Efficacy of ozonated olive oil against peri-implant microbes isolated from peri-implantitis.

This study aimed to investigate the antibacterial and antimicrobial activity of ozone gel against oral biofilms grown on titanium dental implant discs. The experiment used medical grade five titanium discs on which peri-implant isolated biofilms were grown. The experimental groups were control, Streptococcus mutans (S. mutans) and Granulicatella adiacens (G. adiacens), (n = 6). The oral microbes grown on titanium discs were exposed to ozone gel for 3 minutes and the antibacterial activity was assessed by turbidity test and adherence test for the antibiofilm activity test. Bacterial morphology and confluence were investigated by scanning electron microscopy (SEM), (n=3). Two bacterial species were identified from the peri-implant sample, S. mutans and G. adiacens. The results showed that adding ozone to the bacterial biofilm on titanium dental implants did not exhibit significant antibacterial activity against S. mutans. Moreover, there was no significant difference in antibiofilm activity between control and treatment groups. However, significant antibacterial and antibiofilm effect was exhibited by ozone gel against G. adiacens. Ozonated olive oil can be considered as a potential antimicrobial agent for disinfecting dental implant surfaces and treating peri-implantitis.

Evaluation of the restorative effect of ozone and chitosan-hyaluronic acid with and without mesenchymal stem cells on wound healing in rats.

This study evaluated the effect of ozone, chitosan-hyaluronic (Cs-HA) acid and mesenchymal stem cells (MSCs) on wound healing in rats. A total of 64 rats were randomly divided into four groups: control, ozone, Cs-HA + ozone and Cs-HA + ozone + MSCs. A 5 mm full-thickness wound was created on the back of each rat. The wound area was measured macroscopically on days 3, 5, 9 and 14. Tissue sections were prepared for histopathological evaluation of inflammation, collagen arrangement, neovascularization and epithelial tissue rearrangement. Macroscopic assessment showed differences in wound area on days 5, 9 and 14. Histopathological examination showed that the Cs-HA + ozone + MSCs and Cs-HA + ozone groups had significantly higher vascularization on day 3 compared to the ozone-treated and control groups. All treatment groups had significantly better collagen arrangement than the control group. On day 5, no significant difference was observed between different groups. On day 9, the inflammation level in the Cs-HA + ozone + MSCs group was significantly lower than in the other groups. All treatment groups had significantly better vascularization compared to the control group. On day 14, the rate of inflammation was significantly lower in the treatment groups than in the control group. Significantly higher collagen arrangement levels were observed in the Cs-HA + ozone and Cs-HA + ozone + MSCs groups compared to the control and ozone groups. All treatment groups had significantly better epithelial tissue rearrangement than the control group. Overall, the results of this study indicated that treatment with ozone, Cs-HA acid, Cs-HA and MSCs accelerated wound healing in rats. The effect of using Cs-HA acid with mesenchymal cells was better than the other types of treatment. Larger clinical trials are needed to assess these factors for improving chronic wound treatment.

Short-time ozone treatment promotes protease-mediated destruction of B cell allergen epitopes by altering the structural characteristics of whey protein.

A novel processing method combining short-time ozone pretreatment with hydrolysis has been developed to reduce whey protein allergenicity. The results showed that ozone treatment altered the whey protein spatial structure, initially increasing the surface hydrophobicity index, and then decreasing due to polymer formation as the time increased. Under the optimized conditions of alkaline protease-mediated hydrolysis, a 10-second pre-exposure to ozone significantly promoted the reduction in the IgE binding capacity of whey protein without compromising the hydrolysis efficiency. Compared with whey protein, the degranulation of KU812 cells stimulated by this hydrolysate decreased by 20.54%, 17.99%, and 22.80% for IL-6, ß-hexosaminidase, and histamine, respectively. In vitro simulated gastrointestinal digestion confirmed increased digestibility and reduced allergenicity. Peptidomics identification revealed that short-time ozonation exposed allergen epitopes, allowing alkaline protease to target these epitopes more effectively, particularly those associated with α-lactalbumin. These findings suggest the promising application of this processing method in mitigating the allergenicity of whey protein.

