Letters in Applied Microbiology 2004, 39, 194–198 doi:10.1111/j.1472-765X.2004.01561.

Virucidal activity of the new disinfectant monopercitric acid

P. Wutzler and A. Sauerbrei

Institute of Virology and Antiviral Therapy, Friedrich-Schiller University, Jena, Germany

2003/0915: received 13 October 2003, revised 4 May 2004 and accepted 11 May 2004

ABSTRACT
P . W U T Z L E R A N D A . S A U E R B R E I . 2004.
Aims: The virucidal efficacy of monopercitric acid (MPCA) was evaluated against the enveloped vaccinia virus as
well as the nonenveloped adenovirus type 2 and poliovirus type 1. The results were compared with that obtained
with peracetic acid (PAA).
Methods and Results: In the virucidal suspension test without and with protein burden, all viruses were
inactivated by 0Æ5% MPCA within 0Æ5 min or by 0Æ1% MPCA within 5 min as measured by a >104-fold reduction
in virus titres. For MPCA, there was a better virucidal efficacy than for PAA which inactivated all viruses
included in the test within 15–30 min at a concentration of 0Æ2%.
Significance and Impact of the Study: The high virucidal activity, short exposure times, and nontoxic
by-products seem to make MPCA suitable as disinfectant for medical use and should warrant further investigation.

Keywords: disinfectant, monopercitric acid, peracetic acid, suspension test, virucidal activity.

severe illnesses in humans than the viruses characterized by

INTRODUCTION
Human pathogenic viruses can be detected in the hospital
environment, on the hands of hospital patients and staff
members, on contaminated surfaces or medical instruments.
The viral transmission to susceptible patients has been
recognized as a significant cause of hospital-acquired
infections (Aitken and Jeffries 2001). Lipophilic viruses
with an envelope, such as the respiratory syncytial virus, the
influenza viruses or the human immunodeficiency virus, are
markedly sensitive to liquid chemical disinfectants (Jülich
et al. 1993). But, they can survive after inadequate disin-
fection, particularly, in the presence of proteins or blood. By
contrast, viruses without envelope have to be regarded as
more resistant. Nonenveloped viruses with capsomeric
lipophilicity such as rotaviruses and adenoviruses have been
reported as moderately sensitive (Prince et al. 1991).
Hydrophilic viruses without envelope as the picornaviruses
only possess a slight sensitivity to chemical disinfectants. In
principle, the more resistant viruses are less associated with
Correspondence to: Andreas Sauerbrei, Friedrich-Schiller University Jena, Institute
of Virology and Antiviral Therapy, Hans-Knöll-Straße 2, D-07745 Jena, Germany
(e-mail: andreas.sauerbrei@med.uni-jena.de).
a marked chemical sensitivity.
The heterogeneous sensitivity of viruses to liquid chem-
icals should be considered for virucidal disinfection, espe-
cially, when inactivation of nonenveloped viruses is
required. Thus, viral prototype strains have been recom-
mended for testing of chemical disinfectants in many
European countries (Anon. 1989; German Association for
the Control of Virus Diseases 1990). Commercially available
liquid disinfectants with activity against the resistant
nonenveloped viruses can generally be classified in three
groups including aldehydes, chlorine-based and peroxygen
compounds (Robert-Koch Institute 2003). Aldehyde disin-
fectants such as glutaraldehyde and formaldehyde, which are
commonly used, need considerably long exposure time
intervals and can sensitize users by causing irritations of the
skin and the mucous membranes (Spicher and Peters 1976;
Favero and Bond 1991; Schnuch et al. 1998). Due to their
toxicity, chlorinaceous compounds are not well suited for
decontamination of the environment, equipment or exposed
persons (Hamouda et al. 1999). The peracetic acid (PAA)
has been known as a reliable disinfectant with a broad
spectrum germicidal activity including nonenveloped vir-
uses (Block 1991).
ª 2004 The Society for Applied Microbiology
 VIRUCIDAL ACTIVITY OF MONOPERCITRIC ACID 195

