Review Reports

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Dear Authors,

After Carefully reviewing your Brief Report entitled "Residual Bacterial Contamination of Surgical Instruments After Chemical Disinfection in Veterinary Clinics in Veracruz, Mexico".

I see that this report deserves attention because it highlights on a very important topic from different perspectives. In fact this work is very important from microbiological, medical, infection prevention and control, and veterinary points of view. It highlights on the prevalence of different bacterial contaminations of Surgical Instruments used in veterinary hospitals which was treated by chemical disinfection. Data were collected from different 10 veterinary facilities, and analyzed to understand the effectiveness of chemical disinfection in veterinary medicine. 

The paper needs a lot of work before being accepted. here are my comments regarding the present report:

01- Why did you choose to submit this manuscript as Brief Report? Why not submitted as Original Research Article? or at list you can submit it as a Pilot Study. Can you please argument your answer? 

02- The Whole paper needs extensive English polishing by a Native English Speaker. Since several sentences are not clear for readers and several typo errors are present in the paper.

03- The Paper lacks Authors' names and affiliations? Can you please add this information to the manuscript.

04- In the Whole manuscript you are invited to put the name of bacterial species in italic.

05- Concerning your list of Keywords, you are kindly invited to add 2 keywords, I suggest to add the follow "Hospital Acquired Infection" and "Surgical Site Infection".

06- Concerning The Introduction section, I invite you to start broadly by talking about Hospital Acquired Infection general, then you can talk briefly about it in Human medicine, and finally you can go to talking about it in veterinary medicine. Since HAIs are still a big issue in Human Medicine. You can refer to the following paper concerning this point:

-- Healthcare – associated infections: A public health problem

07- In your Introduction section, when you are talking about main bacteria causing Hospital Acquired Infections, you are kindly invited to use the following references concerning these bacteria:

-- The intricacies of Acinetobacter baumannii: a multifaceted comprehensive review of a multidrug-resistant pathogen and its clinical significance and implications

-- General Overview of Klebsiella pneumonia: Epidemiology and the Role of Siderophores in Its Pathogenicity

-- Occurrence of Healthcare-Associated Infections (HAIs) by Escherichia coli and Klebsiella spp. Producing Extended-Spectrum β-lactamases (ESBL) and/or Carbapenemases in Portuguese Long-Term Care Facilities

-- The Key Element Role of Metallophores in the Pathogenicity and Virulence of Staphylococcus aureus: A Review

-- Hospital-acquired fungal infections by Candida auris and Aspergillus species: Pathogenesis, resistance mechanisms, prevention and control measures

-- Prevalence and antimicrobial resistance of Acinetobacter baumannii isolates at Lebanese Hospital Geitaoui-University Medical Center: A Five-year study (2017–2021)

-- A Review of Pseudomonas aeruginosa Metallophores: Pyoverdine, Pyochelin and Pseudopaline

08- In your Materials and Methods section, concerning "Statistical Analysis" sub-section, Can you please indicate the name of Statistical tool you used in your work?

09- You Results section is very weak, you can make it more clear for readers if you add more figures and tables, for example a figure illustration the prevalence of each detected bacterium in the cohort.

10- What is the importance of your Figure 1 in the manuscript!! I suggest to remove it.

11- Why you did not apply AST (Antimicrobial Susceptibility Test) on your isolates to see if they are, or not, resistant to antibiotics. It will increase the significance of your results.

12- In your Discussion, I invite to you talk about the importance of fighting HAIs by increasing the knowledge among healthcare workers and among community people towards Infection Prevention and Control, about AMR and MDR. You can refer to the following papers concerning this point:

-- Assessment of Knowledge, Attitude, and Practice on Hospital-Acquired Infection Prevention Among Lebanese Healthcare Workers

-- Caretaker knowledge, attitudes, and practices (KAP) and carriage of extended-spectrum beta-lactamase-producing E. coli (ESBL-EC) in children in Quito, Ecuador

-- Assessment of Methicillin-Resistant Staphylococcus aureus (MRSA) knowledge and awareness among healthcare workers in South-Lebanon

-- Assessment of Knowledge, Attitude and Practice (KAP) Regarding Antimicrobial Usage and Resistance Among Animal Health Professionals of East Wallaga Zone, Oromiya, Ethiopia