The effect of intra-articular ozone injection combined with home-based exercise on pain and function in daily living activities of patients with mild to moderate knee osteoarthritis, a randomized double-blinded controlled clinical trial.

BACKGROUND: Knee osteoarthritis is the most common arthritis. Various treatments such as analgesics, exercise therapy, and surgery in high-grade OA have been shown to reduce pain and improve patients' function; however, determining the optimal treatment remains a challenge. Ozone therapy is one of the injection techniques used for symptom relief in these patients. Therefore, this study aimed to evaluate the effect of ozone injection in mild to moderate knee osteoarthritis. METHODS: Thirty-three patients with grade II-III knee osteoarthritis based on the Kellgren-Lawrence classification were involved in the study, by block randomisation. Totally 42 knees were included. All patients received exercise therapy, 500 mg of acetaminophen tablets (up to 2 g per day as needed), and healthy nutrition. In a double-blinded method, the intervention group received Ozone injections, but the control group received placebo injections. Functional tests, including timed-up-and-go and 6-min walk tests, were assessed at baseline and immediately after the 6-week intervention. In addition, the pain was measured by VAS score, and stiffness and activity of daily living (ADL) were evaluated by KOOS questionnaire before and after a 6-week intervention and then one and six months afterwards. FINDINGS: Improvements in pain and KOOS scores were seen in both groups in the 6th week of injections (p < 0.05), with significant differences between groups. However, the effects on pain and KOOS scores disappeared in the 1st and 6th months of follow-ups in the control group. Nevertheless, the effects persisted in the intervention group compared to the baseline and control group, which means that in the mentioned time points intervention group showed significant improvement compared to the control group (p < 0.05). In addition, functional tests showed significant differences between the two groups in the 6th week of injections (p < 0.001). INTERPRETATION: Ozone injection is a non-surgical treatment for mild to moderate knee osteoarthritis that could decrease pain and improve function and ADL of patients in the short to mid-term (3-6 months), so it seems that adding Ozone injection to the routine exercise therapy in management of patients with knee OA could improve outcomes.

Effects of systemic ozone administration on the fresh extraction sockets healing: a histomorphometric and immunohistochemical study in rats.

OBJECTIVE: Studies have highlighted numerous benefits of ozone therapy in the field of medicine and dentistry, including its antimicrobial efficacy against various pathogenic microorganisms, its ability to modulate the immune system effectively, reduce inflammation, prevent hypoxia, and support tissue regeneration. However, its effects on dental extraction healing remain to be elucidated. .Therefore, this study aimed to evaluate the effects of systemically administered ozone (O3) at different doses in the healing of dental extraction sockets in rats. METHODOLOGY: To this end, 72 Wistar rats were randomly divided into four groups after extraction of the right upper central incisor: Group C - control, no systemic treatment; Group OZ0.3 - animals received a single dose of 0.3 mg/kg O3; Group OZ0.7 - a single dose of 0.7 mg/kg O3; and Group OZ1.0 - a single dose of 1.0 mg/kg O3, intraperitoneally. In total, six animals from each group were euthanized at 7, 14, and 21 days after the commencement of treatment. Bone samples were harvested and further analyzed by descriptive histology, histomorphometry, and immunohistochemistry for osteocalcin (OCN) and tartrate-resistant acid phosphatase (TRAP) protein expression. RESULTS: All applied doses of O3 were shown to increase the percentage of bone tissue (PBT) after 21 days compared to group C. After 14 days, the OZ0.7 and OZ1.0 groups showed significantly higher PBT when compared to group C. The OZ1.0 group presented the most beneficial results regarding PBT among groups, which denotes a dose-dependent response. OCN immunostaining was higher in all groups at 21 days. However, after seven and 14 days, the OZ1.0 group showed a significant increase in OCN immunostaining compared to C group. No differences in TRAP+ osteoclasts were found between groups and time points. CONCLUSION: Therefore, O3 therapy at higher doses might be beneficial for bone repair of the alveolar socket following tooth extraction.

