wcm.01.2025.105.112

RESEARCH ON THE PROCESS OF DISINFECTING AND PURIFYING PATHOGENIC
BACTERIA AND VIRUSES IN DRINKING WATER USING OZONE TECHNOLOGY

Journal: Water Conservation and Management (WCM)
Author: Askar Abdykadyrov, Palvan Kalandarov, Aidar Kuttybayev,, Mukhit Abdullayev, Daria Zhumakhanova, Anar Khabay, Maral Abulkhanova, Sunggat Marxuly,* and Arailym Madelbekova
Print ISSN : 2523-5664
Online ISSN : 2523-5672

This is an open access article distributed under the Creative Commons Attribution License CC BY 4.0, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited

Doi: 10.26480/wcm.01.2025.105.112

Abstract

Keywords

Ozone technology, pathogenic microorganisms, bacteria and viruses, disinfection process, oxidizing agent, combination methods, drinking water purification process.

1. INTRODUCTION

In Table 1, the key differences between chlorination and ozone treatment are compared; while chlorination is effective in eliminating microbes, it can produce harmful residues, whereas ozone is environmentally friendly but comes with higher operational costs; ozone enhances the taste and odor of water and does not negatively affect the ecosystem, making it one of the most efficient modern disinfection methods; one of the main drawbacks of ozone is its rapid decomposition in water, which limits its disinfection effect to a short duration; therefore, researchers are exploring ways to enhance ozone’s effectiveness by combining it with other technologies; for instance, when used in combination with ultraviolet (UV) radiation or ultrasound, the disinfection effect of ozone is significantly increased; thus, combining ozone with other methods can provide more effective protection of water from pathogens (Draginsky et al., 2007; Hossen et al., 2090; Beltrán et al., 2019); the benefits of integrating ozone with other methods are summarized in Table 2 below.

Table 2 outlines the advantages of combining ozone with other disinfection methods; these combined approaches enhance disinfection efficiency, reduce harmful residues, and improve water quality; such combinations also help minimize environmental impact and improve the removal of pollutants; research is also investigating the potential of combining ozone with nanotechnology; nanomaterials can stabilize ozone molecules, allowing them to remain active for longer periods; this approach helps improve the effectiveness of ozone in water disinfection and encourages its broader use in water treatment plants; additionally, reducing the cost of ozone disinfection is one of the critical challenges, and ongoing studies aim to make this technology more affordable by using new methods (Li et al., 2008; Amin et al., 2014); Table 3 below presents the potential of combining ozone with nanotechnology.

Table 3 highlights the potential of combining ozone with nanotechnology; nanomaterials stabilize ozone molecules, allowing them to remain active for longer, thus enhancing the efficiency of water disinfection; additionally, research aimed at reducing the cost of ozone disinfection is focused on introducing new methods that expand its use in water treatment plants; another advantage of using ozone for water disinfection is its ability to eliminate unpleasant tastes and odors in the water (as shown in Table 4); this is particularly beneficial for improving the quality of drinking water (Kizi et al., 2021; Camel and Bermond, 1998); currently, research is focusing on the ability of ozone to address virological threats, including the destruction of new viral strains; advances in technology and scientific breakthroughs continue to expand the potential applications of ozone as an effective method for water purification (Costa and Féris, 2023; Kokkinos et al., 2021).

