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Influence of possible disinfectant transfer on Staphylococcus aureus plate counts after agar contact sampling.

Asbestos-vinyl tile floor panels were mopped with three types of chemical disinfectant product, as well as after contact, with the untreated control panel were prepared according to the manufacturer's label instructions. Similar floor panels were inoculated artificially with Staphylococcus aureus. RODAC (replicate organism detection and counting) surface sampling plates were pressed to the disinfectant-treated panels or to the untreated control panel and then immediately pressed to sampling sites on the artificially inoculated floor panels. Plate counts were determined after contact with panels treated with each type of disinfectant, product, as well as after contact with the untreated control panel. Results indicate that disinfectant residues on environmental surfaces do not alter the average plate counts obtained by RODAC samplings.

Agar↗

Effect of methodology, dilution, and exposure time on the tuberculocidal activity of glutaraldehyde-based disinfectants.

The Association of Official Analytical Chemists (AOAC) test for assessing the tuberculocidal activity of disinfectants has been shown to be variable. A modified AOAC test, which substituted Middlebrook 7H9 broth as the primary subculture medium and used neutralization by dilution, was compared with the standard AOAC method to assess the mycobactericidal activity of three glutaraldehyde-based disinfectants at 20 degrees C and various exposure times. These changes had a marked effect on results, with the modified AOAC test providing more positive penicylinders per 10 replicates in 12 of the 13 comparisons that provided positive results. These differences were observed with both Mycobacterium bovis (ATCC 35743) and a clinical isolate of Mycobacterium tuberculosis. The effects of various exposure times to and dilutions of the glutaraldehyde-based disinfectants were also examined. The minimum exposure time needed to inactivate reliably M. bovis or M. tuberculosis with 2% glutaraldehyde was 20 min at 20 degrees C. Diluting 2% glutaraldehyde caused a significant decline in mycobactericidal activity. Modification of the standard AOAC test to improve its sensitivity in detecting the failure of disinfectants to inactivate mycobacteria is indicated.

Aldehydes↗

Bacterial spores survive treatment with commercial sterilants and disinfectants.

This study compared the activity of commercial liquid sterilants and disinfectants on Bacillus subtilis spores deposited on three types of devices made of noncorrodible, corrodible, or polymeric material. Products like Renalin, Exspor, Wavicide-01, Cidexplus, and cupric ascorbate were tested under conditions specified for liquid sterilization. These products, at the shorter times indicated for disinfection, and popular disinfectants, like Clorox, Cavicide, and Lysol were also studied. Data obtained with a sensitive and quantitative test suggest that commercial liquid sterilants and disinfectants are less effective on contaminated surfaces than generally acknowledged.

Bacillus subtilis↗

A spore counting method and cell culture model for chlorine disinfection studies of Encephalitozoon syn. Septata intestinalis.

The microsporidia have recently been recognized as a group of pathogens that have potential for waterborne transmission; however, little is known about the effects of routine disinfection on microsporidian spore viability. In this study, in vitro growth of Encephalitozoon syn. Septata intestinalis, a microsporidium found in the human gut, was used as a model to assess the effect of chlorine on the infectivity and viability of microsporidian spores. Spore inoculum concentrations were determined by using spectrophotometric measurements (percent transmittance at 625 nm) and by traditional hemacytometer counting. To determine quantitative dose-response data for spore infectivity, we optimized a rabbit kidney cell culture system in 24-well plates, which facilitated calculation of a 50% tissue culture infective dose (TCID(50)) and a minimal infective dose (MID) for E. intestinalis. The TCID(50) is a quantitative measure of infectivity and growth and is the number of organisms that must be present to infect 50% of the cell culture wells tested. The MID is as a measure of a system's permissiveness to infection and a measure of spore infectivity. A standardized MID and a standardized TCID(50) have not been reported previously for any microsporidian species. Both types of doses are reported in this paper, and the values were used to evaluate the effects of chlorine disinfection on the in vitro growth of microsporidia. Spores were treated with chlorine at concentrations of 0, 1, 2, 5, and 10 mg/liter. The exposure times ranged from 0 to 80 min at 25 degrees C and pH 7. MID data for E. intestinalis were compared before and after chlorine disinfection. A 3-log reduction (99.9% inhibition) in the E. intestinalis MID was observed at a chlorine concentration of 2 mg/liter after a minimum exposure time of 16 min. The log(10) reduction results based on percent transmittance-derived spore counts were equivalent to the results based on hemacytometer-derived spore counts. Our data suggest that chlorine treatment may be an effective water treatment for E. intestinalis and that spectrophotometric methods may be substituted for labor-intensive hemacytometer methods when spores are counted in laboratory-based chlorine disinfection studies.

