Multiple Disinfectants Research Articles

Identification and Quantification of Aqueous Disinfectants Using an Array of Carbon Nanotube-Based Chemiresistors.

Disinfection of water is essential to prevent the growth of pathogens, but at high levels, it can cause harm to human health. Therefore, accurate monitoring of disinfectant concentrations in water is essential to ensure safe drinking water. The use of multiple disinfectants at different stages in water treatment plants makes it necessary to also identify the type and concentrations of all of the disinfectant species present. Here, we demonstrate an effective approach to identify and quantify multiple disinfectants (using the example of free chlorine and potassium permanganate) in water using single-walled carbon nanotube (SWCNT)-based reagent-free chemiresistive sensing arrays. Facile fabrication of chemiresistive devices makes them a popular choice for the implementation of sensor arrays. Our sensing array consists of functionalized and unfunctionalized (blank) SWCNT sensors to distinguish the disinfectants. The distinct responses from the different sensors at varying concentrations and pH can be fitted to the mathematical model of a Langmuir adsorption isotherm separately for each sensor. Blank and functionalized sensors respond through different mechanisms that result in varying responses that are concentration- and pH-dependent. Chemometric techniques such as principal component analysis (PCA) and partial least-squares-discriminant analysis (PLS-DA) were used to analyze the sensor data. PCA showed an excellent separation of the analytes over five different pHs (5.5, 6.5, 7.5, 8.5, and 9.5). PLS-DA provided excellent separability as well as good predictability with a Q2 of 94.26% and an R2 of 95.67% for the five pH regions of the two analytes. This proof-of-concept solid-state chemiresistive sensing array can be developed for specific disinfectants that are commonly used in water treatment plants and can be deployed in water distribution and monitoring facilities. We have demonstrated the applicability of chemiresistive devices in a sensor array format for the first time for aqueous disinfectant monitoring.

Single-walled Carbon Nanotube Chemiresistive Sensors for the Identification and Quantification of Disinfectants

Disinfectants are essential to keep water safe as they kill pathogens by oxidizing the cell membrane. Free chlorine, potassium permanganate, monochloramine, hydrogen peroxide, chlorine dioxide, ozone, and hypobromous acid are the most commonly used disinfectants for the disinfection of water.1 Monitoring the concentration of the disinfectant is crucial as the effectiveness mainly depends on the amount of disinfectants present in water.2 Insufficient levels of disinfectant in drinking water could impose health risks as pathogens could regrow in the distribution system. Standard methods for measuring the disinfectant levels are colorimetric and titrimetric.3 For free chlorine, DPD colorimetric method is well established.1 , 4 For potassium permanganate (KMnO4), colorimetric methods (persulfate and periodate) and spectroscopic methods are available.1 , 5 Currently, the monitoring of disinfectants is done on samples collected at the treatment plant and in different locations throughout the distribution system. This reagent-based measurement of discrete samples requires reagents and spectroscopic readout devices and can suffer from interference. Therefore, these methods are not suitable for the continuous monitoring of the disinfectant.6 Furthermore, there is no known method that can differentiate between multiple disinfectants in real samples without any previous knowledge.Two of the most important water quality parameters are pH and oxidation-reduction potential (ORP). Disinfectants undergo different reactions at different pH, resulting in different active species for different pH. ORP measurement can be used to monitor whether the disinfection was successful as oxidants’ reactivity in water depends on redox conditions.7 ORP of a solution is associated with the pH and the concentration of the disinfectant. At a fixed pH, the ORP of a disinfectant will depend on the concentration. Therefore, just by knowing the ORP, we cannot distinguish disinfectant unless the pH or concentration of the oxidant is known.8 , 9 To sort out these parameters, we propose to introduce a chemiresistive sensing array capable of distinguishing and quantifying disinfectants. Due to simpler fabrication techniques, lower cost and the ability to sense various analytes through appropriate ligand, chemiresistors have great promise in water quality sensing.6 , 10 Previously reported chemiresistive sensor for continuous measurement of free chlorine used carbon nanotube (CNT) substrate functionalized with a redox-active aniline oligomer named phenyl capped aniline tetramer (PCAT).11 Oxidation of PCAT attached to the surface of CNT by free chlorine leads to a change in the oxidation state of the oligoaniline; this change in oxidation state in the molecule changes the doping characteristics of CNT leading to a change in resistance of the CNT film. This resistance change was used to quantify the concentration of free chlorine.11 In this work, we have constructed an array of single-walled carbon nanotubes (SWCNT) sensors functionalized with five redox-active molecules. Structurally different from each other, these molecules will create different active sites. Upon interaction with analytes, each molecule will give different magnitudes of responses. Sensor responses are collected for free chlorine and KMnO4 over a range of concentrations for pH 6.5 and 7.5. In general, sensors give increasing responses to the increasing concentration and decreasing pH of the solution. Though the responses from the sensors have a similar trend to the change of the parameters their varied magnitudes make them suitable for an array. Sensor responses are then analyzed with principal component analysis (PCA). PCA analysis shows clear separation between the analytes and as well as to different pH and concentrations of the solutions (figure). We have therefore demonstrated an array of chemiresistive sensors that can distinguish and quantify disinfectants.

