A systematic review on chlorine tolerance among bacteria and standardization of their assessment protocol in wastewater.

Though chlorine is a cost-effective disinfectant for water and wastewaters, the bacteria surviving after chlorination pose serious public health and environmental problems. This review critically assesses the mechanism of chlorine disinfection as described by various researchers; factors affecting chlorination efficacy; and the re-growth potential of microbial contaminations in treated wastewater post chlorination to arrive at meaningful doses for ensuring health safety. Literature analysis shows procedural inconsistencies in the assessment of chlorine tolerant bacteria, making it extremely difficult to compare the tolerance characteristics of different reported tolerant bacteria. A comparison of logarithmic reduction after chlorination and the concentration-time values for prominent pathogens led to the generation of a standard protocol for the assessment of chlorine tolerance. The factors that need to be critically monitored include applied chlorine doses, contact time, determination of chlorine demands of the medium, and the consideration of bacterial counts immediately after chlorination and in post chlorinated samples (regrowth). The protocol devised here appropriately assesses the chlorine-tolerant bacteria and urges the scientific community to report the regrowth characteristics as well. This would increase the confidence in data interpretation that can provide a better understanding of chlorine tolerance in bacteria and aid in formulating strategies for effective chlorination.

Assessment of physiological responses of bacteria to chlorine and UV disinfection using a plate count method, flow cytometry and viability PCR.

AIMS: This study aimed to investigate the physiological responses of two gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) and two gram-positive bacteria (Enterococcus faecalis and Bacillus sphaericus) to ultraviolet (UV) and chlorine disinfection. METHODS AND RESULTS: Bacterial inactivation by UV and chlorine disinfection were evaluated with a plate count method for culturability, FCM and PMA-qPCR for membrane integrity and DyeTox13-qPCR for enzymatic activity, respectively. Both UV and chorine disinfection caused complete loss of culturability while membrane integrity remained intact after UV disinfection. Both DyeTox13-qPCR and PMA-qPCR showed high ΔCt values up to 8.9 after chlorine disinfection, indicating that both methods were able to distinguish non-treated from chlorine-treated cells. Although PMA-qPCR could not differentiate membrane integrity of cells on UV exposure, DyeTox13-qPCR showed significant differences in ΔCt values of 5.05 and 10.4 for gram-negative (E. coli) and gram-positive (Enterococcus) bacteria, respectively. However, DyeTox13-qPCR for gram-negative bacteria displayed relatively small differences in ΔCt values compared with gram-positive bacteria. CONCLUSION: UV and chlorine disinfection led to changes in physiological state of gram-negative and gram-positive bacteria. Particularly, UV disinfection could induce active but non-culturable (ABNC) for gram-negative bacteria and dormant cell for gram-positive bacteria where intact cells no longer showed the enzymatic activity. SIGNIFICANCE AND IMPACT OF THE STUDY: UV and chlorine are commonly used to disinfect water, food and fomites to inactivate pathogenic bacteria. However, a viable but non-culturable (VBNC) state of bacteria induced by disinfection may underestimate the health risks because of the potential resuscitation of VBNC cells. This study highlighted that bacteria could undergo different physiological (ABNC or dormant) states during UV and chlorine disinfection. In addition, viability PCR techniques could provide insight into the changes in physiological states during disinfection processes.

Flow cytometric assessment of the chlorine/chloramine efficacy of particle-associated bacteria in drinking water.

Chlorine and chloramine are widely used to maintain the microbial safety after drinking water treatment plants. Particles existing in the treated water may react with these chemical disinfectants, and impact the efficacy of disinfection. However, the protective effects of particles without-disinfectant demand on bacteria in the chlorination/chloramination are not well known. In this study, two laboratory-derived bacteria (Staphylococcus aureus and Escherichia coli) and two no-disinfectant demand particles (Fe2O3 and kaolin) in drinking water were selected to build particle-associated bacteria (PAB) systems, and their resistance to chlorine/chloramine was further assessed. Flow cytometry (FCM) was employed to image PAB systems and assess the removal rate of bacteria. The results were that particles showed protective effects on bacteria in half of chlorine experiments and 90% of chloramination. The protection was related to the combination form of particles and bacteria tied to neither particle species nor size, and there was no positive relationship between the protection effect and water turbidity. S. aureus attached to Fe2O3 had stronger resistance than kaolin, and kaolin protected E. coli better than Fe2O3. The same trend was observed in both chemical disinfectants, and more significant resistance had been shown in chloramination than chlorination. FCM images which gave a qualitative description on the combination states of different PAB systems may be a clue to explain the strength of the resistance. Environmental bacterial strains and particles are recommended in the future to explore practical applications.