Animals↗

Chlorine disinfection of atypical mycobacteria isolated from a water distribution system.

We studied the resistance of various mycobacteria isolated from a water distribution system to chlorine. Chlorine disinfection efficiency is expressed as the coefficient of lethality (liters per minute per milligram) as follows: Mycobacterium fortuitum (0.02) > M. chelonae (0.03) > M. gordonae (0.09) > M. aurum (0.19). For a C.t value (product of the disinfectant concentration and contact time) of 60 mg.min.liter(-1), frequently used in water treatment lines, chlorine disinfection inactivates over 4 log units of M. gordonae and 1.5 log units of M. fortuitum or M. chelonae. C.t values determined under similar conditions show that even the most susceptible species, M. aurum and M. gordonae, are 100 and 330 times more resistant to chlorine than Escherichia coli. We also investigated the effects of different parameters (medium, pH, and temperature) on chlorine disinfection in a chlorine-resistant M. gordonae model. Our experimental results follow the Arrhenius equation, allowing the inactivation rate to be predicted at different temperatures. Our results show that M. gordonae is more resistant to chlorine in low-nutrient media, such as those encountered in water, and that an increase in temperature (from 4 degrees C to 25 degrees C) and a decrease in pH result in better inactivation.

Chlorine↗

Cell culture-Taqman PCR assay for evaluation of Cryptosporidium parvum disinfection.

Cryptosporidium parvum represents a challenge to the water industry and a threat to public health. In this study, we developed a cell culture-quantitative PCR assay to evaluate the inactivation of C. parvum with disinfectants. The assay was validated by using a range of disinfectants in common use in the water industry, including low-pressure UV light (LP-UV), ozone, mixed oxidants (MIOX), and chlorine. The assay was demonstrated to be reliable and sensitive, with a lower detection limit of a single infectious oocyst. Effective oocyst inactivation was achieved (>2 log(10) units) with LP-UV (20 mJ/cm(2)) or 2 mg of ozone/liter (for 10 min). MIOX and chlorine treatments of oocysts resulted in minimal effective disinfection, with <0.1 log(10) unit being inactivated. These results demonstrate the inability of MIOX to inactivate Cryptosporidium. The assay is a valuable tool for the evaluation of disinfection systems for drinking water and recycled water.

Animals↗

Reduction of Norwalk virus, poliovirus 1, and bacteriophage MS2 by ozone disinfection of water.

Norwalk virus and other human caliciviruses (noroviruses) are major agents of gastroenteritis, and water is a major route of their transmission. In an effort to control Norwalk virus in drinking water, Norwalk virus reduction by bench-scale ozone disinfection was determined using quantitative reverse transcription (RT)-PCR for virus assays. Two other enteric viruses, poliovirus 1 and coliphage MS2, were included for comparison, and their reductions were assayed by infectivity assays as well as by RT-PCR. Virus reductions by ozone were determined using a dose of 0.37 mg of ozone/liter at pH 7 and 5 degrees C for up to 5 min. Based on two RT-PCR assays, the reductions of Norwalk virus were >3 log(10) within a contact time of 10 s, and these were similar to the reductions of the other two viruses determined by the same assay methods. Also, the virus reductions detected by RT-PCR assays were similar to those detected by infectivity assays, indicating that the RT-PCR assay is a reliable surrogate assay for both culturable and nonculturable viruses disinfected with ozone. Overall, the results of this study indicate that Norwalk virus as well as other enteric viruses can be reduced rapidly and extensively by ozone disinfection and that RT-PCR is a useful surrogate assay for both culturable and nonculturable viruses disinfected with ozone.

