Effects of biocides on the transduction of Pseudomonas aeruginosa PAO by F116 bacteriophage

The transduction of Pseudomonas aeruginosa PAO by the F116 bacteriophage was monitored after phage treatment with seven biocides commonly used in disinfection processes. Ethanol (70% and 100%), isopropanol (100%) and phenol (2%) did not affect the ability of the phage to transduce. However, chlorhexidine diacetate (1%), cetylpyridinium chloride (0·05%), glutaraldehyde (2%), peracetic acid (1%) and sodium hypochlorite (0·1%) significantly inhibited F116 transduction. The transduction process was more sensitive to biocides than the phage itself. Inactivation of F116 and inhibition of the transduction process appeared to involve two distinct mechanisms implying that the process depends on cellular target sites not affected by some biocides.     

The effects of disinfectant foam on microbial biofilms

This investigation examined the effects of common aqueous biocides and disinfectant foams derived from them on Pseudomonas aeruginosa biofilms. Biofilms were grown on stainless steel coupons under standardised conditions in a reactor supplemented with low concentrations of organic matter to simulate conditions prevalent in industrial systems. Five-day-old biofilms formed under ambient conditions with continuous agitation demonstrated a low coefficient of variation (5.809%) amongst viable biofilm bacteria from independent trials. Scanning electron microscopy revealed biofilms on coupons with viable biofilm bacteria observed by confocal microscopy. An aqueous solution of a common foaming agent amine oxide (AO) produced negligible effects on bacterial viability in biofilms (p>0.05). However, significant biofilm inactivation was noted with aqueous solutions of common biocides (peracetic acid, sodium hypochlorite, sodium ethylenediaminetetraacetic acid) with or without AO (p<0.05). Aereation of a mixture of AO with each of these common biocides resulted in significant reductions in the viability of biofilm bacteria (p<0.05). In contrast, limited effects were noted by foam devoid of biocides. A relationship between microbial inactivation and the concentration of biocide in foam (ranging from 0.1-0.5%) and exposure period were noted (p<0.05). Although, lower numbers of viable biofilm bacteria were recovered after treatment with the disinfectant foam than by the cognate aqueous biocide, significant differences between these treatments were not evident (p>0.05). In summary, the studies revealed significant biofilm inactivation by biocidal foam prepared with common biocides. Validation of foam disinfectants in controlled trials at manufacturing sites may facilitate developments for clean in place applications. Advantages of foam disinfectants include reductions in the volumes of biocides for industrial disinfection and in their disposal after use.     

The effects of disinfectant foam on microbial biofilms

This investigation examined the effects of common aqueous biocides and disinfectant foams derived from them on Pseudomonas aeruginosa biofilms. Biofilms were grown on stainless steel coupons under standardised conditions in a reactor supplemented with low concentrations of organic matter to simulate conditions prevalent in industrial systems. Five-day-old biofilms formed under ambient conditions with continuous agitation demonstrated a low coefficient of variation (5.809%) amongst viable biofilm bacteria from independent trials. Scanning electron microscopy revealed biofilms on coupons with viable biofilm bacteria observed by confocal microscopy. An aqueous solution of a common foaming agent amine oxide (AO) produced negligible effects on bacterial viability in biofilms (p?>?0.05). However, significant biofilm inactivation was noted with aqueous solutions of common biocides (peracetic acid, sodium hypochlorite, sodium ethylenediaminetetraacetic acid) with or without AO (p?<?0.05). Aereation of a mixture of AO with each of these common biocides resulted in significant reductions in the viability of biofilm bacteria (p?<?0.05). In contrast, limited effects were noted by foam devoid of biocides. A relationship between microbial inactivation and the concentration of biocide in foam (ranging from 0.1?–?0.5%) and exposure period were noted (p?<?0.05). Although, lower numbers of viable biofilm bacteria were recovered after treatment with the disinfectant foam than by the cognate aqueous biocide, significant differences between these treatments were not evident (p?>?0.05). In summary, the studies revealed significant biofilm inactivation by biocidal foam prepared with common biocides. Validation of foam disinfectants in controlled trials at manufacturing sites may facilitate developments for clean in place applications. Advantages of foam disinfectants include reductions in the volumes of biocides for industrial disinfection and in their disposal after use.     

