids (MWF).
The method was conducted
by inoculating once a week
during 10 weeks mixed
bacteria and optionally
mixed fungi into water
based MWF emulsions
containing various amounts
of the test substance
diluted in 3% synthetic oil.
After each week a sample
of the emulsion was plated.
After plate incubation the
number of surviving
organisms was compared
to the control The criteria
for measuring good
microbial count of <10
cfu/ml after 10 inoculations
for all microorganisms.
Not Disinfectant for 1-5 Using a growth inhibition Manufacturer
available swimming pools for test method for recording
control of bacteria, fungi MIC values to test the
and algae. efficacy of the product at
Determination of various diluted
Minimum Inhibitory concentrations. MIC value
Concentration Against was determined using
Bacteria, Fungi, Algae microdilution technique in
and Cyanobacteria (in microtiter plates
house method) Tests were carried out in
quadruplicate. The ppm
level of the product in the
171
INFORMATION
PROVIDED FROM
ELSEWHERE)
last microtiter well
demonstrating no growth
was determined as the MIC
for the product against that
microorganism.
Not Disinfectant treatment 1-5 Several concentrations of Manufacturer
available of waste water. A the product were added to
laboratory study river water and allowed to
involving disinfection of react for 30 mins. Total
untreated river water, as counts and coliforms were
a surrogate for a full determined. Efficacy was
field trial. (in-house determined based on
methods) reduction in CFU counts.
Not Disinfectant treatment 1-5 A product was used to treat Manufacturer
available of waste water. Full waste water at 3
scale field trial concentrations under real
field conditions involving a
waste water treatment
plant; variable flow and
bacterial load, and
operating temperatures.
Efficacy was measured
based on a log removal of
bacteria after 15 and 30
mins.
Not Disinfectant treatment 1-5 Experiments were Manufacturer
available of waste water. conducted to investigate in
Inactivation of enteric pilot scale the disinfection
micro-organisms in efficiency against enteric
tertiary treated micro-organisms in tertiary
municipal waste waters treatment waste waters and
(in-house methods) the effect of dose and
contact time on disinfection
efficiency. After incubation
bacterial colonies were
counted and microbial
numbers calculated as cfu/
100 ml.
percentage reductions of
microorganisms after 8 and
18 minutes contact time.
Not Disinfectant treatment 1-5 A purpose built disinfection, Manufacturer
available of waste water. Pilot continuous flow, Pilot Plant
studies to assess the was specially constructed
disinfection and installed at a
performance of on a functioning wastewater
physicochemical treatment plant. Prior to
wastewater effluent. installation the wastewater
was treated only by
chemically assisted
172
INFORMATION
PROVIDED FROM
ELSEWHERE)
sedimentation, making
disinfection particularly
difficult. As part of this pilot
study, modifications to the
Plant and process were
explored and adopted on
the basis of experience and
experimental data using
standard methodologies.
Disinfection efficiency
under a number of
conditions and wastewater
loads were explored.
Method for recording
faecal coliforms: American
Public Health association;
American Water Works
Association and Water
Environment federation
(1998) Standard Methods
for the Examination of
Water and Wastewater
20th Ed.; Washington DC,
USA; Method 9222D.
Method for recording
Enterococcus, coliphage
Clostridium
according to laboratory
methods. Efficacy was
determined by measured
Log reductions (or removal)
in counts. Colonies were
counted before and after
disinfection.
Not Field Evaluation of in a 18 Structures in this study Manufacturer
available Termite Control Baiting were supplied by
System cooperating pest control
companies. A number of
the structures had been
identified as "problem
houses" by the pest control
operator.
Stations were inspected at
approximately monthly
(when active), and bi-
monthly (when not active)
intervals for termite activity,
with the condition of the
station (active, inactive and
baited) being noted on an
inspection form. When
173
INFORMATION
PROVIDED FROM
ELSEWHERE)
activity was found in a
station, bait was placed in
the station.
The thin wood strips of the