Cell Line↗

EVALUATION OF DISINFECTANTS FOR HOSPITAL HOUSEKEEPING USE.

A use-dilution procedure and screening method are proposed to aid the hospital bacteriologist in selecting the most effective disinfectants. Sixteen disinfectants were tested with and without organic material on six different organisms. Sporadic results usually obtained with quaternaries tested by the procedure of the Association of Official Agricultural Chemists (AOAC) were eliminated by the methods of this study. The use-dilution procedure employs the material upon which the disinfectant is to be used, rather than stainless steel, as in the AOAC use-dilution test. The importance of testing each disinfectant against several organisms and in the presence of organic material is discussed.

Anti-Infective Agents↗

Inactivation of vesicular stomatitis virus by disinfectants.

Twenty-four chemical disinfectants considered to be viricidal were tested. Ten disinfectants were not viricidal for vesicular stomatitis virus within 10 min at 20 C when an LD(50) titer of 10(8.5) virus units per 0.1 ml were to be inactivated. Quantitative inactivation experiments were done with acid, alkaline, and a substituted phenolic disinfectant to determine the kinetics of the virus inactivation. Substituted phenolic disinfectants, halogens, and cresylic and hydrochloric acids were viricidal. Basic compounds such as lye and sodium metasilicate were not viricidal.

Animals↗

Factors affecting comparative resistance of naturally occurring and subcultured Pseudomonas aeruginosa to disinfectants.

A strain of Pseudomonas aeruginosa was isolated in pure culture from the reservoir of a hospital mist therapy unit by an extinction-dilution technique; its natural distilled water environment was used as a growth and maintenance medium. After a single subculture on Trypticase soy agar, the strain showed a marked decrease in resistance to inactivation by acetic acid, glutaraldehyde, chlorine dioxide, and a quaternary ammonium compound when compared with naturally occurring cells grown in mist therapy unit water. The following factors were observed to affect the relative resistances of naturally occurring and subcultured cells of the P. aeruginosa strain: (i) temperature at which the cultures were incubated prior to exposure to disinfectants, (ii) growth phase of the cultures at the time of exposure to disinfectants, (iii) nature of the suspending menstruum for disinfectants, and (iv) exposure to fluorescent light during incubation of inocula prior to testing. The applied significance of these findings may alter the present concepts of disinfectant testing as well as routine control procedures in the hospital environment.

Acetates↗

Inactivation of hepatitis B virus by intermediate-to-high-level disinfectant chemicals.

In five separate tests, hepatitis B virus in dried human plasma was exposed for 10 min at 20 degrees C to disinfectant chemicals having activity levels ranging from intermediate (e.g., 70% isopropyl alcohol) to high (e.g., 2% aqueous glutaraldehyde at pH 8.6). Five chimpanzees (one animal per disinfectant chemical) received treated material intravenously, and none showed signs of infection after post-inoculation periods of 9 months. Two animals were rechallenged with inoculum treated in the same manner, except that saline was used instead of a disinfectant chemical; both were infected within 4 weeks. Our results showed that hepatitis B virus was not as resistant to disinfectant chemicals as once thought and suggested that chemicals with similar activity levels (intermediate to high) might possibly be used on hepatitis B virus contamination with a margin of safety.

Animals↗

In-use testing of disinfectants in hospitals.

One hundred and five samples of clear, soluble phenolic disinfectants were obtained from varying sites in the wards of six hospitals. The concentration of disinfectant in each sample was measured by a colorimetric method and bacterial contamination was measured by an ;in-use' test and a membrane filter technique. The concentrations of disinfectant in 24/105 (23%) samples were at the recommended level and 53 (50.5%) were below. Bacterial contamination with Gram-negative bacilli was found in 26/49 (53%) samples containing less than 0.8% of disinfectant and 5/86 (8.9%) samples containing more than 0.8%. The concentrations in two of the heavily contaminated samples were 1.5% and 1.6% respectively.

