Degradation of Azithromycin from aqueous solution using Chlorine-ferrous- oxidation: ANN-GA modeling and Daphnia magna biotoxicity test assessment.

Azithromycin (AZM), an antibacterial considered one of the most consumed drugs, especially during the period against the Covid 19 pandemic, and it is one of the persistent contaminants that can be released into aquatic ecosystems. The purpose of this study is to determine the efficacy of a Fenton-like process (chlorine/iron) for the degradation of AZM in an aqueous medium by determining the impact of several factors (the initial concentration of (FeSO4, NaClO, pollutant), and the initial pH) on the degradation rate. The Response Surface Methodology (RSM) based on the Box-Wilson design as well as the Artificial Neural Network (ANN) modeling combined with a genetic algorithm (GA) approaches were used to determine the optimal levels of the selected variables and the optimal rate of degradation. The quadratic model of multi-linear regression developed indicated that the optimal conditions were a concentration of chlorine of 600 µM, the concentration of AZM is 32.8 mg/L, the mass of the catalyst FeSO4 is 3.5 mg and a pH of 2.5, these optimal values gave a predicted and experimental yield of 64.05% and 70% respectively, the lack of fit test in RSM modeling (F0 = 3.31 which is inferior to Fcritic (0.05, 10.4) = 5.96) indicates that the true regression function is not linear therefore, the ANN-GA modeling as non-linear regression indicated that the optimal conditions were a concentration of chlorine of 256 µM, the concentration of AZM is 5 mg/L, the mass of the catalyst FeSO4 is 9.5 mg and a pH of 2.8, these optimal values gave a predicted and experimental yield of 79.69% and close to 80% respectively, Furthermore, biotoxicity tests were conducted to confirm the performance of our process using bio-indicators called daphnia (Daphnia magna), which demonstrated the efficacy of the like-Fenton process after 4 h of degradation.

Reaction of diazepam and related benzodiazepines with chlorine. Kinetics, transformation products and in-silico toxicological assessment.

In this work, the reaction of four benzodiazepines (diazepam, oxazepam, nordazepam and temazepam) during water chlorination was studied by means of liquid chromatography-quadrupole-time of flight-mass spectrometry (LC-QTOF-MS). For those compounds that showed a significant degradation, i.e. diazepam, oxazepam and nordazepam, parameters affecting to the reaction kinetics (pH, chlorine and bromide level) were studied in detail and transformation products were tentatively identified. The oxidation reactions followed pseudofirst-order kinetics with rate constants in the range of 1.8-42.5 M-1 s-1, 0.13-1.16 M-1 s-1 and 0.04-20.4 M-1 s-1 corresponding to half-life values in the range of 1.9-146 min, 1.8-87 h and 2.5-637 h for oxazepam, nordazepam and diazepam, respectively, depending of the levels of studied parameters. Chlorine and pH affected significantly the reaction kinetics, where an increase of the pH resulted into a decrease of the reaction rate, whereas higher chlorine dosages led to faster kinetics, as expected in this case. The transformation of the studied benzodiazepines occurs mainly at the 1,4-diazepine 7-membered-ring, resulting in ring opening to form benzophenone derivatives or the formation of a 6-membered pyrimidine ring, leading to quinazoline derivatives. The formation of these by-products was also tested in real surface water samples observing kinetics of oxazepam degradation slower in river than in creek water, while the degradation of the two other benzodiazepines occurred only in the simpler sample (creek water). Finally, the acute and chronical toxicity and mutagenicity of precursors and transformation products were estimated using quantitative structure-activity relationship (QSAR) software tools: Ecological Structure Activity Relationships (ECOSAR) and Toxicity Estimation Software Tool (TEST), finding that some transformation products could be more toxic/mutagenic than the precursor drug, but additional test would be needed to confirm this fact.

Chlorination-induced genotoxicity in the mussel Perna viridis: assessment by single cell gel electrophoresis (comet) assay.

Mussels are important fouling organisms in the cooling water systems of coastal power plants. Continuous low-dose chlorination (CLDC) is being practiced as an effective method to control mussel biofouling in power plant cooling water systems. CLDC effectively controls mussel fouling by discouraging larval settlement rather than by killing the larvae or adults. Mussels are an integral part of the natural benthic community in the receiving water body where the coolant water is discharged. Hence, from a toxicological point of view, they can serve as both target and non-target organisms. Previous researchers have indicated that chlorine residual, rather than elevated temperature, can be the major stress factor in the effluents released from coastal power plants. However, very little data are available on the sub-lethal effects of low level chlorination on representative benthic fauna. In this study, we used native and transplanted mussels (Perna viridis) to study lethal and sub-lethal effects of chlorination in the cooling water circuit of an operating power plant. Experiments involving comet assay suggested that CLDC can cause DNA damage in treated mussels. However, activation of DNA repair appeared to get initiated after the accrued damage reached a threshold. The results indicate that, at chlorine residual levels observed at the discharge point, exposure to chlorinated effluents is unlikely to cause significant genetic damage to mussels in the recipient water body.