Efflux Pump-Driven Antibiotic and Biocide Cross-Resistance in Pseudomonas aeruginosa Isolated from Different Ecological Niches: A Case Study in the Development of Multidrug Resistance in Environmental Hotspots.

Pseudomonas aeruginosa is an opportunistic pathogen displaying high intrinsic antimicrobial resistance and the ability to thrive in different ecological environments. In this study, the ability of P. aeruginosa to develop simultaneous resistance to multiple antibiotics and disinfectants in different natural niches were investigated using strains collected from clinical samples, veterinary samples, and wastewater. The correlation between biocide and antimicrobial resistance was determined by employing principal component analysis. Molecular mechanisms linking biocide and antimicrobial resistance were interrogated by determining gene expression using RT-qPCR and identifying a potential genetic determinant for co- and cross-resistance using whole-genome sequencing. A subpopulation of P. aeruginosa isolates belonging to three sequence types was resistant against the common preservative benzalkonium chloride and showed cross-resistance to fluoroquinolones, cephalosporins, aminoglycosides, and multidrug resistance. Of these, the epidemiological high-risk ST235 clone was the most abundant. The overexpression of the MexAB-OprM drug efflux pump resulting from amino acid mutations in regulators MexR, NalC, or NalD was the major contributing factor for cross-resistance that could be reversed by an efflux pump inhibitor. This is the first comparison of antibiotic-biocide cross-resistance in samples isolated from different ecological niches and serves as a confirmation of laboratory-based studies on biocide adapted isolates. The isolates from wastewater had a higher incidence of multidrug resistance and biocide-antibiotic cross-resistance than those from clinical and veterinary settings.

Effect of Different Disinfectants on Bacterial Aerosol Diversity in Poultry Houses.