Microbial and physicochemical assessment of irrigation water treatment methods.

AIMS: The presence of foodborne pathogens in preharvest agricultural water has been identified as a potential contamination source in outbreak investigations, driving markets and auditing bodies to begin requiring water treatment for high-risk produce. Therefore, it is essential that we identify water treatment methods which are effective as well as practical in their application on farm. METHODS AND RESULTS: In this work, we evaluated two sanitizers which are most prominent in preharvest agricultural water treatment (calcium hypochlorite (free chlorine: 3-5 ppm) and peracetic acid (PAA: 5 ppm)), an EPA registered antimicrobial device (ultraviolet light (UV)), in addition to a combination approach (chlorine + UV, PAA + UV). Treatments were evaluated for their ability to inactivate total coliforms and generic Escherichia coli and consistency in treatment efficacy over 1 h of operation. Physicochemical variables were measured along with microbial populations at 0, 5, 15, 30, 45 and 60 min of operation. Escherichia coli and coliform counts showed a significant (P < 0·05) reduction after treatment, with combination and singular treatments equally effective at inactivating E. coli and coliforms. A significant increase (P < 0·05) in oxidation-reduction potential was seen during water treatment (Chlorine; UV + Chlorine), and a significant reduction (P < 0·05) in pH was seen after PAA and PAA + UV treatments (60 min). CONCLUSION: Overall, the results indicate that all treatments evaluated are equally efficacious for inactivating E. coli and coliforms present in surface agricultural water. SIGNIFICANCE AND IMPACT OF THE STUDY: This information when paired with challenge studies targeting foodborne pathogens of interest can be used to support grower decisions when selecting and validating a preharvest agricultural water treatment programme.

Diversity Assessment of Toxic Cyanobacterial Blooms during Oxidation.

Fresh-water sources of drinking water are experiencing toxic cyanobacterial blooms more frequently. Chemical oxidation is a common approach to treat cyanobacteria and their toxins. This study systematically investigates the bacterial/cyanobacterial community following chemical oxidation (Cl2, KMnO4, O3, H2O2) using high throughput sequencing. Raw water results from high throughput sequencing show that Proteobacteria, Actinobacteria, Cyanobacteria and Bacteroidetes were the most abundant phyla. Dolichospermum, Synechococcus, Microcystis and Nostoc were the most dominant genera. In terms of species, Dolichospermum sp.90 and Microcystis aeruginosa were the most abundant species at the beginning and end of the sampling, respectively. A comparison between the results of high throughput sequencing and taxonomic cell counts highlighted the robustness of high throughput sequencing to thoroughly reveal a wide diversity of bacterial and cyanobacterial communities. Principal component analysis of the oxidation samples results showed a progressive shift in the composition of bacterial/cyanobacterial communities following soft-chlorination with increasing common exposure units (CTs) (0-3.8 mg·min/L). Close cyanobacterial community composition (Dolichospermum dominant genus) was observed following low chlorine and mid-KMnO4 (287.7 mg·min/L) exposure. Our results showed that some toxin producing species may persist after oxidation whether they were dominant species or not. Relative persistence of Dolichospermum sp.90 was observed following soft-chlorination (0.2-0.6 mg/L) and permanganate (5 mg/L) oxidation with increasing oxidant exposure. Pre-oxidation using H2O2 (10 mg/L and one day contact time) caused a clear decrease in the relative abundance of all the taxa and some species including the toxin producing taxa. These observations suggest selectivity of H2O2 to provide an efficient barrier against toxin producing cyanobacteria entering a water treatment plant.

An evaluation of buffered peracetic acid as an alternative to chlorine and hydrogen peroxide based disinfectants.