常用消毒剂次氯酸钠和二氧化氯对小林三代虫的杀灭效果

为了有效控制三代虫病, 实验以寄生于金鱼的小林三代虫(Gyrodactylus kobayashii)为动物模型, 研究了两种常用消毒剂次氯酸钠溶液(NaClO)和二氧化氯(ClO2)的杀虫效果。结果表明: 在离体(in vitro)条件下, 当NaClO的有效浓度0.2 mg/L或ClO2的有效浓度0.15 mg/L 时, 小林三代虫的平均存活时间均少于2h, 而对照组中小林三代虫的平均存活时间是20.8h。当ClO2的有效浓度0.15 mg/L时, 70%以上的虫体发黑, 而其他浓度处理组, 大部分虫体即使死亡, 虫体依然保持透明。在在体(in vivo)条件下, 当 NaClO的有效浓度0.2 mg/L或ClO2的有效浓度0.5 mg/L 时, 驱虫率都几乎达到100%, 并且驱虫率随着药物浓度的增加而提高,但当ClO2的有效浓度为0.6 mg/L时, 养殖水体出现了白色絮状物。在在体条件下, NaClO的驱虫效果好于ClO2。在金鱼的急性毒性实验中, NaClO和ClO2的安全浓度分别是0.18和0.48 mg/L, 仅稍低于其在在体条件下完全驱除小林三代虫的最小浓度(0.2、0.5 mg/L), 说明次氯酸钠溶液和二氧化氯在驱除三代虫时对金鱼不太安全, 因此, 在治疗金鱼的三代虫病时要慎使次氯酸钠溶液和二氧化氯。然而, 这两种消毒剂能否适用于其他鱼类三代虫病的治疗则有待进一步研究。     

Analysis of Cell Wall Constituents of Biocide-Resistant Isolates from Dental-Unit Water Line Biofilms

In past years, the significance of microbial resistance to biocides has increased. Twenty biocide-resistant bacterial strains were isolated from dental-unit water line biofilm. All strains resisted high biocide concentrations (up to 100 mug ml(-)1): sodium dodecyl sulphate, hydrogen peroxide, sodium hypochlorite, phenol, Tween 20, ethylenediaminetetraacetic acid, chlorohexidine gluconate, and povidine iodine. Among bacteria, biocide sensitivity is based on permeability of biocides through the cell wall. Gram-positive bacteria are more permeable and susceptible to biocides, whereas Gram-negative bacteria have a more complex cell wall and are the least sensitive bacteria. The present study was designed to study the effect of biocides on the cell wall of biocide-resistant bacteria. Peptidoglycan (PG), diaminopimelic acid (DAP), and teichoic acid contents of the cell wall were determined in L-broth and L-broth supplemented with biocides at different temperatures (37 degrees C and 45 degrees C) and pH levels (7 and 9). In general and Gram staining-specific comparison, a significant increase (p < 0.05) in the DAP content of biocide-resistant bacteria was observed at pH 7 and at both temperatures. In tubing-specific comparison, a significant increase in the amount of teichoic acid in air water tubing (37 degrees C at pH 9) and DAP in patient tubing (pH 7 at both temperatures) was observed. In main water pipe, a significant decrease (p > 0.05) in PG content was noticed at 45 degrees C and pH 9. Overall, a significant increase in DAP content may be an important constituent in the manifestation of isolate resistance against various biocides.     