Table 4 outlines the advantages of ozone in water disinfection; ozone improves the quality of drinking water by removing unpleasant tastes and odors, thus enhancing the overall consumption experience; in addition, research is being conducted to address the destruction of new viral strains, with technological and scientific advancements expanding the potential for using ozone as an effective method; experimental studies on ozonation of drinking water contaminated with typhoid-causing bacteria have been previously conducted in both domestic and international research; however, their findings primarily confirmed the possibility of disinfecting water contaminated with typhoid bacteria using ozone; there is comparatively less data on the effect of ozone on bacterial dysentery pathogens (Schoenen, 2002; Bitton, 2014); the antiviral properties of ozone in water treatment have only recently begun to be thoroughly studied; research has focused on ozone’s inactivation of the poliomyelitis virus and its effect on other viruses found in drinking water that play a significant role in human pathology; these studies aim to prevent intestinal infections through preventative measures; based on these findings, scientists have begun exploring the efficiency of ozonating drinking water and the inactivation of poliomyelitis and Coxsackie viruses with ozone (Chen et al., 2021; Chen et al., 2021; Kholikulov and Matkarimov, 2021); in line with these issues, we conducted experimental work in the laboratory using a specially designed ETRO-03 ozonator, based on an electric discharge principle, to study the impact of ozone on various bacterial contaminants in water; the research findings are presented in the following sections.

2.1 Mathematical Model of the Disinfection Process of Pathogenic
Microorganisms Based on Ozone Technology

To develop a mathematical model for the disinfection process of pathogenic microorganisms based on ozone technology, it is essential to consider the mechanism of ozone action, the response of pathogenic microorganisms to ozone, and the concentration of ozone during the disinfection process; the following factors can be considered when constructing the mathematical model: key parameters of the model include ozone concentration (C(t)), the number of pathogenic microorganisms (N(t)), disinfection rate (r), and exposure time (t); the disinfection process can be described using logarithmic equations that model the change in the number of pathogenic microorganisms over time: dN(t)dt=−k⋅C(t)⋅N(t)\frac{dN(t)}{dt} = -k \cdot C(t) \cdot N(t) (1), where N(t) is the number of viable microorganisms at time t, C(t) is the ozone concentration at time t, and k is the disinfection constant (which characterizes the sensitivity of pathogenic microorganisms to ozone); the concentration of ozone can vary over time, but for simplicity, if we assume the ozone concentration is constant, then C(t)=C0C(t) = C_0; if C(t)C(t) is constant, solving the differential equation yields: N(t)=N0⋅e−k⋅C0⋅tN(t) = N_0 \cdot e^{-k \cdot C_0 \cdot t} (2), where N0N_0 is the initial number of microorganisms, C0C_0 is the constant ozone concentration, and t is the exposure time; disinfection efficiency (E) can be defined as the fraction of the initial number of microorganisms that have been inactivated over time: E=N0−N(t)N0=1−e−k⋅C0⋅tE = \frac{N_0 – N(t)}{N_0} = 1 – e^{-k \cdot C_0 \cdot t} (3); this formula demonstrates the relationship between disinfection efficiency, time, and ozone concentration; as the ozone concentration C0C_0 increases or the exposure time t lengthens, the number of microorganisms N(t)N(t) decreases, meaning that disinfection efficiency increases; this model can be used to determine the necessary ozone concentration and exposure time for effective disinfection; the parameter k should be determined empirically for each microorganism based on its sensitivity to ozone; such a mathematical model helps evaluate the effectiveness of ozone technology in disinfecting pathogenic microorganisms.

3.RESULTS AND DISCUSSIONS

The scientific research plan is as follows: The use of ozone technology for the disinfection of pathogenic bacteria and viruses in drinking water is investigated. First, water samples are collected, and microbiological contamination is identified. Then, the water is treated with ozone, and the concentration of pathogens is measured again. The purification results and the effectiveness of ozone are evaluated comparatively.

3.1 Experimental Research

In this scientific research, 12 strains of typhoid bacteria and 36 strains of dysentery bacteria, including Flexner, Sonne, and Shigella, were used. Bacterial suspensions were prepared from 24-hour agar cultures. Before each experiment, the concentration of microorganisms in the contaminated isotonic water was determined, filtered through a No. 2 membrane filter, and final samples were cultured on Endo agar. All studies, except for those aimed at determining the disinfection efficiency of water quality, were conducted in water purified through coagulation, sedimentation, and filtration in accordance with the sanitary regulations 3.02.002.04 SanPiN and GOST 2874 standards.