Deflamin Attenuated Lung Tissue Damage in an Ozone-Induced COPD Murine Model by Regulating MMP-9 Catalytic Activity.

Chronic obstructive pulmonary disease (COPD) is comprised of histopathological alterations such as pulmonary emphysema and peribronchial fibrosis. Matrix metalloproteinase 9 (MMP-9) is one of the key enzymes involved in both types of tissue remodeling during the development of lung damage. In recent studies, it was demonstrated that deflamin, a protein component extracted from Lupinus albus, markedly inhibits the catalytic activity of MMP-9 in experimental models of colon adenocarcinoma and ulcerative colitis. Therefore, in the present study, we investigated for the first time the biological effect of deflamin in a murine COPD model induced by chronic exposure to ozone. Ozone exposure was carried out in C57BL/6 mice twice a week for six weeks for 3 h each time, and the treated group was orally administered deflamin (20 mg/kg body weight) after each ozone exposure. The histological results showed that deflamin attenuated pulmonary emphysema and peribronchial fibrosis, as evidenced by H&E and Masson's trichrome staining. Furthermore, deflamin administration significantly decreased MMP-9 activity, as assessed by fluorogenic substrate assay and gelatin zymography. Interestingly, bioinformatic analysis reveals a plausible interaction between deflamin and MMP-9. Collectively, our findings demonstrate the therapeutic potential of deflamin in a COPD murine model, and suggest that the attenuation of the development of lung tissue damage occurs by deflamin-regulated MMP-9 catalytic activity.

Use of Ozone as a Substrate Treatment for the Control of Trichoderma in the Production of Pleurotus ostreatus.

Pleurotus ostreatus is one of the most widely cultivated species in the world. It can be produced in many lignocellulosic substrates after carrying out a treatment to eliminate competing microorganisms. The most commonly used is pasteurization by steam or by immersion in hot water. The aim of this work is to evaluate if ozone can be employed as treatment for decontamination of the substrate used for the production of the edible mushroom P. ostreatus to control of green mold Trichoderma. Wheat straw was employed as a substrate. We used two different methodologies: bubbling ozone into a tank with water and the substrate, and injecting ozone into a closed tank with the substrate inside. Ten treatments were carried out including two treatments with inoculation by a spray of conidia of Trichoderma. The effect of ozone on the conidia was also evaluated. We found that the treatment of the substrate with ozone in immersed water resulted more effective (lower growth of Trichoderma) than injecting ozone into a closed tank. Anyway, we found that the contaminant fungi could grow on the substrate in both treatments with ozone. We observed that although ozone affected the conidia when it was bubbled into water, some of them still managed to survive and could germinate 72 h later. P. ostreatus could grow and produce fruiting bodies on a substrate that was previously treated with ozone and yields were not affected. Based on the results obtained, we conclude that ozone may not be an effective agent to control Trichoderma in highly contaminated substrates, at least in the experimental conditions that we used, for the production of P. ostreatus.

Extraction of homogeneous lignin oligomers by ozonation of Miscanthus giganteus and vine shoots in a pilot scale reactor.

Lignin, a complex phenolic polymer crucial for plant structure, is mostly used as fuel but it can be harnessed for environmentally friendly applications. This article explores ozonation as a green method for lignin extraction from lignocellulosic biomass, aiming to uncover the benefits of the extracted lignin. A pilot-scale ozonation reactor was employed to extract lignin from Miscanthus giganteus (a grass variety) and vine shoots (a woody biomass). The study examined the lignin extraction and modification of the fractions and identified the generation of phenolic and organic acids. About 48 % of lignin was successfully extracted from both biomass types. Phenolic monomers were produced, vine shoots yielding fewer monomers than Miscanthus giganteus. Ozonation generated homogeneous lignin oligomers, although their molecular weight decreased during ozonation, with vine shoot oligomers exhibiting greater resistance to ozone. Extracted fractions were stable at 200 °C, despite the low molecular weight, outlining the potential of these phenolic fractions.

