Low Ozone Doses Research Articles

Low ozone concentrations do not exert cytoprotective effects on tamoxifen-treated breast cancer cells in vitro.

Medical treatment with low ozone concentrations proved to exert therapeutic effects in various diseases by inducing a cytoprotective antioxidant response through the nuclear factor erythroid derived-like 2 (Nrf2) transcription factor pathway. Low ozone doses are increasingly administered to oncological patients as a complementary treatment to mitigate some adverse side-effects of antitumor treatments. However, a widespread concern exists about the possibility that the cytoprotective effect of Nrf2 activation may confer drug resistance to cancer cells or at least reduce the efficacy of antitumor agents. In this study, the effect of low ozone concentrations on tamoxifen-treated MCF7 human breast cancer cells has been investigated in vitro by histochemical and molecular techniques. Results demonstrated that cell viability, proliferation and migration were generally similar in tamoxifen-treated cells as in cells concomitantly treated with tamoxifen and ozone. Notably, low ozone concentrations were unable to overstimulate the antioxidant response through the Nfr2 pathway, thus excluding a possible ozone-driven cytoprotective effect that would lead to increased tumor cell survival during the antineoplastic treatment. These findings, though obtained in an in vitro model, support the hypothesis that low ozone concentrations do not interfere with the tamoxifen-induced effects on breast cancer cells.

N-Nitrosodimethylamine formation from anthropogenic nitrogenous compounds during preozonation and post-chloramination with characteristic low treatment dose.

One effective option to minimize N-nitrosodimethylamine (NDMA) in finished drinking water is to identify and control its precursors. However, previous works to identify significant precursors use formation potential (FP) tests using high doses to assure the maximum NDMA formation rather than the NDMA formation in finished waters. In this study, we applied characteristic low treatment doses of ozone (O3)-to-dissolved organic carbon (DOC) of target compounds of 0.8mg/mg and NH2Cl of 2.5 ± 0.2mg Cl2/L to evaluate the NDMAFP yields of organic compounds bearing N,N-dimethylamine (DMA) and N,N-dimethylhydrazine (DMH) during preozonation and post-chloramination. The results in pH-buffered Milli-Q water showed a significant decrease from ≤ 52% to non-detectable levels in the O3-NDMAFP yields of O3-reactive precursors (i.e., DMH-like compounds) after preozonation and post-chloramination. Similarly, a significant decrease from 0.5 to 12% to nonquantifiable levels was observed for the NH2Cl-NDMAFP yields of NH2Cl-reactive precursors; however, the NH2Cl-NDMAFP yields of N,N-dimethylbenzylamine (DMBzA)-like compounds only decreased from ~ 110 to ≤ 43%, suggesting that these compounds could contribute to NH2Cl-NDMAFPs even after preozonation. The effect of the matrix in sewage-effluent and lake water samples varied and was specific for precursors; for example, the O3-NDMAFP yield of 1,1,1',1'-tetramethyl-4,4'-(methylene-di-p-phenylene) disemicarbazide (TMDS), an important O3-reactive NDMA precursor, did not significantly decrease when tested in sewage-effluent samples. Based on the previous occurrence concentration of TMDS in sewage samples, we estimated an NDMAFP of ~ 315 ng/L. This estimate exceeds the guidance concentrations of NDMA (3-100ng/L), highlighting the importance of TMDS and its related compounds for NDMA formation.

Accumulation of Alpha-Synuclein and Increase in the Inflammatory Response in the substantia nigra, Jejunum, and Colon in a Model of O3 Pollution in Rats.