Preliminary experiments showed that sublethal doses of ozone altered the morphological, cultural, tintorial, biochemical, antigenic, and virulent properties of the bacteria, making it difficult to assess the bactericidal effect of ozone. Therefore, before infecting the water, we sterilized it with ultraviolet rays.

The research was conducted at the Kazakh National Research Technical University named after K.I. Satbayev, at the Department of Electrical Technology and Space Technologies (ET&ST). It was carried out using the ETRO – 03 ozonator device, based on a pilot electric discharge. Figure 1 below shows the laboratory setup and its electrical schematic. This setup allows for ozone treatment of water under conditions close to those used in water pipelines.

In Figure 1, the main elements of the ozone treatment device consist of the ozonator, capacitors, diodes, and a transformer. The ETRO-03 ozonator is used to enrich the water with ozone, effectively disinfecting bacteria and viruses in the water. K75 capacitors store the necessary energy for the ozonator and provide a stable direct current. KC201E diodes regulate the flow of current in the correct direction, while the 220/10kV transformer supplies high voltage to the ozonator, ensuring efficient operation. Together, these components form the ozone generation system essential for disinfecting drinking water.

The technological scheme of the equipment used for conducting the research is shown in Figure 2 below. Ozone treatment of the water is carried out through barbotage (mixing with air bubbles): the output transformer allows voltage to be adjusted from 1,000 to 18,000 (18kV) volts. By varying the voltage at the output electrodes, the ozone concentration in the ozone-air mixture can be regulated from 0.5 to 30 g/m³.

In Figure 2, the technological process of the ozone treatment system is shown step-by-step. First, in stage 1, the air is dried (Air dryer), ensuring the efficient operation of the device. In stage 2, the air passes through a filter, removing dust and particles from the air. In stage 3, ozone is generated through an ozone-producing tube, where materials like copper (Cu), tungsten (W), salt (NaCl), and glass are involved. In stage 4, ozone oxide is absorbed by the ozone oxide absorber, enhancing the effect of ozone on the purified water. In stage 5, the sterilization process of the water takes place in the sterilizing column, and in stage 6, unused ozone is removed through the unused ozone absorber. In the final stages, the ozone flow is measured using a rheometer, and the process is completed by distributing it through the blower.

Due to the variation in ozone utilization efficiency, which can range from 20% to 95% depending on the experimental conditions and the method of mixing water with the ozone-air mixture, we considered not only the total amount of ozone introduced into the water but also the actual amount used during the ozonation process. The amount of ozone for treating 1 liter of water was measured in milligrams and conditionally referred to as ozone per liter of water. By subtracting the amount of ozone retained in the absorber, we obtain the “pure ozone” amount (mg/L).

It is assumed that a portion of the pure ozone is used to oxidize organic substances in the water, including microbial dust, while the remainder breaks down into free radicals, with only a small portion detectable in the water (for a brief period, up to about 1 minute, after the water comes into contact with ozone). This phenomenon is referred to as the “short ozone zone.” The nature of this process is unknown, but it is believed to involve rapidly degrading radicals or unstable compounds. The decomposition rate of ozone in water depends on factors such as temperature, pH, the characteristics and concentration of contaminants, some of which are present in trace concentrations. These factors catalyze the ozone decomposition reaction.

After completing the disinfection process, the remaining ozone was determined using the iodometric method, and 400 ml of water was passed through membrane filters. Final samples were placed on Endo agar and incubated at 37°C for 5 days in a thermostat. These studies showed the survivability of bacteria, so to check for sterilization, samples were taken immediately after ozonation, every 30 minutes for 6 hours, and after 12, 24, and 48 hours. Each experiment was repeated at least 8-10 times under similar conditions. The microbiological results were processed statistically (see Table 5).

