Journal of Applied Microbiology 1999, 87, 718–725

A comparison of the bactericidal efficacy of 18 disinfectants

used in the food industry against Escherichia coli O157:H7
and Pseudomonas aeruginosa at 10 and 20 °C
J.H. Taylor, S.J. Rogers and J.T. Holah
Food Hygiene Department, Campden and Chorleywood Food Research Association, Chipping Campden, UK
7272/06/99: received 21 June 1999 and accepted 5 July 1999

J .H . T A YL OR , S. J. R OG ER S AN D J . T. HO L AH . 1999. A number of proprietary disinfectant

products (18) used in the food industry were tested for their bactericidal efficacy against
Pseudomonas aeruginosa and Escherichia coli O157:H7 at 20 and 10 °C according to the
BS EN 1276 (1997) quantitative suspension test for the evaluation of bactericidal activity of
chemical disinfectants and antiseptics used in food, industrial, domestic and institutional
areas. At 20 °C, 13 products passed at their in-use concentration (under clean and dirty
conditions) against Ps. aeruginosa and 15 passed against E. coli O157:H7. The number
of products passing the test at 10 °C was 11 and 14 for Ps. aeruginosa and E. coli O157:H7,
respectively. The products exhibiting reduced efficacy at the lower temperature were
amphoterics and quaternary ammonium compounds although some of these types
of products were effective at both temperatures. Products that passed against Ps.
aeruginosa generally also passed against E. coli O157:H7. Taking all the results
together, only 11 of the total of 18 products achieved a pass result under all the
parameters tested. This work demonstrates the need for final verification of
disinfectant efficacy by undertaking field trials in the food-processing environment in
which the product is intended for use.

INTRODUCTION efficacy of disinfectants has not been validated at these tem-

peratures.
Food product contamination may occur from environmental
Low temperature disinfection is often carried out with
routes such as air, people and surfaces, with the surface route
products which were formulated before the advent of the
being the most important to control on a day to day basis by
modern chilled food industry. Certain pathogens, such as
the implementation of a sanitation programme (Holah 1995).
Listeria monocytogenes, have become significant in chilled
Disinfection is the final stage in a sanitation programme that
environments and, in spite of greater hygiene standards, food
is designed to remove product residues and foreign bodies, factory trouble-shooting audits by the authors have identified
in addition to reducing the level of micro-organisms to ensure problems with Listeria spp. surviving on surfaces after cle-
both the safety and quality of food. aning and disinfection. Listeria spp. are able to grow at tem-
In the last few years there have been considerable changes peratures below 4 °C (Walker et al. 1990) and are very tolerant
in eating habits in the UK (Anon. 1995) with increased sales of adverse environmental conditions at low temperatures
of convenience foods such as ready meals, pizza, prepared (Tuncan 1993). Another organism of concern is Escherichia
salads and potted desserts. To optimize product quality and coli O157:H7 and outbreaks involving this organism, along
to ensure food safety, the production of these food types is with other pathogens, are still increasing (Anon. 1997a) even
often carried out at chilled temperatures, yet the biocidal though we have a good understanding about the environ-
mental route of contamination and the physiology of food
pathogens. To date, little, if any, information has been pub-
Correspondence to: J.H. Taylor, Food Hygiene Department, Campden and
Chorleywood Food Research Association, Chipping Campden, lished relating to the resistance of E. coli O157:H7 to com-
Gloucestershire GL55 6LD, UK (e-mail: J—TAYLOR@ mercial disinfectants and thus whether it can be controlled
Campden.co.UK). by current sanitation programmes.
© 1999 The Society for Applied Microbiology
 D IS IN F EC TA N T E FF I CA CY O F 1 8 P R OD UC T S 719