To better understand the effect of different disinfectants on the types and quantities of microorganisms in a broiler chicken house, five different types of disinfectants, including ozone, available chlorine, quaternary ammonium salt, glutaraldehyde, and mixed disinfectant, were used. The broiler house microbial communities were analyzed by high-throughput sequencing combined with air sampling. The results showed that the concentrations of airborne aerobic bacteria in the empty broiler houses after application of different disinfectants were significantly reduced compared to a house untreated with disinfectant (P < 0.05 or P < 0.01), and the number of inhalable particles of airborne aerobic bacteria sharply decreased after disinfection. Of the five disinfectants, the mixed disinfectant had the best disinfection efficacy on the total microbial communities (P < 0.05). A total of 508,143 high-quality sequences were obtained by high-throughput sequencing, which identified 1995 operational taxonomic units. In total, 42 phyla and 312 genera were identified. The structures of airborne microbial communities in the broiler houses after the different disinfectants were applied differed. In the house treated with the mixed disinfectant, the microbial communities containing opportunistic pathogens, such as Escherichia-Shigella, Bacillus, and Pseudomonas, had the lowest abundance, with a significant decrease compared to the house untreated with disinfectant. The alpha diversity index showed low diversity of the microbial communities in the house treated with mixed disinfectant. In contrast to the other four disinfectants, only a small amount of bacteria was detected in the air sample in the house treated with the mixed disinfectant; specifically, only four phyla were found (Proteobacteria, Bacteroidetes, Actinobacteria, and Firmicutes). The mixed disinfectant produced a positive effect on disinfection for four phyla; however, it didn’t thoroughly eliminate them. At genus level, Bacillus, Arenimonas, and Shinella could not be detected in the house treated with the mixed disinfectant, but were detected in houses treated with other disinfectants. The high-throughput sequencing results revealed that the combination of multiple disinfectants exhibited a good disinfection efficacy and that this technique could disinfect the air of broiler houses. These results will help guide the development of a reasonable program for broiler house disinfection.

Phenotypic changes associated with Colistin resistance due to Lipopolysaccharide loss in Acinetobacter baumannii

ABSTRACTAcinetobacter baumannii can acquire resistance to colistin via complete loss of lipopolysaccharide (LPS) biosynthesis due to mutations in the lpxA, lpxC and lpxD genes. However, although colistin is increasingly being used for the treatment of multidrug resistant infections, very few A. baumannii clinical isolates develop colistin resistance through loss of LPS biosynthesis. This may suggest that LPS loss affects virulence traits that play a role in the transmission and pathogenesis of A. baumannii. In this study we characterize multiple virulence phenotypes of colistin resistant, LPS-deficient derivatives of the ATCC 19606 strain and five multidrug resistant clinical isolates and their colistin resistant, LPS-deficient derivatives. Our results indicate that LPS loss results in growth defects compared to the parental strain in vitro both in laboratory media and human serum (competition indices of 0.58 and 7.0 × 10−7, respectively) and reduced ability to grow and disseminate in vivo (competition index 6.7 × 10−8). Infection with the LPS-deficient strain resulted in lower serum levels of pro-inflammatory cytokines TNF-α and IL-6 compared to the parent strain, and was less virulent in a mouse model of disseminated sepsis. LPS loss also significantly affected biofilm production, surface motility, growth under iron limitation and susceptibility to multiple disinfectants used in the clinical setting. These results demonstrate that LPS loss has a significant effect on multiple virulence traits, and may provide insight into the low incidence of colistin resistant strains lacking LPS that have been reported in the clinical setting.

Alcohols as Surface Disinfectants in Healthcare Settings.

Isopropyl alcohol and ethyl alcohol have been used as low-level disinfectants in healthcare settings for many years. Recent studies have found that ethyl alcohol inhibits protein synthesis in Escherichia coli by direct effects on ribosomes and RNA polymerase and that 60%-70% solutions have in vitro efficacy against murine norovirus, Ebola virus, and several coronaviruses. Alcohol prep pads or towelettes containing isopropyl or ethyl alcohol and water have been used primarily for disinfection of small noncritical items due to a concern regarding their rapid evaporation rates and associated short contact times. Sterile alcohol solutions are used mostly for disinfection of compounding pharmacies and controlled areas. One new Environmental Protection Agency (EPA)-registered cleaner/disinfectant formulation differs from other alcohol-based disinfectants by virtue of having a 30-second contact time for multiple pathogens and a toxicity rating of category IV. Multiple disinfectants containing ethyl alcohol and/or isopropyl alcohol combined with other active agents such as quaternary ammonium or phenolic compounds are widely used for disinfecting environmental surfaces in healthcare facilities. Infect Control Hosp Epidemiol 2018;39:323-328.