The results of the research are presented in Table 5, from which it can be observed that, after ozone treatment of 1 liter of water, on average, 2 unstable strains of typhoid bacteria and several microbial cells of stable strains remained. The maximum and minimum concentrations of microorganisms under the ozonation conditions showed significant deviations when calculated based on confidence intervals and average values. The number of typhoid bacteria ranged from 1 to 13, while the concentration of stable bacteria varied from 1 to 13, and the maximum concentration could reach up to 3 microbial cells per liter. Although the effect of ozone on typhoid strains was significant, the resistance of microorganisms to ozone was found to be slightly higher. The level of ozone’s impact on contaminated water sources was studied in relation to the ozone concentration and exposure duration. It was determined that the concentration of ozone in these conditions had a certain effect on the contamination level of the water. Much depended on the initial contamination of the water. For example, when ozone concentration was 4-5 mg/L and the water source was exposed for several hours, the contamination by pathogenic bacteria in the first hours of ozonation was less than 1,000 bacteria per liter, but remained at a high level during the first hours of ozonation (over 100,000 bacteria per liter).

In experiments with ozonation of water contaminated with dysentery bacteria at different concentrations, similar patterns were found as in the disinfection of water contaminated with typhoid bacteria. The bactericidal doses were close to the doses that resulted in the destruction of typhoid pathogens.

The effect of water quality on disinfection efficiency was studied in distilled water (transparency 0°, oxygen acidity 0.8 mg/L), ozone-purified river water (transparency 110°, oxygen acidity 24.3 mg/L, ozone concentration 7.2 mg/L), and untreated river water (transparency 110°, oxygen acidity 24.3 mg/L, ozone concentration 7.2 mg/L).

In Figure 3 below, it is shown that the bactericidal dose of ozone increases significantly with the concentration of organic substances and ozone binding capacity. If 22 mg/L of pure ozone was required to disinfect untreated river water, only about 7 mg/L of ozone was needed to purify the same water according to GOST 2874-54 standards. Therefore, as expected, pre-purifying the water with ozone before disinfection is advisable. If the initial contamination of the water does not exceed 10,000 typhoid or dysentery bacteria per liter, the pH fluctuations of water between 5.8 and 8.0 had little effect on ozone disinfection. Similarly, the data presented in this table show that when disinfecting water that meets GOST 2874-54 standards, the contamination level of pathogenic bacteria ranges from 1,000 to 10,000 per liter, while the remaining ozone concentration is about 0.15–0.2 mg/L, which demonstrates the reliability of the disinfection. The required amount of ozone increases with the contamination level. For instance, when the number of microorganisms in the water reaches 100,000 bacteria per liter, and the residual ozone concentration is 0.2 mg/L, disinfection is not reliable. Disinfection becomes reliable when the number of microorganisms per liter reaches 400,000, but only when the residual ozone concentration exceeds 2 mg/L.

Thus, determining the residual ozone concentration is a reliable method of evaluating water disinfection in real conditions, especially as an alternative to determining residual chlorine. The overall effect of the ozonation process on bacteria and viruses, as well as the dynamics of ozone concentration, is shown in Figure 3 below.

In Figure 3, the impact of the ozonation process on bacteria and viruses and the dynamics of ozone concentration are demonstrated. The initial graphs show how the viability of various bacteria (such as typhoid and dysentery strains) decreases with increasing ozonation time. As the ozone concentration rises, the survival rate of bacteria and viruses significantly decreases, with complete elimination occurring within 5 to 12 minutes. The final graph illustrates how the total and residual ozone concentrations change over time: ozone concentration increases proportionally with the duration of ozonation. This helps determine the required time and concentration to enhance the disinfection efficiency of ozone. Some researchers indicate the possibility of bacterial reactivation in ozonated water, which they explain by the rapid decomposition of residual ozone. Continuing research in this area, we found that, although the disinfection efficiency was high (99.95%), this does not always guarantee sterility, as several types of bacteria in the water, especially microorganisms that were not directly exposed to ozone, may retain their viability. If water samples are not examined immediately after ozonation, bacterial colony growth can be observed (in 400 ml of water). This effect was seen in samples examined immediately and 30 minutes after ozonation. However, after 6 hours and again after 24-48 hours, the samples once again showed sterility. These findings are consistent with the research results on the sterility of ozonated water by Draginsky V. L. and Alekseeva L. P. According to GOST standards, the coliform index for drinking water should not exceed 3. Based on these requirements, we studied the viability of various strains of typhoid and dysentery bacteria, which were present in a 1:10 ratio with ozone-resistant microbes. The average data are shown in Figure 4, which confirms that these trends hold even in the presence of pathogenic bacteria.