Development and assessment of countermeasure formulations for treatment of lung injury induced by chlorine inhalation.

Chlorine is a commonly used, reactive compound to which humans can be exposed via accidental or intentional release resulting in acute lung injury. Formulations of rolipram (a phosphodiesterase inhibitor), triptolide (a natural plant product with anti-inflammatory properties), and budesonide (a corticosteroid), either neat or in conjunction with poly(lactic:glycolic acid) (PLGA), were developed for treatment of chlorine-induced acute lung injury by intramuscular injection. Formulations were produced by spray-drying, which generated generally spherical microparticles that were suitable for intramuscular injection. Multiple parameters were varied to produce formulations with a wide range of in vitro release kinetics. Testing of selected formulations in chlorine-exposed mice demonstrated efficacy against key aspects of acute lung injury. The results show the feasibility of developing microencapsulated formulations that could be used to treat chlorine-induced acute lung injury by intramuscular injection, which represents a preferred route of administration in a mass casualty situation.

Assessment of locomotion in chlorine exposed mice by computer vision and neural networks.

Assessment of locomotion following exposure of animals to noxious or painful stimuli can offer significant insights into underlying mechanisms of injury and the effectiveness of various treatments. We developed a novel method to track the movement of mice in two dimensions using computer vision and neural network algorithms. By using this system we demonstrated that mice exposed to chlorine (Cl(2)) gas developed impaired locomotion and increased immobility for up to 9 h postexposure. Postexposure administration of buprenorphine, a common analgesic agent, increased locomotion and decreased immobility times in Cl(2)- but not air-exposed mice, most likely by decreasing Cl(2)-induced pain. This method can be adapted to assess the effectiveness of various therapies following exposure to a variety of chemical and behavioral noxious stimuli.

Electrochemical disinfection for ballast water management: technology development and risk assessment.

Ballast water is essential in maintaining the balance and structural integrity of ships during voyage. However, it has created biological invasion threats to the ocean environment. An innovative electrochemical technology was developed in this study. The microorganisms regulated by the International Maritime Organization (D2) were used as the target organisms. It was found that the required energy to meet the D2 was below 0.006 kWh/m3. The size of disinfector (m3) was about 0.5% of treatment flow rate (m3/h). The complete disappearance of chlorine in seawater was achieved after three days. The ballast tank corrosion was not worsened due to the application of technology. The ecotoxicity studies showed no toxic effect on fish, invertebrate, and algae. Finally, the environmental risk assessment showed the treated water did not pose threats to the environment. It can therefore be concluded that the technology provides a cost-effective and environmental friendly solution to ballast water management.

Chlorine truck attack consequences and mitigation.

We develop and apply an integrated modeling system to estimate fatalities from intentional release of 17 tons of chlorine from a tank truck in a generic urban area. A public response model specifies locations and actions of the populace. A chemical source term model predicts initial characteristics of the chlorine vapor and aerosol cloud. An atmospheric dispersion model predicts cloud spreading and movement. A building air exchange model simulates movement of chlorine from outdoors into buildings at each location. A dose-response model translates chlorine exposures into predicted fatalities. Important parameters outside defender control include wind speed, atmospheric stability class, amount of chlorine released, and dose-response model parameters. Without fast and effective defense response, with 2.5 m/sec wind and stability class F, we estimate approximately 4,000 (half within ∼10 minutes) to 30,000 fatalities (half within ∼20 minutes), depending on dose-response model. Although we assume 7% of the population was outdoors, they represent 60-90% of fatalities. Changing weather conditions result in approximately 50-90% lower total fatalities. Measures such as sheltering in place, evacuation, and use of security barriers and cryogenic storage can reduce fatalities, sometimes by 50% or more, depending on response speed and other factors.

Follow-up assessment of health consequences after a chlorine release from a train derailment--Graniteville, SC, 2005.