This work aimed to study the effect of repeated exposure to low doses of ozone on alpha-synuclein and the inflammatory response in the substantia nigra, jejunum, and colon. Seventy-two male Wistar rats were divided into six groups. Each group received one of the following treatments: The control group was exposed to air. The ozone groups were exposed for 7, 15, 30, 60, and 90 days for 0.25 ppm for four hours daily. Afterward, they were anesthetized, and their tissues were extracted and processed using Western blotting, immunohistochemistry, and qPCR. The results indicated a significant increase in alpha-synuclein in the substantia nigra and jejunum from 7 to 60 days of exposure and an increase in NFκB from 7 to 90 days in the substantia nigra, while in the jejunum, a significant increase was observed at 7 and 15 days and a decrease at 60 and 90 days for the colon. Interleukin IL-17 showed an increase at 90 days in the substantia nigra in the jejunum and increases at 30 days and in the colon at 15 and 90 days. Exposure to ozone increases the presence of alpha-synuclein and induces the loss of regulation of the inflammatory response, which contributes significantly to degenerative processes.

Assessment of an hybrid process coupling ozonation and anodic oxidation in a monophasic configuration

This prospective study assessed a hybrid process that combines ozonation with anodic oxidation (AO/O3) with two boron-doped diamond (BDD) electrodes to treat a model aqueous solution containing 30 ppm of alachlor at pH 5 in a phosphate buffer. An innovative monophasic configuration, operated batch-wise, in which an ozone-stock solution was injected at t0 in the electrochemical reactor, was proposed in order to avoid introduction of ozone-enriched gas bubbles. The alachlor parent molecule was almost entirely degraded within around 150 min of reaction time by anodic oxidation alone, whatever the applied current intensity (200, 500 and 800 mA). The mineralization efficiency varied from 50 % at 200 mA to 80 % at 800 mA after 120 min. The results emphasized that the anodic oxidation was a diffusion-controlled process. Addition of ozone at a low ozone dose of 1.9–2.3 mol of ozone per mol of alachlor allowed to decrease the ozone half-life time 2.7 times with the hybrid process compared to ozonation alone. This faster ozone consumption was concomitant with an enhanced alachlor degradation rate, with a corresponding half-life time around two-times lower. However, owing to the low ozone dose applied and the short ozone lifetime in solution (around 25–30 min), similar mineralization efficiencies were noticed for both AO/O3 and AO processes. Thus, the application of pulse ozone-stock solution re-injections every 20 min to prolong the ozone exposure was assessed to overcome this limitation. However, the effect on the mineralization rate remained small, even if it allows to enhance even more the alachlor parent molecule degradation rate. The low influence of the hybrid process on the mineralization rate was attributed to the production of aliphatic fatty acids by-products that are poorly reactive with hydroxyl radicals, and whose the degradation by AO is the rate limiting step.

Removal characteristics of 53 micropollutants during ozonation, chlorination, and UV/H2O2 processes used in drinking water treatment plant

The removal of 53 emerging micropollutants (MPs), including 10 per- and polyfluorinated substances (PFASs), 25 pharmaceuticals and personal care products (PPCPs), 7 pesticides, 5 endocrine disrupters (EDCs), 3 nitrosamines, and 3 taste and odor compounds (T&Os), by chlorination, ozonation, and UV/H2O2 treatment was examined in deionized water and surface waters used as the raw waters in drinking water treatment plants (DWTPs) in South Korea. The UV/H2O2 treatment was effective in the removal of most MPs, whereas chlorination was selectively effective for 19 MPs, including EDCs (>70 %). MPs containing aromatic ring with electron-donating functional group, or primary and secondary amines were effectively removed by chlorination immediately upon reaction initiation. The removal of MPs by ozonation was generally lower than that of the other two processes at a low ozone dose (1 mg L−1), but higher than chlorination at a high ozone dose (3 mg L−1), particularly for 16 MPs, including T&Os. Compared in deionized water, the removals of MPs in the raw water samples were lower in all three processes. The regression models predicting the rate constants (kobs) of 53 MPs showed good agreement between modeled and measured value for UV/H2O2 treatment (R2 = 0.948) and chlorination (R2 = 0.973), despite using only dissolved organic carbon (DOC) and oxidant concentration as variables, whereas the ozonation model showed a variation (R2 = 0.943). Our results can provide the resources for determining which oxidative process is suitable for treating specific MPs present in the raw waters of DWTPs.