This figure illustrates the decrease in the survival rate of intestinal and typhoid bacteria in water as the ozone dose (mg/l) increases. Intestinal bacteria are more susceptible to ozone than typhoid bacteria, as they are rapidly eliminated at lower doses. When the ozone dose exceeds 4 mg/l, the concentration of intestinal bacteria approaches zero. Although typhoid bacteria survive at higher ozone doses, their concentration also declines significantly. This demonstrates that ozone is an effective tool for water disinfection.

The coliform index, set at 3, along with the presence of intestinal bacteria, can serve as an indicator of reliable disinfection using ozone. Instances were observed where bacteria and intestinal bacilli remained alive at residual concentrations, indicating that some portions of typhoid and dysentery bacteria retained their viability even after disinfection. In studies on the virulence properties of ozone, three serotypes of Coxsackie B virus strains were used. Similarly, foreign scientific studies have shown that high ozone concentrations significantly reduce viral impact. As depicted in the graph, after 60 minutes of ozone exposure, virus viability dropped from 70% to 10%, highlighting ozone’s strong antiviral properties. Ozone’s effects on various infectious diseases have been studied in this context.

The dynamics of enterovirus inactivation during ozone treatment of water indicate that the virulence effect of ozone depends on the treatment duration, residual ozone amount, concentration, and the characteristics of substances (both easily and difficultly oxidized) present in the water (Figure 4). No virus inactivation was observed in the first 5 minutes, as no residual ozone was detected—it was entirely consumed in oxidizing organic matter. During this time, the oxidation capacity of the water decreased by 10%. Easily oxidized substances were eliminated within the first minute. Over the next two minutes, by the 7th minute of ozonation, 90% of poliovirus and 97% of Coxsackie viruses were inactivated, with approximately 0.1 mg/l of residual ozone found in the water (Figure 5).

In Figure 5, the inactivation levels of poliovirus and Coxsackie virus are presented as a function of ozonation duration. Poliovirus inactivation rises sharply after 5 minutes, reaching 100% at the 6-minute mark. Coxsackie virus begins to inactivate between 4 and 5 minutes and is fully eliminated by the 7th minute. This demonstrates that the duration of ozonation significantly influences virus inactivation, with the sensitivity of both poliovirus and Coxsackie virus to ozone increasing over time.

Each experiment used 50 ml samples for virus testing. In the first and third trials, virus concentrations were measured before and after ozone treatment, while in the second trial, residual ozone concentration (lysogenic mixture) was determined using a general method. Inactivation effects of ozone were studied over seven days in multiple cases. The quantity of viruses, measured by TCID50/ml, was monitored throughout this period. Virus inactivation depended on ozone’s antimicrobial properties and stability.

In new experiments, ozone’s effect on viruses was observed within 3 to 5 minutes. In some cases, ozone did not completely eliminate the virus, while in others, it partially inactivated them. After this period, the inactivation rate slowed, explaining the resistance observed in certain virus strains. Over 15 to 16 minutes, inactivation levels ranged from 99.7% to 99.99%. The slow inactivation process was closely related to ozone concentration. These findings suggest that Coxsackie-type viruses exhibit high resistance to ozone. While ozone effectively inactivated viruses within 15 minutes, its effect did not persist for an extended period.