Effects of the peracetic acid and sodium hypochlorite on the colour stability and surface roughness of the denture base acrylic resins polymerised by microwave and water bath methods

Objective: This study evaluated the surface roughness (Ra) and color stability of acrylic resin colors (Lucitone 550, QC‐20 and Vipi‐Wave) used for fabricating bases for complete, removable dentures, overdentures and prosthetic protocol after immersion in chemical disinfectants (1% sodium hypochlorite and 2% peracetic acid) for 30 and 60 minutes. Material and Methods: Sixty specimens were made of each commercial brand of resin composite, and divided into 2 groups according to the chemical disinfectants. Specimens had undergone the finishing and polishing procedures, the initial color and roughness measurements were taken (t=0), and after this, ten test specimens of each commercial brand of resin composite were immersed in sodium hypochlorite and ten in peracetic acid, for 30 and 60 minutes, with measurements being taken after each immersion period. These data were submitted to statistical analysis. Results: There was evidence of an increase in Ra after 30 minutes immersion in the disinfectants in all the resins, with QC‐20 presenting the highest Ra values, and Vipi‐Wave the lowest. After 60 minutes immersion in the disinfectants all the resins presented statistically significant color alteration. Conclusions: Disinfection with 1% sodium hypochlorite and peracetic acid altered the properties of roughness and color of the resins.     

Impact of biocides on biofilm formation by methicillin-resistant Staphylococcus aureus (ST239-SCCmecIII) isolates

Procedures of sterilization and disinfection are essential to ensure that medical and surgical instruments will not transmit infectious pathogens to patients. In the present paper, we tested the residual effect of these compounds on biofilm formation and its efficiency in disrupting preformed biofilms using methicillin-resistant Staphylococcus aureus (MRSA) isolates of the lineage ST239-SCCmecIII. All compounds examined, except 70% alcohol, caused a significant impairment in biofilm formation with concomitant inhibition of cell growth. Among the compounds examined, 10% povidone-iodine (PVP-I) was the only antiseptic that exhibited more than 90% reduction of both biofilm formation and dispersion. In the group of sterilants and disinfectants, a formulation containing 7% hydrogen peroxide and 0.2% peracetic acid (HP-PA), and sodium hypochlorite with 1% active chlorine (NaOCl) were equally effective.     

Fungicidal effects of chemical disinfectants, UV light, desiccation and heat on the amphibian chytrid Batrachochytrium dendrobatidis

The efficacy of a number of disinfection treatments was tested on in vitro cultures of the fungus Batrachochytrium dendrobatidis, the causative agent of chytridiomycosis in amphibians. The aim was to evaluate the fungicidal effects of chemical disinfectants, sterilising ultraviolet (UV) light, heat and desiccation, using methods that were feasible for either disinfection in the field, in amphibian husbandry or in the laboratory. The chemical disinfectants tested were: sodium chloride, household bleach (active ingredient: sodium hypochlorite), potassium permanganate, formaldehyde solution, Path-X agricultural disinfectant (active ingredient: didecyl dimethyl ammonium chloride, DDAC), quaternary ammonium compound 128 (DDAC), Dithane, Virkon, ethanol and benzalkonium chloride. In 2 series of experiments using separate isolates of B. dendrobatidis, the fungicidal effect was evaluated for various time periods and at a range of chemical concentrations. The end point measured was death of 100% of zoospores and zoosporangia. Nearly all chemical disinfectants resulted in 100%, mortality for at least one of the concentrations tested. However, concentration and time of exposure was critical for most chemicals. Exposure to 70% ethanol, 1 mg Virkon ml(-1) or 1 mg benzalkonium chloride ml(-1) resulted in death of all zoosporangia after 20 s. The most effective products for field use were Path-X and the quaternary ammonium compound 128, which can be used at dilutions containing low levels (e.g. 0.012 or 0.008%, respectively) of the active compound didecyl dimethyl ammonium chloride. Bleach, containing the active ingredient sodium hypochlorite, was effective at concentrations of 1% sodium hypochlorite and above. Cultures did not survive complete drying, which occurred after <3 h at room temperature. B. dendrobatidis was sensitive to heating, and within 4 h at 37 degrees C, 30 min at 47 degrees C and 5 min at 60 degrees C, 100% mortality occurred. UV light (at 1000 mW m(-2) with a wavelength of 254 nm) was ineffective at killing B. dendrobatidis in culture.     