There is a range of test methods for evaluating disinfectant Table 1 Summary of product types and in-use concentration
efficacy throughout Europe (Reybrouk 1982) and products —
–––––––––––––––––––––––––––––––––––––––––––––––––––––
may have to undergo a range of different tests to satisfy Recommended
legislative requirements in different countries. Consequently, Product in-use
code Product type concentration
products available on the market are likely to exhibit various
—
–––––––––––––––––––––––––––––––––––––––––––––––––––––
biocidal efficacies because they have been tested under dif- 1 Quat 1·0%
ferent conditions (level and type of organic load, type of test 2 Quat 1·0%
micro-organism, hard water, etc.) against different methods 3 Quat 1·0%
and to different standards. To take into account these factors 4* Quat/amphoteric 1·0%
and to harmonize testing across Europe, the European Tech- 5* Quat/amphoteric 1·0%
nical Committee CEN/TC216/WG3 has developed sus- 6* Quat/glutaraldehyde 1·0%
pension tests (bactericidal and fungicidal) specifically for the 7 Amphoteric 1·0%
food, industrial and domestic markets. These tests will be 8* Amphoteric 1·0%
recognized as being appropriate for use within the European 9 Amphoteric 1·0%
Biocide Directive (Anon. 1997b), whereby the efficacy claims 10 Chlorine dioxide 1·0%
11 Potassium hydroxide/sodium 5·0%
of disinfectant products have to be supported by data from
hypochlorite
recognized test methods. 12 Sodium hypochlorite 500 ppm
This paper describes work which was undertaken to inves- 13 Peracetic acid/hydrogen peroxide 0·3%
tigate the disinfection efficacy of proprietary products used 14 Iodophor 1·0%
in the food industry. Studies were undertaken to examine the 15 Biguanide 1·0%
effect of lower temperatures on disinfection efficacy. Pseudo- 16* Biguanide/quat 1·0%
monas aeruginosa was used as the test organism because of its 17 Sodium dichloroisocyanate 333 pm
known resistance to disinfectant action. Disinfectant efficacy 18 Acid detergent/sanitizer 1·0%
was also tested against E. coli O157:H7 to establish whether —
–––––––––––––––––––––––––––––––––––––––––––––––––––––
proprietary products are capable of killing this organism * These disinfectants have been specifically formulated and
under normal sanitation conditions. Finally, this study marketed for use in the UK chilled food industry ‘high care’
areas operating at low temperatures.
enabled a practical evaluation of the new European bac-
Quat, Quaternary ammonium compound.
tericidal suspension test BS EN 1276 (1997) (Anon. 1997c).

MATERIALS AND METHODS

Commercially available products (18) were chosen to rep- concentrations of 0·5%, 1·0% and 2% was added to 1·0 ml
resent a range of products with respect to price and product bovine albumen (Fraction V; BDH, Poole, UK) serum at
type (encompassing both oxidative and non-oxidative bioc- concentrations of 0·3% (w/v) and 0·03% (w/v) (to represent
ides) and markets throughout Europe. The product types dirty and clean conditions, respectively) to which 1·0 ml of
included in the study are shown in Table 1. the bacterial suspension (1·5–5·0 × 108 cfu ml−1, grown in
Products were tested according to the BS EN 1276 (1997) high nutrient conditions) had been added. After a contact
quantitative suspension test for the evaluation of the bac- time of 5 min, 1·0 ml of the test mixture was pipetted into
tericidal activity of chemical disinfectants and antiseptics used 8·0 ml neutralizer (comprising polysorbate 80 3·0% (v/v),
in food, industrial, domestic and institutional areas against E. saponin 3·0% (w/v) and lecithin 0·3% (w/v)) and 1·0 ml de-
coli O157:H7 (CCFRA reference code CRA 4497) and Ps. ionized water. After 5 min neutralization time, duplicate 1·0-
aeruginosa (NCIMB 10421) at two different temperatures ml volumes were pour plated with tryptone soya agar and
(20 °C is the standard temperature in the test method and incubated at 37 °C for 48 h prior to counting.
10 °C an additional test temperature which also reflects the Three validation (control) procedures were undertaken in
temperature encountered in chilled food-processing environ- parallel for each disinfection test on all occasions as follows.
ments) using temperature-controlled water-baths (10 and Validation A. The test was undertaken with the addition of
20 °C). The products were tested at three concentrations, 8·0 ml water of standard hardness in place of the disinfectant
including their recommended in-use concentration (usually to ensure that there was no biocidal action of the other
1% for both liquids (v/v) and solids (w/v), unless specified, experimental parameters.
see Table 1), in both clean and dirty conditions. Validation B. Neutralizer (8·0 ml) and 1·0 ml water were
An 8·0-ml sample of disinfectant diluted in water of stan- added to the bacterial suspension and then plated out to
dard hardness (300 mg kg−1 CaCO3) at concentrations of ensure that the neutralizer had no disinfectant activity.
0·63% (v/v), 1·25% (v/v) and 2·5% (v/v) to give final test Validation C. Bacterial suspension (1·0 ml) was added to neu-
© 1999 The Society for Applied Microbiology, Journal of Applied Microbiology 87, 718–725
720 J .H . T A YL OR E T A L .