Bacteria↗

Influence of phosphate and disinfection on the composition of biofilms produced from drinking water, as measured by fluorescence in situ hybridization.

Biofilms were grown in annular reactors supplied with drinking water enriched with 235 microg C/L. Changes in the biofilms with ageing, disinfection, and phosphate treatment were monitored using fluorescence in situ hybridization. EUB338, BET42a, GAM42a, and ALF1b probes were used to target most bacteria and the alpha (alpha), beta (beta), and gamma (gamma) subclasses of Proteobacteria, respectively. The stability of biofilm composition was checked after the onset of colonization between T = 42 days and T = 113 days. From 56.0% to 75.9% of the cells detected through total direct counts with DAPI (4'-6-diamidino-2-phenylindole) were also detected with the EUB338 probe, which targets the 16S rRNA of most bacteria. Among these cells, 16.9%-24.7% were targeted with the BET42a probe, 1.8%-18.3% with the ALF1b probe, and <2.5% with the GAM42a probe. Phosphate treatment induced a significant enhancement to the proportion of gamma-Proteobacteria (detected with the GAM42a probe), a group that contains many health-related bacteria. Disinfection with monochloramine for 1 month or chlorine for 3 days induced a reduction in the percentage of DAPI-stained cells that hybridized with the EUB338 probe (as expressed by percentages of EUB338 counts/DAPI) and with any of the ALF1b, BET42a, and GAM42a probes. The percentage of cells detected by any of the three probes (ALF1b+BET42a+GAM42a) tended to decrease, and reached in total less than 30% of the EUB338-hybridized cells. Disinfection with chlorine for 7 days induced a reverse shift; an increase in the percentage of EUB338 counts targeted by any of these three probes was noted, which reached up to 87%. However, it should be noted that the global bacterial densities (heterotrophic plate counts and total direct counts) tended to decrease over the duration of the experiment. Therefore, those bacteria that could be considered to resist 7 days of chlorination constituted a small part of the initial biofilm community, up to the point at which the other bacterial groups were destroyed by chlorination. The results suggest that there were variations in the kinetics of inactivation by disinfectant, depending on the bacterial populations involved.

Alphaproteobacteria↗

[Inactivation of mycobacteria by disinfectants with a tuberculocidal label].

INTRODUCTION: Disinfectants available in Venezuela, registered as tuberculocidal, and used in clinical and laboratory settings were tested for their mycobactericidal activity. METHODS: The European Standard Quantitative Suspension test prEN14348 was used in simulated clean and dirty conditions. The disinfectants tested included Gerdex and K-ller (both containing the quaternary ammonium compound dimethyl benzyl lauryl ammonium bromide in concentrations of 10% and 0.16% respectively), Microplus-Action (5% polymethylene urea), Cidex (2% glutaraldehyde) and Cidex OPA (0.55% ortho-phthalaldehyde). The mycobactericidal activity was evaluated with M. smegmatis ATCC 19420 and in clinical isolates of M. fortuitum, M. chelonae and M. abscessus. The tuberculocidal activity was assessed with M. tuberculosis H37Rv. RESULTS: Cidex and Cidex OPA were effective against all the test organisms, showing a > 5-log 10 reduction in CFU after 5 minutes of exposure under both clean and dirty conditions. Gerdex and K-ller only achieved a 2-log reduction in M. tuberculosis, M. abscessus and M. chelonae CFU, even after 60 minutes of exposure. Micro Plus Action showed a 4-log reduction in M. tuberculosis viability but less than a 1-log reduction in M. abscessus and M. chelonae CFUs after 60 minutes of incubation. CONCLUSION: The tuberculocidal activity claimed by the manufacturers of several disinfectants sold in Venezuela, Gerdex, K-ller and Microplus-Action is inaccurate when tested by European norms. The use of these products for disinfecting material used in critical and semi-critical patients could result in iatrogenic infections.

