Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital

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The urgency in this research is ozone water as a disinfectant is still limited and no research has been found on the use of ozone water on tooth extraction instruments. However, ozone has antibacterial properties so researchers are interested in studying the effectiveness of ozone water as a disinfectant when compared to the commonly used 4.8% chloroxylenol. This study aimed to determine the effectiveness of disinfection of ozone water and 4.8% chloroxylenol in reducing the number of bacterial colonies on dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital. Methods The samples used were mandibular molar forceps have been used in tooth extraction procedures. This study was experimental and used 30 sample, where each group consisted of 10 tooth extraction instruments. The treatment group used ozone water and 4.8% chloroxylenol and the negative control group was cleaned with distilled water. The ozone water group was produced with an ozone generator with a concentration of 15 mg/L for 20 minutes. Then the active side of tooth extraction forceps was soaked for 30 minutes. In the 4.8% chloroxylenol (dettol®️) group, the active side of the mandibular molar tooth extraction forceps was soaked in 250 mL for 60 minutes. In the control group, the mandibular molar tooth extraction forceps were rinsed with distilled water. After that, the sample was counted using a bactery colony counter. Results The results of the data analysis showed a p-value ≤0.001, which means there was a significant difference in the disinfection using ozone water and 4.8% chloroxylenol on the number of bacterial colonies on dental extraction instruments. Conclusions From this study it can be said that ozone water is more effective in disinfecting than 4.8% chloroxylenol. 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Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.12688/f1000research.132941.4 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. Close Copy Citation Details Export Export Citation Sciwheel EndNote Ref. Manager Bibtex ProCite Sente EXPORT Select a format first Track Share ▬ ✚ Research Article Revised Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] Previously titled: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the USU Dental and Oral Hospital Hendry Rusdy https://orcid.org/0009-0004-7182-4901 1 , Rahmi Syaflida Dalimunthe 1 , Ahyar Riza 1 , Ismahani Akilah D 1 Hendry Rusdy https://orcid.org/0009-0004-7182-4901 1 , Rahmi Syaflida Dalimunthe 1 , Ahyar Riza 1 , Ismahani Akilah D 1 PUBLISHED 31 Jul 2025 Author details Author details 1 Universitas Sumatera Utara, Medan, North Sumatra, Indonesia Hendry Rusdy Roles: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Investigation, Methodology, Project Administration, Resources, Software, Supervision, Validation, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing Rahmi Syaflida Dalimunthe Roles: Data Curation, Investigation, Methodology, Project Administration, Resources, Validation, Writing – Review & Editing Ahyar Riza Roles: Data Curation, Investigation, Project Administration, Validation, Writing – Original Draft Preparation, Writing – Review & Editing Ismahani Akilah D Roles: Data Curation, Formal Analysis, Investigation, Resources, Visualization, Writing – Review & Editing OPEN PEER REVIEW DETAILS REVIEWER STATUS Abstract Background The application of disinfectants on dental instruments is one way to prevent cross-infection. The urgency in this research is ozone water as a disinfectant is still limited and no research has been found on the use of ozone water on tooth extraction instruments. However, ozone has antibacterial properties so researchers are interested in studying the effectiveness of ozone water as a disinfectant when compared to the commonly used 4.8% chloroxylenol. This study aimed to determine the effectiveness of disinfection of ozone water and 4.8% chloroxylenol in reducing the number of bacterial colonies on dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital. Methods The samples used were mandibular molar forceps have been used in tooth extraction procedures. This study was experimental and used 30 sample, where each group consisted of 10 tooth extraction instruments. The treatment group used ozone water and 4.8% chloroxylenol and the negative control group was cleaned with distilled water. The ozone water group was produced with an ozone generator with a concentration of 15 mg/L for 20 minutes. Then the active side of tooth extraction forceps was soaked for 30 minutes. In the 4.8% chloroxylenol (dettol®️) group, the active side of the mandibular molar tooth extraction forceps was soaked in 250 mL for 60 minutes. In the control group, the mandibular molar tooth extraction forceps were rinsed with distilled water. After that, the sample was counted using a bactery colony counter. Results The results of the data analysis showed a p-value ≤0.001, which means there was a significant difference in the disinfection using ozone water and 4.8% chloroxylenol on the number of bacterial colonies on dental extraction instruments. Conclusions From this study it can be said that ozone water is more effective in disinfecting than 4.8% chloroxylenol. READ ALL READ LESS Keywords Ozone water, Chloroxylenol, Bacterial Colony Corresponding Author(s) Hendry Rusdy ( [email protected] ) Close Corresponding author: Hendry Rusdy Competing interests: No competing interests were disclosed. Grant information: The author(s) declared that no grants were involved in supporting this work. Copyright: © 2025 Rusdy H et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The author(s) is/are employees of the US Government and therefore domestic copyright protection in USA does not apply to this work. The work may be protected under the copyright laws of other jurisdictions when used in those jurisdictions. How to cite: Rusdy H, Dalimunthe RS, Riza A and D IA. Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.12688/f1000research.132941.4 ) First published: 21 Jun 2023, 12 :726 ( https://doi.org/10.12688/f1000research.132941.1 ) Latest published: 31 Jul 2025, 12 :726 ( https://doi.org/10.12688/f1000research.132941.4 ) Revised Amendments from Version 3 In the previous 3rd version, there was still an errors in writing the abbreviation of USU which was written in English, so after consultation the naming of USU still remain in Bahasa. In the previous 3rd version, there was still an errors in writing the abbreviation of USU which was written in English, so after consultation the naming of USU still remain in Bahasa. See the authors' detailed response to the review by Dario Gaudioso See the authors' detailed response to the review by Beni Jequicene Mussengue Chaúque READ REVIEWER RESPONSES Introduction Dental health workers are a group that is vulnerable to infection. 1 In dental practice, microorganisms can spread through blood, saliva or droplets through direct or indirect contact. One route of cross-infection due to indirect contact is tooth extraction instruments that have been contaminated with pathogenic microorganisms. 2 Tooth extraction procedures have a high risk of infection transmission due to contact of the instruments with the patient’s blood and saliva. 3 These must be aseptic at the time of tooth extraction to avoid bacterial infection, which is one of the complications that can occur as a result of tooth extraction. 4 If the tooth extraction instrument is contaminated, microorganisms can be transmitted to dentists, nurses, or other patients who will be infected with diseases. 4 According to the American Dental Association (ADA), it is estimated that dentists and patients may be exposed to around 40 types of infectious diseases when carrying out dental treatment procedures. Therefore, it is necessary to exercise prevention by disinfecting dental extraction instruments. 2 Alternative materials for disinfection in dentistry have been developed. Ozone water can function as a disinfectant with the ability to oxidize amino acids and destroy proteins in the cellular membranes of microorganisms. 5 Ozone water has the ability to kill pathogenic microorganisms and the resulting wastewater is safe to enter waterways. 6 Chloroxylenol or para-chloro-meta-xylenol (PCMX) is a chemical-level disinfectant commonly used to disinfect skin and surgical instruments in dentistry. 7 4.8% chloroxylenol has a broad-spectrum antibacterial ability that can kill most bacteria and fungi. 4.8% chloroxylenol works by denaturing proteins, changing the permeability of cell walls and causing cell leakage. 4.8% chloroxylenol is one of the most widely used disinfectants in dentistry. 4.8% chloroxylenol is one of the phenol groups, and works by denaturing proteins, changing the permeability of cell walls and causing cell leakage. 8 Research conducted by Ochie K and Ohagwu C in 2009 demonstrated that 3.5% sodium hypochlorite was the most effective in disinfecting x-ray equipment and accessories, followed by methylated spirit, 4.8% chlorocylenol, and 2% dichlorocylenol. 9 A study by Igizeneza et al. in 2020 revealed that 4.8% chlorocylenol was highly efficient in eradicating Staphylococcus sp. and Streptococcus sp. strains by 100%. Guther's research in 2020, focusing on skin surfaces and hospital environments, discovered that 4.8% chlorocylenol could inhibit the metabolic activity of 38.9% of gram-positive bacteria and 60.7% of gram-negative bacteria. Research on the effectiveness of ozone water (10 mg/L) as a disinfection agent on diamond burs that have been contaminated by Streptococcus aureus, Escherichia coli, Candida albicans and spores of Bacillus atrophaeus showed a reduction of microorganisms by 90.15-99.33%. 9 Research on the effectiveness of 4.8% chloroxylenol as a disinfecting agent showed its ability to interfere with the metabolic activity of Gram-positive bacteria at 38.9% concentration and Gram-negative at 60.7% concentration. 10 There are many disinfection agents that can be used in dentistry. Hypochlorous acid (HOCl) was evaluated as an antimicrobial disinfectant for alginate impression materials using a spraying technique. Research conducted by Bayan et al. HOCl at a concentration of 200 ppm for 5 and 10 minutes was compared with a negative control (no treatment) and a positive control using 0.5% sodium hypochlorite. Antimicrobial activity against Candida albicans, Staphylococcus aureus, and Pseudomonas aeruginosa was assessed using colony forming unit tests, alongside tests for dimensional stability and reproduction of details. Results indicated that HOCl exhibited significant antimicrobial activity against all tested microorganisms. Importantly, HOCl did not affect the dimensional stability of the alginate impression material. Therefore, HOCl has the potential to be an effective antimicrobial agent for alginate impression materials without compromising their surface details or dimensional stability. 