baiting station were fan
sprayed with product
solution using a
compressed air sprayer.
Product was mixed in the
acetone such that when the
solution was applied to the
strips, various
concentrations of product
were imparted to the wood
strips .A tumble technique
was employed to prevent
uneven concentrations of
active ingredient in the
sawdust matrix. Efficacy
was measured by
infestation
Not Field trial in a Plant 11 The antimicrobial Manufacturer
available recirculating cooling effectiveness of biocide
system was determined by
monitoring chemical and
physical cooling water
characteristics,
microbiological parameters
such as viable counts of
bacteria, fungi and algae,
and operational and
engineering parameters of
the tested cooling system
Analysis methods used for
microbial counting were
from:
Standard methods for the
Waste Water (APHA 1992),
and in-house Laboratory
SOPs. Efficacy was
determined by a reduction
in microbial counts
Not In house Laboratory 18 Formulation, applied as a Manufacturer
available Study of the Efficacy residual spray, was
against Reticulitennes assessed for efficacy
fIavipes against cockroaches (Blatta
orientalis, Blatella
germanica and Periplaneta
americana) under
174
INFORMATION
PROVIDED FROM
ELSEWHERE)
laboratory conditions. The
spray deposits were
assessed at intervals up to
24 weeks post application.
Efficacy of the formulation
at each interval, in terms of
knockdown and mortality,
was evaluated over a
period of 96 hours post
treatment application
Efficacy was measured
when all termites in a
container had died, the
length of time of
consumption by the
termites of the treated
sawdust was recorded
Not Insecticidal activity of an 18 10 German cockroaches Manufacturer
available oil based aerosol (Blattella germanica) were
against German released into a plastic
cockroach, Blattella container within a glass
germanica (in-house cylinder and the aerosol
laboratory study) was sprayed into the
cylinder. Knocked down
Not Insecticidal efficacy of 18 6 adult American Manufacturer
available an oil based aerosol cockroaches (Periplaneta
against American Americana) were released
cockroach, Periplaneta into plastic container which
Americana was placed in the centre at
the bottom of the glass
cylinder and 1013mg of
aerosol was sprayed into
the cylinder and was then
covered by a glass lid.
Knocked down insects
were counted at intervals
up to 20 minutes.
Not Insecticidal efficacy of 18 100 houseflies were Manufacturer
available an oil-based aerosol released into a Peet-Grady
against Housefly, chamber and aerosol was
Musca domestica. sprayed into the chamber.
up to 20 minutes.
Not Insecticidal efficacy of 18 50 mosquitoes (Culex Manufacturer
available an oil-based aerosol pipiens pallens) were
against mosquito, Culex released into a Peet-Grady
pipiens pallens chamber and aerosol was
sprayed into the chamber.
175
INFORMATION
PROVIDED FROM
ELSEWHERE)
up to 20 minutes. The
chamber was then
ventilated by an exhaust
fan and all insects were
transferred to a clean
recovery container with diet
and water within 20
minutes
Not Insecticide primarily for 18 Various concentrations of Manufacturer
available the control of termites: product were impregnated
Laboratory Study in a sawdust matrix readily
against Reticulitennes consumed by termites.
fIavipes Product was incorporated
into the sawdust by placing
each batch of solution and
one batch of sawdust in the
mixing bowl of a Kitchenaid
Model K5SS mixer. One
untreated batch of sawdust
served as a control.
Sterile sand and distilled
water was mixed to form a
moistened sand substrate
to sustain the termites as
they fed upon the samples.
Eight replications of each
concentration plus the
control were tested.
Termite activity was
observed in each of the
containers for four days.
infestation.
Not Laboratory studies to 18 Product was applied as a Manufacturer
available assess products applied direct spray, was assessed
as direct sprays for for efficacy against
efficacy against a range cockroaches (Blatta
of crawling insects orientalis, Blatella
germanica and Periplaneta
americana) under
laboratory conditions.
Efficacy criteria based on
mortality and knocked
down insects
Not Other in can 6 The method was conducted Manufacturer
available preservatives. In-house by inoculating mixed