This short report documents an in-use evaluation of three disinfectant solutions that was conducted within the operating theatre of a South Australian hospital to address a high occurrence of Clostridium difficile Infection (CDI). The disinfectants were all registered by the Therapeutic Goods Administration (TGA) and included a buffered peracetic acid, a chlorine-based disinfectant used at 1000 ppm, and a hydrogen peroxide-based disinfectant. The use of the chlorine and hydrogen peroxide disinfectants both caused a number of adverse staff reactions and increased safe-work related incident reporting. The peracetic acid-based product met all criteria for use, including staff acceptance, cleaning expectation, cost and efficacy requirements.

Assessment of the bacterial viability of chlorine- and quaternary ammonium compounds-treated Lactobacillus cells via a multi-method approach.

AIMS: The assessment of the bacterial viability of chlorine- and quaternary ammonium compounds (QACs)-treated Lactobacillus cells by culture-dependent and -independent methods. METHODS AND RESULTS: Lactobacillus isolates (Lactobacillus plantarum G1, Lactobacillus plantarum B1, Lactobacillus brevis S1 and Lactobacillus paracasei W1) in biofilm and planktonic cell suspensions were treated with chlorine-based (0·018 and 0·18%) and QACs-based (0·2 and 2·0%) disinfectants for 5 min and then analysed by plate counting, flow cytometry (FCM) and fluorescence activated cell sorting (FACS). The reaction of sessile cells to disinfectants was assessed with the confocal laser scanning microscopy (CLSM). Plate counts revealed L. paracasei W1 to be substantially inactivated by both disinfectants, while counts of the other isolates to be significantly reduced only by QACs, with L. plantarum B1 and L. brevis S1 showing a greater difference between QACs concentrations and cell types. In several cases, the disinfectants caused slightly higher inactivation of planktonic than biofilm cells, with L. plantarum B1 being significantly less sensitive to QACs in biofilm cells (P < 0·05). Following FCM with a Syto® 9/PI assay, which addresses cell membrane integrity, the emergence of damaged (Syto® 9- PI+ ) and injured (Syto® 9+ PI+ ) subpopulations was often observed in cells when they were treated with QACs, whereas intact (Syto® 9+ PI- ) and unstained (Syto® 9- PI- ) subpopulations were mostly encountered in chlorine-treated cells. Except Syto® 9- PI+ , all subpopulations were recovered on agar plates following FACS, with biofilm cells showing higher culturability irrespective of conditions, probably because of the residues of the biofilm matrix which serve as a protective cover for the bacteria. The CLSM revealed a substantial cell membrane damage within the QACs-treated biofilms, however, some cells deep in the biofilm were still intact and thus remained protected against this disinfectant. CONCLUSION: We found that FCM/FACS proved useful in the analysis of lactobacilli membrane integrity in disinfection experiments as well as in recovery evaluation of planktonic-biofilm cell subpopulations. In turn, CLSM was particularly useful in investigating the resistance mechanism when Lactobacillus cells were embedded in biofilms. SIGNIFICANCE AND IMPACT OF THE STUDY: This study highlights the need for treatment optimization on a case-by-case basis to avoid the emergence of cells in intermediate states with recovery potential and to reach and, thus, kill all bacteria in already developed lactobacilli biofilms.

Assessment of an anti-scale low-frequency electromagnetic field device on drinking water biofilms.

Low intensity and very low-frequency electromagnetic fields (EMF) used for preventing scaling in water distribution systems were tested for the first time for their potential impact on drinking water biofilms. The assays were carried out in laboratory-scale flow-through reactors that mimic water distribution systems. The drinking water biofilms were not directly exposed to the core of the EMF generator and only subjected to waterborne electromagnetic waves. The density and chlorine susceptibility of nascent or mature biofilms grown under exposure to EMF were evaluated in soft and hard water. This EMF treatment was able to modify CaCO3 crystallization but it did not significantly affect biofilms. Indeed, over all the tested conditions, there was no significant change in cell number, or in the integrity of the cells (membrane, culturability), and no measurable effect of chlorine on the biofilm.

Chlorination disinfection by-products and comparative cost analysis of chlorination and UV disinfection in sewage treatment plants: Indian scenario.