11 Sodium hypochlorite is one of the original and most widely used disinfectants, effective against a broad spectrum of microorganisms including Human Immunodeficiency Virus, viruses, fungi, bacterial species, and their spores. A study conducted by Aseel M. Al-khafaji, Shorouq M. Abass, and Bayan S. Khalaf explored the use of SOLO and sodium hypochlorite disinfectant solutions for the disinfection of dental casts by spraying the stone specimen. This research aimed to assess the surface hardness, dimensional accuracy, reproduction of details, and surface porosity of type III, type IV, and type IV extra hard dental stone. The findings revealed no significant differences in dimensional accuracy, reproduction of details, surface hardness, and surface porosity among all types of tested stone samples after immersion or spraying with SOLO and sodium hypochlorite, except for type IV extra hard stone, which showed significant differences in surface hardness after spraying with SOLO and in surface porosity after immersion in both SOLO and sodium hypochlorite solutions. 12 Improper cleaning and maintenance of dentures can lead to contamination by various microbial pathogens, contributing to oral health issues. In a study by Maha A. Mahmood, Bayan S. Khalaf, and Shorouq M. Abass, HOCl was investigated to compare its antimicrobial effectiveness with oxalic, tartaric, citric acids, and alkaline peroxide, along with microwave irradiation, on the growth of Candida albicans and Staphylococcus aureus, respectively. Microwave irradiation was used to disinfect specimens of heat-activated acrylic resin and soft liner material. Microwave treatment or tartaric acid achieved sterilization of both heat-cured acrylic resin and soft lining material contaminated with Staphylococcus aureus, while microwave energy, oxalic acid, tartaric acid, and alkaline peroxide achieved complete sterilization against Candida albicans. Microwave irradiation was found to be the most effective method for sterilizing both heat-cured acrylic resin and soft lining material contaminated with either Staphylococcus aureus or Candida albicans, with tartaric acid ranking second. 13 This study aimed to compare the effectiveness of ozone water and 4.8% chloroxylenol disinfectants on tooth extraction instruments, because these critical instruments have a high risk of causing infection as they penetrate the mucous membranes of the oral cavity. 9 Methods Study type This research was a laboratory experimental research with post-test-only control group designs. The treatment group used ozone water and 4.8% chloroxylenol (dettol ® ) and the negative control group was cleaned with distilled water (aquadest). Location Universitas Sumatera Utara Dental and Oral Hospital, Laboratory of Microbiology, Faculty of Pharmacy, Mathematics and Natural Science University of Sumatera Utara, Medan, Indonesia. Duration The study duration was about 3 months, from October to December. Sampling The sampling technique used in this study was purposive sampling with 30 mandibular molar tooth extraction forceps as the sample. There were three treatment groups: the ozone water treatment group with 15 mg/L concentration, the 4.8% chloroxylenol treatment group, and the negative control (Aquadest) with 10 molar tooth extraction forceps in each treatment group. 7 Ozone water production Ozone water was prepared using an ozone generator with an ozone water concentration of 15 mg/L. The sterile nierbeken container is filled with 250 mL of distilled water, the container is inserted with a hose from the ozone generator. The timer on the ozone generator machine is set for 20 minutes to change the mineral water into ozone water. 14 Disinfection procedure In the ozone water and 4.8% chloroxylenol treatment group, the mandibular molar tooth extraction forceps were rinsed under running water, brushed with an antiseptic solution and rinsed again until the forceps were free of blood and saliva before soaking in a disinfecting solution. After that, the mandibular molar tooth extraction forceps were soaked in ozonized water for 30 minutes. 15 In the 4.8% chloroxylenol treatment group, mandibular molar extraction forceps were soaked for 60 minutes in 250 mL 4.8% chloroxylenol. 7 In the negative control group (Aquadest) the forceps were rinsed with distilled water without using soap or antiseptic solution at all. After disinfection, the forceps were removed from the disinfection container using tool tongs and dried with sterile gauze. The mandibular molar tooth extraction forcep is treating individually. Taking the samples Sampling was carried out by immersing mandibular molar extraction forceps in a container containing 50 mL of saline solution for five minutes, then covered with aluminum foil. The forceps are removed from the sample container and the container is closed again with plastic wrap and labeled for each sample. Group label for ozonized water was Z, 4.8% chloroxylenol was K, and distilled water was A. 10 Counting bacterial colonies The samples were then taken to the Faculty of Pharmacy, Mathematics and Natural Science, University of Sumatera Utara, to count the number of bacterial colonies. Samples were diluted using NaCl as much as 10 3 , then implanted in a petri dish containing plant count agar (PCA) media, using the spread plate method. A spread method plate is a technique for growing microorganisms in agar media isolate or count the bacteria present. 14 The sample was spread using a sterile spreader, and the petri dish was rotated at an angle of 45° above a Bunsen burner. After that, the petri dishes were incubated in the incubator using an autoclave at a temperature of 37°C for 24 hours. Following this, the number of bacterial colonies was counted using a bacterial colony counter. Statistical analysis Data analysis was performed using the SPSS version 22.0 software. The normality of the data was tested using the Shapiro-Wilk test, 9 since the number of samples was less than 50. The results of the data normality test showed that the data were not normally distributed, with p-values with a degree of significance < 0.05; therefore the next test used was the non-parametric Kruskal Wallis test followed up by the Mann-Whitney U test with a significance degree of p ≤ 0.05, to find out which treatment group had the best effectiveness as a disinfecting agent ( Figure 1 ). 16 Figure 1. Research design. Results This study consisted of 30 mandibular molar extraction forceps with three treatment groups, namely the mandibular molar extraction forceps group which was disinfected with ozonized water (Z), the mandibular molar forceps group which was disinfected with 4.8% chloroxylenol solution (K), and the mandibular molar forceps group which was rinsed with distilled water (A) as a negative control. 17 Each group consisted of 10 mandibular molar extraction forceps. 18 The 10 mandibular molar-removing forceps that were disinfected with ozonized water had a mean bacterial count of 4.00 ± 4.32 CFU/mL, showing they were still contaminated with pathogenic microorganisms with the highest number of bacterial colonies being 11·10 3 CFU/mL. 19 The 10 forceps which were disinfected with 4.8% chloroxylenol had a mean bacterial count of 16.00 ± 6.65 CFU/mL, showing they were still contaminated with pathogenic microorganisms, with the highest number of bacterial colonies being 27·10 3 CFU/mL. 20 Meanwhile, the mean bacterial count in the negative control group (Aquades) the 10 forceps extracting mandibular molars showed 217.50 ± 39.24 CFU/mL still forming bacterial colonies, with the highest number of bacterial colonies being 292·10 3 CFU/mL ( Table 1 ). 14 Table 1. Number of bacterial colonies (10 3 × CFU/mL). Code Ozone water 4.8% Chloroxylenol Aquadest 1 0 14 292 2 0 15 230 3 0 5 257 4 4 11 236 5 0 13 195 6 5 22 253 7 11 12 270 8 9 27 191 9 2 24 205 10 9 17 173 Based on the Kruskal Wallis statistical test results the effectiveness of ozone water, 4.8% chloroxylenol and negative control (Aquadest) on the number of bacterial colonies in mandibular tooth extraction forceps was significantly different, with a significance value of p = 0.000 (p < 0.05) ( Table 2 ). 4 Table 2. Kruskal Wallis test results for ozone water, 4.8% chloroxylenol, and negative control (Aquadest). Treatment Mean ± SD p-value Ozone water 4.00 ± 4.32 0.000 4.8% Chloroxylenol 16.00 ± 6.65 Negative control (Aquadest) 217.50 ± 39.24 Based on the results of a further using the Mann-Whitney U test comparing disinfection effectiveness between groups treated with ozone water disinfection and negative control (Aquadest), the significance value was p = 0.000 (p < 0.05). 21 Ozone water was more effective as a disinfection agent compared to negative control on the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital, as indicated by the lower average value of bacterial counts in the ozone water group, which was 4.00 ± 4.32 CFU/mL. In contrast, for the negative control group (Aquadest) the value was 217.50 ± 39.24 ( Table 3 ). Table 3. Test results of the Mann-Whitney U comparison of ozone water and negative control (Aquadest). Treatment Mean p-value Ozone water 4.00 ± 4.32 0.000 Negative control (Aquadest) 217.50 ± 39.24 Further Mann-Whitney U test results showed that there was a significant difference between the effectiveness of disinfection between groups treated with 4.8% chloroxylenol and negative control with a significance value of p = 0.000 (p < 0.05). 4.8% chloroxylenol was more effective as a disinfecting agent compared to negative control on the number of bacterial colonies in dental extraction instruments. 22 The Universitas Sumatera Utara indicated that the average bacterial count value of the 4.8% chloroxylenol group was lower with 16 ± 6.65 CFU/mL, while the value for the negative control group was 217.50 ± 39.24 ( Table 4 ). Table 4. Test results of the Mann-Whitney U comparison of 4.8% chloroxylenol, and negative control (Aquadest). Treatment Mean ± SD p-value 4.8% Chloroxylenol 16.00 ± 6.65 0.000 Negative control (Aquadest) 217.50 ± 39.24 Further Mann-Whitney U test results showed that there was a significant difference in effectiveness between ozone water and 4.8% chloroxylenol with a significance value of p = 0.000 (p < 0.05). 23 Ozone water was more effective as a disinfection agent compared to 4.8% chloroxylenol against bacterial colonies on dental extraction instruments, as indicated by the lower average bacterial count value of the ozone water group which was 4.00 ± 4.32 CFU/mL, while for the disinfection treatment group with 4.8% chloroxylenol, the value was 16 ± 6.65 CFU/mL ( Table 5 ). 24 Table 5. Mann-Whitney U test results comparing ozone water and chloroxylenol 4.8%. Treatment Mean p-value Ozone water 4.00 ± 4.32 0.000 4.8% chloroxylenol 16.00 ± 6.65 Based on the results of data analysis from each treatment group, it was shown that ozone water was significantly more effective in disinfecting mandibular molar tooth extraction forceps compared to chloroxylenol 4.8% and negative control. 25 Instruments in the 4.8% chloroxylenol group showed a smaller number of bacterial colonies than the negative control ( Figure 2 ). Figure 2. Total plate count 4.8% chloroxylenol, aquadest, and ozone water. Discussion This study compared two disinfection solutions, namely ozone water and 4.8% chloroxylenol. Ozone water is a strong alternative to disinfectants and effectively kills pathogenic microorganisms such as bacteria, viruses, fungi, protozoa, and endospores. 