Systematic investigation and modeling prediction of virus inactivation by ozone in wastewater: Decoupling the matrix effects.

Water disinfection is undoubtedly regarded as a critical step in ensuring the water safety for human consumption, and ozone is widely used as a highly effective disinfectant for the control of pathogenic microorganisms in water. Although the diminished ozone efficiencies in complex water matrices have been widely reported, the specific extent to which individual components of matrix act on the virus inactivation by ozone remains unclear, and effective methodologies to predict the comprehensive effects of various factors are needed. In this study, the decoupled impact of the intricate water matrix on the ozone inactivation of viruses was systematically investigated and assessed from a simulative perspective. The concept of "equivalent ozone depletion rate constant" (k') was introduced to quantify the influence of different species, and a kinetic model was developed based on the k' values for simulating the ozone inactivation processes in complex matrix. The mechanisms through which diverse species influenced the ozone inactivation effectiveness were identified: 1) competition effects (k' = 105∼107 M-1s-1), including organic matters and reductive ions (SO32-, NO2-, and I-), which were the most influential species inhibiting the virus inactivation; 2) shielding effects (k' = 103∼104 M-1s-1), including Ca2+, Mg2+, and kaolin; 3) insignificant effects (k' = 0∼1 M-1s-1), including Cl-, SO42-, NO3-, NH4+, and Br-; 4) promotion effects (k' = ∼-103 M-1s-1), including CO32- and HCO3-. Prediction of ozone disinfection efficiency and evaluation of species contribution under complex aquatic matrices were successfully realized utilizing the model. The systematic understanding and methodologies developed in this research provide a reliable framework for predicting ozone inactivation efficiency under complex matrix, and a potential tool for accurate disinfectant dosage determination and interfering factors control in actual wastewater treatment processes.

Divergent responses of ascorbate and glutathione pools in ozone-sensitive and ozone-tolerant wheat cultivars under elevated ozone and carbon dioxide interaction.

Crop plants face complex tropospheric ozone (O3) stress, emphasizing the need for a food security-focused management strategy. While research extensively explores O3's harmful effects, this study delves into the combined impacts of O3 and CO2. This study investigates the contrasting responses of O3-sensitive (PBW-550) and O3-resistant (HUW-55) wheat cultivars, towards elevated ozone (eO3) and elevated carbon dioxide (eCO2), both individually and in combination. The output of the present study confirms the positive effect of eCO2 on wheat cultivars exposed to eO3 stress, with more prominent effects on O3-sensitive cultivar PBW-550, as compared to the O3-resistant HUW-55. The differential response of the two wheat cultivars can be attributed to the mechanistic variations in the enzyme activities of the Halliwell-Asada pathway (AsA-GSH cycle) and the ascorbate and glutathione pool. The results indicate that eCO2 was unable to uplift the regeneration of the glutathione pool in HUW-55, however, PBW-550 responded well, under similar eO3 conditions. The study's findings, highlighting mechanistic variations in antioxidants, show a more positive yield response in PBW-550 compared to HUW-55 under ECO treatment. This insight can inform agricultural strategies, emphasizing the use of O3-sensitive cultivars for sustained productivity in future conditions with high O3 and CO2 concentrations.

Virus inactivation in low ozone exposure water reuse applications.