Evaluation of Cryopreserved Ram Sperm with Nano-Ozone Solution and Post-Thaw Life Span by Flow Cytometric Analysis.

Ozone has been used as a therapy tool in medical science for conditions such as ulcers, peritonitis, wounds, and mostly joint problems. Ozone therapy strengthens the resistance to infections by kick-starting antioxidant, anti-inflammatory, and immune modulation systems. Ozone creates a defensive response against oxidative stress in membranes and protects metabolism against reactive oxygen species (ROS). Sperm membranes are one of ROS's main targets; therefore, the cells' cryopreservation process requires more defensive elements for better results. This study aimed to investigate the protective effect of nano-ozone solution (NOS) on ram sperm cryopreservation and the influence of the process on various sperm parameters for post-thaw (0 hour) and postincubation (6 hours) time points. Samples were collected from six Merino rams in the breeding season by electroejaculation five times at 3-day intervals. The study was conducted by cryopreservation of the samples using a tris citric acid-egg yolk-based extender. The samples were subjected to freezing in control and NOS (0.5, 1, and 2 μg/mL nano-ozone supplemented). Post-thaw motility, hypo-osmotic swelling test, acrosome (fluorescein isothiocyanate-conjugated Pisum sativum agglutinin [PSA-FITC]), and DNA integrities (terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL]) were evaluated with a phase-contrast microscope. Mitochondrial membrane potential (MMP) assessments were conducted by JC1-PI dual staining with a flow cytometer. Malondialdehyde and glutathione (GSH) levels were measured by a spectrophotometer. Sperm kinematics were investigated by a computer-assisted sperm analyzer (CASA) at the post-thaw time point. Compared with the control, relatively low doses of NOS (0.5 and 1 μg/mL) yielded better results in many parameters (motility, membrane and acrosomal integrities, MMP, various sperm kinematics, and GSH levels) (p < 0.05). The addition of low ozone doses to cryopreservation extenders improved the results compared with the control group at post-thaw and postincubation time points. Despite the valuable potential of nano-ozone supplementation in ram sperm cryopreservation, this subject requires further investigations with fertility trials soon.

The Role of Ozone as an Nrf2-Keap1-ARE Activator in the Anti-Microbial Activity and Immunity Modulation of Infected Wounds.

Ozone is an allotrope of oxygen, widely known to exert an anti-oxidant potential. The ability of low, controlled and standardized doses of ozone in the ozone adjunct treatment of bacterial infections, which occur in wounds, is engaging clinical research to deepen the role of ozone in eradicating even multidrug-resistant bacteria. Ozone activates the nuclear factor erythroid 2-related factor 2 (Nrf2), and this activation triggers a complex cascade of events, which ultimately leads to macrophage training and an improvement in their ability to operate a clearance of bacteria in the patient's anatomical districts. In this review, we try to elucidate the recent evidence about the mechanisms with which ozone can actually remove bacteria and even multi-drug-resistant (MDR) bacteria, accounting on its complex ability in modulating immunity.

Removal of contaminants of emerging concern from the supernatant of anaerobically digested sludge by O3 and O3/H2O2: Ozone requirements, effects of the matrix, and toxicity

Digestate is a rich source of nutrients that can be applied in agricultural fields as fertilizer or irrigation water. However, most of the research about application of digestate have focused on its agronomic properties and neglected the potential harm of the presence of contaminants of emerging concern (CECs). Aadvanced oxidation processes (AOPs) have proved to be effective for removing these compounds from drinking water, yet there are some constrains to treat wastewater and digestate mainly due to their complex matrix. In this study, the feasibility to remove different CECs from digestate using O3 and O3/H2O2 was assessed, and the general effect of the matrix in the oxidation was explained. While the lab-scale ozonation provided an ozone dose of 1.49 mg O3/mg DOC in 5 h treatment, almost all the compounds were removed at a lower ozone dose of maximum 0.48 mg O3/mg DOC; only ibuprofen required a higher dose of 1.1 mg O3/mg DOC to be oxidized. The digestate matrix slowed down the kinetic ozonation rate to approximately 1% compared to the removal rate in demineralized water. The combined treatment (O3/H2O2) showed the additional contribution of H2O2 by decreasing the ozone demand by 59–75% for all the compounds. The acute toxicity of the digestate, measured by the inhibition of Vibrio fisheries luminescence, decreased by 18.1% during 5 h ozonation, and by 34% during 5 h O3/H2O2 treatment. Despite the high ozone consumption, the ozone dose (mg O3/mg DOC) required to remove all CECs from digestate supernatant was in the range or lower than what has been reported for other (waste-)water matrix, implying that ozonation can be considered as a post-AD treatment to produce cleaner stream for agricultural purposes.