The present study reports the virucidal efficacy of
monopercitric acid (MPCA), a new potential disinfectant
on the basis of peroxy acids. The virucidal activity was
evaluated against enveloped and nonenveloped viruses
recommended by the German guidelines for testing the
virucidal effectiveness of chemical disinfectants. Virucidal
efficacy of MPCA was compared with that of PAA.
MATERIALS AND METHODS
Chemicals
The MPCA was obtained at a concentration of 20% from
1 Kesla Pharma Wolfen GmbH (Greppin, Germany). As
PAA, the preparation Wofasteril
(Kesla Pharma Wolfen)
was used. Wofasteril
contains 40 ± 2% (w/v) PAA. Both
concentrated MPCA and Wofasteril
were stored at 4C.
Immediately before testing, the concentrations of MPCA
and PAA were prepared by dilution with distilled water and
subsequently measured using the standard iodometric
titration method. In brief, peroxygen compounds oxidize
iodide to iodine which is titrated with thiosulphate solution
(Mücke 1973).
Viruses and cells
Following viruses were included in this study: (i) the
enveloped vaccinia virus strain Elstree, (ii) the nonenveloped
adenovirus type 2 strain Adenoid 6, and (iii) the nonenvel-
oped poliovirus type 1 strain Mahoney (German Association
for the Control of Virus Diseases 1990). The viruses were
grown and titrated using monolayers of human embryo lung
fibroblasts (HEF, vaccinia virus), Vero 76 cells (adenovirus)
and HEp-2 cells (poliovirus), respectively, as described
previously (Wutzler and Sauerbrei 2000). For virucidal
testing, all viruses were used in titres of 107–108 tissue
culture infective dose 50% (TCID50).
The mixtures were serially diluted 10-fold using ice-cold
cell culture medium and, thereafter, 200 ll of each were
seeded into micro-plate wells of the appropriate cell culture
for measuring the TCID50. Each experiment was performed
twice. The virucidal effect expressed as the log10 reduction
in virus titres was calculated by the difference between the
virus titres following exposure to biocide and the virus titres
following exposure to control medium. The criterion used
for assessment of the virucidal activity was a log10 reduction
of ‡4 in virus titre corresponding to 99Æ99% inactivation. To
compare the virucidal effectiveness of MPCA with that of
PAA, the concentration-time values (Weavers and Wickra-
manayake 2001) for 99Æ99% were calculated. For evaluation
of virucidal efficacy in the presence of proteins, 10% FCS
was preferred to 2% serum albumin which caused in
principle a lower inhibition of MPCA and PAA.
To differentiate between virus-induced cytopathogenic
changes and the cytotoxic effect caused by the
test substances, the cell monolayers were monitored for
morphological changes after exposure to the disinfectant
solution.
RESULTS
All test viruses were 99Æ99% inactivated by 0Æ5% MPCA
within 0Æ5 min or by 0Æ1% MPCA within 5 min (Tables
1–3). These values correspond to the virucidal efficacy
without and with 10% FCS against poliovirus type 1
(Table 1) which has to be considered the most resistant test
virus. For inactivation of adenovirus type 2 by >4 log10,
Table 1 Virucidal activity of monopercitric acid (MPCA) and
peracetic acid (PAA) against poliovirus type 1 strain Mahoney in
suspension tests without (in bold) and with 10% FCS (in italic)
Reduction of virus titre by log10/exposure time (min)
Biocides (%) 0Æ5 1 1Æ5 2 5 15 30 60

MPCA
Virucidal assays 0Æ01 0Æ3 0Æ3 0Æ3 0Æ7 0Æ7 0Æ7 0Æ7 0Æ7
0 0 0Æ2 0Æ2 0Æ2 0Æ2 0Æ2 0Æ2
Virucidal assays were performed according to the Guidelines 0Æ025 0Æ8 0Æ8 0Æ8 0Æ8 0Æ8 0Æ8 1Æ0 1Æ2
of the German Federal Health Office and the German 0Æ1 0Æ3 0Æ3 0Æ3 0Æ3 0Æ3 0Æ5 0Æ5
Association for the Control of Virus Diseases for testing the 0Æ05 0Æ9 0Æ9 0Æ9 1Æ0 1Æ0 1Æ0 1Æ2 2Æ1
effectiveness of chemicals disinfectants against viruses 0Æ5 0Æ7 0Æ7 0Æ7 0Æ7 0Æ7 0Æ9 1Æ5
(German Association for the Control of Virus Diseases 0Æ075 0Æ9 0Æ9 0Æ9 1Æ0 1Æ0 1Æ2 1Æ7 3Æ8
1990). After exposure times of 0Æ5, 1, 2, 5, 15, 30, and 0Æ5 0Æ8 0Æ9 0Æ9 0Æ9 0Æ9 1Æ2 2Æ3
60 min without and with 10% foetal calf serum (FCS), 0Æ1 0Æ9 1Æ6 1Æ9 1Æ9 >4Æ0 – – –
MPCA and PAA were subsequently neutralized by mixing 0Æ6 1Æ5 1Æ6 1Æ6 >4Æ0 – – –
with equal volume of phosphate-buffered saline containing 0Æ5 >4Æ0 – – – – – – –
>4Æ0 – – – – – – –
cysteine hydrochloride at concentrations corresponding to
PAA
that of MPCA or PAA (Wutzler and Sauerbrei 2000). This
0Æ2 0Æ8 0Æ9 1Æ1 1Æ2 2Æ0 >4Æ0 – –
neutralizer quenched completely the activity of MPCA and 0 0 0Æ5 0Æ6 0Æ9 2Æ0 >4Æ0 –
PAA by reduction to citric acid and acetic acid, respectively.
ª 2004 The Society for Applied Microbiology, Letters in Applied Microbiology, 39, 194–198, doi:10.1111/j.1472-765X.2004.01561.x
196 P . W U T Z L E R A N D A . S A U E R B R E I