In drinking water applications, an ozone exposure (Ct) based framework has been historically used to validate ozone disinfection. However, significant viral inactivation can be achieved with little to no measurable ozone exposure. Additionally, ozone exposure depends on multiple water quality variables as well as the calculation/ozone measurement method used. In this study, we evaluated alternative ozone monitoring frameworks as well as the impact of water quality variables on ozone decay kinetics and virus/coliform inactivation. Here we show that both change in UV254 absorbance and applied O3:TOC were well correlated with viral inactivation and these frameworks were resilient to changes in water quality. Both increasing temperature (12-30 °C) and pH (5.5-8.4) was shown to significantly increase the ozone decay rate and decreased the resulting ozone exposure by as much as ∼90% in the case of pH. However, due to the increased reaction rate of ozone with viruses at elevated temperature and pH, there was only a minor impact (∼20% in the case of pH) in overall disinfection performance for a given O3:TOC. These frameworks were also considered for variable source water with TOC (5-11 mg/L) and TSS (1.2-5.8 mg/L). Change in UV254 absorbance or applied ozone dose (mg/L) were the strongest indicators of disinfection performance for source waters of variable TOC, however site-specific testing may be needed to apply this framework. Challenge testing with influent nitrite indicated that ozone disinfection performance is significantly impacted (>50% reduction in inactivation) in the presence of nitrite thus enforcing the importance of accounting for this value in the applied ozone dose. Multi-point ozone dissolution was investigated as an alternative ozone application method that may present a benefit with respect to overall disinfection performance especially if nitrite was present. Developing and validating these alternative monitoring frameworks and ozone application methods is imperative in water reuse applications where unnecessary elevated ozone exposure may lead to harmful byproduct formation.

MnO2-based capacitive system enhances ozone inactivation of bacteria by disrupting cell membrane.

The application of ozone (O3) disinfection has been hindered by its low solubility in water and the formation of disinfection by-products (DBPs). In this study, capacitive disinfection is applied as a pre-treatment for O3 oxidation, in which manganese dioxide with a rambutan-like hollow spherical structure is used as the electrode to increase the charge density on the electrode surface. When a voltage is applied, the negative-charged microbes are attracted to the electrodes and killed by electrical interactions. The contact between microbes and capacitive electrodes leads to changes in cell permeability and burst of reactive oxygen species, thereby promoting the diffusion of O3 into the cells. After O3 penetrates the cell membrane, it can directly attack the cytoplasmic constituents, accelerating fatal and irreversible damage to pathogens. As a result, the performance of the capacitance-O3 process is proved better than the direct sum of the two individual process efficiencies. The design of capacitance-O3 system is beneficial to reduce the ozone dosage and DBPs with a broader inactivation spectrum, which is conducive to the application of ozone in primary water disinfection.

Gaseous ozone and ozonized mist in the control of Escherichia coli on 'Rama Forte' persimmon.

This study aimed to evaluate the effectiveness of using two ozone applications (gaseous and mist) as a disinfection method for fresh persimmon. To test these sanitizers, in vitro and in vivo assays were performed, and the Escherichia coli was selected because it is a pathogen that causes foodborne diseases in humans. For in vitro experiments, a plate was inoculated with Escherichia coli strain ATCC 25922 and treated. For in vivo assays, persimmon fruit surface was inoculated with the bacteria and treated. For both assays, it was used 10,15,20,30,40 and 50 µL L-1 of gaseous ozone or ozonized mist for five minutes. The results demonstrated that the gas ozone application significantly reduced the growth of E. coli on the plate surface in vitro at doses of 30, 40 and 50 µL L-1 (with 0.83, 0.89 and 0.95 log CFU mL-1, respectively). The application of ozonized mist showed a significant reduction for 50 µL L-1 (with 1.28 log CFU g-1). And, for the in vivo assays, ozonized mist significantly reduced the number of bacteria on the persimmon surface, with a 1.57 log reduction, which was the largest for 40 µL L-1. Therefore, it is possible to conclude that the ozone application can contribute to the control of microorganisms present on fruit surfaces.

In Vitro Antibacterial Activity of Ozonated Olive Oil against Bacteria of Various Antimicrobial Resistance Profiles Isolated from Wounds of Companion Animals.