6 Ozone is more effective against large numbers of microorganisms because of its ability to oxidize microorganisms without causing resistance. Ozone can inhibit the control of enzymes and therefore act on bacterial cell metabolism and damage bacterial cell membranes. Ozone water disrupts the integrity of the bacterial cell envelope through the oxidation of phospholipids and lipoproteins. In this study, the concentration of ozone water used was 15 mg/L, with a soaking time of 30 minutes. The use of ozone water as a material for disinfecting dental instruments was recommended at a concentration of 10-20 mg/L. As reported by previous research, a concentration of ozone water that is too low showed a reduction in the antibacterial effectiveness of ozone water, while a concentration that is too high and exposure for too long could cause toxicity. Ozone maintained its antibacterial properties for the first 20 minutes, and after 30 minutes the stability of ozone water slowly decreased. After eight hours, there was no more ozone in the ozonized water. Based on the data on the number of bacterial colonies in the treatment group with ozone water samples, ozone water was more effective in disinfecting the mandibular molar forceps. It was observed that four out of 10 mandibular molar extraction forceps were successfully decontaminated, with six samples showing bacterial growth with the highest number of bacteria being 11·10 3 CFU/mL. The number of bacterial colonies in the treatment group with ozone water was the lowest when compared to the other two treatment groups, with an average value of 4.00 ± 4.32. This is due to the greater efficiency of ozone water in killing bacterial endospores by oxidizing and damaging the bacterial membrane which contains lipoproteins and fatty acids, so that the bacteria experience failure in the germination process. In contrast, 4.8% chloroxylenol is unable to kill bacterial endospores, therefore 4.8% chloroxylenol is still classified as medium-level chemical disinfection. This makes ozone water more effective in killing microorganisms when compared to 4.8% chloroxylenol according to César et al. , 2012, who stated that the antimicrobial activity of ozone water against B. atrophaeus spores resulted in a decrease in growth of 90.15% and 98.74% after 10 and 30 minutes of exposure to ozone water, respectively. In this study, the average number of bacterial colonies in the ozone water treatment group was lower due to the ability of the ozone water to kill bacterial endospores. Endospora is very resistant to high temperatures, extreme environmental conditions, or chemicals. The ability of ozone water to kill bacterial endospores was likely one of the causes of the fewer bacterial colonies formed in the ozone water treatment group. Data from César et al. also shows that the use of ozonated water (10 mg/L) for 30 minutes was effective for disinfection of diamond burs that have been contaminated by S. aureus , E. coli , C. albicans , and spores of B. atrophaeus , reducing bacterial contamination by 90.15-99.33%. In addition, based on Bezirtzoglou et al. found the research of César et al. 10 states that a few teeth that have been soaked for 30 minutes with ozone water showed no bacterial colonies formed on the toothbrush. In the research by Nielsen et al. , 2007, the addition of ozone to water that has been inoculated with S. aureus showed a reduction of 98.9%; S. faecalis , P. aeruginosa were respectively reduced by 64.2% and 57.4%. 26 According to Xiao Hu et al. , 2021, ozone water can eliminate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with an ozone concentration above 18 mg/L within one minute. 15 Wood et al. stated that ozone water can reduce fungal spores by 50.7-91.2%. Thus, ozone water can kill microorganisms in the form of bacteria, viruses, fungi, and bacterial endospores. 27 In the 4.8% chloroxylenol treatment group, there was bacterial growth in all mandibular molar tooth extraction forceps with a lower value compared to ozone water, but the number of bacterial colonies formed was less than the negative control. The mean bacterial count value in the 4.8% chloroxylenol group was 16.00 ± 6.65, while for the negative control group, the value was 217.50 ± 39.24. The research by Xiang et al. in 2018 showed that 11 out of 18 mandibular molar tooth extraction forceps which were cleaned using 4.8% chloroxylenol did not form bacterial colonies, and seven mandibular molar tooth extraction forceps had a maximum bacterial count of 812·10 3 CFU/mL and the average number of bacterial colonies formed was 82.5. This is by the results of this study: in all research samples bacterial colonies still formed although with fewer bacterial colonies on average. 28 Liu et al. , 2020, who researched the skin and the hospital environment, found that 4.8% chloroxylenol could interfere with the metabolic activity of Gram-positive bacteria by 38.9% and Gram-negative by 60.7%. 29 Mohammed AL-jaleel Khalil et al. , 2023, showed that 4.8% chloroxylenol was effective in killing 100% isolates of Staphylococcus sp. and Streptococcus sp. Thus, 4.8% chloroxylenol was able to kill pathogenic microorganisms. Still, its effectiveness is lower when compared to ozone water. The -OH hydroxyl group of the 4.8% chloroxylenol molecule binds to proteins in the bacterial cell membrane to disrupt it, thereby allowing the contents of the bacterial cell to leak. This allows 4.8% chloroxylenol to enter the bacterial cell to bind more proteins and enzymes and deactivate cell functions. Meanwhile, the negative control group (Aquadest) had a lethal effect on microorganisms because distilled water is a neutral organic compound used as a pure solvent and does not have antibacterial activity. 20 Differences in the number of bacterial colonies can also be caused by several factors, one of which is the number of microorganisms and contaminants, the condition of the patient’s oral cavity, the number and type of normal flora in the oral cavity of each patient which can differ due to severe caries, periodontal disease, pulpal necrosis or abscess at the time of tooth extraction. 4 Disinfectants have different abilities and susceptibility depending on the number and type of bacteria present in the patient’s oral cavity; the duration of exposure and the concentration of the disinfecting agents also affect the ability to kill microorganisms. 8 The decrease in the number of different bacterial colonies in each treatment is also influenced by open environmental conditions, environmental temperature, and humidity both during sampling and when processing samples in the laboratory, ozone generator output, and operator negligence. Conclusions Ozone water was significantly more effective in disinfecting dental extraction instruments at Universitas Sumatera Utara Dental and Oral Hospital than 4.8% chloroxylenol with a significance value of p = 0.000 (p < 0.05). Data availability Underlying data Zenodo: The Data Set “Ozone Water And 4.8% Chloroxylenol Against The Number Of Bacterial Colonies In Dental Extraction Instruments at The Universitas Sumatera Utara Dental and Oral Hospital”, https://doi.org/10.5281/zenodo.7950593 . 30 This project contains the following underlying data: • Raw data of total plate count aquades.csv (Raw data of total plate count aquades) • Raw data of total plate count chloroxylenol- Sheet1.csv (Raw data of total plate count 4.8% choloroxylenol) • Raw data of total plate count.ozone water.csv (Raw data of total plate count.ozone water) • Result of Normalitas Test.csv (raw data of result of normalitas test with shapiro wilk) • Result of Kruskal wallis test.csv (raw data of result of kruskal wallis) • Result of comparison ozone water and Choloroxylenol.csv (raw data of result of comparison ozon water and 4,8% chloroxylenol) • Result of Comparison Choloxylenol and Aquades.csv (raw data of result of comparison 4,8% chloroxylenol and aquades) • Result of Comparison ozone water and Aquades.csv (raw data of result of comparison ozone water and aquades) • Z1-10.rar (image of colony bactery plate of ozone water) • Choloroxylenol.jpg (image of colony bactery plate of 4,8% chloroxylenol) • Aquades.jpg (image of colony bactery plate of aaquades) Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0). Acknowledgments An earlier version of this article can be found in the authors’ institutional repository at https://repositori.usu.ac.id/handle/123456789/83488 . References 1. Malsam R, Nienhaus A: Occupational infections among dental health workers in germany—14-year time trends. Int. J. Environ. Res. Public Health . 2021; 18 . 2. Walley S: Essential microbiology for dentistry Br. Dent. J. 4th edition.2012; 212 . : 350. Publisher Full Text 3. Nielsen PV, Xu C: Multiple airflow patterns in human microenvironment and the influence on short-distance airborne cross-infection – A review. Indoor Built Environ. 2022; 31 : 1161–1175. Publisher Full Text 4. Winarti S, Peristiowati Y, Siyoto S: Analysis of Patient Safety Management Implementation Towards the Occurrence of Post-Tooth Extraction Infection in Oral Surgery Clinic at RSGM IIK Bhakti Wiyata Kediri. J. Qual. Public Heal. 2018; 2 : 17–24. Publisher Full Text 5. 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Publisher Full Text Comments on this article Comments (0) Version 4 VERSION 4 PUBLISHED 21 Jun 2023 ADD YOUR COMMENT Comment Author details Author details 1 Universitas Sumatera Utara, Medan, North Sumatra, Indonesia Hendry Rusdy Roles: Conceptualization, Data Curation, Formal Analysis, Funding Acquisition, Investigation, Methodology, Project Administration, Resources, Software, Supervision, Validation, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing Rahmi Syaflida Dalimunthe Roles: Data Curation, Investigation, Methodology, Project Administration, Resources, Validation, Writing – Review & Editing Ahyar Riza Roles: Data Curation, Investigation, Project Administration, Validation, Writing – Original Draft Preparation, Writing – Review & Editing Ismahani Akilah D Roles: Data Curation, Formal Analysis, Investigation, Resources, Visualization, Writing – Review & Editing Competing interests No competing interests were disclosed. Grant information The author(s) declared that no grants were involved in supporting this work. Article Versions (4) version 4 Revised Published: 31 Jul 2025, 12:726 https://doi.org/10.12688/f1000research.132941.4 version 3 Revised Published: 12 May 2025, 12:726 https://doi.org/10.12688/f1000research.132941.3 version 2 Revised Published: 24 May 2024, 12:726 https://doi.org/10.12688/f1000research.132941.2 version 1 Published: 21 Jun 2023, 12:726 https://doi.org/10.12688/f1000research.132941.1 Copyright © 2025 Rusdy H et al . This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The author(s) is/are employees of the US Government and therefore domestic copyright protection in USA does not apply to this work. The work may be protected under the copyright laws of other jurisdictions when used in those jurisdictions. Download Export To Sciwheel Bibtex EndNote ProCite Ref. Manager (RIS) Sente metrics Views Downloads F1000Research - - PubMed Central info_outline Data from PMC are received and updated monthly. - - Citations open_in_new 0 open_in_new 0 open_in_new SEE MORE DETAILS CITE how to cite this article Rusdy H, Dalimunthe RS, Riza A and D IA. Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.12688/f1000research.132941.4 ) NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS track receive updates on this article Track an article to receive email alerts on any updates to this article. TRACK THIS ARTICLE Share Open Peer Review Current Reviewer Status: ? Key to Reviewer Statuses VIEW HIDE Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Version 4 VERSION 4 PUBLISHED 31 Jul 2025 Revised Views 0 Cite How to cite this report: Gaudioso D. Reviewer Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.182380.r405353 ) The direct URL for this report is: https://f1000research.com/articles/12-726/v4#referee-response-405353 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 23 Aug 2025 Dario Gaudioso , Università degli Studi di Firenze, Sesto Fiorentino, Firenze, Italy Approved VIEWS 0 https://doi.org/10.5256/f1000research.182380.r405353 Paper's experimental simplicity is very clear. It compares the use of ozonated water with the traditional treatment with chloroxylenol 4.8%. There are some points that would have been interesting to explore further, such as a more thorough screening of the genera ... Continue reading READ ALL Paper's experimental simplicity is very clear. It compares the use of ozonated water with the traditional treatment with chloroxylenol 4.8%. There are some points that would have been interesting to explore further, such as a more thorough screening of the genera of microorganisms that survive the different treatments. In parts of the paper, the genus and species are not italicized. Methodologically, some literature reports that the effect of ozone after 5 minutes is not effective, contrary to what is reported here. Overall, the paper, despite its operational and experimental simplicity, presents some interesting key points. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: Molecular biology and microbiology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Gaudioso D. Reviewer Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.182380.r405353 ) The direct URL for this report is: https://f1000research.com/articles/12-726/v4#referee-response-405353 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 16 Oct 2025 Hendry Rusdy , Universitas Sumatera Utara, Medan, Indonesia 16 Oct 2025 Author Response Thankyou for your feedback and accepting my paper Competing Interests: No competing interests were disclosed. Thankyou for your feedback and accepting my paper Thankyou for your feedback and accepting my paper Competing Interests: No competing interests were disclosed. Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 16 Oct 2025 Hendry Rusdy , Universitas Sumatera Utara, Medan, Indonesia 16 Oct 2025 Author Response Thankyou for your feedback and accepting my paper Competing Interests: No competing interests were disclosed. Thankyou for your feedback and accepting my paper Thankyou for your feedback and accepting my paper Competing Interests: No competing interests were disclosed. Close Report a concern COMMENT ON THIS REPORT Version 2 VERSION 2 PUBLISHED 24 May 2024 Revised Views 0 Cite How to cite this report: Chaúque BJM. Reviewer Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.158749.r319597 ) The direct URL for this report is: https://f1000research.com/articles/12-726/v2#referee-response-319597 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 20 Sep 2024 Beni Jequicene Mussengue Chaúque , Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Universidade Rovuma, Niassa Branch, Lichinga, Mozambique Not Approved VIEWS 0 https://doi.org/10.5256/f1000research.158749.r319597 The MS titled “Ozone Water or Chloroxylenol: A Comparison of Disinfection Effectiveness Against Bacterial Colonies on Dental Extraction Instruments at the USU Dental and Oral Hospital” explores the effectiveness of ozone water and chloroxylenol in inactivating bacteria on dental instruments. ... Continue reading READ ALL The MS titled “Ozone Water or Chloroxylenol: A Comparison of Disinfection Effectiveness Against Bacterial Colonies on Dental Extraction Instruments at the USU Dental and Oral Hospital” explores the effectiveness of ozone water and chloroxylenol in inactivating bacteria on dental instruments. General comments While the topic is of considerable interest, the study suffers from several key issues, particularly in the writing and methodology sections, which are presented in an unstructured way. Moreover, the study's overall contribution to the existing literature appears limited. The sampling procedures are unclear, and the authors do not provide sufficient detail on how they minimized the risk of bias in the allocation of instrument samples. With only 10 samples, the study’s findings are not robust enough to justify any broad generalizations. Another major limitation is the lack of identification and/or isolation of microorganisms, which would have allowed for a more standardized evaluation of disinfectant performance against comparable inoculum sizes. Additionally, the washing processes used prior to instrument handling, to remove blood and saliva, could introduce bias especially given the small sample size. Specific comments Title 1- It is not advisable to use abbreviations in the title. I suggest either writing "USU" in full or omitting it entirely. Abstract 2 -The rationale for conducting the study is not clearly established. Please elaborate further. For example, isn't the use of ozone water and 4.8% chloroxylenol already a proven and standardized aseptic practice? If so, why is this study necessary? 3 -Additionally, the abbreviation "USU" appears in the abstract without being explained. 4 -“This study aimed to evaluate the effectiveness of ozone water and 4.8% chloroxylenol in reducing bacterial colonies on dental extraction instruments at the USU Dental and Oral Hospital between October and December 2022.” Note: The phrase "between October and December 2022" seems unnecessary here and can be removed. 5 -The methods section of the abstract should be written more formally, detailing the sampling, transportation, treatment (including the form, application, and exposure), and the plate inoculation process. 6 -The study is based on only 10 different instruments. Why such a small sample size? How representative is this sample? 7 -The results are reported in a confusing manner. The p-value analysis is presented before the colony count values. The authors present two values for the number of colonies in the treatment group: 4.00 ± 4.32 and 16.00 ± 6.65. Which value is correct? If the values refer to ozone water and chloroxylenol, respectively, please specify this clearly. Introduction 8 -Paragraphs 1, 2, and 3 contain redundant information. Please consider rewriting to avoid repetition. 9 - Note: Only separate paragraphs when necessary for clarity. 10 - In paragraph 5, ensure the names of microorganisms are spelled correctly. Methods 11 - The sentence "The patient’s oral cavity state, the kind and quantity of bacteria present, the surrounding circumstances, and the instrument settings before tooth extraction are all non-parametric" is unclear. This sounds more like a result rather than a method. Please consider rewriting 12 - The following information needs to be placed in the appropriate section: "The study parameters include the concentration of ozone water (15 mg/L), the concentration of chloroxylenol (4.8%), the immersion time in ozone water (30 minutes), the immersion time in chloroxylenol solution (60 minutes), and the pre-treatment procedures." This information is also repeated in the "Disinfection procedure" section. 13 - Instead of simply stating that " The sampling technique used in this study was purposive sampling" provide a detailed, step-by-step explanation of the sampling process to ensure reproducibility by other researchers. 14 - The subsections “Disinfection procedure” and “Taking the samples” are unclear and need to be rewritten in a more straightforward, didactic manner. Clearly distinguish between the different types of samples used in the study, such as instruments and inoculum. Include specific details regarding their identification, selection, and collection. 15 - It is also unclear whether the instruments were treated individually or as a group. 16 - In the “Counting bacterial colonies” subsection, specify the volume of the sample plated and the incubation temperature. Results 17 - The results in Tables 2–5 would be easier to understand if presented in a graph, allowing readers to visualize and compare the data more effectively. Conclusion 18 - The statement “Ozone water can kill bacterial endospores while 4.8% chloroxylenol cannot” should not be included in the conclusions, as this was not assessed in the study. Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Partly Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? No Are the conclusions drawn adequately supported by the results? Partly Competing Interests: No competing interests were disclosed. Reviewer Expertise: Microbiology and Pharmacology I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Chaúque BJM. Reviewer Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.158749.r319597 ) The direct URL for this report is: https://f1000research.com/articles/12-726/v2#referee-response-319597 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Author Response 12 May 2025 Hendry Rusdy , Universitas Sumatera Utara, Medan, Indonesia 12 May 2025 Author Response General comments The amount of sample is 30 Specific comments Title Change USU to University of South Sumatera Abstract 2 -The rationale for conducting the study is not clearly established. ... Continue reading General comments The amount of sample is 30 Specific comments Title Change USU to University of South Sumatera Abstract 2 -The rationale for conducting the study is not clearly established. Please elaborate further. For example, isn't the use of ozone water and 4.8% chloroxylenol already a proven and standardized aseptic practice? If so, why is this study necessary? answer : Research on ozone water as a disinfectant is still limited and no research has been found on the use of ozone water on tooth extraction instruments. However, ozone has antibacterial properties so researchers are interested in studying the effectiveness of ozone water as a disinfectant when compared to the commonly used 4.8% chloroxylenol 3 - "USU" appears in the abstract without being explained. 4 -The phrase "between October and December 2022" seems unnecessary here and can be removed. 