Table 2 Virucidal activity of monopercitric acid (MPCA) and Table 4 Comparison of concentration-time values (mg min l)1) for
peracetic acid (PAA) against adenovirus type 2 strain Adenoid 6 in 99Æ99% inactivation of poliovirus type 1 strain Mahoney, adenovirus
suspension tests without (in bold) and with 10% FCS (in italic) type 2 strain Adenoid 6, and vaccinia virus strain Elstree by
monopercitric acid and peracetic acid without (in bold) and with 10%
Reduction of virus titre by log10/exposure time (min) FCS (in italic)
Biocides (%) 0Æ5 1 1Æ5 2 5 15 30 60 Virus Monopercitric acid Peracetic acid
MPCA Poliovirus type 1 £0Æ25–0Æ5 3
0Æ01 0Æ5 0Æ5 0Æ5 0Æ5 0Æ6 0Æ6 0Æ7 1Æ4 £0Æ25–0Æ5 6
0 0 0Æ2 0Æ2 0Æ2 0Æ2 0Æ2 0Æ6 Adenovirus type 2 0Æ05–0Æ25 3
0Æ025 0Æ5 0Æ6 0Æ6 0Æ6 0Æ6 1Æ0 1Æ0 1Æ4 0Æ1–0Æ75 6
0Æ2 0Æ2 0Æ3 0Æ3 0Æ3 0Æ3 0Æ3 0Æ6 Vaccinia virus 0Æ025–0Æ15 £0Æ1
0Æ05 2Æ6 3Æ3 3Æ7 3Æ7 >4Æ0 – – – 0Æ025–0Æ15 £0Æ1
1Æ4 1Æ7 2Æ0 2Æ2 2Æ9 >4Æ0 – –
0Æ075 3Æ4 >4Æ0 – – – – – –
1Æ5 3Æ8 >4Æ0 – – – – – The concentration-time values for 99Æ99% inactivation of
0Æ1 >4Æ0 – – – – – – –
poliovirus type 1, adenovirus type 2 and vaccinia virus by
3Æ5 >4Æ0 – – – – – –
MPCA and PAA are summarized in the Table 4. The lower
0Æ5 >4Æ0 – – – – – – –
>4Æ0 – – – – – – –
values of MPCA in comparison with PAA indicate a higher
PAA efficacy against the viruses included in this study.
0Æ2 1Æ2 1Æ4 1Æ6 1Æ7 2Æ5 >4Æ0 – –
0Æ8 0Æ8 1Æ0 1Æ3 2Æ1 3Æ6 >4Æ0 –
DISCUSSION
The increase in hospital-acquired infections with viral
aetiology can require disinfectants which are capable of
Table 3 Virucidal activity of monopercitric acid (MPCA) and
peracetic acid (PAA) against vaccinia virus strain Elstree in suspension
inactivating both enveloped as well as nonenveloped viruses
tests without (in bold) and with 10% FCS (in italic) in dependence on application. To date, only a small number
of disinfectants used in hospitals fulfils this demand. For
Reduction of virus titre by log10/exposure this reason, a main objective is to search for new broad-
time (min) spectrum biocides which act within short time intervals and
Biocides (%) 0Æ5 1 1Æ5 2 5 15 are approved for hospital use.
The present study demonstrates the new peroxygen
MPCA compound MPCA to be an excellent virucidal disinfectant
0Æ01 0Æ1 0Æ1 0Æ2 0Æ6 1Æ3 >4Æ0 as evaluated against the enveloped vaccinia virus as well as
0 0 0Æ1 0Æ2 1Æ0 >4Æ0
the nonenveloped adenovirus type 2 and poliovirus type 1.
0Æ025 2Æ1 2Æ1 3Æ1 >4Æ0 – –
The virucidal efficacy was slightly reduced in the presence
1Æ6 1Æ6 2Æ0 >4Æ0 – –
0Æ05 >4Æ0 – – – – –
of 10% FCS. In comparative investigations, the virucidal
>4Æ0 – – – – – effectiveness without and with protein burden was better
PAA than that of PAA. These findings suggest a higher oxidizing
0Æ2 >4Æ0 – – – – – activity of MPCA. The strong germicidal including viru-
>4Æ0 – – – – – cidal properties of PAA have been accepted for many years
(Sprößig and Mücke 1968; Baldry 1983; Block 1991). Thus,
the Centers for Disease Control of the United States have
lower concentrations of MPCA, e.g. 0Æ1% for 0Æ5–1 min or listed this compound as chemical sterilant and high-level
0Æ05% for 5–15 min, were necessary (Table 2). For vaccinia disinfectant (Block 1991).
virus, a virucidal effect could be achieved without and with For MPCA, we have also found an excellent activity
protein burden when 0Æ05% MPCA was used for 0Æ5 min against spores of different clostridial species (unpublished
(Table 3). A comparable efficacy had 0Æ025% MPCA after data). This sporicidal efficacy was at least comparable with
2 min and 0Æ01% MPCA after 15 min. that of PAA. Due to the high resistance of bacterial spores to
In comparison with MPCA, PAA at a concentration of chemical and physical agents, the sporicidal activity is
0Æ2% inactivated the most resistant poliovirus type 1 and the commonly used to evaluate disinfecting and sterilizing
adenovirus type 2 within 15 min (Tables 1 and 2). When agents (Beloian 1990). In general, concentrations of biocides
10% calf serum was added, the exposure time was extended with sporicidal activity also include the virucidal efficacy.
to 30 min. We found, however, that 0Æ05% MPCA, to be able to kill the
ª 2004 The Society for Applied Microbiology, Letters in Applied Microbiology, 39, 194–198, doi:10.1111/j.1472-765X.2004.01561.x