Apart from numerous other well-known drawbacks of chlorination, viz. on-site operational hazards and residual chlorine toxicity, trihalomethane (THM) formation is the major factor that came into limelight in the last 40 years, primarily in drinking water treatment industry. Treated effluent from wastewater treatment plants is also chlorinated and then discharged, indirectly coming in human contact, so there is need to consider THM as a potable as well as wastewater parameter. In this study, THMs were identified in seven sewage treatment plants (STPs) in North India. STPs were selected based on treatment technology employed, viz., up-flow anaerobic sludge blanket (UASB), activated sludge process (ASP), sequential batch reactor (SBR), and oxidation pond (OP). THM concentrations obtained at all the seven STPs were below BIS standards of drinking water (0-40 µg L-1). UASB plant shows considerably higher concentration of THM. UV followed by chlorination is suggested as an alternative to chlorination. Per million liter per day (MLD) capital and operation and maintenance (O&M) costs of UV disinfection were analyzed revealing decreasing per MLD capital cost of UV with increasing plant capacity. The comparative annual O&M cost analysis of chlorination, dechlorination, and UV disinfection shows that there is up to 63% reduction of the total annual O&M cost by UV in comparison to chlorination, whereas in the case of chlorination followed by dechlorination, total reduction is 71%.

Assessment of chlorine tolerance profile of Citrobacter species recovered from wastewater treatment plants in Eastern Cape, South Africa.

This present study assessed the chlorine tolerance of some Citrobacter species recovered from secondary effluents from the clarifiers of two wastewater treatment plants in the Eastern Cape, South Africa. The bacterial survival, chlorine lethal dose and inactivation kinetics at lethal doses were examined. Inactivation of the test bacteria (n = 20) at the recommended dose of 0.5 mg/l for 30 min exposure showed a progressive reduction in bacterial population from 4 to 5 log reduction and residuals ranged between 0.12 and 0.46 mg/l. The bactericidal activity of chlorine increased at higher dosages with a substantial reduction in viability of the bacteria and complete inactivation of the bacterial population at a lethal dose of 0.75 and 1.0 mg/l in 30 min. For the inactivation kinetics, bactericidal activity of chlorine increased with time showing a 3.67-5.4 log reduction in 10 min, 4.0-5.6 log reduction in 20 min and above 6.3 log reductions to complete sterilization of bacterial population over 30 min for all the entire test Citrobacter isolates used in this study. Furthermore, there was a strong correlation (R 2 > 0.84) between bacteria inactivation and increase in contact time. This study appears to have provided support for laboratory evidence of bacterial tolerance to chlorine disinfection at current recommended dose (0.5 mg/l for 30 min), and chlorine concentration between 0.75 and 1.0 mg/l was found to have a better disinfecting capacity to check tolerance of Citrobacter species.

Quantitative assessment of the impact of cross-contamination during the washing step of ready-to-eat leafy greens on the risk of illness caused by Salmonella.

The aim of this study was to develop a quantitative microbial risk assessment (QMRA) model to estimate the risk of illness caused by Salmonella in ready-to-eat (RTE) leafy greens, based on common practices in Brazilian processing plants. The risk assessment model considered five modules: in field, washing step, retail storage, home storage and dose-response. Fifty thousand iterations of a @Risk model built in Excel were run for each of sixty scenarios. These scenarios considered different initial pathogen concentrations, fractions of contaminated produce and chlorine concentrations. For chlorine, seven pre-set concentrations (0, 5, 10, 25, 50, 150 and 250mg/L) and three triangular distributions were considered [RiskTriang(0,5,10mg/L), RiskTriang(0,80,250mg/L) and RiskTriang(10,120,250mg/L)]. The outputs were risk of infection, estimated number of illnesses and estimated percent of illnesses arising from cross-contamination. The QMRA model indicated quantitatively that higher chlorine concentrations resulted in lower risk of illness. When simulation was done with <5mg/L of chlorine, most (>96%) of the illnesses arose from cross-contamination, but when a triangular distribution with 10, 120 and 250mg/L of chlorine was simulated, no illnesses arising from cross-contamination were predicted. Proper control of the sanitizer in the washing step is essential to reduce initial contamination and avoid cross-contamination.

Quantitative assessment of human exposure to extended spectrum and AmpC ß-lactamases bearing E. coli in lettuce attributable to irrigation water and subsequent horizontal gene transfer.