Cross-Resistance of UV- or Chlorine Dioxide-Resistant Echovirus 11 to Other Disinfectants

The emergence of waterborne viruses with resistance to disinfection has been demonstrated in the laboratory and in the environment. Yet, the implications of such resistance for virus control remain obscure. In this study we investigate if viruses with resistance to a given disinfection method exhibit cross-resistance to other disinfectants. Chlorine dioxide (ClO2)- or UV-resistant populations of echovirus 11 were exposed to five inactivating treatments (free chlorine, ClO2, UV radiation, sunlight, and heat), and the extent of cross-resistance was determined. The ClO2-resistant population exhibited cross-resistance to free chlorine, but to none of the other inactivating treatments tested. We furthermore demonstrated that ClO2 and free chlorine act by a similar mechanism, in that they mainly inhibit the binding of echovirus 11 to its host cell. As such, viruses with host binding mechanisms that can withstand ClO2 treatment were also better able to withstand oxidation by free chlorine. Conversely, the UV-resistant population was not significantly cross-resistant to any other disinfection treatment. Overall, our results indicate that viruses with resistance to multiple disinfectants exist, but that they can be controlled by inactivating methods that operate by a distinctly different mechanism. We therefore suggest to utilize two disinfection barriers that act by different mechanisms in order to control disinfection-resistant viruses.

In vitro antibiofilm efficacy of Piper betle against quorum sensing mediated biofilm formation of luminescent Vibrio harveyi

Vibrio harveyi is a potent biofilm former, which confers resistance to multiple antimicrobials, disinfectants, chemicals and biocides. The prevalence of biofilm mediated antibiotic resistance among aquatic bacterial pathogens stresses the search for novel alternative approach to treat vibriosis in aquaculture. Exploring suitable therapeutics from natural resources could be a novel area of research. Therefore, this work was executed to evaluate the inhibitory effect of Piper betle ethyl acetate extract (PBE) on bioluminescence production and biofilm formation of V. harveyi. Minimal inhibitory concentration (MIC) of PBE against planktonic V. harveyi was found to be 1600 μg ml−1; furthermore, PBE inhibited the quorum sensing (QS) mediated bioluminescence production and biofilm formation in V. harveyi upto 98 and 74% respectively, at its sub-MIC concentration of 400 μg ml−1 without affecting their cell viability. Similar results were obtained for exopolysaccharides production and swimming motility related to biofilm formation of V. harveyi, where PBE reduced EPS production upto 64%. Light and confocal laser scanning microscopic analyses further confirmed that the PBE effectively prevented the initial attachment as well as microcolonies formation of V. harveyi biofilm, when compared to their untreated controls. This study demonstrates the promising antibiofilm activity of PBE and confirms the ethnopharmacological potential of this plant against V. harveyi infections.

Microbots Decorated with Silver Nanoparticles Kill Bacteria in Aqueous Media.

Water contamination is one of the most persistent problems of public health. Resistance of some pathogens to conventional disinfectants can require the combination of multiple disinfectants or increased disinfectant doses, which may produce harmful byproducts. Here, we describe an efficient method for disinfecting Escherichia coli and removing the bacteria from contaminated water using water self-propelled Janus microbots decorated with silver nanoparticles (AgNPs). The structure of a spherical Janus microbot consists of a magnesium (Mg) microparticle as a template that also functions as propulsion source by producing hydrogen bubbles when in contact with water, an inner iron (Fe) magnetic layer for their remote guidance and collection, and an outer AgNP-coated gold (Au) layer for bacterial adhesion and improving bactericidal properties. The active motion of microbots increases the chances of the contact of AgNPs on the microbot surface with bacteria, which provokes the selective Ag+ release in their cytoplasm, and the microbot self-propulsion increases the diffusion of the released Ag+ ions. In addition, the AgNP-coated Au cap of the microbots has a dual capability of capturing bacteria and then killing them. Thus, we have demonstrated that AgNP-coated Janus microbots are capable of efficiently killing more than 80% of E. coli compared with colloidal AgNPs that killed only less than 35% of E. coli in contaminated water solutions in 15 min. After capture and extermination of bacteria, magnetic properties of the cap allow collection of microbots from water along with the captured dead bacteria, leaving water with no biological contaminants. The presented biocompatible Janus microbots offer an encouraging method for rapid disinfection of water.