method based on bacteria and mixed fungi
‘Antimicrobial into a interior paint
176
INFORMATION
PROVIDED FROM
ELSEWHERE)
Preservative formulation containing
Effectiveness Challenge various concentrations of
Test’ and DAB 10 the active substance which
guidelines was repeated on a weekly
basis. After 0, 7 14, 21 and
28 days samples of the
formulations were placed in
a preservative neutralizer
compared to the control.
good preservation efficacy
was a microbial count of
<10 cfu/ml at day 7, 14 , 21
and 28 days
Not Preservative for 6 The method was conducted Manufacturer
available detergents. In-house by inoculating mixed
method based on bacteria and mixed fungi
‘Antimicrobial into a washing up liquid
Preservative formulation containing
Effectiveness Challenge various concentrations of
Test’ and DAB 10 the active substance which
guidelines was repeated on a weekly
basis. After 0, 7 14, 21 and
28 days samples of the
formulations were placed in
a preservative neutralizer
compared to the control .
good preservation efficacy
was a microbial count of
<10 cfu/ml at day 7, 14 , 21
and 28 days.
Not Repellency of Two 19 Vinyl floor tiles were Manufacturer
available Formulations against sprayed with the repellent
Ants (in-house solution and placed in
laboratory study) cages. A food attractant
was placed in the centre of
each tile. The number of
ants crossing each tile was
counted at 30 and 60
minutes after tiles were
placed into cages. Data for
30 and 60 min was
combined and averaged.
Repellency was also
177
INFORMATION
PROVIDED FROM
ELSEWHERE)
measured the day after the
fourth consecutive
application. Sufficient
>65%.
Not Repellency of Two 19 The cockroaches had a Manufacturer
available product formulations choice of two shelters, one
against the German treated with the product
Cockroach (in-house and one untreated.
laboratory study) Repellency after 1, 3, 7
and 14 days was recorded.
Percent repellency was
calculated. Sufficient
>65%, excellent repellency
was defined as >85%.
Not Swimming pool 1-5 A residential spa was used Manufacturer
available disinfectant: to test the efficacy of the
Residential spa field product in typical field
test conditions. The
maintenance pattern used
was considered to be
typical of the “average spa
owner”, i.e. conforming to
minimum and not optimum
use pattern and
maintenance procedures.
Bacteriological analysis
was conducted on 150
samples were taken over
the 90 day test period.
Efficacy was determined by
a reduction in cfu/100ml.
testing of wood
preservatives.
Mycological test.
"Jordburk" method - A
NWPC soil block test with
1.4.1.1./7 wood-rotting
0 Basidiomycetes.
testing of wood
preservatives.
NWPC Mycological test.
Standard "Mullåde" method - A
1.4.1.2./7 soil block test in
0 unsterile soil.
NWPC Mycological testing of 8 Test institute
Standard anti-stain preservatives
1.4.1.3./7 for freshly sawn timber.
178
INFORMATION
PROVIDED FROM
ELSEWHERE)
9 The Miniboard method.
(Mykologisk provning av
träskyddsmedel mot
blånad och mögel på
nysågat virke.
Minibrädmetoden.)
NWPC wood preservatives
Standard Mycological test. Field
1.4.2.1./7 test - A field test with
1 stakes.
wood preservatives
NWPC Marine test - A test
Standard against marine
1.4.2.2./7 woodboring organisms
3 in sea water.
OECD Proposed development 1-5 Own searches
(ENV/JM/ of an OECD Guidance
BCID(200 Document for
7)2) establishing the efficacy
of biocides used in
swimming pools and
spas
OECD Update on work to 10 Own searches
(ENV/JM/ validate efficacy
BCID(200 methods for
7)3) antimicrobials used on
hard surfaces
OECD Guidance document on 9 Own searches
(ENV/JM/ the evaluation of the
BCID(200 efficacy of antimicrobial
7)5) treated articles with
claims for external
effects
203 with UK (HSE)
(1992) Fish, Acute toxicity test
204 Fish, Prolonged toxicity with UK (HSE)
(1984) test: 14-day study
OPPTS General Considerations 18 General guide Manufacturer
810.3000 for Efficacy of
(1999) Invertebrate Control
Agents
UK guidelines
179
INFORMATION
PROVIDED FROM
ELSEWHERE)
OPPTS Soil treatments for 18 Own searches
810.3100 imported fire ants
OPPTS Livestock, poultry, fur- 18 Own searches
810.3200 and wool-bearing
animal treatments
OPPTS Treatments to control 18 UK guidelines
810.3300 pests of humans and
pets
OPPTS Mosquito, black fly, and 18 Test of insecticides against UK guidelines