The contribution of the fresh produce production environment to human exposure with bacteria bearing extended spectrum ß-lactamases and AmpC ß-lactamases (ESBL/AmpC) has not been reported. High prevalence of ESBLs/AmpC bearing E. coli as well as a high gene transfer efficiency of lettuce and irrigation water E. coli isolates was previously reported. This stochastic modeling was aimed at quantitatively assessing human exposure to ESBL/AmpC bearing E. coli through lettuce attributable to irrigation water and subsequent horizontal gene transfer. Modular process risk approach was used for the quantitative exposure assessment and models were constructed in Ms. Excel spreadsheet with farm to consumption chain accounted for by primary production, processing, retail and consumer storage. Probability distributions were utilised to take into account the variability of the exposure estimates. Exposure resulting from ESBL/AmpC positive E. coli and gene transfer was taken into account. Monte Carlo simulation was carried out using @Risk software followed by sensitivity and scenario analysis to assess most effective single or combinations of mitigation strategies for the ESBL/AmpC positive E. coli events from farm to fork. Three percent of South African lettuce consumers are exposed to lettuce contaminated with about 106.4±106.7 (95% CI: 105.1-107) cfu of ESBL/AmpC positive E. coli per serving. The contribution of originally positive isolates and conjugative genetic transfer was 106±106.7 (95% CI: 105-107) and 105.2±105.6 (95% CI: 103.9-105.8) cfu per serving respectively. Proportion of ESBL/AmpC positive E. coli (Spearman's correlation coefficient (ρ)=0.85), conjugative gene transfer (ρ=0.05-0.14), washing in chlorine water (ρ=0.18), further rinsing (ρ=0.15), and prevalence of E. coli in irrigation water (ρ=0.16) had highest influence on consumer exposure. The most effective single methods in reducing consumer exposure were reduction in irrigation water microbial quality variation (87.4% reduction), storage period (49.9-87.4% reduction) and growth rate reduction by 75% (90% reduction). Reduction in growth rate together with storage time (92.1-99.4%) and reduction in storage time combined with E. coli concentration in irrigation water (95-96% reduction) were most effective combinations of mitigation measures. The high variation in exposure reflected the high irrigation water quality variation. The exposure levels may impose higher consumer risk than acceptable for irrigation water risk. E. coli contamination and growth related measures, as well as measures to reduce contamination with antimicrobial resistant E. coli from lettuce production environment are recommended. This exposure model could form a basis for the development of similar models assessing the impact of contaminated irrigation water and gene transfer in other microbial hazards, antimicrobial resistance types and fresh produce types.

Efficacy of various sanitizers against Salmonella during simulated commercial packing of tomatoes.

Chemical sanitizers are usually added to dump tank water to minimize cross-contamination during tomato packing. However, the efficacy of sanitizers continues to be questioned. This study assessed the ability of six commonly used sanitizers (40 ppm of peroxyacetic acid, 40 ppm of mixed peracid, 40 ppm of available chlorine alone or acidified to pH 6.0 with citric acid or T-128, and electrolyzed water containing 40 ppm of available chlorine at pH 6.7) to reduce Salmonella on tomatoes, in wash water, and on equipment surfaces using a pilot-scale processing line. Red round tomatoes (11.3 kg) were dip inoculated to contain Salmonella at ∼6 log CFU/g, air dried for 2 h, treated for 2 min in a 3.3-m-long dump tank and then dried on a roller conveyor, with sanitizer-free water serving as the control. Tomato and water samples were collected at 15-s intervals during washing with additional dump tank, water tank, and roller conveyor surface samples collected after washing. All samples were appropriately neutralized, diluted, and surface plated on Trypticase soy agar containing 0.6% yeast extract, 0.05% ferric ammonium citrate, and 0.03% sodium thiosulfate with or without membrane filtration to enumerate Salmonella. All six sanitizer treatments were more efficacious than the water control (P ≤ 0.05), with chlorine plus citric acid yielding the greatest Salmonella reduction on tomatoes (3.1 log CFU/g). After processing, all sanitizer wash solutions contained significantly lower (P ≤ 0.05) levels of Salmonella than the water control (3.0 log CFU/ml). The four chlorine-based sanitizer treatments yielded significantly lower Salmonella populations (P ≤ 0.05) in the wash solution compared with peroxyacetic acid and mixed peracid. After processing with sanitizers, Salmonella populations decreased to nondetectable levels (<0.2 log CFU/100 cm(2) ) on the equipment surfaces.

Comparative study of disinfectants for use in low-cost gravity driven household water purifiers.