-- Knowledge, Attitude, and Practices of Healthcare Workers Towards Tuberculosis, Multidrug-Resistant Tuberculosis, and Extensively Drug-Resistant Tuberculosis

-- Knowledge, attitudes, and practices on antimicrobial use and antimicrobial resistance among poultry practitioner veterinarians

-- Knowledge, Attitude, and Practices (KAP) Survey among Veterinarians, and Risk Factors Relating to Antimicrobial Use and Treatment Failure in Dairy Herds of India

13- In your Discussion, I suggest to highlight on the importance of finding new strategies to fight antimicrobial resistance is pathogenic bacteria, such as in the case bacteria causing HAIs, which are usually MDR strains. Here I invite you to talk about different new strategies such as talking about the use of Trojan Horse technique to introduce the antibiotic inside a bacterial cell, or by targeting new bacterial ultra-structures such as targeting bacterial metallophores which can impair the bacterial virulence, or even the use of specific bacteriophage to destroy MDR strains. You can use the following papers as references for this section:

-- Trojan Horse Antibiotics—A Novel Way to Circumvent Gram-Negative Bacterial Resistance?

-- Towards new antibiotics classes targeting bacterial metallophores

-- Emerging roles of bacteriophage-based therapeutics in combating antibiotic resistance

 

Best Regards,

Author Response

We sincerely thank the reviewer for the careful evaluation of our manuscript and for the constructive comments provided. Following the reviewer’s recommendations, the manuscript was substantially revised to improve methodological clarity, language quality, organization of results, and interpretation of findings.

Comment 01:

“Why did you choose to submit this manuscript as Brief Report? Why not submitted as Original Research Article? or at list you can submit it as a Pilot Study. Can you please argument your answer?”

Response:

We thank the reviewer for this important observation. The manuscript was submitted as a Brief Report because of the preliminary exploratory nature of the study, the limited sample size, and the observational cross-sectional design. Although the study provides relevant preliminary findings regarding residual bacterial contamination after routine instrument reprocessing in veterinary clinics, we considered that the available data were more appropriately presented as a concise exploratory report rather than a full original research article. To further clarify this aspect, the terms “preliminary exploratory” were incorporated throughout the revised manuscript.

Comment 02:

“The Whole paper needs extensive English polishing by a Native English Speaker. Since several sentences are not clear for readers and several typo errors are present in the paper.”

Response:

We appreciate the reviewer’s observation. The manuscript underwent extensive English revision and stylistic editing to improve grammar, readability, sentence structure, scientific clarity, and overall language quality throughout the text.

Comment 03:

“The Paper lacks Authors' names and affiliations? Can you please add this information to the manuscript.”

Response:

We thank the reviewer for identifying this omission. Complete author names, affiliations, and corresponding author information were added to the revised manuscript.

Comment 04:

“In the Whole manuscript you are invited to put the name of bacterial species in italic.”

Response:

We thank the reviewer for this observation. Scientific names of bacterial species were revised and formatted in italics throughout the manuscript according to journal formatting guidelines.

Comment 05:

“Concerning your list of Keywords, you are kindly invited to add 2 keywords, I suggest to add the follow "Hospital Acquired Infection" and "Surgical Site Infection".”

Response:

We appreciate the reviewer’s recommendation. The keywords “Healthcare-Associated Infections” and “Surgical Site Infection” were added to improve indexing and manuscript visibility.

Comment 06:

“Concerning The Introduction section, I invite you to start broadly by talking about Hospital Acquired Infection general, then you can talk briefly about it in Human medicine, and finally you can go to talking about it in veterinary medicine. Since HAIs are still a big issue in Human Medicine. You can refer to the following paper concerning this point:

-- Healthcare – associated infections: A public health problem”

Response:

We thank the reviewer for this valuable suggestion. The Introduction section was substantially revised to provide broader context regarding healthcare-associated infections, including their relevance in human medicine and veterinary healthcare settings. The suggested reference was incorporated into the revised manuscript.

