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[Standardization of the testing of disinfectants in Europe].

Several European countries have adopted the evaluation of the microbicidal properties of disinfectants in two stages: after some preliminary tests, tests under practical conditions or main tests are carried out, as was described originally in the German guidelines for the testing of disinfectants. Nevertheless in the Anglosaxon countries, the use dilution of products intended for surface and floor disinfection is determined by suspension tests, capacity tests, or carrier tests. In some European countries, however, tests under practical conditions are developed for this purpose, but if these Belgian (B), German (D), French (F) and Dutch (NL) techniques are analysed and compared in their composing elements, great differences are found (Tables 1 to 3). Therefore the Belgian and Dutch tests must be the more severe. Another example of discordance is given in the preoperative disinfection of the surgeons' hand. Between 1973 and 1984 23 different testing techniques were described (Table 4). The differences in technique explain the differences in results obtained. In order to achieve a greater unification of the testing techniques, it is desirable to agree firstly on the requirements of disinfectants. Perhaps it is time to discuss it again.

Anti-Infective Agents, Local↗

Assessment of viral disinfection by means of bacteriophage OX 174.

A method was developed for evaluating the viricidal effectiveness of chemical disinfectants. The method uses the DNA bacteriophage phi X 174 (ATCC 13706-B1), whose host is Escherichia coli C (ATCC 13706-1), as the model virus. The bacteriophage has particles of icosahedral symmetry, 26 nm in diameter, and resembles animal entero- and parvoviruses by its resistance to physicochemical factors. Disinfection effectiveness was evaluated in suspension medium and on carriers of different materials that are disinfected in practice. The Horvath-Alföldi method on plexiglass panels containing 21 dishes of 40-mm diameter and 2-mm depth was used. A volume of 0.2 ml of phage suspension or one flat carrier 25 mm X 25 mm was put per panel dish and overlaid with 3 ml of dissolved soft agar medium containing at 37 degrees C, quantitatively as VE = log (No/Nd) in suspension tests and semiquantitatively on a 0-5 scale in tests on carrier materials. The viricidal effect of disinfectant-containing washing agents was tested on textile carriers during washing in an experimental washing machine. The method can be adjusted to evaluate the viricidal effectiveness of disinfection under different environmental conditions, e.g. in sanitary, agricultural, veterinary or food-industry facilities.

Bacteriophage phi X 174↗

Testing disinfectants for efficacy.

Testing under laboratory conditions is undoubtedly useful in assessing the activity of disinfectants. However, such testing must be regarded as no more than a preliminary to field trials. This not only indicates the unreliability of laboratory tests, but also poses the wider problem of how any laboratory evaluation can be correlated with the field requirements. Many attempts have therefore been made to devise tests which are suitable to evaluate the in-use requirements of disinfection (e.g. on farms). The aim of these techniques is to record the end-point of a disinfection procedure on the surfaces in animal houses. In view of the need to standardise testing conditions in the field, the author provides details of the optimum time and place for taking samples under field conditions. Samples for culture media should be taken from the floor when dry. The various techniques (i.e. swabbing, agar cylinder, agar carrier, 'ready-to-use' set) should provide information on the end-point of the disinfection. The desired end-points is approximately one viable bacterium per cm2 of surface area. The technique used should demonstrate whether the disinfection has been effective.

Animals↗

Cleaning and disinfection practice in the meat industries of Europe.

The application and efficacy of cleaning and disinfection methods are reviewed, together with the relevant European and French legislation. European Commission Hygiene Directive 93/43/EEC of 14 June 1993 proposes the adoption of hazard analysis and critical control points (HACCP) for the meat industry, and this includes cleaning and disinfection. It is necessary to organise a team for washing, cleaning, rinsing, disinfection and final rinsing; three different types of organisation are compared. Application of HACCP and its contribution to the shelf life of products and their contamination with Listeria monocytogenes is discussed in the light of practical experience with poultry meat and cured pork products. Various means of verifying the efficacy of cleaning and disinfection (turbidimetry, adenosine triphosphate assay and macroscopic observation) are compared with the techniques of conventional microbiology. The authors conclude that cleaning and disinfection are essential for application of HACCP to the meat industry.