Assaying the Stability and Inactivation of AAV Serotype 1 Vectors.

Adeno-associated virus (AAV) vectors are a commonplace tool for gene delivery ranging from cell culture to human gene therapy. One feature that makes AAV a desirable vector is its stability, in regard to both the duration of transgene expression and retention of infectivity as a viral particle. This study examined the stability of AAV serotype 1 (AAV1) vectors under different conditions. First, transducibility after storage at 4°C decreased 20% over 7 weeks. Over 10 freeze-thaw cycles, the resulting transduction efficiency became variable at 60-120% of a single thaw. Using small stainless steel slugs to mimic a biosafety cabinet or metal lab bench surface, it was found that an AAV1 vector can be reconstituted after 6 days of storage at room temperature. The stability of AAV is a desired feature, but effective decontamination procedures must be available for safety and experimental integrity. Multiple disinfectants commonly used in the laboratory for ability to inactivate an AAV1 vector were tested, and it was found that autoclaving, 0.25% peracetic acid, iodine, or 10% Clorox bleach completely prevented AAV-mediated transgene expression. These data suggest that peracetic acid should be used for inactivating AAV1 vectors on metal-based surfaces or instruments in order to avoid inadvertent transgene expression in human cells or cross-contamination of instruments.

Development of a New &amp;lt;i&amp;gt;Pseudomonas&amp;lt;/i&amp;gt; Agar Medium Containing Benzalkonium Chloride in Cetrimide Agar

Members of the Pseudomonas family are commonly found in nature, some species are pathogenic for humans, as well as being resistant to multiple disinfectants. Various studies have revealed that benzalkonium chloride (BKC) has an inhibitory effect on many bacteria but it has no significant effect on Pseudomonas aeruginosa. Cetrimide agar medium is recommended for the isolation and enumeration of Ps. aeruginosa in food and environmental samples. However, there are claims that for some food factories and in particular the bottled water industry, the selectivity of this medium is not sufficient. The aim of the current research is the creation of a more selective medium for Ps. aeruginosa with BKC. A total of 28 isolates were isolated with Cetrimide agar from raw water samples and identified using biochemical tests and commercial identification kits. All the bacteria were inoculated in Cetrimide agar plates containing 0 - 625 μg/mL BKC. The Petri dishes were incubated at 37°C and 42°C for 24 h. The results showed that 375 μg/mL BKC was sufficient to suppress Burk. pseudomallei at both incubation temperatures. Ps. fluorescens-35 could not grow at 42°C at any concentration, including the control, and was suppressed at 500 μg/mL BKC. All the Ps. aeruginosa isolates and control strain were grown at both incubation temperatures at 375 μg/mL BKC concentration. In conclusion, the analysis of Ps. aeruginosa showed that the growth of accompanying flora may be suppressed by adding 375-μg/mL BKC into Cetrimide agar and incubating at an elevated temperature of 42°C.