Comment 07:

“In your Introduction section, when you are talking about main bacteria causing Hospital Acquired Infections, you are kindly invited to use the following references concerning these bacteria:

-- The intricacies of Acinetobacter baumannii: a multifaceted comprehensive review of a multidrug-resistant pathogen and its clinical significance and implications

-- General Overview of Klebsiella pneumonia: Epidemiology and the Role of Siderophores in Its Pathogenicity

-- Occurrence of Healthcare-Associated Infections (HAIs) by Escherichia coli and Klebsiella spp. Producing Extended-Spectrum β-lactamases (ESBL) and/or Carbapenemases in Portuguese Long-Term Care Facilities

-- The Key Element Role of Metallophores in the Pathogenicity and Virulence of Staphylococcus aureus: A Review

-- Hospital-acquired fungal infections by Candida auris and Aspergillus species: Pathogenesis, resistance mechanisms, prevention and control measures

-- Prevalence and antimicrobial resistance of Acinetobacter baumannii isolates at Lebanese Hospital Geitaoui-University Medical Center: A Five-year study (2017–2021)

-- A Review of Pseudomonas aeruginosa Metallophores: Pyoverdine, Pyochelin and Pseudopaline”

Response:

We appreciate the reviewer’s detailed recommendations regarding additional literature. Several suggested references were incorporated into the Introduction and Discussion sections to strengthen the contextual background regarding healthcare-associated pathogens, antimicrobial resistance, and environmental persistence. However, some highly specialized references focusing on molecular virulence mechanisms and antimicrobial strategies beyond the scope of the present exploratory observational study were not included in order to maintain the manuscript’s focus and conciseness.

Comment 08:

“In your Materials and Methods section, concerning "Statistical Analysis" sub-section, Can you please indicate the name of Statistical tool you used in your work?”

Response:

We thank the reviewer for this observation. The statistical software used for data analysis was specified in the revised manuscript as IBM SPSS Statistics version 25.0 (IBM Corp., Armonk, NY, USA).

Comment 09:

“You Results section is very weak, you can make it more clear for readers if you add more figures and tables, for example a figure illustration the prevalence of each detected bacterium in the cohort.”

Response:

We appreciate the reviewer’s recommendation. The Results section was reorganized and expanded to improve clarity and descriptive presentation. Additional information regarding bacterial distribution and contamination patterns was incorporated, and Figure 1 was modified to better summarize the recovered bacterial genera detected across sampled clinics.

Comment 10:

“What is the importance of your Figure 1 in the manuscript!! I suggest to remove it.”

Response:

We respectfully appreciate the reviewer’s opinion regarding Figure 1. However, the original MALDI-TOF spectrum figure was removed and replaced with a figure illustrating the distribution of recovered bacterial genera. We believe the revised figure provides a clearer visual summary of contamination patterns and complements the descriptive findings presented in the Results section.

Comment 11:

“Why you did not apply AST (Antimicrobial Susceptibility Test) on your isolates to see if they are, or not, resistant to antibiotics. It will increase the significance of your results.”

Response:

We thank the reviewer for this important observation. We agree that antimicrobial susceptibility testing would increase the microbiological significance of the study. However, due to logistical and resource limitations, AST was beyond the scope of this preliminary exploratory investigation. This limitation was explicitly acknowledged in the revised Discussion section, and future studies incorporating resistance profiling were recommended.

Comment 12:

“In your Discussion, I invite to you talk about the importance of fighting HAIs by increasing the knowledge among healthcare workers and among community people towards Infection Prevention and Control, about AMR and MDR. You can refer to the following papers concerning this point:

-- Assessment of Knowledge, Attitude, and Practice on Hospital-Acquired Infection Prevention Among Lebanese Healthcare Workers

-- Caretaker knowledge, attitudes, and practices (KAP) and carriage of extended-spectrum beta-lactamase-producing E. coli (ESBL-EC) in children in Quito, Ecuador

-- Assessment of Methicillin-Resistant Staphylococcus aureus (MRSA) knowledge and awareness among healthcare workers in South-Lebanon

-- Assessment of Knowledge, Attitude and Practice (KAP) Regarding Antimicrobial Usage and Resistance Among Animal Health Professionals of East Wallaga Zone, Oromiya, Ethiopia

-- Knowledge, Attitude, and Practices of Healthcare Workers Towards Tuberculosis, Multidrug-Resistant Tuberculosis, and Extensively Drug-Resistant Tuberculosis

-- Knowledge, attitudes, and practices on antimicrobial use and antimicrobial resistance among poultry practitioner veterinarians

-- Knowledge, Attitude, and Practices (KAP) Survey among Veterinarians, and Risk Factors Relating to Antimicrobial Use and Treatment Failure in Dairy Herds of India”

Response:

We appreciate the reviewer’s suggestion. The Discussion section was revised to briefly address the importance of infection prevention practices, antimicrobial resistance awareness, and standardized instrument reprocessing procedures in veterinary healthcare settings. However, because the present study did not directly evaluate knowledge, attitudes, or practices, discussion of these aspects was intentionally kept concise in order to maintain the primary focus of the manuscript.