Animals↗

Evaluation of soft contact lens disinfection in the home environment.

Recent reports of bacterial corneal ulcers in cosmetic soft contact lens wearers prompted an evaluation of one possible source of contamination, inadequate home disinfection of the lenses. Twenty-nine patients were asked to come for an early morning examination. They were told not to wear their lenses so their usual home disinfection procedure could be observed. On arrival, extensive bacterial and fungal cultures were taken of the solution in the previously unopened cases. Ten of the 29 patients (34.5%) had contaminated cases. Some patients did not follow the manufacturer's disinfection guidelines. The current disinfection procedure has repeatedly proven adequate in the laboratory. However, the practical fact is that a substantial percentage (34.5% in this study) of soft lens wearers are inadequately disinfecting their lenses. This serves as a possible source of contamination leading to bacterial corneal ulcers.

Activities of Daily Living↗

Comparative efficacies of soft contact lens disinfectant solutions against microbial films in lens cases.

Biofilms of Pseudomonas aeruginosa, Serratia marcescens, Staphylococcus epidermidis, Streptococcus pyogenes, and Candida albicans, established in the wells of a polyethylene contact lens case, retained viability to certain soft contact lens disinfectant solutions after exposure for the manufacturer's minimum recommended disinfection times. The relative order of resistance of bacterial biofilms was as follows: S marcescens was greater than P aeruginosa, which was greater than S epidermidis, which was greater than S pyogenes. Air drying of biofilms for 10 hours increased the efficacy of the disinfectant solutions, but drying was not enough to decrease the incidence of recovery to 0% for all solutions. Hydrogen peroxide was more effective against biofilms than disinfectant solutions formulated with chlorhexidine gluconate or polyquaternium-1 or polyaminopropyl biguanide. We recommend that determination of efficacy of contact lens disinfectant solutions should include challenges against biofilms.

Bacteria↗

Glutaraldehyde retains its disinfectant properties in presence of hydroxypropylmethyl cellulose (HPMC) gel.

Explanted medical devices are routinely sent to laboratories at the Center for Devices and Radiological Health for analysis. The shipping of these devices presents potential hazards to personnel as well as an opportunity for damage to the devices. In an effort to address these concerns, a viscous disinfecting shipping medium that would limit splashing and cushion a suspended device was proposed. Consequently, we investigated the disinfectant properties of adding a gelling agent, hydroxypropylmethyl cellulose (HPMC) to common disinfectants. We found that the germicidal effectiveness of 2.5% glutaraldehyde in 0.05 M borax when tested against Bacillus subtilis spores was not changed by the addition of 2% HPMC. In addition, HPMC appears to be compatible with 70% ethanol and at least one commercial disinfectant containing a quaternary ammonium compound. Preliminary experiments indicate that an HPMC-disinfectant gel is a potentially useful packaging agent for minimizing the hazards to personnel and materials during shipping of explanted medical devices. The use of such a medium would be subject to guidelines within the context of a program for handling biologically contaminated materials.

Bacillus subtilis↗

Vaginal disinfection during labour for reducing the risk of mother-to-child transmission of HIV infection.

BACKGROUND: Mother-to-child transmission (MTCT) of HIV infection is one of the most tragic consequences of the HIV epidemic, especially in resource-limited countries, resulting in about 650 000 new paediatric HIV infections each year world-wide. The paediatric HIV epidemic threatens to seriously undermine decade-old child survival programmes. This is one of several reviews assessing the available evidence for preventing HIV transmission from an HIV-infected woman to her child. The other reviews assess the effects of antiretroviral therapy, Caesarean section delivery, breast feeding, and vitamin A supplementation. OBJECTIVES: To estimate the effect of vaginal lavage on the risk of MTCT of HIV and infant and maternal mortality and morbidity, as well as tolerability of vaginal lavage in HIV infected women. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, Cochrane Pregnancy and Childbirth Register, PubMed, EMBASE, AIDSLINE, LILACS, AIDSTRIALS, and AIDSDRUGS, using standardised methodological filters for identifying trials. We also searched reference lists of identified articles, relevant editorials, expert opinions and letters to journal editors, and abstracts and proceedings of relevant conferences, and contacted subject experts and pharmaceutical companies. There were no language restrictions. SELECTION CRITERIA: Randomised trials or clinical trials comparing vaginal disinfection during labour with placebo or no treatment, in known HIV infected pregnant women. Trials had to include an estimate of the effect of vaginal disinfection on MTCT of HIV and/or any other pre-specified adverse pregnancy outcome to be included. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial eligibility and quality, and extracted data. MAIN RESULTS: Only one low quality trial included an estimate of the effect of vaginal disinfection on at least one pre-specified outcome. There was no evidence of an effect of vaginal disinfection on MTCT of HIV (odds ratio 0.93, 95% confidence interval 0.63 to 1.38), and no information was available on the other pre-specified outcomes. REVIEWER'S CONCLUSIONS: There is a need for well-designed randomised controlled trials to estimate the effects of vaginal disinfection on MTCT of HIV.