most resistant spores of Clostridium septicum after 60 min,
was not effective within this time interval against poliovirus
type 1 (Table 1). This illustrates that the sporicidal
effectiveness of a potential disinfectant does not have to
include its virucidal activity. Therefore, disinfectants should
be evaluated separately for their virucidal as well as
sporicidal efficacy.
Among the test viruses, only the nonenveloped poliovirus
type 1 and adenovirus type 2 have a high or moderate
resistance to chemical biocides. The enveloped vaccinia
virus and the nonenveloped papovavirus SV40, which is also
recommended by the German guidelines (German Associ-
ation for the Control of Virus Diseases 1990) and was not
included in this study, have generally to be regarded as
sensitive. On the basis of these data, the European standard
of the virucidal suspension test for chemical disinfectants
and antiseptics used in human medicine (European Com-
mittee for Standardization 1996) only considers poliovirus
type 1 and adenovirus type 5. In addition, the hepatitis B
virus of ducks has been selected as one of the future test
viruses (Wang et al. 2002). Because of the intensive efforts
to eradicate polio, polioviruses may no longer be available to
test disinfectants. In our experience, adenovirus serotypes 5
and 44 might be suitable as model viruses for testing the
broad-spectrum virucidal activity of disinfectants and could
replace the resistant poliovirus type 1 (Sauerbrei et al. 2004).
Recently, poliovirus wild-type 1 and adenovirus type 2 were
replaced by poliovirus vaccine-type 1 and adenovirus type 5
in the German guidelines (Fachausschuss ‘Virusdesinfek-
tion’ der Deutschen Vereinigung zur Bekämpfung der
Viruskrankheiten und des Robert-Koch Instituts 2003).
The suspension test used in this study has widely been
accepted for the determination of virucidal activity.
Although this test only furnishes the minimum require-
ments for virus inactivation, it allows the evaluation of
virucidal chemical disinfectants under standardized condi-
tions. Attempts to standardize virucidal testing have been
made in the member countries of the European Union
(European Committee for Standardization 1996). The main
problem when measuring the virucidal activity is the
cytotoxicity of the chemical test compounds. In the case of
peroxy acids, the toxic changes of cells are mainly caused by
the low pH. But, the problem of cytotoxicity can be
effectively circumvented by the preparation of viral suspen-
sions with a high number of infectious particles or by gel
filtration technique for separating disinfectants from the
virus before titration (Valot et al. 2000). However, tech-
niques separating peroxygen compounds have not been
reported to date.
The mechanism of virucidal action of the new peroxy acid
MPCA should be regarded as comparable with that of PAA.
By its nonspecific oxidizing effect, PAA likely acts on SH-,
OH- and NH-groups of amino acids, nucleotides, and
ª 2004 The Society for Applied Microbiology, Letters in Applied Microbiology, 39, 194–198, doi:10.1111/j.1472-765X.2004.01561.x
VIRUCIDAL ACTIVITY OF MONOPERCITRIC ACID 197
unsaturated fatty acids. Thus, it is able to interact with lipid