Comment 13:

“In your Discussion, I suggest to highlight on the importance of finding new strategies to fight antimicrobial resistance is pathogenic bacteria, such as in the case bacteria causing HAIs, which are usually MDR strains. Here I invite you to talk about different new strategies such as talking about the use of Trojan Horse technique to introduce the antibiotic inside a bacterial cell, or by targeting new bacterial ultra-structures such as targeting bacterial metallophores which can impair the bacterial virulence, or even the use of specific bacteriophage to destroy MDR strains. You can use the following papers as references for this section:

-- Trojan Horse Antibiotics—A Novel Way to Circumvent Gram-Negative Bacterial Resistance?

-- Towards new antibiotics classes targeting bacterial metallophores

-- Emerging roles of bacteriophage-based therapeutics in combating antibiotic resistance”

Response:

We thank the reviewer for these interesting suggestions regarding emerging antimicrobial strategies. A brief discussion regarding the growing interest in alternative approaches to combat antimicrobial-resistant pathogens was initially considered during manuscript revision. However, because the present study focused on residual bacterial contamination after routine instrument reprocessing rather than antimicrobial resistance mechanisms or therapeutic interventions, detailed discussion of metallophore-targeting therapies, Trojan horse antibiotics, and bacteriophage-based therapeutics was intentionally omitted in the final revised version in order to maintain the scientific focus and conciseness of the manuscript.

Reviewer 2 Report

Comments and Suggestions for Authors

I think the manuscript addresses an important and practical topic in veterinary infection control. The presence of residual bacterial contamination on surgical instruments after chemical disinfection is a relevant issue, especially in clinics with limited resources. I also believe that the study has practical value because it draws attention to a real problem that may affect animal health, veterinary personnel, and the clinical environment.

However, I think the manuscript still needs major improvement before it can be considered scientifically strong. The main limitation is the study design. The number of clinics is small, and only one clinic used a high-level disinfectant. Because of this, I believe the comparison between low-level and high-level disinfectants is not strong enough. The authors should present the work more clearly as a preliminary or exploratory study and avoid making conclusions that are too broad.

I also think the methodology needs to be described in more detail. Important information is missing, such as the exact sampling area, the number of samples taken from each type of instrument, the real concentration of disinfectants used, contact time, dilution procedures, frequency of disinfectant replacement, and whether neutralizing agents were used after sampling. These details are very important because the bacterial contamination observed could be due to poor cleaning, incorrect disinfectant use, contamination during handling, or true disinfectant failure.

In my opinion, the results are interesting but need to be presented more clearly. Table 1 contains useful information, but it is difficult to read in its current form. I think the authors should reorganize it to show, for each clinic, the number of samples collected, the number of positive samples, the contamination percentage, the type of instrument, the disinfectant used, and the bacterial species identified. This would make the results much easier to understand.

I believe Figure 1 has limited value in its current form. It shows a representative MALDI-TOF spectrum, but it does not really help the reader understand the contamination pattern or the effectiveness of disinfection. The manuscript would benefit more from a figure showing contamination frequency by clinic, instrument type, or disinfectant category.

I think the discussion is relevant, but it is sometimes too general and repetitive. The One Health perspective is important, but it should be more directly linked to the actual results of this study. The authors should also discuss the isolated bacterial species more critically, especially their possible origin, clinical importance, and resistance to chemical disinfection.

I also believe the conclusions should be softened. The study shows that bacterial contamination remained after routine chemical disinfection, but it does not prove causality and does not provide strong evidence to compare disinfectant types. Therefore, the conclusions should be more cautious and should clearly mention that larger and better-controlled studies are needed.