5 -The methods has been updated 6 -The study is based on only 10 different instruments. Why such a small sample size? How representative is this sample? answer : The amount of sample is 30 7 -The results are reported in a confusing manner. The p-value analysis is presented before the colony count values. The authors present two values for the number of colonies in the treatment group: 4.00 ± 4.32 and 16.00 ± 6.65. Which value is correct? If the values refer to ozone water and chloroxylenol, respectively, please specify this clearly. answer : value refer to ozone water, which is 4.00 ± 4.32 Introduction 8 -Paragraphs 1, 2, and 3 has been updated 9 - updated 10 - In paragraph 5, ensure the names of microorganisms are spelled correctly has been updated Methods 11 - The sentence "The patient’s oral cavity state, the kind and quantity of bacteria present, the surrounding circumstances, and the instrument settings before tooth extraction are all non-parametric" is unclear. This sounds more like a result rather than a method. Please consider rewriting have been updated 12 - The following information needs to be placed in the appropriate section: "The study parameters include the concentration of ozone water (15 mg/L), the concentration of chloroxylenol (4.8%), the immersion time in ozone water (30 minutes), the immersion time in chloroxylenol solution (60 minutes), and the pre-treatment procedures." This information is also repeated in the "Disinfection procedure" section have been updated 13 - Instead of simply stating that " The sampling technique used in this study was purposive sampling" provide a detailed, step-by-step explanation of the sampling process to ensure reproducibility by other researchers. have been updated 14 - The subsections “Disinfection procedure” and “Taking the samples” are unclear and need to be rewritten in a more straightforward, didactic manner. Clearly distinguish between the different types of samples used in the study, such as instruments and inoculum. Include specific details regarding their identification, selection, and collection. have been updated 15 - It is also unclear whether the instruments were treated individually or as a group. have been updated 16 - In the “Counting bacterial colonies” subsection, specify the volume of the sample plated and the incubation temperature. have been updated Results 17 - The results in Tables 2–5 would be easier to understand if presented in a graph, allowing readers to visualize and compare the data more effectively. have been updated Conclusion 18 - The statement “Ozone water can kill bacterial endospores while 4.8% chloroxylenol cannot” should not be included in the conclusions, as this was not assessed in the study. have been updated General comments The amount of sample is 30 Specific comments Title Change USU to University of South Sumatera Abstract 2 -The rationale for conducting the study is not clearly established. Please elaborate further. For example, isn't the use of ozone water and 4.8% chloroxylenol already a proven and standardized aseptic practice? If so, why is this study necessary? answer : Research on ozone water as a disinfectant is still limited and no research has been found on the use of ozone water on tooth extraction instruments. However, ozone has antibacterial properties so researchers are interested in studying the effectiveness of ozone water as a disinfectant when compared to the commonly used 4.8% chloroxylenol 3 - "USU" appears in the abstract without being explained. 4 -The phrase "between October and December 2022" seems unnecessary here and can be removed. 5 -The methods has been updated 6 -The study is based on only 10 different instruments. Why such a small sample size? How representative is this sample? answer : The amount of sample is 30 7 -The results are reported in a confusing manner. The p-value analysis is presented before the colony count values. The authors present two values for the number of colonies in the treatment group: 4.00 ± 4.32 and 16.00 ± 6.65. Which value is correct? If the values refer to ozone water and chloroxylenol, respectively, please specify this clearly. answer : value refer to ozone water, which is 4.00 ± 4.32 Introduction 8 -Paragraphs 1, 2, and 3 has been updated 9 - updated 10 - In paragraph 5, ensure the names of microorganisms are spelled correctly has been updated Methods 11 - The sentence "The patient’s oral cavity state, the kind and quantity of bacteria present, the surrounding circumstances, and the instrument settings before tooth extraction are all non-parametric" is unclear. This sounds more like a result rather than a method. Please consider rewriting have been updated 12 - The following information needs to be placed in the appropriate section: "The study parameters include the concentration of ozone water (15 mg/L), the concentration of chloroxylenol (4.8%), the immersion time in ozone water (30 minutes), the immersion time in chloroxylenol solution (60 minutes), and the pre-treatment procedures." This information is also repeated in the "Disinfection procedure" section have been updated 13 - Instead of simply stating that " The sampling technique used in this study was purposive sampling" provide a detailed, step-by-step explanation of the sampling process to ensure reproducibility by other researchers. have been updated 14 - The subsections “Disinfection procedure” and “Taking the samples” are unclear and need to be rewritten in a more straightforward, didactic manner. Clearly distinguish between the different types of samples used in the study, such as instruments and inoculum. Include specific details regarding their identification, selection, and collection. have been updated 15 - It is also unclear whether the instruments were treated individually or as a group. have been updated 16 - In the “Counting bacterial colonies” subsection, specify the volume of the sample plated and the incubation temperature. have been updated Results 17 - The results in Tables 2–5 would be easier to understand if presented in a graph, allowing readers to visualize and compare the data more effectively. have been updated Conclusion 18 - The statement “Ozone water can kill bacterial endospores while 4.8% chloroxylenol cannot” should not be included in the conclusions, as this was not assessed in the study. have been updated Competing Interests: No competing interests were disclosed. Close Report a concern Respond or Comment COMMENTS ON THIS REPORT Author Response 12 May 2025 Hendry Rusdy , Universitas Sumatera Utara, Medan, Indonesia 12 May 2025 Author Response General comments The amount of sample is 30 Specific comments Title Change USU to University of South Sumatera Abstract 2 -The rationale for conducting the study is not clearly established. ... Continue reading General comments The amount of sample is 30 Specific comments Title Change USU to University of South Sumatera Abstract 2 -The rationale for conducting the study is not clearly established. Please elaborate further. For example, isn't the use of ozone water and 4.8% chloroxylenol already a proven and standardized aseptic practice? If so, why is this study necessary? answer : Research on ozone water as a disinfectant is still limited and no research has been found on the use of ozone water on tooth extraction instruments. However, ozone has antibacterial properties so researchers are interested in studying the effectiveness of ozone water as a disinfectant when compared to the commonly used 4.8% chloroxylenol 3 - "USU" appears in the abstract without being explained. 4 -The phrase "between October and December 2022" seems unnecessary here and can be removed. 5 -The methods has been updated 6 -The study is based on only 10 different instruments. Why such a small sample size? How representative is this sample? answer : The amount of sample is 30 7 -The results are reported in a confusing manner. The p-value analysis is presented before the colony count values. The authors present two values for the number of colonies in the treatment group: 4.00 ± 4.32 and 16.00 ± 6.65. Which value is correct? If the values refer to ozone water and chloroxylenol, respectively, please specify this clearly. answer : value refer to ozone water, which is 4.00 ± 4.32 Introduction 8 -Paragraphs 1, 2, and 3 has been updated 9 - updated 10 - In paragraph 5, ensure the names of microorganisms are spelled correctly has been updated Methods 11 - The sentence "The patient’s oral cavity state, the kind and quantity of bacteria present, the surrounding circumstances, and the instrument settings before tooth extraction are all non-parametric" is unclear. This sounds more like a result rather than a method. Please consider rewriting have been updated 12 - The following information needs to be placed in the appropriate section: "The study parameters include the concentration of ozone water (15 mg/L), the concentration of chloroxylenol (4.8%), the immersion time in ozone water (30 minutes), the immersion time in chloroxylenol solution (60 minutes), and the pre-treatment procedures." This information is also repeated in the "Disinfection procedure" section have been updated 13 - Instead of simply stating that " The sampling technique used in this study was purposive sampling" provide a detailed, step-by-step explanation of the sampling process to ensure reproducibility by other researchers. have been updated 14 - The subsections “Disinfection procedure” and “Taking the samples” are unclear and need to be rewritten in a more straightforward, didactic manner. Clearly distinguish between the different types of samples used in the study, such as instruments and inoculum. Include specific details regarding their identification, selection, and collection. have been updated 15 - It is also unclear whether the instruments were treated individually or as a group. have been updated 16 - In the “Counting bacterial colonies” subsection, specify the volume of the sample plated and the incubation temperature. have been updated Results 17 - The results in Tables 2–5 would be easier to understand if presented in a graph, allowing readers to visualize and compare the data more effectively. have been updated Conclusion 18 - The statement “Ozone water can kill bacterial endospores while 4.8% chloroxylenol cannot” should not be included in the conclusions, as this was not assessed in the study. have been updated General comments The amount of sample is 30 Specific comments Title Change USU to University of South Sumatera Abstract 2 -The rationale for conducting the study is not clearly established. Please elaborate further. For example, isn't the use of ozone water and 4.8% chloroxylenol already a proven and standardized aseptic practice? If so, why is this study necessary? answer : Research on ozone water as a disinfectant is still limited and no research has been found on the use of ozone water on tooth extraction instruments. However, ozone has antibacterial properties so researchers are interested in studying the effectiveness of ozone water as a disinfectant when compared to the commonly used 4.8% chloroxylenol 3 - "USU" appears in the abstract without being explained. 4 -The phrase "between October and December 2022" seems unnecessary here and can be removed. 5 -The methods has been updated 6 -The study is based on only 10 different instruments. Why such a small sample size? How representative is this sample? answer : The amount of sample is 30 7 -The results are reported in a confusing manner. The p-value analysis is presented before the colony count values. The authors present two values for the number of colonies in the treatment group: 4.00 ± 4.32 and 16.00 ± 6.65. Which value is correct? If the values refer to ozone water and chloroxylenol, respectively, please specify this clearly. answer : value refer to ozone water, which is 4.00 ± 4.32 Introduction 8 -Paragraphs 1, 2, and 3 has been updated 9 - updated 10 - In paragraph 5, ensure the names of microorganisms are spelled correctly has been updated Methods 11 - The sentence "The patient’s oral cavity state, the kind and quantity of bacteria present, the surrounding circumstances, and the instrument settings before tooth extraction are all non-parametric" is unclear. This sounds more like a result rather than a method. Please consider rewriting have been updated 12 - The following information needs to be placed in the appropriate section: "The study parameters include the concentration of ozone water (15 mg/L), the concentration of chloroxylenol (4.8%), the immersion time in ozone water (30 minutes), the immersion time in chloroxylenol solution (60 minutes), and the pre-treatment procedures." This information is also repeated in the "Disinfection procedure" section have been updated 13 - Instead of simply stating that " The sampling technique used in this study was purposive sampling" provide a detailed, step-by-step explanation of the sampling process to ensure reproducibility by other researchers. have been updated 14 - The