and protein components of the viral envelope, to denature
viral capsid proteins and to inactivate nucleic acids of the
viral genome (Maillard et al. 1996a,b). As shown by electron
microscopic observations, PAA can completely destroy the
structure of virus particles (Maillard et al. 1995; Wutzler
and Sauerbrei 2000). Like PAA, the use of MPCA is
environment-friendly as the compound is decomposed to
citric acid and water. Disadvantages of peroxygen disinfect-
ants are the high hydrolysis paralleled with storage at room
temperature, the strong dependence of the germicidal
activity on pH, and the possibility of partial inactivation
by the haemoglobin of erythrocytes in the presence of blood
(Jülich et al. 1993). Nevertheless, the high virucidal activity
without and with protein burden, short exposure times and
nontoxic by-products of MPCA should warrant further
investigation as potential disinfectant for medical use.
REFERENCES
Aitken, C. and Jeffries, D.J. (2001) Nosocomial spread of viral disease.
Clinical Microbiology Reviews 14, 528–546.
Anon. (1989) Normalisation Française T 72-180, T 72-181. Antiseptiques
et désinfectants utilisés à l‘état liquide, miscibles à l‘eau. Détermination de
l‘activité virucide vis-à-vis des virus de vertébrés. AFNOR, Paris.
Baldry, M.G.C. (1983) The bactericidal, fungicidal and sporicidal
properties of hydrogen peroxide and peracetic acid. Journal of
Applied Bacteriology 54, 417–423.
Beloian, A. (1990) Disinfectants. In AOAC Official Methods of Analysis.
ed. Helrich, K. pp. 133–146. Arlington, TX: Association of Official
2 Analytical Chemists.
Block, S.S. (1991) Peroxygen compounds. In Disinfection, Sterilisation,
and Preservation ed. Block, S.S. pp. 167–181. Philadelphia, PA: Lea
& Febiger.
European Committee for Standardization (1996) European Standard of
Virucidal Suspension Test for Chemical Disinfectants and Antiseptics
Used in Human Medicine. Brussels: European Committee for
3 Standardization.
Fachausschuss ‘Virusdesinfektion’ der Deutschen Vereinigung zur
Bekämpfung der Viruskrankheiten und des Robert Koch-Instituts
(2003) Richtlinie des Bundesgesundheitsamtes und der Deutschen
Vereinigung zur Bekämpfung der Viruskrankheiten zur Prüfung
von chemischen Desinfektionsmitteln auf Wirksamkeit gegen
Viren. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz
4 46, 619.
Favero, M.S. and Bond, W.W. (1991) Chemical disinfection of medical
and surgical materials. In Disinfection, Sterilisation, and Preservation
ed. Block, S.S. pp. 617–641. Philadelphia, PA: Lea & Febiger.
German Association for the Control of Virus Diseases (1990)
Guidelines of Bundesgesundheitsamt (BGA; German Federal
Health Office) and Deutsche Vereinigung zur Bekämpfung der
Viruskrankheiten e.V. (DVV; German Association for the Control of
Virus Diseases) for testing the effectiveness of chemical disinfectants
against viruses. Zentralblatt für Hygiene und Umweltmedizin 189,
554–556.
 198 P . W U T Z L E R A N D A . S A U E R B R E I
Hamouda, T., Hayes, M.M., Cao, Z., Tonda, R., Johnson, K., Wright,
D.C., Brisker, J. and Baker, J.R. Jr (1999) A novel surfactant
nanoemulsion with broad-spectrum sporicidal activity against
Bacillus species. Journal of Infectious Diseases 180, 1939–1949.