810.3400 biting midge (sand fly) flying insects: Mosquito
treatments Black Fly
Biting Midge (Sand Fly)
OPPTS Premises treatments 18 General guideline Manufacturer
810.3500
UK guidelines
OPPTS Insect Repellents For 19 Own search

810.3700 Human Skin and
“public Outdoor Premises
draft”
OPPTS Methods for efficacy 18 Own searches
810.3800 testing of termite baits
Powers et Biocidal Efficacy of a 1-5 Biocidal emergency water Manufacturer
al. Applied Flocculating Emergency purification tablets were
and Water Purification tested for bactericidal,
Environm Tablet. virucidal and cysticidal
ental efficacy in water at
Microbiolo temperatures ranging from
gy, 60: 5 to 25ºC to validate and
2316 – verify the effectiveness of
2323 the tablets for the
(1994) destruction and removal of
microorganisms. Counts of
surviving organisms were
compared with the initial
counts to calculate the log-
reduction. Replicate
results were averaged
prEN Chemical disinfectants 1-5 Own searches
Quantitative suspension
bactericidal activity
against Legionella
pneumophila of
for aqueous systems -
Test method and
step 1)
180
INFORMATION
PROVIDED FROM
ELSEWHERE)
prEN Chemical disinfectants 1-5 Manufacturer
Quantitative carrier test
for the evaluation of TM II05 (Fr)
mycobactericidal or
tuberculocidal activity of
used for instruments in
method and
step 2)
prEN Chemical disinfectants 1-5 Industry
06 Application of European
Standards for chemical Manufacturer
disinfectants and
antiseptics
TM II05 (Fr)
prEN 807 Wood preservatives - 8 Own searches