I think the English language is understandable, but it still needs improvement. Some sentences are long, repetitive, or awkwardly written, and several formatting issues reduce the clarity of the manuscript.

I believe the manuscript has an important topic and useful preliminary observations, but it requires major revision. The authors should improve the methodology, reorganize the results, clarify the tables and figures, update and correct the references, and moderate the conclusions according to the real limits of the study.

Comments on the Quality of English Language

I think the English language is generally understandable, and the main message of the manuscript can be followed. However, I believe the text would benefit from careful language revision to improve clarity, fluency, and scientific precision. Some sentences are too long or repetitive, and there are several awkward expressions and formatting issues that interrupt the reading flow. I suggest a thorough English editing to make the manuscript clearer, more polished, and more suitable for publication.

Author Response

We sincerely thank the reviewer for the careful evaluation of our manuscript and for the constructive comments provided. We appreciate the recognition of the clinical and microbiological relevance of the study. Following the reviewer’s recommendations, the manuscript was substantially revised to improve methodological clarity, organization of the results, discussion focus, language quality, and interpretation of findings.

Comment:
“I think the manuscript addresses an important and practical topic in veterinary infection control. The presence of residual bacterial contamination on surgical instruments after chemical disinfection is a relevant issue, especially in clinics with limited resources. I also believe that the study has practical value because it draws attention to a real problem that may affect animal health, veterinary personnel, and the clinical environment.”

Response:
We sincerely thank the reviewer for these positive comments and for recognizing the practical and clinical relevance of our study.

Comment:
“However, I think the manuscript still needs major improvement before it can be considered scientifically strong. The main limitation is the study design. The number of clinics is small, and only one clinic used a high-level disinfectant. Because of this, I believe the comparison between low-level and high-level disinfectants is not strong enough. The authors should present the work more clearly as a preliminary or exploratory study and avoid making conclusions that are too broad.”

Response:
We thank the reviewer for this important observation. We fully agree that the limited number of participating clinics, particularly those routinely using high-level disinfectants, restricts the strength of comparisons between disinfectant categories. In response, the manuscript was revised to consistently emphasize the preliminary exploratory nature of the study. Statements implying definitive conclusions regarding disinfectant performance were softened throughout the Abstract, Results, Discussion, and Conclusions sections. Additional limitations regarding sample size and observational design were also incorporated into the Discussion section.

Comment:
“I also think the methodology needs to be described in more detail. Important information is missing, such as the exact sampling area, the number of samples taken from each type of instrument, the real concentration of disinfectants used, contact time, dilution procedures, frequency of disinfectant replacement, and whether neutralizing agents were used after sampling. These details are very important because the bacterial contamination observed could be due to poor cleaning, incorrect disinfectant use, contamination during handling, or true disinfectant failure.”

Response:
We appreciate the reviewer’s valuable comments regarding methodological clarity. Additional methodological details were incorporated into the revised manuscript, including the approximate sampled surface area (~5 cm²), number of collected samples per clinic, and clarification regarding the use of neutralizing agents after sampling. We also explicitly stated that disinfectant concentrations, dilution procedures, contact times, and disinfectant replacement frequencies could not be independently standardized or experimentally verified under field conditions because the study was designed as an observational assessment of routine clinical practices under real-world conditions. Furthermore, the potential influence of residual disinfectant activity on bacterial recovery was acknowledged as a limitation in the revised Discussion section.

Comment:
“In my opinion, the results are interesting but need to be presented more clearly. Table 1 contains useful information, but it is difficult to read in its current form. I think the authors should reorganize it to show, for each clinic, the number of samples collected, the number of positive samples, the contamination percentage, the type of instrument, the disinfectant used, and the bacterial species identified. This would make the results much easier to understand.”

Response:
We thank the reviewer for this helpful recommendation. Table 1 was reorganized to improve readability and clarity. The revised table now presents, for each clinic, the number of collected samples, number and percentage of positive samples, contaminated instruments or surfaces, disinfectant strategy used, disinfectant category, and bacterial species identified. The Results section was also revised to improve organization and descriptive clarity.

Comment:
“I believe Figure 1 has limited value in its current form. It shows a representative MALDI-TOF spectrum, but it does not really help the reader understand the contamination pattern or the effectiveness of disinfection. The manuscript would benefit more from a figure showing contamination frequency by clinic, instrument type, or disinfectant category.”