97. Stability and Inactivation of AAV Serotype 1 Vectors

Adeno-associated viral (AAV) vectors have become a commonplace tool for gene delivery ranging from cell lines to human gene therapy. One feature that makes AAV desirable for gene delivery is its relative safety owing to no known disease associated with wild-type AAV. For this reason, it is often used with BSL1 practices negating the need for dedicated BSL2 facilities. Another useful property of AAV is its stability, both the duration of transgene expression and retention of infectivity as a viral particle. In the current study, we examine the stability of AAV serotype 1 vectors which are capable of transducing human cells and are routinely produced and used in our facility. First, we examined stability at 4° C for up to 7 weeks and found <10% decrease in transduction efficiency compared to a freshly-thawed aliquot. After 10 freeze-thaw cycles, 25% loss in transduction efficiency was observed. These data indicated that AAV serotype 1 is a highly stable viral particle with regards to retaining infectivity. Using small stainless steel slugs to mimic a biosafety cabinet or metal lab bench surface, we demonstrate that infectious AAV vector expressing Cre-recombinase can be reconstituted and used up to 7 days at room temp, although there was a clear decrease in transgene expression over time. The stability of AAV is a desired feature as a tool; however, it is important that the vector can be inactivated to avoid transgene delivery to end users or cross-contamination of non-disposable instruments. AAV decontamination procedures can vary. We tested multiple disinfectants commonly used in the laboratory for their ability to inactivate an AAV serotype 1 vector. The vector was added directly into solution for 5 or 30 minutes at a 1:10 ratio. It was then diluted, dialyzed, concentrated, and tested for transduction on primary neurons. Of the decontamination procedures tested, only autoclaving, 3.9% peracetic acid, iodine or 10% Clorox bleach completely prevented the subsequent detection of transgene expression. Although the nature of the transgene (e.g. proto oncogene or toxin) can elevate an AAV vector to BSL2, our data show that when working with AAV serotype 1 vectors, it's important that safety procedures for handling and decontaminating AAV are effective to avoid inadvertent transgene expression in human cells or cross contamination of AAV vectors used in non-disposable instruments. Other serotypes have not yet been tested.

Suscetibilidade a desinfetantes e perfil de resistência a antimicrobianos em isolados de Escherichia coli

A colibacilose, causada por Escherichia coli, é a enfermidade entérica de maior impacto na produção de suínos, podendo levar à morte do animal. Esta bactéria possui grande capacidade de desenvolver resistência a múltiplos antimicrobianos e a desinfetantes. Desta forma, estudos que abordem mecanismos de resistência e perfil de amostras de campo tornam-se necessários. E. coli é amplamente utilizada como modelo de estudos que exploram a resistência intrínseca e extrínseca a multidrogas. Neste trabalho, buscou-se verificar o perfil de sensibilidade de 62 isolados de E. coli de suínos frente a três desinfetantes e a 13 antimicrobianos. Ainda, em 31 destes isolados foi pesquisada a presença de mecanismo de efluxo. Dos três desinfetantes avaliados, o cloreto de alquil dimetil benzil amônio+poliexietilenonilfenileter foi o que se mostrou mais eficaz (100%), seguido do glutaraldeído+cloreto de alquil dimetil benzil amônio (95,2%) e do cloreto de alquil dimetil benzil amônio (88,8%). Dentre os antimicrobianos testados, observou-se maior resistência para a tetraciclina (62,2%) e maior sensibilidade para o florfenicol (88,6%). A alta sensibilidade dos isolados frente aos desinfetantes pode estar relacionada à ausência de mecanismo de efluxo. O índice de resistência múltipla médio aos antimicrobianos foi de 0,52, o que demonstra um perfil multirresistente dos isolados, conduzindo para a necessidade do uso racional destas drogas em suinocultura.

Correlation between reduced susceptibility to disinfectants and multidrug resistance among clinical isolates of Acinetobacter species