INTRODUCTION: After a train derailment released chlorine gas in Graniteville, South Carolina, in 2005, a multiagency team performed an epidemiologic assessment of chlorine exposure and resulting health effects. Five months later, participants were resurveyed to determine their health status and needs and to assist in planning additional interventions in the community. METHODS: Questionnaires were mailed to 279 patients interviewed in the initial assessment; follow-up telephone calls were made to nonresponders. The questionnaire included questions regarding duration of symptoms experienced after exposure and a posttraumatic stress disorder (PTSD) assessment tool. RESULTS: Ninety-four questionnaires were returned. Seventy-six persons reported chronic symptoms related to the chlorine exposure, 47 were still under a doctor's care, and 49 were still taking medication for chlorine-related problems. Agreement was poor between the first and second questionnaires regarding symptoms experienced after exposure to the chlorine (κ=0.30). Forty-four respondents screened positive for PTSD. PTSD was associated with post-exposure hospitalization for three or more nights [relative risk (RR) = 1.7; 95% confidence interval (CI)=1.1-2.6] and chronic symptoms (RR=9.1; 95% CI=1.3-61.2), but not with a moderate-to-extreme level of chlorine exposure (RR=1.2; 95% CI=0.8-1.8). CONCLUSIONS: Some victims of this chlorine exposure event continued to experience physical symptoms and continued to require medical care 5 months later. Chronic mental health symptoms were prevalent, especially among persons experiencing the most severe or persistent physical health effects. Patients should be interviewed as soon as possible after an incident because recall of acute symptoms experienced can diminish within months.

Spatial variations of human health risk associated with exposure to chlorination by-products occurring in drinking water.

During disinfection, chlorine reacts with organic matter present in drinking water and forms various undesirable chlorinated by-products (CBPs). This paper describes a study of the spatial variability of human health risk (i.e., cancer effects) from CBP exposure through drinking water in a specific region. The region under study involves nine drinking water distribution systems divided into several zones based on their characteristics. The spatial distribution of cancer risk (CR) was estimated using two years of data (2006-2008) on various CBP species. In this analysis, trihalomethanes (THMs) and haloacetic acids (HAAs) served as surrogates for CBPs. Three possible routes of exposure (i.e., via ingestion, inhalation and dermal contact) were considered for each selected compound. The cancer risk assessment involved estimating a unit risk (R(T)) in each zone of the selected distribution systems. A probabilistic analysis based on Monte Carlo simulations was employed. Risk assessment results showed that cancer risk varied between systems, but also within individual systems. As a result, the population of the same region was not exposed to the same risk associated with CBPs in drinking water. Unacceptable levels (i.e., R(T) > 10(-4)) for the estimated CR were determined for several zones in the studied region. This study demonstrates that a spatial-based analysis performed to represent the spatial distribution of risk estimates can be helpful in identifying suitable risk management strategies. Suggestions for improving the risk analysis procedure are also presented.

What's in the pool? A comprehensive identification of disinfection by-products and assessment of mutagenicity of chlorinated and brominated swimming pool water.

BACKGROUND: Swimming pool disinfectants and disinfection by-products (DBPs) have been linked to human health effects, including asthma and bladder cancer, but no studies have provided a comprehensive identification of DBPs in the water and related that to mutagenicity. OBJECTIVES: We performed a comprehensive identification of DBPs and disinfectant species in waters from public swimming pools in Barcelona, Catalonia, Spain, that disinfect with either chlorine or bromine and we determined the mutagenicity of the waters to compare with the analytical results. METHODS: We used gas chromatography/mass spectrometry (GC/MS) to measure trihalomethanes in water, GC with electron capture detection for air, low- and high-resolution GC/MS to comprehensively identify DBPs, photometry to measure disinfectant species (free chlorine, monochloroamine, dichloramine, and trichloramine) in the waters, and an ion chromatography method to measure trichloramine in air. We assessed mutagenicity with the Salmonella mutagenicity assay. RESULTS: We identified > 100 DBPs, including many nitrogen-containing DBPs that were likely formed from nitrogen-containing precursors from human inputs, such as urine, sweat, and skin cells. Many DBPs were new and have not been reported previously in either swimming pool or drinking waters. Bromoform levels were greater in brominated than in chlorinated pool waters, but we also identified many brominated DBPs in the chlorinated waters. The pool waters were mutagenic at levels similar to that of drinking water (approximately 1,200 revertants/L-equivalents in strain TA100-S9 mix). CONCLUSIONS: This study identified many new DBPs not identified previously in swimming pool or drinking water and found that swimming pool waters are as mutagenic as typical drinking waters.

Ecotoxicological assessment of surfactants in the aquatic environment: combined toxicity of docusate sodium with chlorinated pollutants.