rev Determination of the
and other soil inhabiting
micro-organisms
Rawlinson A recirculating test rig 13 TNsG on Prod
and for the investigation of Eval
Shennan, metal-working fluid
1987. spoilage. In Industrial
microbiological testing.
Edited by Hopton and
Hill, Blackwell Scientific
Publications, Oxford.
ISBN 0 632 01793 7. pp
227-231
RENAULT Evaluation of the 13 TNsG on Prod
D551721 Biostability of Aqueous Eval
(1987) Metal Working Fluids
Report Biological Test Method: 12 Efficacy test against Alga: Communication
EPS Growth Inhibition Test Selenastrum capriconutum with UK (HSE)
1/RM/25 Using the Freshwater
Environm Alga Selenastrum Used in the process of oil
ent capriconutum recovery
Canada.
1992
SABS Bacterial efficacy of 6 Efficacy test for in can TNsG on Prod
1102 biocides used in water- preservatives in paints Eval
(1987) based emulsion paints (emulsion) against bacteria.
SABS South African standard 13 TNsG on Prod
181
INFORMATION
PROVIDED FROM
ELSEWHERE)
1435- specification for Eval
1987 biocides for use in
emulsions of aqueous
metal working fluid and
aqueous hydraulic fluid.
SABS 233 Pesticides: Biological 18 Manufacturer
st
1 rev evaluation of mists and
fogs - first revision
SABS 303 Pesticides – Rearing 18 Manufacturer
and handling of the
human body louse
(Pediculus humanus
humanus L.) - first
revision
and handling of the
common clothes moth
(Tineola bisseliella
Hummel) - second
revision
and handling of the
German cockroach
(Blatella germanica (L.))
- second revision
SABS 576 Pesticides – Biological 18 Manufacturer
evaluation of
insecticidal oil-based
space spray in low-
pressurized dispensers
- first revision
evaluation of the
contact efficacy of liquid
residual insecticides -
first revision
6136 evaluation of materials
(2003) that release an
insetticide upon heating
rd
3 ed evaluation of knock-
(2002) down and killing
proprieties of liquid and
aerosol formulation (al
posto di Standard
methods SABS Method
8689-first revision)
SABS 690 Pesticides: biological 18 Manufacturer
(DRAFT) evaluation of the
proprerties of solid fly
182
INFORMATION
PROVIDED FROM
ELSEWHERE)
baits - DRAFT
evaluation of the
efficacy of mosquito
repellents - first
revision
SABS 807 Methods for testing 18 TNsG on Prod
insecticides against Eval
flying and crawling
insects. Manufacturer
UK guidelines
SABS 899 Insecticidal space spray 18 Manufacturer