tralized disinfectant to ensure that the disinfectant had been showed a temperature effect (i.e. passed at 20 °C but failed
neutralized. at 10 °C).
Full details of the disinfection and validation procedures
are described in the test method (Anon. 1997c). For safety Escherichia coli O157:H7 at 10 °C
reasons, all E. coli O157:H7 testing was carried out in a Class
The results in Table 3 show that 14 products passed under
II safety cabinet.
clean and dirty conditions and that products passing against
The reduction in viability was calculated by subtracting
Ps. aeruginosa passed against E. coli O157:H7.
the log of the viable count after disinfection (Na) from the
The products that failed were an amphoteric (no. 7), chlor-
log of the initial count in the test chamber (N × 10−1). To
ine dioxide (no. 10), a biguanide (no. 15) and an acid deter-
pass the test, products must achieve a five log reduction in
gent/sanitizer (no. 18). Only one product (amphoteric (no.
viable counts. In this study, tests were undertaken twice and,
7)) showed a temperature effect (i.e. passing at 20 °C but
where necessary, a third decisive test was carried out with
failing at 10 °C) for E. coli O157:H7.
the overall performance of products assessed at their in-use
concentration.
DISCUSSION

RESULTS Evaluation of disinfection efficacy

Only 11 of the 18 products gave a pass result at the rec-
Pseudomonas aeruginosa at 20 °C
ommended in-use concentration against the two organisms
The results in Table 2 show that 13 of the 18 products tested for both temperatures on at least two occasions for both clean
in this study achieved pass results at 20 °C for both clean and and dirty conditions. Work by Jacquet and Reynaud (1994)
dirty conditions against Ps. aeruginosa. The products that also showed that, of eight disinfectants tested (at rec-
failed under both dirty and clean conditions were an ampho- ommended in-use concentration) using the French Standard
teric (no. 7), chlorine dioxide (no. 10), a biguanide (no. 15) AFNOR NFT 721701989 at 20 °C, only two products
and an acid detergent/sanitizer (no. 18). A quaternary achieved a five log reduction in viable counts against the
ammonium product (no. 1) passed under clean conditions test organisms (L. monocytogenes, Staphylococcus aureus and
but failed (on two occasions) under dirty conditions. Enterococcus faecium strains). These results show that the
efficacy of some products may be compromised by certain
conditions, such as low temperature and organic residues,
Escherichia coli O157:H7 at 20 °C which may be encountered in food-processing environments.
With factories operating at chilled temperatures (10 °C)
The results in Table 2 show that 15 of the 18 products passed
several chemical suppliers have formulated their disinfectants
for both clean and dirty conditions at 20 °C. Products which
specifically to be used in these environments; these are
passed against Ps. aeruginosa also passed against E. coli
marked with an asterisk in Table 1 (other products are gen-
O157:H7 but products failing against Ps. aeruginosa did not
erally formulated for use at ambient temperatures). As
necessarily fail against E. coli O157:H7. The biguanide (no.
expected, all five of these products passed the tests for both
15) failed under dirty conditions but passed under clean
dirty and clean conditions at this temperature and at 20 °C.
conditions for E. coli O157:H7. The quaternary ammonium
Such formulation of products would mask temperature
product (no. 1), which failed under dirty conditions against
effects and may explain why there were only two types of
Ps. aeruginosa, passed against E. coli O157:H7. Chlorine diox-
product giving a clear difference in activity with temperature.
ide (no. 10) and the acid detergent/sanitizer (no. 18) failed
Other workers have found that temperature has a marked
under both clean and dirty conditions.
effect on biocidal efficacy. Tuncan (1993) found that a quat-
ernary ammonium and an iodophor can be ineffective against
some Listeria spp. at cold temperatures (ranging from 2 to
Pseudomonas aeruginosa at 10 °C
15 °C), whilst being effective at 25 °C. However, it was found
The results in Table 3 show that 11 products achieved a pass that efficacy was improved by increasing the exposure time
under both clean and dirty conditions against Ps. aeruginosa. or concentration. The concentrations used for the quaternary
The products that failed were two quaternary ammonium ammonium were up to 200 ppm (0·02%) and for the iodophor
products (nos 1 and 2), two amphoterics (nos 7 and 8), 50 ppm (0·005%), which are much lower than those used in
chlorine dioxide (no. 10), a biguanide (no. 15) and an acid this study. Tuncan (1993) also found that the various species
detergent/sanitizer (no. 18). Of these, two products (an of Listeria demonstrated different sensitivities to the dis-
amphoteric (no. 8) and a quaternary ammonium (no. 2)) infectants. Gelinas et al. (1984) studied temperature effects
© 1999 The Society for Applied Microbiology, Journal of Applied Microbiology 87, 718–725
 D IS IN F EC TA N T E FF I CA CY O F 1 8 P R OD UC T S 721