Frequent colonization and bacterial infection of skin wounds in small animals prevent or impair their healing. However, the broadly applied antimicrobial therapy of wounds is not always necessary and promotes the spread of bacterial resistance. Thus, alternatives to antimicrobial therapy, including preventive measures in the form of wound dressings with antibiotic properties, should be searched for. The aim of this study was to develop a new, efficient, cost-effective and non-toxic formulation with antimicrobial properties to serve as an alternative to antibiotic administration in wound-healing stimulation in companion animals. Nano/microencapsulated ozonated olive oil in a hyaluronan matrix was developed, with ozone concentration high enough to prevent bacterial growth. The presence and size of nano- and microcapsules were determined with scanning electron microscopy (SEM). Antibacterial activity of developed formulations was examined in vitro on 101 Gram-positive and Gram-negative bacteria isolated from the wounds of companion animals. The highest ozone concentration in the developed formulations inhibited the growth of 40.59% bacteria. Species and genus-specific differences in reactions were observed. Enterococcus spp. proved the least susceptible while non-pathogenic Gram-positive bacteria were the most susceptible to the examined formulations. Changes in the bacterial morphology and cell structure of Psychrobacter sanguinis suspension mixed with Ca-stabilized formulations with nano/microencapsulated ozonized olive oil were revealed during SEM observations. The combination of compounds that promote wound healing (hyaluronic acid, olive oil, ozone and calcium) with the antibacterial activity of the developed formula makes it a promising bionanocomposite for use as a topical dressing.

In vitro effects of different ozone preparations on microorganisms responsible for endometritis in the mare.

Infectious endometritis is considered one of the major causes of infertility and it can affect up to 60% of barren mares. It is characterized by the presence of one or more microorganisms in the reproductive tract and it is treated with the administration of antibiotics, ecbolic agents and uterine lavages. Ozone, thanks to its antimicrobial properties that are based on its high oxidative potential, could represent an effective alternative treatment for endometritis. The aim of this study was to test in vitro the bactericidal and fungicidal properties of different ozone formulations, either as gas (experiment 1) or dissolved in two liquid matrices (experiment 2), specifically distilled water or oil (Neozone 4000, Cosmoproject, Parma, Italy), onto 6 different species of microorganisms isolated from mares with clinical endometritis, namely Escherichia coli, Staphylococcus aureus, Streptococcus equi subsp. Zooepidemicus, Pseudomonas aeruginosa, Klebsiella pneumoniae and Candida albicans. In the first experiment, 3 clinical antibiotic-resistant strains per each species were exposed to different conditions: to O2O3 gas mixtures (15 and 40 µg/ml for 1, 3 and 5 min), to 100 % O2 or left untreated. The results showed a reduction of the microbial count of over 99,9% for every pathogen, time and concentration of O2O3 gas mixtures tested. Furthermore, gaseous ozone showed both a time-dependant effect (5 vs 3 vs 1 min of exposure) and a concentration-dependant effect (40 vs 15 µg/ml) at 1 and 3 min, while after 5 min no differences were observed. In the second experiment, minimum inhibitory concentration (MIC), and minimum bactericidal/fungicidal concentration (MBC, MFC) of ozonated distilled water and ozonated oil were evaluated. Ozonated oil showed a bactericidal/fungicidal activity against all the strains tested (MIC range 12.5-25 % v/v, MBC/MFC range 12.5-50 % v/v) while ozonated distilled water didn't show an observable antimicrobial effect, discouraging its use as an antimicrobial agent for the treatment of endometritis. The results of this in vitro study indicate that both gaseous ozone and ozonated oil exerted remarkable antimicrobial activities and are promising alternative treatments for infectious endometritis, even when caused by antibiotic-resistant bacteria, and encourage further experiments in an effort to scale down or even prevent the use of antibiotics in equine reproduction.

Effect of tropospheric ozone and its protectants on gas exchange parameters, antioxidant enzymes and quality of Garlic (Allium sativum. L).