A conductance-based surface disinfection test for food hygiene

A surface disinfectant test method for food hygiene is described in which biocidal action was assessed using bacterial biofilms developed on stainless steel. The viability of control and surface-bound treated biofilms was assessed, using a Malthus microbiological growth analyser. The surface test was compared with the European Suspension Test on three bacteria, Pseudomonas aeruginosa, Staphylococcus aureus and Proteus mirabilis , for 12 commonly used food surface disinfectants. In all cases the bacteria were shown to be up to 10 times more resistant to biocides, and in seven cases up to 100 times more resistant, when surface-attached. Disinfectants that performed well against bacteria in suspension did not necessarily perform well against the same bacteria when attached to surfaces.     

Nitrogen removal via nitrite in a sequencing batch reactor treating sanitary landfill leachate

The present paper reports the results of the application of a control system, based on artificial intelligence concepts, for the automation of a bench-scale SBR treating leachate generated in old landfills. Attention was given to the nitritation and denitritation processes in order to enhance the nitrogen removal efficiency. Nitrification and nitrogen removal were usually higher than 98% and 95%, respectively, whereas COD removal was approximately 20-30% due to the low biodegradability of organic matter in the leachate from old landfills; therefore, external COD was added to accomplish the denitrification process. Adjusting the length of the oxic phase, almost complete inhibition of the nitrite oxidizing organisms was observed. The results confirm the effectiveness of the nitrite route for nitrogen removal optimisation in leachate treatment. A significant saving of approximately 35% in external COD addition was achieved.     

Wide variation in effectiveness of laboratory disinfectants against bacteriophages

Aims: The purpose of this study was to identify an effective disinfectant for the inactivation of the bacteriophages (phages) being used in our laboratory, as published studies on phage inactivation are far from unanimous in their conclusions. Methods and Results: The phages studied were three closely related strains of Myoviridae and three strains of Siphoviridae. Three disinfectants which are used commonly in microbiology laboratories were evaluated: Virkon (1%), ethanol (75%) and sodium hypochlorite (2500 ppm available chlorine). The most effective of these was Virkon, which inactivated all six phages rapidly. Ethanol was effective against the Myoviridae but had little effect on the Siphoviridae. Sodium hypochlorite was the least effective of the disinfectants evaluated. Conclusions: The findings of this study demonstrate a wide diversity in the effectiveness of disinfectants tested for inactivation of phages. Significance and Impact of the Study: Of the disinfectants tested Virkon is the most suitable choice for those unable to carry out disinfection validation studies, or where a broad spectrum disinfectant against phages is required. All of the phages in this study showed resilience to inactivation by sodium hypochlorite, and therefore this disinfectant is an unwise choice for use against phage without first assessing its effectiveness.     

Process monitoring the inactivation of ricin and model proteins by disinfectants using fluorescence and biological activity