Table 2 Results at 20 °C
––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––
—
Pseudomonas aeruginosa Escherichia coli
—
–––––––––––––––––––––– —
––––––––––––––––––––––
Disinfectant Disinfectant
in-use concentration in-use concentration
Product Clean/ —
–––––––––––––––––––––– —
––––––––––––––––––––––
Product type code dirty × 0·5 × 1·0 × 2·0 × 0·5 × 1·0 × 2·0
––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––
—
Amphoteric 7 Clean F F P P P P
Clean F F P F P P
Dirty F F P P P P
Dirty F F F F F P
Clean P P P
Dirty P P P
Amphoteric 8 Clean P P P P P P
Clean P P P P P P
Dirty P P P F P P
Dirty P P P P P P
Amphoteric 9 Clean P P P P P P
Clean P P P P P P
Dirty P P P P P P
Dirty F P P P P P
Chlorine dioxide 10 Clean F F P F F P
Clean F F P F F P
Dirty F F P F F P
Dirty F F P F F P
Potassium hydroxide/sodium hypochlorite 11 Clean P P P P P P
Clean P P P P P P
Dirty P P P P P P
Dirty P P P P P P
Sodium hypochlorite 12 Clean P P P P P P
Clean P P P P P P
Dirty F P P P P P
Dirty P P P F P P
Peracetic acid/hydrogen peroxide 13 Clean P P P P P P
Clean P P P P P P
Dirty F P P P P P
Dirty P P P P P P
Iodophor 14 Clean P P P P P P
Clean P P P F P P
Dirty P P P F P P
Dirty P P P F P P
Biguanide 15 Clean F F P P P P
Clean F F P F P P
Dirty F F F F F F
Dirty F F P F F F
Biguanide/quat 16 Clean P P P P P P
Clean P P P P P P
Dirty P P P P P P
Dirty P P P P P P
Sodium dichloroisocyanate 17 Clean P P P P P P
Clean F P P P P P
Dirty F P P F P P
Dirty F P P P P P
Acid detergent/sanitizer 18 Clean F F F F F F
Clean F F F F F F
Dirty F F F F F F
Dirty F F F F F F
––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––
—
P (pass), 5-log reduction or greater in viable counts; F (fail), less than 5-log reduction in viable counts; Quat, quaternary ammonium
compound.
722 J .H . T A YL OR E T A L .