Disinfection↗

Genotoxicity of drinking water disinfectants in plant bioassays.

The genotoxicity of two widely used drinking water disinfectants, sodium hypochlorite (NaClO) and chlorine dioxide (ClO(2)), and a new disinfectant, peracetic acid (PAA, CH(3)-CO-COOH), was evaluated in three short-term plant tests: (1) induction of anaphase chromosome aberrations in the root cells of Allium cepa, (2) micronucleus induction in the root cells of Vicia faba, and (3) micronucleus induction in Tradescantia pollen cells. The study was carried out in the laboratory by directly exposing the plants to several concentrations of the disinfectants in redistilled water at unadjusted (acid) and adjusted (neutral) pHs. Both 0.1 and 0.2 mg/l NaClO induced chromosome aberrations in the Allium cepa test at acid pH, but concentrations up to 0.5 mg/l of all the disinfectants were negative at neutral pH. Concentrations ranging from 0.1 to 0.5 mg/l NaClO, ClO(2,) and PAA induced micronuclei in Vicia faba at acid pH, while 1-2 mg/l NaClO and ClO(2) and 0.5-2 mg/l PAA gave positive responses at neutral pH. Most of concentrations of ClO(2) produced positive responses in the Tradescantia micronucleus test. In general, the highest levels of genotoxicity were observed under acid conditions; at acid pH, significant effects were induced by low concentrations of ClO(2) and PAA. Since the test concentrations of disinfectants are typical of those encountered in the biocidal treatment of tap water and similar concentrations are consumed daily by a large number of people, the genotoxicity of these compounds may constitute a significant public health concern.

Biological Assay↗

Evaluation of in vitro efficacy of the disinfectant Virkon.

A study was conducted on a new acid peroxygen system based disinfectant (Virkon), in order to assess its in vitro efficacy. The chemical was tested on different bacteria (Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli), spores (Bacillus subtilis) and on the Hepatitis B surface antigen (HBsAg), and compared in its activity with phenol and glutaraldehyde (calculation of the 'phenol coefficient' and the 'glutaraldehyde coefficient'). The constancy of speed of disinfection, the coefficient of concentration, the minimal inhibitory concentration (MIC) were also determined, and the destruction of the HBsAg antigenic activity was studied using an ELISA kit. The sporicidal efficacy of Virkon was assessed by cultivating spores in agar nutrient after contact with different dilutions of the disinfectant. The results of the tests showed that Virkon has a high concentration coefficient (mean value of k: 0.374/min) and a wide range of action. The low MIC demonstrates how little concentrations of Virkon can inactivate all studied bacteria. The disinfectant was also able to destroy the hepatitis B surface antigen, and it demonstrated good activity against spores, especially if used in physiologic solution. These characteristics, coupled with the absence of initiation or toxic effects on animals showed by other studies, make wide fields of application for the new disinfectant foreseeable.

Animals↗

Toxocara canis larvae viability after disinfectant-exposition.