It is important to develop rapid and reliable processes to monitor the decontamination of toxins released to the environment. The inactivation of the protein toxin ricin by the disinfectants bleach (sodium hypochlorite) and monochloramine was measured by the effect on mammalian cell cytotoxicity. The effect of the disinfectants on the native fluorescence (due mainly to tryptophan and to a lesser extent tyrosine) of ricin was also measured in parallel. Reactions of the disinfectants resulted in a decrease in the native fluorescence that was measured in real time in a noninvasive manner. We compared the inactivation of two well-characterized model enzymes to the behavior of ricin. The model enzymes studied were lysozyme, a small basic enzyme stabilized with internal disulfide bonds, and heart-muscle-type lactate dehydrogenase (LDH), a large protein composed of four subunits. The biological activities of the model enzymes were measured in parallel with their fluorescence. Gel electrophoresis showed a large number of modifications of the proteins caused by the disinfectants reflected in changes in mobility and the formation of higher-order aggregates. Size-exclusion chromatography showed that the disinfectants did not break down the subunit structure of ricin but instead resulted in an increased size and heterogeneity of the protein. Size-exclusion chromatography of LDH indicated that the subunits were dissociated and that higher-order aggregates were also formed. Bleach caused a rapid inactivation of biological activity correlated with a rapid decrease in the fluorescence. Monochloramine required much higher concentrations for significant effects and the kinetics of the reactions were slow, with half-life values of the decrease on the order of minutes. Each protein showed individual differences in responses to the disinfectants, but there was a consistent correlation between the loss of fluorescence and the decrease in biological activity. These results indicate that the monitoring the fluorescence is a useful process with limitations that can be used to monitor the inactivation of toxins using disinfectants.     

Nitrogen removal performance of anaerobic ammonia oxidation (ANAMMOX) in presence of organic matter

A sequencing batch reactor (SBR) was used to test the nitrogen removal performance of anaerobic ammonium oxidation (ANAMMOX) in presence of organic matter. Mesophilic operation (30 ± 0.5 °C) was performed with influent pH 7.5. The results showed, independent of organic matter species, ANAMMOX reaction was promoted when COD was lower than 80 mg/L. However, specific ANAMMOX activity decreased with increasing organic matter content. Ammonium removal efficiency decreased to 80% when COD of sodium succinate, sodium potassium tartrate, peptone and lactose were 192.5, 210, 225 and 325 mg/L, respectively. The stoichiometry ratio resulting from different OM differed largely and R₁ could be as an indicator for OM inhibition. When COD concentration was 240 mg/L, the loss of SAA resulting from lactose, peptone, sodium potassium tartrate and sodium succinate were 28, 36, 50 and 55%, respectively. Sodium succinate had the highest inhibitory effect on SAA. When ANAMMOX process was used to treat wastewater containing OM, the modified Logistic model could be employed to predict the NRE_max.     

低温条件下常用环境消毒剂对新冠病毒灭活效果的CFD模拟和分析

新冠肺炎疫情以来,冷链食品外包装上的新冠病毒频繁引起局部疫情,对现有的环境消杀体系带来了巨大挑战。为了探究低温环境消毒剂灭活效果下降的内在机制,并为冷链过程中合理选用消毒剂提供依据,对低温条件下消毒剂的灭活过程进行计算流体力学(CFD)建模。以环境消杀常用的75%乙醇溶液和次氯酸钠溶液为工作体系,探讨了不同温度下两种溶液对新冠病毒灭活特性的影响规律。结果表明,75%乙醇溶液的灭活特性在低温环境下受温度的影响较小,而次氯酸钠溶液在环境温度为253.15 K时会发生凝固现象使得其灭活效率显著降低。该模型的模拟结果与实验结果趋势一致,可为冷链运输中疫情防控提供参考。     

Contact inactivation of orthopoxviruses by household disinfectants

AIMS: The aim of this study is to identify common household disinfectants that combine significant activity against the type orthopoxvirus, vaccinia virus with minimal impact in terms of potential toxicity and/or damage to household or personal items. METHODS AND RESULTS: Laboratory scale experiments assessed common disinfectants containing anionic and nonionic detergents, oxygen-based bleach, potassium peroxomonosulfate, chloroxylenol or halogenated phenols. Disinfectants were assessed for their ability to inactivate the virus on contact or after a short incubation period in the presence and absence of foetal bovine serum as a potential interferant. Significant differences were observed ranging from negligible effect of detergents to complete inactivation on contact with chloroxylenol. CONCLUSIONS: At least one chloroxylenol-based household disinfectant is available, which inactivates vaccinia virus on contact. SIGNIFICANCE AND IMPACT OF THE STUDY: In the event of a release or major outbreak of a pathogenic orthopoxvirus there is likely to be significant public demand for disinfectants with activity against these viruses. The identification of common household disinfectants with such activity obviates any requirement to stockpile or distribute laboratory/industrial disinfectants for this purpose.     