Table 3 Results at 10 °C
––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––
—
Pseudomonas aeruginosa Escherichia coli
—
–––––––––––––––––––––– —
––––––––––––––––––––––
Disinfectant Disinfectant
in-use concentration in-use concentration
Product Clean/ —
–––––––––––––––––––––– —
––––––––––––––––––––––
Product type code dirty × 0·5 × 1·0 × 2·0 × 0·5 × 1·0 × 2·0
––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––
—
Quat 1 Clean F F P P P P
Clean F F P P P P
Dirty F F P F P P
Dirty F F P F P P
Quat 2 Clean F F F P P P
Clean F F F P P P
Dirty F F F F P P
Dirty F F F F P P
Quat 3 Clean F P P P P P
Clean F P P P P P
Dirty F P P P P P
Dirty F P P F P P
Quat/amphoteric 4 Clean P P P P P P
Clean P P P P P P
Dirty F P P P P P
Dirty F P P F P P
Quat/amphoteric 5 Clean P P P P P P
Clean F P P P P P
Dirty F P P P P P
Dirty F P P F P P
Quat/glutaraldehyde 6 Clean F P P P P P
Clean F P P P P P
Dirty F P P P P P
Dirty F P P P P P
Amphoteric 7 Clean F F F F F P
Clean F F F F F P
Dirty F F F F F P
Dirty F F F F F F
Amophoteric 8 Clean F F P P P P
Clean F F P P P P
Dirty F F P P P P
Dirty F F P P P P
Amphoteric 9 Clean P P P P P P
Clean F P P F P P
Dirty P P P P P P
Dirty F P P F P P
Chlorine dioxide 10 Clean F F P F F P
Clean F F P F F F
Dirty F F F F F F
Dirty F F F F F F
Potassium hydroxide/sodium hypochlorite 11 Clean P P P P P P
Clean P P P P P P
Dirty P P P P P P
Dirty P P P P P P
Sodium hypochlorite 12 Clean P P P P P P
Clean P P P P P P
Dirty P P P P P P
Dirty P P P P P P
––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––
—

© 1999 The Society for Applied Microbiology, Journal of Applied Microbiology 87, 718–725
 D IS IN F EC TA N T E FF I CA CY O F 1 8 P R OD UC T S 723

Table 3 Continued.
––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––
—
Pseudomonas aeruginosa Escherichia coli
—
–––––––––––––––––––––– —
––––––––––––––––––––––
Disinfectant Disinfectant
in-use concentration in-use concentration
Product Clean/ —
–––––––––––––––––––––– —
––––––––––––––––––––––
Product type code dirty × 0·5 × 1·0 × 2·0 × 0·5 × 1·0 × 2·0
––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––
—
Peracetic acid/hydrogen peroxide 13 Clean P P P P P P
Clean P P P P P P
Dirty P P P P P P
Dirty P P P P P P
Iodophor 14 Clean P P P P P P
Clean P P P P P P
Dirty P P P F F P
Dirty P P P F P P
Clean P P P
Dirty F P P
Biguanide 15 Clean F F P F F P
Clean F F P F F F
Dirty F F F F F F
Dirty F F P F F F
Biguanide/quat 16 Clean F F F P P P
Clean P P P P P P
Clean F P P F P P
Dirty F F F P P P
Dirty F P P
Dirty F P P
Sodium dichloroisocyanate 17 Clean P P P P P P
Clean P P P P P P
Dirty F P P P P P
Dirty F P P P P P
Acid detergent/sanitizer 18 Clean F F F F F F
Clean F F F F F F
Dirty F F F F F F
Dirty F F F F F F
––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––––
—
P (pass), 5-log reduction or greater in viable counts; F (fail), less than 5-log reduction in viable counts; Quat, quaternary ammonium
compound.

on eight products and found that the efficacy was greatly It is not clear why some disinfectant products exhibit
influenced by temperature (4–50 °C). Glutaraldehyde, temperature effects, although it may be related to the reduced
chlorhexidine and amphoteric products were particularly temperatures lowering the metabolic activity of the test
affected, whilst sodium hypochlorite was found to be least micro-organisms or potentially the onset of the cold shock
affected by temperature. The results were expressed as mini- response which may enhance resistance to disinfection. These
mum concentrations obtained by the Association of Official topics are discussed in a comprehensive review article by
Analytical Chemists (Anon. 1980) use-dilution method Berry and Foegeding (1997) on cold temperature adaptation
against Ps. aeruginosa. It is difficult to make direct com- and growth of micro-organisms.
parisons because different methods were used and there are The 5-min contact time in the disinfectant test was chosen
no data at 10 °C. They noted a synergistic effect with glu- because it is representative of the time taken for disinfectant
taraldehyde and a quaternary ammonium and indeed in this to run off equipment surfaces after application. In practice,
study a five log reduction in viable counts was obtained for a however, the effectiveness of products may be enhanced by
product combining glutaraldehyde and a quaternary prolonged contact times, e.g. the use of soak ranks or repeat
ammonium (for the tests undertaken). applications. The presence of organic residues after cleaning
© 1999 The Society for Applied Microbiology, Journal of Applied Microbiology 87, 718–725
724 J .H . T A YL OR E T A L .