An experimental study was conducted to assess the detrimental effect of ground-level ozone (O3) on garlic physiology and to find out appropriate control measures against ground-level O3, at TNAU-Horticultural Research farm, Udhagamandalam. Elevated ground ozone levels significantly decreased garlic leaf chlorophyll, photosynthetic rate, stomatal conductance, total soluble solids and pungency. The garlic chlorophyll content was highest in ambient ozone level and lowest in elevated ozone@200 ppb, highest stomatal conductance was recorded in ambient ozone with foliar spray of 3%Panchagavya, and the lowest was observed in elevated ozone@200 ppb. Since the elevated O3 had reduced in garlic photosynthetic rate significantly the lowest was observed in elevated O3@200 ppb and the highest photosynthetic rate was observed in ambient Ozone with foliar spray 3% of panchagavya after a week. The antioxidant enzymes of garlic were increased with increased concentration of tropospheric ozone. The highest catalase (60.97 µg of H2O2/g of leaf) and peroxidase (9.13 ΔA/min/g of leaf) concentration was observed at 200 ppb elevated ozone level. Garlic pungency content was highest in ambient ozone with foliar spray of 0.1% ascorbic acid and the lowest was observed under elevated O3@200 ppb. Highest total soluble solids were observed in ambient ozone with foliar spray of 3%Panchagavya and the lowest observed in elevated ozone@200 ppb. Thus, tropospheric ozone has a detrimental impact on the physiology of crops, which reduced crop growth and yield. Under elevated O3 levels, ascorbic acid performed well followed by panchagavya and neem oil. The antioxidant such as catalase and peroxidase had positive correlation among themselves and had negative correlation with chlorophyll content, stomatal conductance, photosynthetic rate, pungency and TSS. The photosynthetic rate has high positive correlation with chlorophyll content, pungency and TSS. Correlation analysis confirmed the negative effects of tropospheric ozone and garlic gas exchange parameters and clove quality. The ozone protectants will reduce stomatal opening by which the entry of O3 in to the cell will be restricted and other hand they also will alleviate ROS and allied stresses.

Aqueous Ozone Exposure Inhibits Sporulation in the Cyclospora cayetanensis Surrogate Eimeria acervulina.

Ozone is a potent disinfecting agent used to treat potable water and wastewater, effectively clearing protozoa such as Giardia and Cryptosporidium spp. It is unclear whether ozone treatment of water or fresh produce can reduce the spread of the emerging parasite Cyclospora cayetanensis, which causes cyclosporiasis in humans. Obtaining viable C. cayetanensis oocysts to evaluate inactivation methods is challenging because we lack the means to propagate them in vitro, because of delays in case reporting, and because health departments typically add inactivating fixatives to clinical specimens. Research in various surrogate organisms has sought to bolster understanding of the biology of C. cayetanensis. Among these surrogates is the poultry parasite Eimeria acervulina, a closely related and easily cultured parasite of economic significance. We used this surrogate to evaluate the consequences of ozone treatment, using the sporulation state as an indicator of infectious potential. Treating with ozonated water acidified with citric acid reduced sporulation ability in a dose-dependent manner; treatment with up to 4.93 mg/L initial concentration of ozone resulted in a 93% inactivation of sporulation by 7 days posttreatment. This developmental arrest was accompanied by transcriptional changes in genes involved in regulating the response to reactive oxygen species (ROS) in a time course that is consistent with the production of oxygen free radicals. This study shows that ozone is highly effective in preventing sporulation of E. acervulina, a model coccidian used as a surrogate for Cyclospora. Furthermore, ozone exposure induced molecular responses to general oxidative stress, documented with several well-characterized antioxidant enzymes.

Transcriptomic analysis reveals ozone treatment delays kiwifruit postharvest softening by modulating cell wall metabolism.