The effect of three routinely used disinfectants on the embryonary development of Toxocara canis eggs was evaluated both in vivo and in vitro. In the in vitro experiment, T. canis eggs were treated with the ethanol, sodium hypochlorite, and one commercial mix of benzalconium chloride and formaldehyde, and the embryonary development was assessed. After a period of 24 days incubation, ethanol was the best disinfectant because it prevented the development of the T. canis larvae 2 in the eggs, and sodium hypochlorite caused degeneration in 50% eggs. By using the commercial mix, 25% T. canis eggs developed to 2nd stage larvae. In the in vivo experiment, the embryonated eggs treated with the disinfectants were inoculated to mice, and their brain tissues were examined for larval presence on the 24th day postinfection. In addition, a control group was set up for comparison with the infected groups. No injury or T. canis larvae were observed in mice infected with sodium hypochlorite-treated eggs, opposite to that recorded in the animals infected with the commercial disinfectant-treated eggs. These results showed that both ethanol and sodium hypochlorite are very appropriate because of their full efficacy against infective T. canis eggs. Disinfection of kennels, animal shelters, cages, and veterinary clinics with one of these products to eliminate T. canis eggs and to avoid contamination is strongly recommended.

Animals↗

An easy and safe method to store and disinfect explanted skull bone.

In our department extensive decompression craniectomies became the treatment of choice for patients with massive cerebral oedema following either trauma or acute cerebral infarction. The remarkable survival rates of this neurosurgical technique created the problem of adequate vault defect reconstruction. To evaluate the biological safety of using stored autologous skull flaps for this purpose, we compared three different disinfection methods. Skull bone fragments stored at -21 degrees C for different periods of time were artificially contaminated with clinically relevant strains of Serratia marcescens, Enterococcus faecium and Staphylococcus aureus. As potential methods for disinfection we tested immersion in 3% H2O2, boiling in normal saline for 15 and 30 minutes and a special process of steam disinfection at a temperature of 75 degrees C for 20 minutes. We were able to demonstrate that only steam disinfection completely eliminated the bacterial strains tested. Refrigeration plus steam disinfection of autologous skull bone prior to re-implantation seems to offer reliable safety for its use for defect closure. It is available at reasonable cost in many hospitals and does not require a bone bank.

Bone Transplantation↗

Inactivation of Mycobacterium tuberculosis and Mycobacterium bovis by 14 hospital disinfectants.

Epidemics of mycobacteria due to contamination of medical devices continue to occur. For this reason, we assessed the ability of disinfectants, generally used in hospitals for disinfecting noncritical and semicritical patient care items, to inactivate mycobacteria. A modified Association of Official Analytical Chemists' (AOAC) Tuberculocidal Activity Test, using Middlebrook 7H9 broth as the primary subculture medium and neutralization by dilution, was used to assess the ability of 14 hospital disinfectants to inactivate about 10(6) Mycobacterium tuberculosis and about 10(5) Mycobacterium bovis at 20 degrees C using 10- or 20-minute exposure. All products were tested for each organism using 10 penicylinders (P) and were prepared at the manufacturers' recommended use-dilution. Chlorine dioxide, 0.80% hydrogen peroxide plus 0.06% peroxyacetic acid, and an iodophor achieved complete inactivation (0 + P) of both M. tuberculosis and M. bovis. One quaternary ammonium compound with a tuberculocidal label claim, a quaternary ammonium compound without a tuberculocidal label claim, chlorine (approximately 100 ppm) and 0.13% glutaraldehyde/0.44% phenol/0.08% phenate were not effective (10 + P) against both M. tuberculosis and M. bovis. Another quaternary ammonium compound with a tuberculocidal label claim was tested against only M. bovis and found ineffective (10 + P). Glutaraldehydes (2% alkaline and 2% acid), a phenolic and chlorine (approximately 1,000 ppm) demonstrated complete inactivation of M. tuberculosis (0 + P) and good inactivation of M. bovis (1-3 + P). Two disinfectants, hydrogen peroxide and ethyl alcohol, provided differing results against M. tuberculosis and M. bovis. These studies have important implications for disinfecting semicritical patient care items.

Chlorine↗

The effect of glutaraldehyde base disinfectants on denture base resins.

The effect of two alkaline glutaraldehyde base disinfectants on a heat-cured denture base resin was evaluated by the flexural strength, the repair flexural strength, and the surface morphology of the material that had been immersed up to 12 hours in the disinfecting solutions. The flexural strength of the material was not significantly affected by either disinfectant. The disinfectant with phenolic buffer caused surface pitting of the material after 10 minutes of immersion, and softening and swelling of the surface after 2 hours of immersion. No apparent surface change was observed with the regular alkaline formulation, however. Between the two repair resins, the autopolymerized resin yielded greater repair flexural strength than the light-cured repair resin. However, the repair flexural strength of autopolymerized resin seemed to be influenced by either disinfecting solution whereas the light-cured repair resin was not affected.