Removal of micropollutants and ecotoxicity during combined biological activated carbon and ozone (BO3) treatment

Ozonation is a viable option to improve the removal of micropollutants (MPs) in wastewater treatment plants (WWTPs). Nevertheless, the application of ozonation is hindered by its high energy requirements and by the uncertainties regarding the formation of toxic transformation products in the process. Energy requirements of ozonation can be reduced with a pre-ozone treatment, such as a biological activated carbon (BAC) filter, that removes part of the effluent organic matter before ozonation. This study investigated a combination of BAC filtration followed by ozonation (the BO3 process) to remove MPs at low ozone doses and low energy input, and focused on the formation of toxic organic and inorganic products during ozonation. Effluent from a WWTP was collected, spiked with MPs (approximately 1 µg/L) and treated with the BO3 process. Different flowrates (0.25–4 L/h) and specific ozone doses (0.2–0.6 g O3/g TOC) were tested and MPs, ecotoxicity and bromate were analyzed. For ecotoxicity assessment, three in vivo (daphnia, algae and bacteria) and six in vitro CALUX assays (Era, GR, PAH, P53, PR, andNrf2 CALUX) were used.Results show that the combination of BAC filtration and ozonation has higher MP removal and higher ecotoxicity removal than only BAC filtration and only ozonation. The in vivo assays show a low ecotoxicity in the initial WWTP effluent samples and no clear trend with increasing ozone doses, while most of the in vitro assays show a decrease in ecotoxicity with increasing ozone dose. This suggests that for the tested bioassays, feed water and ozone doses, the overall ecotoxicity of the formed transformation products during ozonation was lower than the overall ecotoxicity of the parent compounds. In the experiments with bromide spiking, relevant formation of bromate was observed above specific ozone doses of approximately 0.4 O3/g TOC and more bromate was formed for the samples with BAC pre-treatment. This indirectly indicates the effectivity of the pre-treatment in removing organic matter and making ozone more available to react with other compounds (such as MPs, but also bromide), but also underlines the importance of controlling the ozone dose to be below the threshold to avoid formation of bromate. It was concluded that treatment of the tested WWTP effluent in the BO3 process at a specific ozone dose of 0.2 g O3/g TOC, results in high MP removal at limited energy input while no increase in ecotoxicity, nor formation of bromate was observed under this condition. This indicates that the hybrid BO3 process can be implemented to remove MPs and improve the ecological quality of this WWTP effluent with a lower energy demand than conventional MP removal processes such as standalone ozonation.

Mitochondrial Features of Mouse Myoblasts Are Finely Tuned by Low Doses of Ozone: The Evidence In Vitro.

The mild oxidative stress induced by low doses of gaseous ozone (O3) activates the antioxidant cell response through the nuclear factor erythroid 2-related factor 2 (Nrf2), thus inducing beneficial effects without cell damage. Mitochondria are sensitive to mild oxidative stress and represent a susceptible O3 target. In this in vitro study, we investigated the mitochondrial response to low O3 doses in the immortalized, non-tumoral muscle C2C12 cells; a multimodal approach including fluorescence microscopy, transmission electron microscopy and biochemistry was used. Results demonstrated that mitochondrial features are finely tuned by low O3 doses. The O3 concentration of 10 μg maintained normal levels of mitochondria-associated Nrf2, promoted the mitochondrial increase of size and cristae extension, reduced cellular reactive oxygen species (ROS) and prevented cell death. Conversely, in 20 μg O3-treated cells, where the association of Nrf2 with the mitochondria drastically dropped, mitochondria underwent more significant swelling, and ROS and cell death increased. This study, therefore, adds original evidence for the involvement of Nrf2 in the dose-dependent response to low O3 concentrations not only as an Antioxidant Response Elements (ARE) gene activator but also as a regulatory/protective factor of mitochondrial function.