Jülich, W.D., von Rheinbaben, F., Steinmann, J. and Kramer, A.
(1993) On the virucidal efficacy of chemical and physical disinfect-
ants or disinfection procedures. Hygiene und Medizin 18, 303–326.
Maillard, J.Y., Hann, A.C., Beggs, T.S., Day, M.J., Hudson, R.A. and
Russell, A.D. (1995) Electron-microscopic investigation of the effect
of biocides on Pseudomonas aeruginosa PAO bacteriophage F116.
Journal of Medical Microbiology 42, 415–420.
Maillard, J.Y., Beggs, T.S., Day, M.J., Hudson, R.A. and Russell,
A.D. (1996a) The effect of biocides on proteins of Pseudomonas
aeruginosa PAO bacteriophage F116. Journal of Applied Bacteriology
80, 291–295.
Maillard, J.Y., Beggs, T.S., Day, M.J., Hudson, R.A. and Russell, A.D.
(1996b) Damage of Pseudomonas aeruginosa PAO1 bacteriophage
F116 DNA by biocides. Journal of Applied Bacteriology 80, 540–544.
Mücke, H. (1973) Zur Bestimmung und zum Spurennachweis von
Peressigsäure in Gegenwart von Wasserstoffperoxid für die Praxis
der Kaltsterilisation und Desinfektion. Zeitschrift für Medizinische
5 Labortechnik 14, 319–323.
Prince, H.N., Prince, D.L. and Prince, R.N. (1991) Principles of viral
control and transmission. In Disinfection, Sterilisation, and Preserva-
tion ed. Block, S.S. pp. 411–444. Philadelphia, PA: Lea & Febiger.
Robert-Koch Institute (2003) Liste der vom Robert-Koch-Institut
geprüften und anerkannten Desinfektionsmittel und – verfahren.
Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz 46,
72–95.
ª 2004 The Society for Applied Microbiology, Letters in Applied Microbiology, 39, 194–198, doi:10.1111/j.1472-765X.2004.01561.x
Sauerbrei, A., Sehr, K., Brandstädt, A., Heim, A., Reimer, K. and
Wutzler, P. (2004) Sensitivity of human adenoviruses to different
groups of chemical biocides. Journal of Hospital Infection 57,
59–66.
Schnuch, A., Uter, W., Geier, J., Frosch, P.J. and Rustemeyer, T.
(1998) Contact allergies in healthcare workers. Results from the
IVDK. Acta Dermato-Venerologica 78, 358–363.
Spicher, G. and Peters, J. (1976) Resistenz mikrobieller Keime
gegenüber Formaldehyd. I. Vergleichende quantitative Untersuc-
hungen an einigen ausgewählten Arten vegetativer Bakterien,
bakteriellen Sporen, Pilze, Bakteriophagen und Viren. Zentralblatt
Bakteriologie (Orig B) 163, 486–508.
Sprößig, M. and Mücke, H. (1968) On the antimicrobial effect of
peracetic acid. 5. Studies on the virucidal effect. Pharmazie 23, 665–
667.
Valot, S., Edert, D. and Le Faou, A. (2000) A simple method for the in
vitro study of the virucidal activity of disinfectants. Journal of
Virological Methods 86, 21–24.
Wang, C.Y., Giambrone, J.J. and Smith, B.F. (2002) Development of
viral disinfectant assays for duck hepatitis B virus using cell culture/
PCR. Journal of Virological Methods 106, 39–50.
Weavers, L.K. and Wickramanayake, G.B. (2001) Kinetics of the
inactivation of microorganisms. In Disinfection, Sterilisation, and
Preservation ed. Block, S.S. pp. 65–78. Philadelphia, PA: Williams &
6 Wilkins.
Wutzler, P. and Sauerbrei, A. (2000) Virucidal efficacy of a
combination of 0Æ2% peracetic acid and 80% (v/v) ethanol (PAA-
ethanol) as a potential hand disinfectant. Journal of Hospital Infection
46, 304–308.