The aim of this study was to investigate the susceptibility profiles to disinfectants and antimicrobial agents of 283 non-repetitive Acinetobacter clinical isolates obtained in 97 Japanese hospitals in March 2002. Susceptibility profiles of the above isolates to four disinfectants, six antimicrobial agents and two dyes were investigated. MICs were measured by the agar dilution method recommended by the CLSI (formerly NCCLS). MBC measurements and time-kill assays were performed using a slightly modified quantitative suspension test based on the European Standard EN 1040. No evident resistance to disinfectants was seen among the 283 strains of Acinetobacter spp. isolated in 2002, but the MIC(90)s of chlorhexidine gluconate, benzalkonium chloride and alkyldiaminoethylglycine hydrochloride were 50, 50 and 400 mg/L, respectively. Interestingly, the MICs of alkyldiaminoethylglycine hydrochloride and benzethonium chloride for four and three clinical isolates, respectively, reached 800 mg/L (approximately half the in-use concentration). The MBCs for the 28 disinfectant reduced susceptibility (DRS) isolates, for which the MICs of at least one of the four disinfectants tested were higher than the MIC(90), were comparable to those for susceptible isolates, in general; however, significant differences (P < 0.01) were observed between disinfectant-susceptible and DRS isolates in the time-kill assays of chlorhexidine gluconate, benzalkonium chloride and benzethonium chloride. Furthermore, DRS isolates tended to demonstrate multiresistance profiles to ceftazidime, ciprofloxacin and amikacin (P < 0.05). Since several Acinetobacter clinical isolates have developed augmented resistance to multiple antimicrobials and disinfectants, it is worth checking the susceptibility to disinfectants if multidrug-resistant Acinetobacter spp. are recurrently isolated clinically.

Structure and Filament Dynamics of the pSK41 Actin-like ParM Protein: IMPLICATIONS FOR PLASMID DNA SEGREGATION

Type II plasmid partition systems utilize ParM NTPases in coordination with a centromere-binding protein called ParR to mediate accurate DNA segregation, a process critical for plasmid retention. The Staphylococcus aureus pSK41 plasmid is a medically important plasmid that confers resistance to multiple antibiotics, disinfectants, and antiseptics. In the first step of partition, the pSK41 ParR binds its DNA centromere to form a superhelical partition complex that recruits ParM, which then mediates plasmid separation. pSK41 ParM is homologous to R1 ParM, a known actin homologue, suggesting that it may also form filaments to drive partition. To gain insight into the partition function of ParM, we examined its ability to form filaments and determined the crystal structure of apoParM to 1.95 A. The structure shows that pSK41 ParM belongs to the actin/Hsp70 superfamily. Unexpectedly, however, pSK41 ParM shows the strongest structural homology to the archaeal actin-like protein Thermoplasma acidophilum Ta0583, rather than its functional homologue, R1 ParM. Consistent with this divergence, we find that regions shown to be involved in R1 ParM filament formation are not important in formation of pSK41 ParM polymers. These data are also consonant with our finding that pSK41 ParM forms 1-start 10/4 helices very different from the 37/17 symmetry of R1 ParM. The polymerization kinetics of pSK41 ParM also differed from that of R1 ParM. These results indicate that type II NTPases utilize different polymeric structures to drive plasmid segregation.

Application of non-biological surrogates for analysis of sequential disinfection continuous flow systems

ABSTRACT Brannon RichardsJoel Ducoste (corresponding author)Department of Civil, Construction, andEnvironmental Engineering,North Carolina State University,208 Mann Hall CB 7908,Raleigh, NC 27695-7908,USAE-mail: jducoste@eos.ncsu.edu Fluorescent YG-microspheres (Polysciences Inc.) were evaluated to simulate Cryptosporidiuminactivation in a continuous flow system that utilizes multiple disinfectants. Experiments wereperformed in a disinfection process consisting of an ozone primary stage and a secondary freechlorine stage. Impacts of the chemical disinfectant exposure were calculated by tracking thechanges in fluorescence distribution with a flow cytometer. Tests were performed at two flowrates (11- and 15.5-ml/s) and a target concentration-time (Ct) product of 1.4mg/L-min for ozoneand 510mg/L-min for chlorine. Analysis of the results suggest that the fluorescence decay ofYG-fluorescent microspheres does display synergistic effects when free chlorine is usedsequentially with ozone in a continuous flow system. The study also included the use of a simpleSegregated Flow Reactor (SFR) model to simulate the sequential disinfection process. The modelwas not effective at predicting fluorescent intensity changes at different intermediate pointswithin the disinfection process stream due to the complexity of the paths taken by themicrospheres through the ozone primary disinfectant chamber and its eventual influence onchlorine secondary disinfectant kinetics. An ozone Ct distribution, which utilized the fluorescencemicrospheres experimental data and was created from the range of paths traveled by themicrospheres, displayed a range of Ct values between 0.5 to 3.2mg/L-min for the low flowcondition and 0.8 to 2.2mg/l-min for the high flow condition. A new model structure wasproposed that may improve the simulation of sequential disinfection systems.