may be reduced by more thorough cleaning. The food indus- the standard method dictates that the three validation tests
try normally requires products to pass under dirty conditions, must recover viable cells in the range 2·0–3·0 × 103 cfu ml−1.
as poor hygienic design of equipment may result in crevices The contact time in the test procedure is 5 min 2 10 s
and dead areas that may protect food soil from cleaning and and, for economic reasons, laboratories may undertake the
there are occasions when equipment has not been adequately test for one organism at three concentrations of disinfectant
cleaned. under both clean and dirty conditions (six reactions) along
The relative strengths and weaknesses of laboratory dis- with the three validation controls (one-test reaction for vali-
infectant tests have recently been reviewed by Holah et al. dation B and two-test reactions each for validations A and
(1998) and ways in which they can be improved to more C). This means that one test procedure with validations
closely simulate in-use conditions in the future are discussed. involves 11 reactions, which have to be manipulated in a
In the short term, and especially for high-risk chilled food- sequence allowing for the contact times and the 10 s tolerance.
processing, final assurance of product disinfectant per- If the procedure is staggered with too large intervals then all
formance can only be validated by undertaking studies in a the 11 reactions cannot be carried out in time. This sequence
food-processing environment using a comprehensive micro- of manipulations, undertaken to maximize time output,
biological verification programme with environmental requires training to ensure competency.
sampling. It is not clear at this stage how repeatable and reproducible
The results for E. coli O157:H7 show that, under suspension this suspension test is. Repeatability studies have been under-
test conditions, it is not a particularly resistant organism. taken on previous suspension tests on which BS EN 1276
Where a five log reduction in viable counts was achieved with (1997) is based (Bloomfield et al. 1991, 1995; Holah 1995) but
Ps. aeruginosa it was also achieved against E. coli 0157:H7. it is not known whether this test is more or less repeatable.
This work shows that the control of E. coli O157: H7 in the To ascertain this, a European Committee for Standardisation
factory environment should be effective with a thoroughly (CEN)-funded European ring trial is currently being under-
designed sanitation programme incorporating an appropriate taken through the auspices of CEN/TC 216 which hopes to
disinfectant. report the results at a later date.
In conclusion, E. coli O157 does not seem to be any more
difficult to kill using food industry disinfectants than Ps.
aeruginosa. Products that passed at 10 °C also passed at 20 °C
Evaluation of BS EN 1276 (1997)
and only three products showed a temperature effect.
The nature of this work with the number of tests undertaken However, of the six parameters examined (two organisms,
has allowed an opportunity to evaluate the test method itself, two temperatures and two soil levels) only 11 of 18 products
particularly in terms of practical ease. The concept of the test achieved the required 5 log reductions. This work dem-
is simple, i.e. the addition of 8·0 ml disinfectant to 1·0 ml onstrates the importance of undertaking laboratory dis-
bacteria and 1·0 ml interfering substance for a 5-min contact infectant tests under appropriate simulations of in-use
time followed by 5 min neutralization prior to pour plating to conditions and of considering final verification with a field
enumerate the number of viable micro-organisms remaining. trial in the food-processing environment by the user. The BS
However, there are a number of issues which make this test EN 1276 (1997) quantitative bacterial suspension test is not
complex. difficult to undertake but it does require training and experi-
The test procedure requires an initial inoculum of 1·5– ence to become fully proficient and should be a useful method
5·0 × 108 cfu ml−1 which may be measured by a variety of to harmonize disinfectant testing across Europe.
means as specified in the test method, including the use of a
spectrophotometer or nephelometer. This laboratory per-
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