The effect of small amounts of sodium hypochlorite on the growth of Earl's "L" cells in suspension growth

Of several candidate disinfectants for use in tissue culture work, especially suspension cultures, sodium hypochlorite solution was selected to test its effect on growing cells. Metabolizing cells reduce, sodium hypochlorite oxidizes ; therefore NaOCl leakage into such systems must be neutralized with no untoward effects on the cells. Dilutions of routine disinfectant-grade sodium hypochlorite were tested against cell cultures. Those exposed to 15.62 to 31.25 ppm of NaOCl grew with no apparent cell damage.     

Comparative susceptibility of Salmonella Typhimurium biofilms of different ages to disinfectants

There is a general consensus that with increasing age a biofilm shows increased resistance to antimicrobials. In this study the susceptibility of 3-, 5- and 7-day-old Salmonella enterica serovar Typhimurium biofilms to disinfectants was evaluated. It was hypothesized that 7-day-old biofilms would be more resistant to disinfectants compared to 3- and 5-day-old biofilms. Biofilms were formed using the MBEC? system and treated with six chemical disinfectants for 1 and 5 min. Four disinfectants at the highest concentration available showed 100% reduction in viable cells from all ages of biofilms after exposure for 5 min, and ethanol at 70% v/v was the least effective against biofilms, followed by chlorhexidine gluconate (CG). At the recommended user concentrations, only sodium hypochlorite showed 100% reduction in viable cells from all ages of biofilms. Benzalkonium chloride and CG were the least effective against biofilms, followed by quaternary ammonium compound which only showed 100% reduction in viable cells from 5-day-old biofilms. Overall, the results from this study do not display enhanced resistance in 7-day-old biofilms compared to 3- and 5-day-old biofilms. It is concluded that under the conditions of this study, the age of biofilm did not contribute to resistance towards disinfectants. Rather, the concentration of disinfectant and an increased contact time were both shown to play a role in successful sanitization.     

Comparative mycobactericidal efficacy of chemical disinfectants in suspension and carrier tests

The efficacy of nine disinfectants on Mycobacterium smegmatis was tested in the presence of sputum, using quantitative suspension and carrier tests. Glutaraldehyde, povidone iodine, and chlorhexidine gluconate produced at least a 6-log10 reduction in CFU in all tests. Four disinfectants (sodium dichloroisocyanurate, phenol, ethanol, and sodium hypochlorite) were not as effective in the carrier tests as in the suspension tests; this difference ranged from a 1- to a 5-log10 reduction in CFU. The efficacy of ethanol and sodium hypochlorite was further reduced (3- and 1-log10 reductions in CFU, respectively) in the presence of sputum. The quaternary ammonium compound and iodophor were ineffective in all tests. The findings of this study demonstrate the need for a quantitative carrier test such as the one presented here.     

Comparative mycobactericidal efficacy of chemical disinfectants in suspension and carrier tests.

The efficacy of nine disinfectants on Mycobacterium smegmatis was tested in the presence of sputum, using quantitative suspension and carrier tests. Glutaraldehyde, povidone iodine, and chlorhexidine gluconate produced at least a 6-log10 reduction in CFU in all tests. Four disinfectants (sodium dichloroisocyanurate, phenol, ethanol, and sodium hypochlorite) were not as effective in the carrier tests as in the suspension tests; this difference ranged from a 1- to a 5-log10 reduction in CFU. The efficacy of ethanol and sodium hypochlorite was further reduced (3- and 1-log10 reductions in CFU, respectively) in the presence of sputum. The quaternary ammonium compound and iodophor were ineffective in all tests. The findings of this study demonstrate the need for a quantitative carrier test such as the one presented here.