The toxicity of perfluorinated surfactants perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA), perfluorobutane sulfonate (PFBS) and PF-656 as well as the sulfosuccinate surfactant docusate sodium has been examined using two bioluminescence inhibition assays based on the marine bacterium Vibrio fischeri and the self-luminescent cyanobacterial recombinant strain Anabaena CPB4337. We also determined multigenerational toxicity towards the growth of the algae Pseudokirchneriella subcapitata. With EC(50) values in the 43-75 mg/L range, docusate sodium exhibited a higher toxicity towards the three organisms than PFOS, PFOA, PF-656 and PFBS. We investigated the toxicological interactions of the most toxic surfactant, docusate sodium, with two chlorinated compounds, triclosan and 2,4,6-trichlorophenol (TCP), in their binary and ternary mixtures using the method of the combination index based on the median-effect equation. In general, the binary mixture of the chlorinated compounds triclosan and TCP exhibited antagonism, which was stronger for the growth test using P. subcapitata. Except for the green alga, the binary mixtures of docusate sodium with TCP or triclosan showed synergism at medium to high effect levels; the synergistic behaviour predominating in the ternary mixture and in the three tested species. This result highlights the potential toxicological risk associated with the co-occurrence of this surfactant with other pollutants.

Single cell level microalgal ecotoxicity assessment by confocal microscopy and digital image analysis.

In ecotoxicological studies involving environmental contaminants, rapid and multi-parametric optical detection based methods have definite advantages over traditional growth inhibition assays. In this context, a confocal laser scanning microscopy (CLSM) based method to assess ecotoxicity arising out of biocide insult to marine microalgae is reported. Using this technique, the effect of in-use concentrations of chlorine (an oxidizing biocide) on a marine diatom (Cocconeis scutellum Ehrenb) was determined based on inhibition of chlorophyll autofluorescence and esterase activity (probed by fluorescein diacetate (FDA) staining). Determination of mean fluorescence intensity (MFI) per cell by collecting auto-fluorescence from single cells in x, y and z dimensions permitted reproducible toxicity evaluation at single-cell level. Chlorine-induced inhibition of autofluorescence in laboratory cultures was dose-dependent. Additional data on metabolic activity of the diatom cells following chlorine exposure was collected by FDA staining. Our results demonstrate that chlorine, an antifouling biocide commonly used in cooling water systems, causes significant reduction in chlorophyll autofluorescence and esterase activity in diatoms in short-term exposure experiments. Tests employing multiple organisms and multiple toxicity endpoints are superior to standard algal growth inhibition assays for they provide a better understanding of algal-algal interactions and real impact in the environment. The combined autofluorescence-FDAtechnique described here is rapid and has clear advantages in terms of using environmentally relevant toxicant and cell concentrations. Additional microalgal species and toxicity end points can be employed in order to develop multi-species and multiparameter bioassay using confocal microscopy.

Application of a virological tracer method for the assessment of pathogen removal by physicochemical treatment and chemical disinfection.

Coxsackie B3 (CoxB3) virus was used as a virological tracer for an assessment of the efficiency of pathogen removal by several typical physicochemical treatment and chemical disinfection processes, such as coagulation-filtration, ultra-filtration, and disinfection using chlorine and ozone, with regard to the pathogenic quality of the treated domestic wastewater for reuse purposes. The CoxB3 virus was seeded to sterilized secondary effluent to make a raw water of known pathogenic level. After applying the raw water to each treatment or disinfection process, the residual virus in the finished water was concentrated, and virus assay was carried out by the Tissue Culture Infectious Dose technique. TCID50 was used as an indicative parameter of CoxB3 virus in the raw and treated water. Parallel experiments were also conducted to evaluate the effectiveness of each process for the removal of coliform bacteria. It was noticed from the experiment that both coagulation-filtration and ultrafiltration could achieve substantial removal of TCID50 at about the same level (2-log removal). However, the effect of the two processes on the removal of coliform bacteria was much different: 2-log removal by coagulation-filtration and 4 to 5-log removal by ultrafiltration. The TCID50 removal correlates more closely with the removal of turbidity than that of coliform bacteria. Chlorine was found to be effective in coliform removal but almost had no effect on TCID50. As ozone was applied, a high removal of both coliform bacteria and TCID50 could be obtained.

Pharmacokinetic-pharmacodynamic models for categorical toxicity data.

We propose a pharmacokinetic-pharmacodynamic (PK/PD) model (with possibly different choices for the PD link) for categorical toxicity data analysis. This is extension of the one-comportment model that applies to toxic endpoints categorised by grades (e.g., benign, mild, severe, and very severe). The model assumes that the area under the curve (AUC) of the internal quantity of the chemical substance is the critical dose-metric that drives the acute toxic phenomenon. That model handles time-varying concentrations and takes into account follow-up time, i.e., time at which effects are observed. Moreover the model bridges mechanistically based dose-response models and standard dose-response models, retaining the advantages of both. We use Markov chain-Monte Carlo (MCMC) simulations to fit the model to mortality data for mice exposed to chlorine, rats exposed to ammonia, and categorical data (different severity levels) from acute exposures of rats and humans to hydrogen sulfide.