subsections “Disinfection procedure” and “Taking the samples” are unclear and need to be rewritten in a more straightforward, didactic manner. Clearly distinguish between the different types of samples used in the study, such as instruments and inoculum. Include specific details regarding their identification, selection, and collection. have been updated 15 - It is also unclear whether the instruments were treated individually or as a group. have been updated 16 - In the “Counting bacterial colonies” subsection, specify the volume of the sample plated and the incubation temperature. have been updated Results 17 - The results in Tables 2–5 would be easier to understand if presented in a graph, allowing readers to visualize and compare the data more effectively. have been updated Conclusion 18 - The statement “Ozone water can kill bacterial endospores while 4.8% chloroxylenol cannot” should not be included in the conclusions, as this was not assessed in the study. have been updated Competing Interests: No competing interests were disclosed. Close Report a concern COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Abass SM. Reviewer Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.158749.r282421 ) The direct URL for this report is: https://f1000research.com/articles/12-726/v2#referee-response-282421 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 06 Jun 2024 Shorouq Majid Abass , Department of Prosthodontics, College of Dentistry, University of Baghdad, Baghdad, Baghdad Governorate, Iraq Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.158749.r282421 Dear Authors, thank you for this interesting article. Here are some of the comments of mine: In the introduction section you have to mention other aspect of disinfection solution, technique, and dental materials in general other than of your ... Continue reading READ ALL Dear Authors, thank you for this interesting article. Here are some of the comments of mine: In the introduction section you have to mention other aspect of disinfection solution, technique, and dental materials in general other than of your specific subject of research. e.g. Khalaf BS, Abass SM, Al-Khafaji AM, and Issa MI, Antimicrobial efficiency of hypochlorous acid and its effect on some properties of alginate impression material. Int J Dent, 2023. 2023: p. 1-7 DOI: 10.1155/2023/8584875. Al-khafaji AM, Abass SM, and Khalaf BS, The effect of SOLO and sodium hypochlorite disinfectant on some properties of different types of dental stone. J Bagh Coll Dent, 2013. 25(2): p. 8-17 Mahmood MA, Khalaf BS, and Abass SM, Efficiency of different denture disinfection methods. Global J Bio-sci Biotech, 2017. 6: p. 439-4. It is only a suggestion for authors to use the above citations because they are related to the subject of disinfection in dentistry in general and the authors can refer to it if they wish. References 1. Khalaf BS, Abass SM, Al-Khafaji AM, Issa MI: Antimicrobial Efficiency of Hypochlorous Acid and Its Effect on Some Properties of Alginate Impression Material. Int J Dent . 2023; 2023 : 8584875 PubMed Abstract | Publisher Full Text 2. SM Abass: The effect of SOLO and sodium hypochlorite disinfectant on some properties of different types of dental stone. University of Baghdad . 1973. 8-17 3. Abass SM: Efficiency of different denture disinfection methods. Global J Bio-sci Biotech . 2017. 439-439 Competing Interests: No competing interests were disclosed. Reviewer Expertise: Prosthodontics, dental materials, disinfection and dental implant. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Abass SM. Reviewer Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.158749.r282421 ) The direct URL for this report is: https://f1000research.com/articles/12-726/v2#referee-response-282421 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Version 1 VERSION 1 PUBLISHED 21 Jun 2023 Views 0 Cite How to cite this report: Palma PJ. Reviewer Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.145897.r207744 ) The direct URL for this report is: https://f1000research.com/articles/12-726/v1#referee-response-207744 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 09 Oct 2023 Paulo J. Palma , Institute of Endodontics, Faculty of Medicine, University of Coimbra, Coimbra, Portugal Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.145897.r207744 The present study aimed to determine the effectiveness of disinfection of ozone water and 4.8% chloroxylenol in reducing the number of bacterial colonies on dental extraction instruments at the USU Dental and Oral Hospital between October-December 2022.. ... Continue reading READ ALL The present study aimed to determine the effectiveness of disinfection of ozone water and 4.8% chloroxylenol in reducing the number of bacterial colonies on dental extraction instruments at the USU Dental and Oral Hospital between October-December 2022.. This research is under the scope of this journal; the topic is relevant for readers, and this research deals with potentially significant knowledge of the field. And It will be important for knowledge. The topic is relevant for readers and this review deals with potentially significant knowledge in the field and opens new ways for future studies. However, there are some aspects that are possibly improved in the manuscript: Keywords Please add more keywords, and order the keywords / Mesh terms alphabetically Introduction What is the importance of this study? Which results are comparable with other articles? What has this study been new? M&M ​​​ A flowchart of all experimental study would be valuable. How many slides per sample obtained, and how many of them were evaluated? How were the sample cells calculated? Did authors performed power analysis to evaluate if this sample size was appropriate? Analysis: How many observers did the scoring? What is the interobserver agreement? How many time was performed done the determinations? Which data are parametric and non-parametric? Please specify? When mentioning materials or devices: for some of them you don't mention the manufacturer at all, for some you mention only the manufacturer, for some the manufacturer and city, for some you mention the manufacturer and city/ country. Discussion Please, clarified more limitations of this study? An objective presentation of the strengths and limitations of the model, study design and methods, must be provided, including any biology/functional variability between the animal model and humans. And also clarified the future perspectives in the discussion. The potential influence of the results on future research plans needs also be discussed. Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Partly Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Partly Are the conclusions drawn adequately supported by the results? Partly Competing Interests: No competing interests were disclosed. Reviewer Expertise: Dentistry, infection control I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Palma PJ. Reviewer Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.145897.r207744 ) The direct URL for this report is: https://f1000research.com/articles/12-726/v1#referee-response-207744 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Views 0 Cite How to cite this report: Abass SM. Reviewer Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.145897.r193467 ) The direct URL for this report is: https://f1000research.com/articles/12-726/v1#referee-response-193467 NOTE: it is important to ensure the information in square brackets after the title is included in this citation. Close Copy Citation Details Reviewer Report 08 Aug 2023 Shorouq Majid Abass , Department of Prosthodontics, College of Dentistry, University of Baghdad, Baghdad, Baghdad Governorate, Iraq Approved with Reservations VIEWS 0 https://doi.org/10.5256/f1000research.145897.r193467 The research needs careful attention to the following: Please reduce the words number of the research title “ The disinfection effectiveness of ozone water and 4.8% chloroxylenol against the number of bacterial colonies in dental extraction ... Continue reading READ ALL The research needs careful attention to the following: Please reduce the words number of the research title “ The disinfection effectiveness of ozone water and 4.8% chloroxylenol against the number of bacterial colonies in dental extraction instruments at the USU Dental and Oral Hospital in October-December 2022” in to at least 12 words number. In introduction you stated that “This study aimed to compare the effectiveness of ozone water and 4.8% chloroxylenol disinfectants on tooth extraction instruments, because these critical instruments have a high risk of causing infection as they penetrate the mucous membranes of the oral cavity” As shown in the bolded text you didn’t specify the concentration of the ozone in the water, so please correct it. In method and in subheading “Sampling” you stated that “The sampling technique used in this study was purposive sampling with 30 mandibular molar tooth extraction pliers as the sample….” As shown in the bolded text you didn’t specify the sample collection technique, please correct it. In method and in subheading “Ozone water production” you stated that “Ozone water was prepared using an ozone generator for 20 minutes with an ozone water concentration of 15 mg/L ” as shown in the bolded text you didn’t specify the ozone generation method, please correct it. In method and in the subheading “Disinfection procedure” you stated that “In the ozone water and 4.8% chloroxylenol treatment group, the mandibular molar tooth extraction pliers were rinsed under running water, brushed with an antiseptic solution and rinsed again until the pliers were free of blood and saliva before soaking in a disinfecting solution… ” as shown in the bolded text you didn’t specify the time and the duration of rinse with water, method of brush, type of antiseptic, please correct it. Also you stated that “After that, the mandibular molar tooth extraction forceps were soaked in ozonized water for 30 minutes and then a sample was taken ..” how you take the sample , please correct it with details. In method and in subheading “Counting bacterial colonies” you stated “……… a bacterial colony counter ” as shown in the yellow highlight you didn’t specify the type of the counter and the type of bacteria or otherwise your test is not specified to specific type of bacteria and you only have a negative or positive growth. In the result section your SD was too high more than 2, please can you state why? Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Partly Are sufficient details of methods and analysis provided to allow replication by others? No If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Partly Are the conclusions drawn adequately supported by the results? Yes Competing Interests: No competing interests were disclosed. Reviewer Expertise: Prosthodontics, dental materials, disinfection and dental implant. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. Close READ LESS CITE CITE HOW TO CITE THIS REPORT Abass SM. Reviewer Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.145897.r193467 ) The direct URL for this report is: https://f1000research.com/articles/12-726/v1#referee-response-193467 NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article. COPY CITATION DETAILS Report a concern Respond or Comment COMMENT ON THIS REPORT Comments on this article Comments (0) Version 4 VERSION 4 PUBLISHED 21 Jun 2023 ADD YOUR COMMENT Comment keyboard_arrow_left keyboard_arrow_right Open Peer Review Reviewer Status info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Reviewer Reports Invited Reviewers 1 2 3 4 Version 4 (revision) 31 Jul 25 read Version 3 (revision) 12 May 25 Version 2 (revision) 24 May 24 read read Version 1 21 Jun 23 read read Shorouq Majid Abass , University of Baghdad, Baghdad, Iraq Paulo J. Palma , University of Coimbra, Coimbra, Portugal Beni Jequicene Mussengue Chaúque , Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Universidade Rovuma, Niassa Branch, Lichinga, Mozambique Dario Gaudioso , Università degli Studi di Firenze, Sesto Fiorentino, Italy Comments on this article All Comments (0) Add a comment Sign up for content alerts Sign Up You are now signed up to receive this alert Browse by related subjects keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2025 Gaudioso D. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 23 Aug 2025 | for Version 4 Dario Gaudioso , Università degli Studi di Firenze, Sesto Fiorentino, Firenze, Italy 0 Views copyright © 2025 Gaudioso D. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Paper's experimental simplicity is very clear. It compares the use of ozonated water with the traditional treatment with chloroxylenol 4.8%. There are some points that would have been interesting to explore further, such as a more thorough screening of the genera of microorganisms that survive the different treatments. In parts of the paper, the genus and species are not italicized. Methodologically, some literature reports that the effect of ozone after 5 minutes is not effective, contrary to what is reported here. Overall, the paper, despite its operational and experimental simplicity, presents some interesting key points. Is the work clearly and accurately presented and does it cite the current literature? Yes Is the study design appropriate and is the work technically sound? Yes Are sufficient details of methods and analysis provided to allow replication by others? Yes If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? Yes Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise Molecular biology and microbiology I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard. reply Respond to this report Responses (1) Author Response 16 Oct 2025 Hendry Rusdy, Universitas Sumatera Utara, Medan, Indonesia Thankyou for your feedback and accepting my paper View more View less Competing Interests No competing interests were disclosed. reply Respond Report a concern Gaudioso D. Peer Review Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.182380.r405353) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-726/v4#referee-response-405353 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2024 Chaúque B. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 20 Sep 2024 | for Version 2 Beni Jequicene Mussengue Chaúque , Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil; Universidade Rovuma, Niassa Branch, Lichinga, Mozambique 0 Views copyright © 2024 Chaúque B. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (1) Not Approved info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The MS titled “Ozone Water or Chloroxylenol: A Comparison of Disinfection Effectiveness Against Bacterial Colonies on Dental Extraction Instruments at the USU Dental and Oral Hospital” explores the effectiveness of ozone water and chloroxylenol in inactivating bacteria on dental instruments. General comments While the topic is of considerable interest, the study suffers from several key issues, particularly in the writing and methodology sections, which are presented in an unstructured way. Moreover, the study's overall contribution to the existing literature appears limited. The sampling procedures are unclear, and the authors do not provide sufficient detail on how they minimized the risk of bias in the allocation of instrument samples. With only 10 samples, the study’s findings are not robust enough to justify any broad generalizations. Another major limitation is the lack of identification and/or isolation of microorganisms, which would have allowed for a more standardized evaluation of disinfectant performance against comparable inoculum sizes. Additionally, the washing processes used prior to instrument handling, to remove blood and saliva, could introduce bias especially given the small sample size. Specific comments Title 1- It is not advisable to use abbreviations in the title. I suggest either writing "USU" in full or omitting it entirely. Abstract 2 -The rationale for conducting the study is not clearly established. Please elaborate further. For example, isn't the use of ozone water and 4.8% chloroxylenol already a proven and standardized aseptic practice? If so, why is this study necessary? 3 -Additionally, the abbreviation "USU" appears in the abstract without being explained. 4 -“This study aimed to evaluate the effectiveness of ozone water and 4.8% chloroxylenol in reducing bacterial colonies on dental extraction instruments at the USU Dental and Oral Hospital between October and December 2022.” Note: The phrase "between October and December 2022" seems unnecessary here and can be removed. 5 -The methods section of the abstract should be written more formally, detailing the sampling, transportation, treatment (including the form, application, and exposure), and the plate inoculation process. 6 -The study is based on only 10 different instruments. Why such a small sample size? How representative is this sample? 7 -The results are reported in a confusing manner. The p-value analysis is presented before the colony count values. The authors present two values for the number of colonies in the treatment group: 4.00 ± 4.32 and 16.00 ± 6.65. Which value is correct? If the values refer to ozone water and chloroxylenol, respectively, please specify this clearly. Introduction 8 -Paragraphs 1, 2, and 3 contain redundant information. Please consider rewriting to avoid repetition. 9 - Note: Only separate paragraphs when necessary for clarity. 10 - In paragraph 5, ensure the names of microorganisms are spelled correctly. Methods 11 - The sentence "The patient’s oral cavity state, the kind and quantity of bacteria present, the surrounding circumstances, and the instrument settings before tooth extraction are all non-parametric" is unclear. This sounds more like a result rather than a method. Please consider rewriting 12 - The following information needs to be placed in the appropriate section: "The study parameters include the concentration of ozone water (15 mg/L), the concentration of chloroxylenol (4.8%), the immersion time in ozone water (30 minutes), the immersion time in chloroxylenol solution (60 minutes), and the pre-treatment procedures." This information is also repeated in the "Disinfection procedure" section. 13 - Instead of simply stating that " The sampling technique used in this study was purposive sampling" provide a detailed, step-by-step explanation of the sampling process to ensure reproducibility by other researchers. 14 - The subsections “Disinfection procedure” and “Taking the samples” are unclear and need to be rewritten in a more straightforward, didactic manner. Clearly distinguish between the different types of samples used in the study, such as instruments and inoculum. Include specific details regarding their identification, selection, and collection. 15 - It is also unclear whether the instruments were treated individually or as a group. 16 - In the “Counting bacterial colonies” subsection, specify the volume of the sample plated and the incubation temperature. Results 17 - The results in Tables 2–5 would be easier to understand if presented in a graph, allowing readers to visualize and compare the data more effectively. Conclusion 18 - The statement “Ozone water can kill bacterial endospores while 4.8% chloroxylenol cannot” should not be included in the conclusions, as this was not assessed in the study. Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Partly Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Yes Are all the source data underlying the results available to ensure full reproducibility? No Are the conclusions drawn adequately supported by the results? Partly Competing Interests No competing interests were disclosed. Reviewer Expertise Microbiology and Pharmacology I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above. reply Respond to this report Responses (1) Author Response 12 May 2025 Hendry Rusdy, Universitas Sumatera Utara, Medan, Indonesia General comments The amount of sample is 30 Specific comments Title Change USU to University of South Sumatera Abstract 2 -The rationale for conducting the study is not clearly established. Please elaborate further. For example, isn't the use of ozone water and 4.8% chloroxylenol already a proven and standardized aseptic practice? If so, why is this study necessary? answer : Research on ozone water as a disinfectant is still limited and no research has been found on the use of ozone water on tooth extraction instruments. However, ozone has antibacterial properties so researchers are interested in studying the effectiveness of ozone water as a disinfectant when compared to the commonly used 4.8% chloroxylenol 3 - "USU" appears in the abstract without being explained. 4 -The phrase "between October and December 2022" seems unnecessary here and can be removed. 5 -The methods has been updated 6 -The study is based on only 10 different instruments. Why such a small sample size? How representative is this sample? answer : The amount of sample is 30 7 -The results are reported in a confusing manner. The p-value analysis is presented before the colony count values. The authors present two values for the number of colonies in the treatment group: 4.00 ± 4.32 and 16.00 ± 6.65. Which value is correct? If the values refer to ozone water and chloroxylenol, respectively, please specify this clearly. answer : value refer to ozone water, which is 4.00 ± 4.32 Introduction 8 -Paragraphs 1, 2, and 3 has been updated 9 - updated 10 - In paragraph 5, ensure the names of microorganisms are spelled correctly has been updated Methods 11 - The sentence "The patient’s oral cavity state, the kind and quantity of bacteria present, the surrounding circumstances, and the instrument settings before tooth extraction are all non-parametric" is unclear. This sounds more like a result rather than a method. Please consider rewriting have been updated 12 - The following information needs to be placed in the appropriate section: "The study parameters include the concentration of ozone water (15 mg/L), the concentration of chloroxylenol (4.8%), the immersion time in ozone water (30 minutes), the immersion time in chloroxylenol solution (60 minutes), and the pre-treatment procedures." This information is also repeated in the "Disinfection procedure" section have been updated 13 - Instead of simply stating that " The sampling technique used in this study was purposive sampling" provide a detailed, step-by-step explanation of the sampling process to ensure reproducibility by other researchers. have been updated 14 - The subsections “Disinfection procedure” and “Taking the samples” are unclear and need to be rewritten in a more straightforward, didactic manner. Clearly distinguish between the different types of samples used in the study, such as instruments and inoculum. Include specific details regarding their identification, selection, and collection. have been