Point-of-use (POU) gravity-driven household water purifiers have been proven to be a simple, low-cost and effective intervention for reducing the impact of waterborne diseases in developing countries. The goal of this study was to compare commonly used water disinfectants for their feasibility of adoption in low-cost POU water purifiers. The potency of each candidate disinfectant was evaluated by conducting a batch disinfection study for estimating the concentration of disinfectant needed to inactivate a given concentration of the bacterial strain Escherichia coli ATCC 11229. Based on the concentration of disinfectant required, the size, weight and cost of a model purifier employing that disinfectant were estimated. Model purifiers based on different disinfectants were compared and disinfectants which resulted in the most safe, compact and inexpensive purifiers were identified. Purifiers based on bromine, tincture iodine, calcium hypochlorite and sodium dichloroisocyanurate were found to be most efficient, cost effective and compact with replacement parts costing US$3.60-6.00 for every 3,000 L of water purified and are thus expected to present the most attractive value proposition to end users.

Risk reduction assessment of waterborne Salmonella and Vibrio by a chlorine contact disinfectant point-of-use device.

Unsafe drinking water continues to burden developing countries despite improvements in clean water delivery and sanitation, in response to Millennium Development Goal 7. Salmonella serotype Typhi and Vibrio cholerae bacteria can contaminate drinking water, causing waterborne typhoid fever and cholera, respectively. Household water treatment (HWT) systems are widely promoted to consumers in developing countries but it is difficult to establish their benefits to the population for specific disease reduction. This research uses a laboratory assessment of halogenated chlorine beads treating contaminated water to inform a quantitative microbial risk assessment (QMRA) of S. Typhi and V. cholerae disease in a developing country community of 1000 people. Laboratory challenges using seeded well water resulted in log10 reductions of 5.44 (± 0.98 standard error (SE)) and 6.07 (± 0.09 SE) for Salmonella serotype Typhimurium and V. cholerae, respectively. In well water with 10% sewage and seeded bacteria, the log10 reductions were 6.06 (± 0.62 SE) and 7.78 (± 0.11 SE) for S. Typhimurium and V. cholerae, respectively. When one infected individual was contributing to the water contamination through fecal material leaking into the water source, the risk of disease associated with drinking untreated water was high according to a Monte Carlo analysis: a median of 0.20 (interquartile range [IQR] 0.017-0.54) for typhoid fever and a median of 0.11 (IQR 0.039-0.20) for cholera. If water was treated, risk greatly decreased, to a median of 4.1 × 10(-7) (IQR 1.6 × 10(-8) to 1.1 × 10(-5)) for typhoid fever and a median of 3.5 × 10(-9) (IQR 8.0 × 10(-10) to 1.3 × 10(-8)) for cholera. Insights on risk management policies and strategies for public health workers were gained using a simple QMRA scenario informed by laboratory assessment of HWT.

Modelling growth of Escherichia coli O157:H7 in fresh-cut lettuce submitted to commercial process conditions: chlorine washing and modified atmosphere packaging.

Fresh-cut iceberg lettuce inoculated with Escherichia coli O157:H7 was submitted to chlorine washing (150 mg/mL) and modified atmosphere packaging on laboratory scale. Populations of E. coli O157:H7 were assessed in fresh-cut lettuce stored at 4, 8, 13 and 16 °C using 6-8 replicates in each analysis point in order to capture experimental variability. The pathogen was able to grow at temperatures ≥8 °C, although at low temperatures, growth data presented a high variability between replicates. Indeed, at 8 °C after 15 days, some replicates did not show growth while other replicates did present an increase. A growth primary model was fitted to the raw growth data to estimate lag time and maximum growth rate. The prediction and confidence bands for the fitted growth models were estimated based on Monte-Carlo method. The estimated maximum growth rates (log cfu/day) corresponded to 0.14 (95% CI: 0.06-0.31), 0.55 (95% CI: 0.17-1.20) and 1.43 (95% CI: 0.82-2.15) for 8, 13 and 16 °C, respectively. A square-root secondary model was satisfactorily derived from the estimated growth rates (R(2) > 0.80; Bf = 0.97; Af = 1.46). Predictive models and data obtained in this study are intended to improve quantitative risk assessment studies for E. coli O157:H7 in leafy green products.

Integrative assessment of chlorine-induced acute lung injury in mice.