Ozonation enables to suppress horizontal transfer of antibiotic resistance genes in microbial communities during swine manure composting

Although aerobic composting process has been proposed as an efficient approach to treat livestock manure, the evolution patterns of extracellular antibiotic resistance genes (eARGs) and intracellular antibiotic resistance genes (iARGs) and their horizontal transfer during composting are unclear. Here, the variations of iARGs and eARGs during swine manure composting were systematically analyzed. We found that the abundance of ARGs rebounded from thermophilic phase to mature phase. This was due to the release of persistent eARGs and the increase of certain iARGs during cell proliferation. The conjugative transfer frequencies of ARGs in composting communities were (1.68 ± 0.2) × 10-1 Transconjugant/Donor + Recipient (TC/D + R) in composting products, which was comparable to the frequency before the composting. Ozone was then optimized to suppress the rebound and spread of ARGs. We found that the low ozone dosage of 0.4 g/kg FW in the cooling phase could not only significantly remove iARGs (2.02-logs less than the control, 8 of 12 targeted iARGs were not detected) by eliminating the potential hosts, but also decrease the possibilities of both conjugation and transformation among the composting products by eliminating both HGT (conjugation and transformation) efficiencies of microbes and microbes themselves. Collectively, this study proposes the optimal ozonation strategy to mitigate the dissemination of ARGs in the composting products.

Experimental Inactivation of Microalgae in Marine Ballast Water by Microbubbles Generated through Hydrodynamic Cavitation

This paper presents a novel approach to microbubble technology for the treatment of aquatic invasive organisms in ship ballast water. The microbubbles are produced by hydrodynamic cavitation with a sudden and dramatic water pressure drop. The air and ozone microbubbles, respectively, verified the bioavailability of ship ballast water treatment using marine microalgae as an indicator. Besides the effects of an ozone injection dose, the morphological changes of cells and the effluent toxicity were investigated. Compared with the ozone microbubble treatment, the inactivation of marine microalgae by air microbubbles required a long treatment time. In the storage experiment, it was found that air microbubbles did not inhibit the growth of microalgae cells, and that the injection of active matter such as ozone was still necessary to ensure the validity of biological invasion. However, even with very low doses of ozone, the inactivation effect of ozone microbubbles was still very evident. Overall, it helps to minimize the use of active matter to reduce the toxicity of treated water, and this has the capability to develop into an environmentally acceptable and practical ballast water treatment technology.

Evaluation of antibacterial oxygen/ozone mixture in vitro activity on bacteria isolated from cervico-vaginal mucus of cows with acute metritis