A Multiple Barrier: The Value and Performance of Multiple Disinfectants in Series for the City of Phoenix

A Multiple Barrier: The Value and Performance of Multiple Disinfectants in Series for the City of Phoenix

Enhanced Bactericidal Effect of O3/H2O2 Followed by Cl2

With the appearance of chlorine resistant microorganisms such as Cryptosporidium parvum and Giardia lamblia in drinking water, significant attention has been drawn to the sequential application of multiple disinfectants including ozone, chlorine dioxide, and UV as a primary disinfectant. However, few studies have reported about the inactivation behavior of ozone-based AOP (advanced oxidation process) or its sequential application combined with other disinfectants. This is especially important since ozone itself experiences difficulty in the inactivation of these pathogens, especially at low temperatures: This study investigates the enhanced inactivation of Bacillus subtilis spores by the presence of an OH radical in the O3/H2O2 system and the synergistically enhanced inactivation in the application of the O3/H2O2 system followed by Cl2. The results suggest that the O3/H2O2 process can be considered as one of the viable alternatives when O3 alone does not satisfy the disinfection requirement.

A non-biological surrogate for sequential disinfection processes

An evaluation of Fluorescent YG-microspheres (Polysciences Inc.) was performed to simulate Cryptosporidium parvum ( C. parvum) oocysts inactivation in treatment systems that utilize multiple disinfectants. Experiments were conducted in batch reactors that included an ozone primary stage and a secondary free chlorine treatment stage. A flow cytometer was used to track changes in the fluorescence intensity distribution due to exposure to the chemical disinfectant. Microsphere ‘survival ratios’ ( N/ No) were calibrated by selecting an appropriate fluorescence intensity threshold to replicate the inactivation of different C. parvum oocysts strains. Results showed that fluorescent microspheres displayed synergistic effects in the presence of two sequential disinfectants. In addition, microsphere structural tests showed that the polystyrene surface was damaged due to exposure to ozone. This polystyrene damage enhanced the diffusion of the secondary disinfectant into the microsphere, where dye was degraded in the opened polymer layer. As a result, YG-fluorescent microspheres is a promising non-biological technique that is capable of producing similar synergistic behavior as with C. parvum oocysts exposed to ozone followed by chlorine.

Association between cyclohexane resistance in Salmonella of different serovars and increased resistance to multiple antibiotics, disinfectants and dyes

A panel of 388 salmonellas of animal and human origin, comprising 35 serotypes, was tested for resistance to cyclohexane and to a range of antibiotics, disinfectants and dyes. Cyclohexane resistance was detected in 41 isolates (10.6%): these comprised members of the serovars Binza (1 of 15), Dublin (1 of 24), Enteritidis (1 of 61), Fischerkietz (4 of 5), Livingstone (9 of 11), Montevideo (1 of 32), Newport (4 of 23), Saint-paul (1 of 3), Senftenberg (10 of 24) and Typhimurium (9 of 93). Most (39 of 41) of the cyclohexane-resistant isolates were from poultry. Statistical analysis showed that the cyclohexane-resistant strains were significantly more resistant than the cyclohexane-susceptible strains to ampicillin, chloramphenicol, ciprofloxacin, erythromycin, nalidixic acid, tetracycline, trimethoprim, cetrimide and triclosan. The multiresistance patterns seen were typical of those caused by efflux pumps, such as AcrAB. The emergence of such resistance may play an important role in the overall antibiotic resistance picture of Salmonella, with particular effect on ciprofloxacin.