Acrylic Resins↗

Sodium hypochlorite disinfection of irreversible hydrocolloid impression material.

Alginate impression material is one of the most frequently used in dentistry. However, this material is susceptible to dimensional distortion during disinfection because of its hydrophilic nature. This study examined the effects of alginate disinfection using a sodium hypochlorite spray or impression immersion. Spray disinfection of an alginate impression did not cause dimensional differences of the poured stone casts when compared with casts from water-rinsed controls. Immersion disinfection created dimensional distortion of the anterior, posterior, and interarch model segments. Both the spray and immersion treatments equally decreased the surface detail reproducibility. The antimicrobial effects of the spray treatment were similar to those of the immersion treatment, while mere water rinsing resulted in inadequate disinfection.

Aerosols↗

The disinfection of baths and shower trolleys in hospitals.

The contamination level of bath tubs and shower trolleys has been studied after use by hospital patients and following either disinfection with a phenolic disinfectant-detergent or cleaning with a detergent. After use the surfaces were contaminated with 0.5 x 10(3) - 1.7 x 10(3) colony-forming units/dm2. Cleaning and disinfection reduced the contamination level approximately tenfold. As disinfection is not more efficient than cleaning the latter procedure should routinely be preferred after each use of a bath or a shower trolley. However, disinfection may be more effective than cleaning after bathing or showering a patient with a major wound or skin infection.

Baths↗

Surgical hand disinfection: effect of sequential use of two chlorhexidine preparations.

The antimicrobial efficacy of three 'two-phase' surgical hand disinfection procedures was compared, in a volunteer study, to 60% n-propanol, applied for 5 min, which is the reference hand-disinfection procedure used in Austria and West Germany (FRG). The procedures involved sequential use of unmedicated soap or a disinfectant-detergent containing 4% chlorhexidine gluconate (CHX; 'Hibiscrub') followed by a handrub preparation containing 70% w/w isopropanol plus 0.5% CHX ('Hibisol'). The immediate and sustained effects (3 h) of washing with unmedicated soap (3 min) followed by rubbing on 'Hibisol' (4 min) were significantly smaller (log10 reductions of 1.72 and 1.12) than with each of the other procedures. Use of 'Hibiscrub' (3 min) and 'Hibisol' (4 min) produced log10 reductions of 2.50 and 1.71, equalling those of the reference procedure with n-propanol (2.49 and 1.78). When 'Hibisol' was used for 5 min rather than 4 min, a considerable, though not significant, increase in effect was achieved (log10 reductions of 2.90 and 2.07). Replacement of unmedicated soap by 'Hibiscrub' could significantly improve the effectiveness of the hand disinfection procedure commonly used by surgeons in German-speaking countries; namely to wash hands first with soap and then disinfect them with an alcoholic preparation. It may also be of additional advantage as this adds another 'layer' of CHX when 'Hibisol' rather than alcohol alone is used.

1-Propanol↗

International standardization of disinfectant testing: is it possible?

At present there is no harmonization of antiseptic and disinfectant testing in the world, not even in Europe. In the case of surface disinfection the bactericidal activity of preparations is evaluated by very different techniques. The Association of Official Analytical Chemists of the USA prescribes a carrier test, the use-dilution test, to determine the maximum dilutions that are effective for practical disinfection. In Germany and in France, the final test for determining the use-dilution of a disinfectant preparation is a practical test mimicking conditions of use. In the UK the Kelsey-Sykes test, a capacity test, is more popular. The Council of Europe prescribes a quantitative suspension test with organic load, the European Suspension Test (EST), as the definitive testing method for disinfectants used in food hygiene. It is to be expected that these varied testing techniques would yield different results. The evaluation of a testing technique should be supported by correlation of the results of the tests with efficacy in practice. The recent founding of a technical committee of the European Committee for Standardization (CEN) raises great hope that, at least in Europe, a harmonization of the testing techniques will be possible in the near future.

Disinfectants↗