Ozone is an oxidating gas showing a strong microbicidal activity on bacteria, fungi, viruses and protozoa. The aim of this study was to test the in vitro bacteriocidal action of an Ozone/Oxygen gas mixture on bacteria isolated from the cervico-vaginal mucus of cows affected by acute metritis. A pilot study was initially carried out on reference strains (Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923 and Mycoplasma bovigenitalium ATCC 19852) that were tested with three different treatments: a control positive baseline group (B-group) was incubated without gas treatment, a control positive oxygen group (O2-group) was treated with pure oxygen 100%, and the treated group (T-group) was exposed to a gaseous constant flow of an Ozone/Oxygen mixture, at 50, 35, 20 μg Ozone/ml and for 5, 3 and 1 min for every different Ozone concentration. In both positive control groups, the number of colony forming units (CFU) per ml was higher than 300 CFU/ml (E. coli and S. aureus) and higher than 30 CFU/ml for M. bovigenitalium, after incubation. The T-groups showed a minimal bacterial growth equal to or lower than 1 CFU/ml per plate.Based on the results of the pilot study, a second phase was performed on bacteria isolated from the cervico-vaginal mucus (Klebsiella pneumoniae, Enterobacter agglomerans, E. coli, Proteus mirabilis and M. bovigenitalium) using the lower concentration of 20 μg/ml of Ozone for the minimum exposure time of 1 min. The E. coli and S. aureus reference strains and the clinical isolates (K. pneumoniae, E. agglomerans, E. coli, P. mirabilis) were incubated at 37 °C for 48 h and the colonies were manually counted at 24 h and 48 h following inoculation. The cultures of M. bovigenitalium (both ATCC and clinical isolate) were incubated in a jar with modified atmosphere conditions with 5% CO2 at 37 °C for 4–7 days and colony counting was performed. The second phase showed a low number of CFUs (equal to or less than 7 CFU/ml) for the clinical isolates K. pneumoniae, E. agglomerans, E. coli and P. mirabilis, and, of note, for M. bovigenitalium, both ATCC and clinical isolate, the growth was completely inhibited. Ozone was demonstrated to have a bacteriocidal activity. This study encourages further research of in vivo application of low doses of gaseous Ozone for the treatment of metritis in cows by using minimal exposure times.

Ozone Pollution, Oxidative Stress, Regulatory T Cells and Antioxidants.

Ozone pollution, is a serious health problem worldwide. Repeated exposure to low ozone doses causes a loss of regulation of the oxidation–reduction systems, and also induces a chronic state of oxidative stress. This fact is of special importance for the regulation of different systems including the immune system and the inflammatory response. In addition, the oxidation–reduction balance modulates the homeostasis of these and other complex systems such as metabolism, survival capacity, cell renewal, and brain repair, etc. Likewise, it has been widely demonstrated that in chronic degenerative diseases, an alteration in the oxide-reduction balance is present, and this alteration causes a chronic loss in the regulation of the immune response and the inflammatory process. This is because reactive oxygen species disrupt different signaling pathways. Such pathways are related to the role of regulatory T cells (Treg) in inflammation. This causes an increase in chronic deterioration in the degenerative disease over time. The objective of this review was to study the relationship between environmental ozone pollution, the chronic state of oxidative stress and its effect on Treg cells, which causes the loss of regulation in the inflammatory response as well as the role played by antioxidant systems in various pathologies.

Acute ecotoxicological effects on daphnids and green algae caused by the ozonation of ibuprofen

Due to its ubiquitous presence in wastewaters, wastewater treatment plant effluents and even surface waters, the removal of the pharmaceutical ibuprofen from water is of special interest. Ozonation is widely applied for the treatment of micropollutants in wastewater treatment plants and is already known to also degrade ibuprofen. However, the formation of a wide range of transformation products during such oxidation steps might affect the aquatic environment. This study focuses on the acute ecotoxicological impact of the ibuprofen ozonation products on the two model organisms Daphnia magna and Desmodesmus subspicatus. For the identification of possibly ecotoxic products, a new workflow combining ecotoxicological testing, analytical methods and toxicity prediction was applied. Examination at different pH conditions with increasing ozone doses can point to respective products for further systematic examination.Seven ozonation products were confirmed in this study, two of them for the first time. Five previously postulated products were rejected. For pH 7 the inhibition of green algae growth was observed for mixtures oxidized with low ozone doses, while at pH 3 the mixtures with higher ozone doses caused toxic effects on the mobility of daphnids. Together with the analytical measurements in combination with ecotoxicity prediction, six products were identified which might have caused the toxic effect on green algae. However, no assignment to the observed toxic effects on daphnids was possible. The gained results indicate that mixture toxicity might play a role in oxidation processes and needs to be considered in ozonation studies concerning the ecotoxicological impact. Furthermore, the different observed toxicity for the two organisms underlines the importance of using multiple test systems for a comprehensive evaluation of the ecotoxicity during ozonation processes.