The genetic basis for the underlying individual susceptibility to chlorine-induced acute lung injury is unknown. To uncover the genetic basis and pathophysiological processes that could provide additional homeostatic capacities during lung injury, 40 inbred murine strains were exposed to chlorine, and haplotype association mapping was performed. The identified single-nucleotide polymorphism (SNP) associations were evaluated through transcriptomic and metabolomic profiling. Using ≥ 10% allelic frequency and ≥ 10% phenotype explained as threshold criteria, promoter SNPs that could eliminate putative transcriptional factor recognition sites in candidate genes were assessed by determining transcript levels through microarray and reverse real-time PCR during chlorine exposure. The mean survival time varied by approximately 5-fold among strains, and SNP associations were identified for 13 candidate genes on chromosomes 1, 4, 5, 9, and 15. Microarrays revealed several differentially enriched pathways, including protein transport (decreased more in the sensitive C57BLKS/J lung) and protein catabolic process (increased more in the resistant C57BL/10J lung). Lung metabolomic profiling revealed 95 of the 280 metabolites measured were altered by chlorine exposure, and included alanine, which decreased more in the C57BLKS/J than in the C57BL/10J strain, and glutamine, which increased more in the C57BL/10J than in the C57BLKS/J strain. Genetic associations from haplotype mapping were strengthened by an integrated assessment using transcriptomic and metabolomic profiling. The leading candidate genes associated with increased susceptibility to acute lung injury in mice included Klf4, Sema7a, Tns1, Aacs, and a gene that encodes an amino acid carrier, Slc38a4.

A nose for trouble--the role of off-flavours in assuring safe drinking water.

A detailed review of drinking water disease outbreaks over the past 30 years in 15 affluent countries produced over 70 case studies, some involving fatalities, which revealed a number of common factors in these disasters. Some of these outbreaks involved off-flavours, either as a driver for reducing disinfection and making the system vulnerable to pathogenic contamination or as an early warning of contamination that was not responded to with sufficient urgency or efficiency to avoid a disease outbreak. The characteristics of these outbreaks are recounted and the important link they reveal between aesthetically pleasing drinking water and safety is documented. Our analysis of common features in drinking water outbreaks also supports an argument that the failure of a water utility to be concerned about aesthetic factors makes such water supplies an inherently greater health risk for their consumers.

Effect of chlorine as substituent on the photochemistry and acid-base properties of beta-carboline alkaloids.

The UV-absorption, fluorescence excitation and emission spectra of the 6-chloro-, 8-chloro-, 6,8-dichloro-derivatives of nor-harmane, harmane and harmine and the 8-chloro-derivative of harmol were studied. These studies were performed in EtOH and in EtOH+1% perchloric acid solutions (pa). Furthermore, fluorescence quantum yields (phi(f)) in both media and in acetonitrile and acetonitrile + 1% perchloric acid solutions at 298 K were measured. The HOMO and LUMO energy, the positions (lambda(max)) and oscillator strength (f) of the 1S1 <-- 1S0 band for all the neutral and protonated beta-carbolines studied were calculated and compared with the experimental data. The pK(a) values in aqueous solution for for 6-chloro-, 8-chloro- and 6,8-dichloro-nor-harmane, harmane and harmine and 8-chloro-harmol were spectrophotometrically measured (pK(a(H2O)). The change of the acid-base character of these compounds on going from the ground state (pK(a)) to the first electronic excited singlet state (pKa*) as DeltapKa = pKa*-pKa, in ethanol solution at 298 K were calculated (DeltapK(a(EtOH))). Ground-state proton affinity (PA) for all the compounds studied defined as minus the enthalpy change of the reaction M + H(+) --> MH+ (gas state) were calculated. Basicity relative to pyridine (DeltaH(rPy)) defined as the enthalpy change of the isodesmic reaction MH(+) + Py --> M + PyH+ in gas state and in water solution, were also calculated (ab initio calculations). The effect of chlorine as substituent on the photochemistry and acid-base properties of the beta-carboline alkaloids is discussed.

Assessment of the ability of the bioelectric effect to eliminate mixed-species biofilms.

Microbes have been able to persist in water distribution systems through the development of multicellular communities known as biofilms. This study evaluated the usefulness of the bioelectric effect for the elimination of water distribution system biofilms from annular reactors. The bioelectric effect did not have any bactericidal action either alone or when coupled with free chlorine.