Antitubercular Agents↗

Antiviral efficacy of disinfectant solution MRI-1.

Disinfectant MRI-1 was prepared by dissolution of non-ionic and ionic detergent in ethanol. The disinfectant inactivated extracellular and intracellular enveloped and non-enveloped viruses including herpes viruses, influenza A and human immunodeficiency disease virus in suspension or on surfaces by pre-exposure or post-exposure to the disinfectant; in addition, cells were disabled as potential hosts for viral infection using concentrations of MRI-1 which were 50-fold less than the operative concentration for disinfection. There was no evidence of in vitro mutagenicity using Salmonella typhimurium or sensitization or other adverse effect in a guinea pig model or in human subjects.

Animals↗

The disinfection of flexible fibre-optic nasendoscopes out-of-hours: confidential telephone survey of ENT units in England.

We present the results of a confidential telephone survey of ENT units in England on the disinfection of flexible fibre-optic nasendoscopes out-of-hours. The on-call residents of 124 units were contacted and questioned. In 35.1 per cent of units surveyed, the on-call resident was primarily responsible for cleaning the scopes after use. Only 46 per cent of these junior doctors had access to a chemical sterilant to allow for high-level disinfection of these scopes. Provision for disinfection of scopes was poorer in teaching hospitals and in units that served inner city populations. Only 12.1 per cent of Senior House Officers (SHOs) received any training in disinfection techniques and only 25.5 per cent of units kept a register of patients nasendoscoped out-of-hours for purposes of contact tracing.

Disinfection↗

Development of a research strategy for integrated technology-based toxicological and chemical evaluation of complex mixtures of drinking water disinfection byproducts.

Chemical disinfection of water is a major public health triumph of the 20th century. Dramatic decreases in both morbidity and mortality of waterborne diseases are a direct result of water disinfection. With these important public health benefits comes low-level, chronic exposure to a very large number of disinfection byproducts (DBPs), chemicals formed through reaction of the chemical disinfectant with naturally occurring inorganic and organic material in the source water. This article provides an overview of joint research planning by scientists residing within the various organizations of the U.S. Environmental Protection Agency Office of Research and Development. The purpose is to address concerns related to potential health effects from exposure to DBPs that cannot be addressed directly from toxicological studies of individual DBPs or simple DBP mixtures. Two factors motivate the need for such an investigation of complex mixtures of DBPs: a) a significant amount of the material that makes up the total organic halide and total organic carbon portions of the DBPs has not been identified; and b) epidemiologic data, although not conclusive, are suggestive of potential developmental, reproductive, or carcinogenic health effects in humans exposed to DBPs. The plan is being developed and the experiments necessary to determine the feasibility of its implementation are being conducted by scientists from the National Health and Environmental Effects Research Laboratory, the National Risk Management Research Laboratory, the National Exposure Research Laboratory, and the National Center for Environmental Assessment.

Chemistry Techniques, Analytical↗

Health effects among newborns after prenatal exposure to ClO2-disinfected drinking water.

Because chlorination of potable surface waters may be associated with increased risk of carcinogenicity, substitute methods for the routine disinfection of public water supplies are being explored. As part of this search, it is especially important that the potential health effects of each alternative method should be considered. Chemical treatment of drinking water by chlorine dioxide (ClO2) is a likely alternative mode of disinfection. Two common by-products of the ClO2 disinfection of surface water are chlorate and chlorite. These oxidants may have negative health effects on certain high risk groups. Newborns, in particular, would seem to be at increased risk to red cell damage from oxidant stress. The historical record study being reported here compares the morbidity and mortality experience of newborns in two similar communities, one of which used chlorination and the other which used high levels of chlorine dioxide for potable water disinfection. A statistically significant positive association was found between exposure of the mother to ClO2-treated water during pregnancy and prematurity of the newborn as assessed by the attending physician and by a greater weight loss after birth. The rates of jaundice, birth defects and fetal and neonatal mortality did not differ significantly between communities. Because of the limitations of the study design, the findings reported here should be considered suggestive rather than definitive.

Adolescent↗