updated 15 - It is also unclear whether the instruments were treated individually or as a group. have been updated 16 - In the “Counting bacterial colonies” subsection, specify the volume of the sample plated and the incubation temperature. have been updated Results 17 - The results in Tables 2–5 would be easier to understand if presented in a graph, allowing readers to visualize and compare the data more effectively. have been updated Conclusion 18 - The statement “Ozone water can kill bacterial endospores while 4.8% chloroxylenol cannot” should not be included in the conclusions, as this was not assessed in the study. have been updated View more View less Competing Interests No competing interests were disclosed. reply Respond Report a concern Chaúque BJM. Peer Review Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.158749.r319597) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-726/v2#referee-response-319597 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2024 Abass S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 06 Jun 2024 | for Version 2 Shorouq Majid Abass , Department of Prosthodontics, College of Dentistry, University of Baghdad, Baghdad, Baghdad Governorate, Iraq 0 Views copyright © 2024 Abass S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions Dear Authors, thank you for this interesting article. Here are some of the comments of mine: In the introduction section you have to mention other aspect of disinfection solution, technique, and dental materials in general other than of your specific subject of research. e.g. Khalaf BS, Abass SM, Al-Khafaji AM, and Issa MI, Antimicrobial efficiency of hypochlorous acid and its effect on some properties of alginate impression material. Int J Dent, 2023. 2023: p. 1-7 DOI: 10.1155/2023/8584875. Al-khafaji AM, Abass SM, and Khalaf BS, The effect of SOLO and sodium hypochlorite disinfectant on some properties of different types of dental stone. J Bagh Coll Dent, 2013. 25(2): p. 8-17 Mahmood MA, Khalaf BS, and Abass SM, Efficiency of different denture disinfection methods. Global J Bio-sci Biotech, 2017. 6: p. 439-4. It is only a suggestion for authors to use the above citations because they are related to the subject of disinfection in dentistry in general and the authors can refer to it if they wish. References 1. Khalaf BS, Abass SM, Al-Khafaji AM, Issa MI: Antimicrobial Efficiency of Hypochlorous Acid and Its Effect on Some Properties of Alginate Impression Material. Int J Dent . 2023; 2023 : 8584875 PubMed Abstract | Publisher Full Text 2. SM Abass: The effect of SOLO and sodium hypochlorite disinfectant on some properties of different types of dental stone. University of Baghdad . 1973. 8-17 3. Abass SM: Efficiency of different denture disinfection methods. Global J Bio-sci Biotech . 2017. 439-439 Competing Interests No competing interests were disclosed. Reviewer Expertise Prosthodontics, dental materials, disinfection and dental implant. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (0) Abass SM. Peer Review Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.158749.r282421) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-726/v2#referee-response-282421 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2023 Palma P. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 09 Oct 2023 | for Version 1 Paulo J. Palma , Institute of Endodontics, Faculty of Medicine, University of Coimbra, Coimbra, Portugal 0 Views copyright © 2023 Palma P. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The present study aimed to determine the effectiveness of disinfection of ozone water and 4.8% chloroxylenol in reducing the number of bacterial colonies on dental extraction instruments at the USU Dental and Oral Hospital between October-December 2022.. This research is under the scope of this journal; the topic is relevant for readers, and this research deals with potentially significant knowledge of the field. And It will be important for knowledge. The topic is relevant for readers and this review deals with potentially significant knowledge in the field and opens new ways for future studies. However, there are some aspects that are possibly improved in the manuscript: Keywords Please add more keywords, and order the keywords / Mesh terms alphabetically Introduction What is the importance of this study? Which results are comparable with other articles? What has this study been new? M&M ​​​ A flowchart of all experimental study would be valuable. How many slides per sample obtained, and how many of them were evaluated? How were the sample cells calculated? Did authors performed power analysis to evaluate if this sample size was appropriate? Analysis: How many observers did the scoring? What is the interobserver agreement? How many time was performed done the determinations? Which data are parametric and non-parametric? Please specify? When mentioning materials or devices: for some of them you don't mention the manufacturer at all, for some you mention only the manufacturer, for some the manufacturer and city, for some you mention the manufacturer and city/ country. Discussion Please, clarified more limitations of this study? An objective presentation of the strengths and limitations of the model, study design and methods, must be provided, including any biology/functional variability between the animal model and humans. And also clarified the future perspectives in the discussion. The potential influence of the results on future research plans needs also be discussed. Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Partly Are sufficient details of methods and analysis provided to allow replication by others? Partly If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Partly Are the conclusions drawn adequately supported by the results? Partly Competing Interests No competing interests were disclosed. Reviewer Expertise Dentistry, infection control I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (0) Palma PJ. Peer Review Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.145897.r207744) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-726/v1#referee-response-207744 keyboard_arrow_left Back to all reports Reviewer Report 0 Views copyright © 2023 Abass S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 08 Aug 2023 | for Version 1 Shorouq Majid Abass , Department of Prosthodontics, College of Dentistry, University of Baghdad, Baghdad, Baghdad Governorate, Iraq 0 Views copyright © 2023 Abass S. This is an open access peer review report distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. format_quote Cite this report speaker_notes Responses (0) Approved With Reservations info_outline Alongside their report, reviewers assign a status to the article: Approved The paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved Fundamental flaws in the paper seriously undermine the findings and conclusions The research needs careful attention to the following: Please reduce the words number of the research title “ The disinfection effectiveness of ozone water and 4.8% chloroxylenol against the number of bacterial colonies in dental extraction instruments at the USU Dental and Oral Hospital in October-December 2022” in to at least 12 words number. In introduction you stated that “This study aimed to compare the effectiveness of ozone water and 4.8% chloroxylenol disinfectants on tooth extraction instruments, because these critical instruments have a high risk of causing infection as they penetrate the mucous membranes of the oral cavity” As shown in the bolded text you didn’t specify the concentration of the ozone in the water, so please correct it. In method and in subheading “Sampling” you stated that “The sampling technique used in this study was purposive sampling with 30 mandibular molar tooth extraction pliers as the sample….” As shown in the bolded text you didn’t specify the sample collection technique, please correct it. In method and in subheading “Ozone water production” you stated that “Ozone water was prepared using an ozone generator for 20 minutes with an ozone water concentration of 15 mg/L ” as shown in the bolded text you didn’t specify the ozone generation method, please correct it. In method and in the subheading “Disinfection procedure” you stated that “In the ozone water and 4.8% chloroxylenol treatment group, the mandibular molar tooth extraction pliers were rinsed under running water, brushed with an antiseptic solution and rinsed again until the pliers were free of blood and saliva before soaking in a disinfecting solution… ” as shown in the bolded text you didn’t specify the time and the duration of rinse with water, method of brush, type of antiseptic, please correct it. Also you stated that “After that, the mandibular molar tooth extraction forceps were soaked in ozonized water for 30 minutes and then a sample was taken ..” how you take the sample , please correct it with details. In method and in subheading “Counting bacterial colonies” you stated “……… a bacterial colony counter ” as shown in the yellow highlight you didn’t specify the type of the counter and the type of bacteria or otherwise your test is not specified to specific type of bacteria and you only have a negative or positive growth. In the result section your SD was too high more than 2, please can you state why? Is the work clearly and accurately presented and does it cite the current literature? Partly Is the study design appropriate and is the work technically sound? Partly Are sufficient details of methods and analysis provided to allow replication by others? No If applicable, is the statistical analysis and its interpretation appropriate? Partly Are all the source data underlying the results available to ensure full reproducibility? Partly Are the conclusions drawn adequately supported by the results? Yes Competing Interests No competing interests were disclosed. Reviewer Expertise Prosthodontics, dental materials, disinfection and dental implant. I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above. reply Respond to this report Responses (0) Abass SM. Peer Review Report For: Ozone water or chloroxylenol: The comparison of disinfection effectiveness against the number of bacterial colonies in dental extraction instruments at the Universitas Sumatera Utara Dental and Oral Hospital [version 4; peer review: 1 approved, 2 approved with reservations, 1 not approved] . F1000Research 2025, 12 :726 ( https://doi.org/10.5256/f1000research.145897.r193467) NOTE: it is important to ensure the information in square brackets after the title is included in this citation. The direct URL for this report is: https://f1000research.com/articles/12-726/v1#referee-response-193467 Alongside their report, reviewers assign a status to the article: Approved - the paper is scientifically sound in its current form and only minor, if any, improvements are suggested Approved with reservations - A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit. Not approved - fundamental flaws in the paper seriously undermine the findings and conclusions Adjust parameters to alter display View on desktop for interactive features Includes Interactive Elements View on desktop for interactive features Competing Interests Policy Provide sufficient details of any financial or non-financial competing interests to enable users to assess whether your comments might lead a reasonable person to question your impartiality. 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