Phytochemical Enhancement in Broccoli Florets after Harvest by Controlled Doses of Ozone.

The objective of this work was to examine the effect of controlled doses of O3 (0, 5 µL L−1 of O3 for 60 min, and 5 µL L−1 of O3 for 720 min) on the quality and phytochemical content of broccoli florets during postharvest storage. The optimal dose was found at 5 µL L−1 of O3 for 60 min, from the color retention of broccoli florets exposed to the gas treatment. Overall, the antioxidant capacity of the florets was significantly affected by both doses of O3 compared to the non-exposed florets. The profile of glucosinolates was determined for up to 14 days in broccoli florets stored at 4 °C by LC-MS. The amount of total glucobrassicins and total hydroxy-cinnamates in florets significantly (p ≤ 0.05) improved by the application of 5 µL L−1 of O3 for 60 min compared to non-treated florets. The up-regulation of genes of the tryptophan-derived glucosinolate pathway was observed immediately after both treatments. The gene expression of CYP79A2 and CYP79B3 in broccoli was significantly higher in broccoli florets exposed to 5 µL L−1 of O3 for 720 min compared to non-exposed florets. Although enhancement of secondary metabolites can be achieved by the fumigation of broccoli florets with low doses of ozone, quality parameters, particularly weight loss, can be compromised.

Chronic exposure to ozone induces cardiac antioxidant response and overexpression of either mitochondrial fision protein DRP1 and hipertrophyc-related proteins.

Pollution is considered a risk factor for cardiovascular disease; however, the mechanisms to explain this relationship are not well understood; ozone is one of the most abundant and studied air contaminants. Our study aimed to evaluate the effect of chronic exposition of rats to controlled low doses of ozone on oxidative stress, apoptosis, mitochondrial dynamics, and cardiac hypertrophy. Male Wistar rats were daily exposed to low ozone doses during 7, 15, 30, and 60days, 4h/day. Hearts were dissected, and homogenates were prepared. Oxidative stress was evaluated by TBARS and protein nitrosylation in addition to Superoxide dismutase 1 (SOD1) and Catalase levels; the apoptosis related-proteins caspase 3, caspase 9, Bax, Bcl-2, and the mitochondrial dynamic-associated proteins Fis1, Drp1, OPA1, and Mfn1 were quantified by western blot among the cardiac hypertrophy indicator alpha-actin (cardiac actin). There were no changes in the oxidative stress markers, however SOD1 expression increases. Caspase 3 expression decreased, whereas caspase 9 increased without changes in Bax or Bcl-2. Mitochondrial fission may be favored according to the increased expression of Drp1 but not changes in fusion-related proteins OPA1 and Mfn1. Finally, the molecular marker for cardiac hypertrophy was overexpressed after 30 and 60days of ozone exposition. The chronic exposition to ozone induces a deleterious effect on cardiac mitochondria. Antioxidant defenses also show changes in relation to exposure time, as well as an apparent pro-hypertrophic effect associated with altered mitochondrial dynamics.

Detrimental Effect of Ozone on Pathogenic Bacteria.

(1) Background: Disinfection of medical devices designed for clinical use associated or not with the growing area of tissue engineering is an urgent need. However, traditional disinfection methods are not always suitable for some biomaterials, especially those sensitive to chemical, thermal, or radiation. Therefore, the objective of this study was to evaluate the minimal concentration of ozone gas (O3) necessary to control and kill a set of sensitive or multi-resistant Gram-positive and Gram-negative bacteria. The cell viability, membrane permeability, and the levels of reactive intracellular oxygen (ROS) species were also investigated; (2) Material and Methods: Four standard strains and a clinical MDR strain were exposed to low doses of ozone at different concentrations and times. Bacterial inactivation (cultivability, membrane damage) was investigated using colony counts, resazurin as a metabolic indicator, and propidium iodide (PI). A fluorescent probe (H2DCFDA) was used for the ROS analyses; (3) Results: No reduction in the count colony was detected after O3 exposure compared to the control group. However, the cell viability of E. coli (30%), P. aeruginosa (25%), and A. baumannii (15%) was reduced considerably. The bacterial membrane of all strains was not affected by O3 but presented a significant increase of ROS in E. coli (90 ± 14%), P. aeruginosa (62.5 ± 19%), and A. baumanni (52.6 ± 5%); (4) Conclusion: Low doses of ozone were able to interfere in the cell viability of most strains studied, and although it does not cause damage to the bacterial membrane, increased levels of reactive ROS are responsible for causing a detrimental effect in the lipids, proteins, and DNA metabolism.