3.2 Analysis of Scientific Research

The results of this scientific study demonstrate that ozone disinfection is highly effective in inactivating pathogenic microorganisms, particularly typhoid and dysentery pathogens. When treating 1 liter of water with an ozone concentration of 14 mg/L, up to 99.95% of pathogenic bacteria were destroyed, highlighting ozone’s strong disinfection capability. Poliovirus was inactivated by 99.99% within 6 minutes, while Coxsackie virus showed an inactivation rate of 99.7% to 99.9% within 15 minutes, indicating varying virus sensitivities to ozone.

Maintaining a residual ozone concentration of 0.15–0.2 mg/L resulted in high disinfection efficiency, emphasizing the crucial role of residual ozone. During ozone application, intestinal bacteria were destroyed more rapidly than typhoid bacteria, with their viability decreasing sharply as ozone exposure increased. The effectiveness of ozone in water disinfection also depended on the composition of organic contaminants in both purified and contaminated water. Research showed that while an ozone concentration of 7 mg/L was sufficient for disinfecting clean water, highly contaminated water required an increased concentration of 22 mg/L.

When ozone was combined with ultraviolet radiation or ultrasound methods, disinfection efficiency significantly improved (Abdykadyrov et al., 2023; Abdykadyrov et al., 2023). This suggests that integrating ozone with other disinfection methods holds promising potential for enhanced water treatment processes (see Figure 6).

In Figure 6, the graph illustrates the inactivation of typhoid and dysentery bacteria as a function of ozonation duration. The data show a significant decline in bacterial viability during ozone treatment. Within 5 minutes, 70% of typhoid bacteria and 75% of dysentery bacteria were inactivated. After 10 minutes, more than 99.9% of both bacterial types were eliminated. This research confirms that ozone is a highly effective disinfection tool, with both exposure duration and ozone concentration playing a crucial role in the process.

4. CONCLUSION

The following key results were achieved in this scientific research: Efficiency of Ozone in Pathogen Elimination – it was demonstrated that ozone disinfection can eliminate up to 99.95% of typhoid and dysentery bacteria. The study used 11 strains of typhoid bacteria and 36 strains of Flexner and Sonne dysentery bacteria. During the treatment of 1 liter of water with an ozone concentration of 14 mg/L, the inactivation level of the bacteria reached 99.95% within 5 minutes; Effect of Ozone on Viruses – inactivation of poliovirus up to 99.99% was achieved when ozone was applied for 6 minutes with a residual concentration of 0.2 mg/L. Although the Coxsackie virus was less sensitive to ozone, its inactivation ranged from 99.7% to 99.9% within 15 minutes; High Ozone Concentration and Virus Viability – the viability of viruses decreased from 70% to 10% within 60 minutes when exposed to high ozone concentrations; Disinfection Efficiency and Residual Ozone Concentration – complete inactivation of typhoid and dysentery bacteria was observed when 4.5 mg/L of ozone was applied per liter of water. The residual ozone concentration was determined to be around 0.15-0.2 mg/L, indicating reliable disinfection. According to the data, an ozone concentration of 22 mg/L was sufficient to completely break down organic contaminants in contaminated water, while approximately 7 mg/L of ozone was required in purified water; Advantages of Ozone in Water Treatment – during water disinfection with ozone, the number of pathogenic microorganisms remained very low after reactivation tests, indicating that water treated with ozone remains safe for an extended period (Abdykadyrov et al., 2024; Abdykadyrov et al., 2024). Although the residual ozone concentration decreased over time, water sterility was maintained for at least the first 6 hours; Efficiency of Ozone in Combination Methods – when ozone was used in combination with ultraviolet radiation or ultrasound, disinfection effectiveness improved, achieving up to 99.99% inactivation of bacteria and viruses. Increasing the ozone concentration and combining it with other methods enhanced the effectiveness of pathogen elimination. This research has proven that ozone technology is an effective method for disinfecting drinking water from pathogens. Ozone’s high oxidative properties and its residue-free decomposition, compared to traditional methods, make water safe and environmentally friendly.

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