Cytotoxicity assessment of chlorinated bacteria in water using the RNA synthesis inhibition method.

Chlorination of drinking water containing organic materials is known to generate toxic by-products. We suggested that such compounds may also be produced by interactions between chlorine and bacteria present in water. To confirm this hypothesis, a method based on RNA synthesis inhibition of HeLa S3 human cells in the presence of toxic compounds was applied. This method is rapid and highly sensitive since the concentration of the samples is not required. Furthermore, it was shown to be a suitable method for measurement of the cytotoxicity of water. Aeromonas hydrophila suspensions, prepared with pyrodistilled water, devoid of any organic material, were chlorinated for a definite contact time. HeLa S3 cells were incubated (20 h, 37 degrees C) in a culture medium prepared with the chlorinated bacteria suspensions. The rate of incorporation of 3H uridine into RNA was used as a measure of RNA synthesis and was evaluated in the presence and absence of chlorinated bacteria suspension. This study showed that chlorinated bacteria suspensions are cytotoxic. We observed that 0.22 microm filters retain cytotoxic compounds but 0.45 microm filters did not. Chlorine concentration and bacteria level influence the cytotoxicity. First, the toxicity level increases with chlorine concentration, then it decreases when chlorine concentration is too high. On another hand, a dose effect relationship between bacteria concentration and cytotoxicity was established.

Monte Carlo uncertainty analysis of a diffusion model for the assessment of halogen gas exposure during dosing of brominators.

Monte Carlo simulation was incorporated into a diffusion-based exposure assessment model for the estimation of worker exposure to halogen gases during dosing of 500-lb sacks of a bromine-based biocide (BCDMH) into brominators. Indoor and outdoor dosing scenarios were modeled for small and large brominators. The diffusion model used describes a concentration gradient of halogen as a function of distance and time from the source instead of ascribing worst-case single point value estimates to the variables used in the diffusion model. Monte Carlo simulation was used to describe a distribution of values for each appropriate model variable. Using a personal computer and Monte Carlo simulation software, 10,000 iterations of the diffusion model were performed for four different dosing scenarios using random and independent samples from the distributions entered. The corresponding output distributions of predicted exposures were then calculated and displayed graphically for each scenario. The results of the Monte Carlo simulation predict that outdoor dosing of either small or large brominators with BCDMH is highly unlikely to result in an exceedance of the working occupational exposure limit for total halogen. In most ambient wind speed conditions, diffusion prevents appreciable airborne exposure to workers in the immediate vicinity of the brominator. Although relatively uncommon, dosing of brominators indoors in the assumed absence of local exhaust ventilation may generate airborne concentrations of total halogen that exceed the working short-term occupational exposure limit. Although very limited and inconclusive, field trial monitoring of BCDMH transfer operations indoors resulted in halogen concentrations well within the distribution of concentrations predicted by the Monte Carlo simulation of the diffusion model.

The chlorine controversy.

Environmental groups have called for a phaseout of the use of chlorine in industrial chemistry on the grounds that such use inevitably leads to the production of persistent, bioaccumulative toxins. This policy prescription is based on an application of the Precautionary Principle which holds that industry should demonstrate that its products and processes are safe before allowing releases of product or waste into the environment. The chemical industry has rejected the call to phaseout chlorine, and claims that available data on either the environmental exposure pathways or the toxicology of chlorinated compounds are inadequate to support responsible decision-making. Industry leaders have called for environmental policy to be based on sound science, comparative risk assessment, and cost-benefit analysis. The controversy over chlorine shows that the U.S. environmental policy-making framework may no longer be capable of producing solutions to environmental problems that satisfy important constituencies within our society.

Drinking water mutagenicity in past exposure assessment of the studies on drinking water and cancer: application and evaluation in Finland.

The assessment of past exposure is a prerequisite to all epidemiological studies on drinking water and cancer. In this study the past exposure assessment of drinking water carcinogenicity was done in terms of the drinking water mutagenicity estimated from historical water parameters and compared with the methods used previously in past exposure assessments in studies on drinking water and cancer. The method was applied in 56 municipalities in Finland. The comparison of different methods in past exposure assessment suggests some advantages for the method presented as it allows a quantitative exposure assessment based on historical information on drinking water mutagenicity. Nevertheless the relevance of the method is with respect to the role of mutagenicity in carcinogenicity and the water type in question.