Elimination of amino trimethylene phosphonic acid (ATMP) antiscalant in reverse osmosis concentrate using ozone: Anti-precipitation property changes and phosphorus removal

Amino trimethylene phosphonic acid (ATMP) was widely used as an antiscalant in reverse osmosis (RO) systems to prevent membrane scaling, and entered RO concentrate at elevated levels. However, phosphonate antiscalants in RO concentrate might aggravate phosphorus pollution, remobilize heavy metals, and adversely affect the sedimentation treatment of RO concentrate. Ozonation was found an efficient method for ATMP treatment. The ATMP removal efficiencies with 8 mg/L ozone were 100% and 86.5% for ultrapure water and RO concentrate, respectively. The ATMP mineralization efficiency reached 46.5% with 8 mg/L ozone. The rate constant for the reaction between ATMP and ozone was 1.92 × 106 M−1 s−1. Increasing the pH from 3 to 9 decreased the ATMP removal efficiency from 90% to 30.9% but increased the orthophosphate formation to ATMP removal ratio from 0.11 to 0.48. The ATMP intermediates generated with low ozone dosages exhibited moderate chelation and anti-precipitation capacity, and their chelation and anti-precipitation capacity could be further attenuated by increasing the ozone dosage. Ozonation alone enhanced the growth potential for microalgae in RO concentrate because orthophosphate formed. Combining ozonation and coagulation effectively removed 83.0% of the total phosphorus from RO concentrate. The maximum algal density of Scenedesmus sp. LX1 decreased by 78.7% by ozonation and coagulation.

Air ozonization for prevention of bacterial and viral infections

Objective. To assess the effectiveness of the low-dose air ozonation for disinfection of the air in the working room.&#x0D; Materials and methods. We investigated 90 air samples (3 samples were taken weekly before and after the production meeting using the automatic sampling device of biological aerosols of air PU-1B). The total bacterial contamination, the content of staphylococci and mold spores were determined. Ozonation of the room (83.3 m3) was carried out for 20 minutes by means of domestic ozonator. The accumulated dose of ozone was 133.3 mg (1.6 mg/m3). Statistical data processing was carried out using the MedStat licensed program. The median, median error (Me me), left and right 95 % confidence intervals (95 % CI) were calculated. Paired comparisons were made using Wilcoxon's T-test.&#x0D; Results. After the meeting, the total bacterial contamination of the air was 56.0 9.3 (47.078.0) CFU. The content of staphylococci and mold spores in the air was 85.5 12.5 (76.0100.0) and 44.5 6.5 (32.054.0) CFU, respectively. After ozonation, the total bacterial contamination of the air was 14.5 3.6 (10.021.0) CFU. The content of staphylococci and mold spores in the air after ozonation was 35.5 6.7 (25.052.0) and 26.0 5.0 (18.032.0) CFU, respectively. Ozonation of the room provided a significant decrease (p 0.001) in all three of the above indicators. The room ozonation carried out promoted a reliable decrease (p 0.001) in all the above mentioned parameters.&#x0D; Conclusions. The above data and analysis of the literature show the possibility of using low doses of ozone for the prevention of bacterial, fungal and viral infections including SARS-CoV-2. Further study and development of reasonable modes of ozone disinfection, including low doses of ozone, is needed, as well as determination of the efficiency degree of air disinfection with non-toxic gas concentrations.