(1987) in pressurized
dispensers
SM019 (in S&M Boko test 11 Preservation of water Manufacturer
house) diluted coolants.
preservatives, inoculated
and streaked on agar
plates.
Test germ: bacteria,
moulds, Yeasts
4 evaluation levels. 12
cycles.
SM020 (in S&M Fako test 6 Preserving effect of Manufacturer
house) chemical preservatives in
water containing coatings,
adhesives and other water
containing technical
emulsions or dispersions
(in can).
plates.
Yeasts
"Well preserved" if after 6w
no microbial growth can be
observed.
Corresponds to 2 years of
4 evaluation levels
SM021 (in S&M KOKO test 6 Preservatives in cosmetic Manufacturer
house) formulations, e g creams,
183
INFORMATION
PROVIDED FROM
ELSEWHERE)
shampoos etc (in can)
plates.
Yeasts
"Well preserved" if after 6w
no microbial growth can be
observed.
Corresponds to 30 months
of microbial stability.
4 evaluation levels
SM022 (in Determining the 10 Resistance of masonry Manufacturer
house) resistance to fungal coatings to fungal growth.
growth Test substrate: Masonry
coatings on paper
Test germ: Aspergillus
niger and Penicillum
funiculosum
"Sufficiently finished
against fungal growth" if
max 1% growth.
SM022a Determining the 10 Resistance of masonry Manufacturer
(in house) resistance to fungal coatings to fungal growth.
growth Test substrate: Masonry
coatings on paper
Test germ: Aspergillus
niger and Penicillum
funiculosum
"Sufficiently finished
against fungal growth" if
max 1% growth.
SM023 (in Determining the 10 Resistance of masonry Manufacturer
house) resistance to algal coatings to algal attack.
growth Test substrate: masonry
coatings on paper.
Test germ: Scenedesmus
vacuolatus.
No algal growth on the test
pieces after 2 weeks:
"Effectively protected
against algal growth"
SM026 (in S&M Betoko test: 6 Preserving effect of Manufacturer
house) Determination of the chemical preservatives in
preserving effect of concrete admixtures (in can
184
INFORMATION
PROVIDED FROM
ELSEWHERE)
chemical preservatives preservation).
in concrete admixtures Test batch: unpreserved
plates.
Yeasts
Free of growth during 4
weeks: "Well preserved"
Corresponds to 1 year of
microbiological stability
SM029 (in S&M FeuTuKo test 6 Preservatives in wet Manufacturer
house) tissues in their original
packaging.
Inoculation of whole
packaging and after 3w of
incubation, pieces of tissue
is placed on agar plates.
Yeasts
3d bacteria, 7d moulds. 4
evaluation levels.
SM036 (in S&M WiWako test 6 Preservation of fountain Manufacturer
house) solutions for offset printing.
plates.
Yeasts
12 cycles
SM037 (in S&M TaBaKo test 6 Preservation of paint baths. Manufacturer
plates.
Yeasts
12 cycles. 4 evaluation
levels.
SM044 (in Determination of the 6 Preserving effect of Manufacturer
house) preserving effect of chemical preservatives in
185
INFORMATION
PROVIDED FROM
ELSEWHERE)
chemical preservatives household formulations, e g
in household softener, washing up
formulations liquids, all purpose cleaner
(in can preservation).
plates.
Yeasts
Free of growth during 6
weeks: "Well preserved". (4
evaluation levels).
Corresponds to 30 months
of microbiological stability
SN 195 Antibacterial Activity,
1994 Test
SN 195 Antimycotic Activity
1994 Test
SN 195 Determination of the 9 Applied to textiles Manufacturer
924 - antibacterial activity,
1983 germ count method
UK MOD Cutting fluid, soluble, 13 TNsG on Prod
91-70 biostable joint service Eval
issue designation ZX-9
(1990)
US Test method for textiles 18 Efficacy test against larvae TNsG on Prod
AATCC to determine resistance Eval
Technical to insects (e.g. moths,
Manual carpet beetles) UK guidelines
Method
24 (1992)
US CSMA Test method for aerosol 18 Test of insecticides against TNsG on Prod
Aerosol space sprays against flying insects: Eval
Guide flying insects
7 th UK guidelines
Edition,
pages Manufacturer
129-134
(1981)
US CSMA Test method for 18 Test of insecticides against TNsG on Prod
Aerosol pressurised spray crawling insects: Eval
Guide products against cockroaches
7 th cockroaches UK guidelines
Edition,
186
INFORMATION
PROVIDED FROM
ELSEWHERE)
pages
135-139
(1991)
US EPA Acute toxicity test for 17 Communication
72-1 freshwater fish with UK (HSE)
US EPA 17 Communication
96-2 Fish control agents with UK (HSE)
US EPA 15 TNsG on Prod
96-5 Avian Toxicants Eval
US EPA Avian Frightening 15 TNsG on Prod
96-7 Agents Eval
US EPA Insect repellents for 19 UK guidelines
Guideline human skin and outdoor
OPPTS premises
810.3700
(1999)
Verwey & Liquid Electric test 18 For testing pyrethroids Manufacturer
Sosa, method (draft method) and natural
2007 actives (Pyrethrum extract)
on mosquitoues
(knockdown). Efficacy
criteria: "effective against
mosquitoes for X hours".
Knockdown is measured
repeatedly for 2h and
mortality after 24h. Control
(no treatment) knockdown:
maximum 10%. 2-4
chamber replicates, 50
organisms in each. Mean
and Standard Deviations
for each time calculated as
well as KT50 and KT80
(Mean time to 50% and
80% knockdown
respectively).
WHO/CD Space spray application 18 Brief description of the TM II05 (Fr)
S/WHOP of insecticides for vector main types of space spray
ES/GCDP and public health pest equipment as well as the
P/2003.5 control – a practitioner’s operational guidelines for
guide space spray application of
insecticides.
cockroaches to
insecticides
187
INFORMATION
PROVIDED FROM
ELSEWHERE)
larvae to insect
susceptibility or
mosquitoes to
organochlorine,
organophosphate and
carbamate insecticides,
- establishment of the
baseline.
susceptibility or
mosquitoes to
organochlorine,
organophosphate and
carbamate insecticides -
diagnostic test
resistance of mosquito UK guidelines
resistance of body or
headlice to insecticides
resistance of adult bed- UK guidelines
bugs to insecticides
blackflies, sandflies and
biting midges to
insecticides
resistance of blackfly UK guidelines
WHO/VB Instructions for 18 UK guidelines
C/81.812 determining the
188
INFORMATION
PROVIDED FROM
ELSEWHERE)
larvae to insect
resistance of houseflies, UK guidelines
tsetse flies, stableflies,
blowflies etc. to
insecticides
resistance of adult ticks UK guidelines
to insecticides
resistance of fleas to UK guidelines
insecticides
189