Kiwifruit ripening and senescence after harvesting are closely related to its economic value. Transcriptome analysis and biochemical parameters were used to investigate the differences in gene expression levels and the potential regulation of cell wall metabolism in kiwifruit treated with ozone, thereby regulating fruit softening and prolonging postharvest life. Compared to the control group, the activities of the cell wall modification enzyme were lower under ozone treatment, the content of polysaccharide in the cell wall of primary pectin and cellulose was higher, and the content of soluble pectin was lower. Meanwhile, ozone treatment delayed the degradation of the cell wall mesosphere during storage. A total of 20 pectinesterase (PE)-related genes were identified by sequencing analysis. The data analysis and quantitative polymerase chain reaction results confirmed that cell wall modifying enzyme genes played an important role in softening and senescence after harvesting, which may reduce or induce the expression of certain genes affecting cell wall metabolism. Ozone treatment not only regulates active genes such as xyloglucan endo glycosyltransferase/hydrolase, cellulose synthase, polygalacturonase, and PE to maintain the quality of fruit after harvest but also acts synergically with cell wall modifying enzymes to inhibit the degradation of cell wall, resulting in changes in the ultrastructure of cell wall, thereby reducing the hardness of kiwifruit. In addition, according to the results of cis-acting elements, cell wall degradation is also related to downstream hormone signaling, especially PE-related genes. These results provide a theoretical basis for studying the mechanism of firmness and cell wall metabolism difference of kiwifruit and also lay a good foundation for further research.

Effect of ozone sterilization on controlling leaf mildew and gray mold tomato disease in a glasshouse and its influence on other growth parameters.

BACKGROUND: In order to explore the effect of ozone sterilization treatment on tomato disease control and increase fruit setting rate, this study took 906 pink fruit tomato as test material, used a small ozone generator to carry out ozone treatment single-factor test, and then selected orthogonal table to guide the ozone treatment combination. The effects of different ozone treatment concentration, ozone treatment duration and ozone treatment times on the growth, disease and fruit setting rate of potted tomato were analyzed. RESULTS: Different ozone treatment had effects on leaf mildew, gray mold and fruit setting rate of tomato. The influence degree of three factors on leaf mildew, gray mold and fruit setting rate was from large to small, a > b > c, a > c > b, b > a > c. A quadratic regression model was established with the control effect of tomato leaf mildew, gray mold and fruit setting rate as response values, and the optimal parameter combination was determined: The ozone treatment concentration was 0.0465 g kg-1, the ozone treatment time was 30 min, and the ozone treatment times were twice a week. In this case, the control efficiency of tomato leaf mildew was 95.02%, the control effect of gray mold was 99.49%, and the fruit setting rate was 76.5%. The test parameters were accurate and reliable. CONCLUSION: The ozone sterilization method proposed in this article is safe and green, and can provide theoretical support for the recovery and reconstruction of tomato disease in a glasshouse. © 2024 Society of Chemical Industry.

Sterilization of Human Amniotic Membrane Using an Ozone Hydrodynamic System.

Human amniotic membrane (hAM) is an important biomaterial for Tissue Engineering, due to its great regenerative properties and potential use as a scaffold. The most used procedure to sterilize biomaterials is gamma-irradiation, but this method can affect several properties, causing damage to the structure and reducing the growth factors. The present work evaluated the efficiency of a new method based on ozonated dynamic water for hAM sterilization. HAM fragments were experimentally contaminated with Staphylococcus aureus, Escherichia coli, Candida albicans, Staphylococcus epidermidis, and Clostridium sporogenes (106 CFU/mL) and submitted to sterilization process for 5, 10 and 15 min. The analyses did not reveal microbial activity after 10 min for S. aureus and C. sporogenes and after 15 min for E. coli and S. epidermidis. The microbial activity of C. albicans was reduced with the exposure time increase, but the evaluated time was insufficient for complete sterilization. The depyrogenation process was investigated for different ozonation times (15, 20, 25, 30, and 35 min) to evaluate the ozone sterilization potential and presented promising results after 35 min. The ozone effect on hAM structure was evaluated by histological analysis. A decrease in epithelium average thickness was observed with the exposure time increase. Furthermore, some damage in the epithelium was observed when hAM was exposed for 10 and 15 min. It can indicate that ozone, besides being effective in sterilization, could promote the hAM sample's de-epithelization, becoming a possible new method for removing the epithelial layer to use hAM as a scaffold.