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October 2012

Efficacy testing of biocidal products

Postadress Besök Faktureringsadress Telefon & fax Internet Org nr

- Overview of available tests

Acronym Full names Web page to organisations (if available)

Efficacy of a biocidal product can be defined as the power to produce an effect,

The numerous ways of testing and measuring efficacy makes comparisons

1 ACRONYMS AND ABBREVIATIONS ................................................................. 1

4 BACKGROUND AND AIM OF THE PRESENT OVERVIEW ............................... 6

4.1 BACKGROUND .................................................................................................. 6

4.1.1 Biocidal Products Directive .............................................................................. 6

4.1.2 Data requirements related to efficacy .............................................................. 6

4.2 AIMS OF THIS OVERVIEW ................................................................................ 8

6.1 PERFORMANCE STANDARDS vs LABEL CLAIMS ....................................... 10

6.2 EXPERIMENTAL DESIGN OF EFFICACY TESTS ........................................... 10

6.3 STANDARDISED AND "IN HOUSE" PROCEDURES ...................................... 11

6.3.1 PT 1-5 ............................................................................................................... 11

6.3.2 PT 6: In-can preservatives .............................................................................. 12

6.3.3 PT 8: Wood preservatives ............................................................................... 13

6.3.4 PT 10: Masonry biocides ................................................................................. 14

6.3.5 PT 13: Metalworking fluid preservatives ........................................................ 14

6.3.6 PT 14: Rodenticides ........................................................................................ 14

6.3.7 PT 18: Insecticides, acaricides and products to control other

6.3.8 PT 21: Antifouling products ............................................................................ 15

7.1.1 International standardisation organisations – own searches ...................... 16

7.1.2 Tests used in other MSs – TM and national guidelines ................................ 17

7.1.3 Methods in use according to manufacturers and industry

7.1.4 Test endpoints ................................................................................................. 19

7.2 LIMITATIONS AND KNOWLEDGE GAPS ........................................................ 19

8.1 EVALUATING EFFICACY – COMPARING TEST RESULTS TO

9 CONCLUSIONS AND SUGGESTIONS............................................................. 22

10 APPENDIX I. TEST PROCEDURES (STANDARDS AND OTHER

4.1.1 Biocidal Products Directive

In the European Community, authorisation of Biocidal Products (BP) is regulated

Efficacy can be defined as the power to produce an effect, and is described in

4.1.2 Data requirements related to efficacy

However, there is no international consensus on what data are required to prove

4.2 AIMS OF THIS OVERVIEW

The TNsG on Product Evaluation includes references to available standard tests

Knowledge gaps should also be identified.

The project was performed during the period of November-December in 2007.

6.1 PERFORMANCE STANDARDS vs LABEL CLAIMS

Performance standards refer to a predetermined efficacy required by the

6.2 EXPERIMENTAL DESIGN OF EFFICACY TESTS

6.3 STANDARDISED AND "IN HOUSE" PROCEDURES

The availability of international standards varies between PTs. In the TNsG on

6.3.2 PT 6: In-can preservatives

6.3.3 PT 8: Wood preservatives

6.3.5 PT 13: Metalworking fluid preservatives

6.3.6 PT 14: Rodenticides

6.3.7 PT 18: Insecticides, acaricides and products to control other arthropods

6.3.8 PT 21: Antifouling products

• Although there are a few standards related to antifouling products, these

7.1 TEST REFERENCES FOUND IN THIS OVERVIEW

About 90 manufacturers, test institutes and industry organisations were contacted.

7.1.1 International standardisation organisations – own searches

EPPO is primarily focused on Plant Protection Products, although some standards

ASTM has several standards applicable to microorganisms and wood

7.1.2 Tests used in other MSs – TM and national guidelines

In Denmark, wood preservatives are efficacy tested according to CEN standards.

7.1.3 Methods in use according to manufacturers and industry organisations

Companies were initially contacted in Sweden but because suitable contact

7.2 LIMITATIONS AND KNOWLEDGE GAPS