Response:
We appreciate the reviewer’s observation regarding Figure 1. In the revised manuscript, Figure 1 was modified to present the distribution of recovered bacterial genera rather than representative MALDI-TOF spectra. We believe the revised figure provides a clearer visual summary of contamination patterns observed across sampled veterinary clinics and better complements the descriptive findings presented in the Results section.

Comment:
“I think the discussion is relevant, but it is sometimes too general and repetitive. The One Health perspective is important, but it should be more directly linked to the actual results of this study. The authors should also discuss the isolated bacterial species more critically, especially their possible origin, clinical importance, and resistance to chemical disinfection.”

Response:
We thank the reviewer for this important suggestion. The Discussion section was substantially revised to improve focus and reduce repetitive general statements. The revised discussion now places greater emphasis on the bacterial species recovered in this study, their possible environmental origin, persistence on surfaces, and clinical relevance in veterinary healthcare settings. In addition, the One Health perspective was condensed and more directly linked to the findings of the present study.

Comment:
“I also believe the conclusions should be softened. The study shows that bacterial contamination remained after routine chemical disinfection, but it does not prove causality and does not provide strong evidence to compare disinfectant types. Therefore, the conclusions should be more cautious and should clearly mention that larger and better-controlled studies are needed.”

Response:
We fully agree with the reviewer regarding the need for cautious interpretation of the findings. The Conclusions section was revised to avoid causal interpretations regarding disinfectant effectiveness and to acknowledge the exploratory observational nature of the study. The revised conclusions now emphasize that residual bacterial contamination was identified after routine chemical disinfection under real-world conditions, while also highlighting the need for larger and better-controlled studies to further evaluate instrument reprocessing practices in veterinary clinical environments.

Comment:
“I think the English language is understandable, but it still needs improvement. Some sentences are long, repetitive, or awkwardly written, and several formatting issues reduce the clarity of the manuscript.”

Response:
We appreciate the reviewer’s recommendation. The manuscript underwent extensive language revision and stylistic editing to improve grammar, readability, sentence structure, and overall clarity. Formatting inconsistencies were also corrected throughout the revised version.

Comment:
“I believe the manuscript has an important topic and useful preliminary observations, but it requires major revision. The authors should improve the methodology, reorganize the results, clarify the tables and figures, update and correct the references, and moderate the conclusions according to the real limits of the study.”

Response:
We sincerely thank the reviewer for the constructive evaluation and recommendations. Following these suggestions, the manuscript was substantially revised to improve methodological transparency, organization of the Results section, clarity of tables and figures, discussion focus, language quality, and interpretation of findings. The exploratory nature and limitations of the study are now more clearly emphasized throughout the revised manuscript.

Reviewer 3 Report

Comments and Suggestions for Authors

This study is particularly timely given the limited attention paid by veterinary clinics to the introduction and spread of microorganisms within clinical environments and to the control of contributing factors. This is especially important considering that animals are commonly colonized by a wide diversity of microorganisms, including potentially pathogenic species.

The findings highlight a pressing need to strengthen infection control practices in veterinary settings, particularly in resource-limited environments. From a One Health perspective, the persistence of microorganisms on surgical instruments may facilitate indirect transmission between animals, humans, and contaminated surfaces. Environmental contamination plays a well-documented role in the transmission of healthcare-associated infections, with estimates suggesting that 20–40% of such infections may result from cross-contamination via surfaces or equipment.

Consistent with previous reports, microbial persistence on medical and surgical instruments is especially evident in settings where low-level disinfectants are used. In the present study, contamination was consistently observed in clinics employing low-level disinfection protocols, whereas no contamination was detected in the clinic using a high-level disinfectant. These findings underscore the importance of appropriate disinfection strategies in reducing microbial burden and limiting transmission risks.

Author Response

We sincerely thank the reviewer for the positive evaluation of our manuscript and for the constructive comments provided. We appreciate the reviewer’s recognition of the clinical relevance and practical value of the study. Following these recommendations, the manuscript was revised to improve methodological clarity, discussion focus, and cautious interpretation of the findings.

Comment:
“This study is particularly timely given the limited attention paid by veterinary clinics to the introduction and spread of microorganisms within clinical environments and to the control of contributing factors. This is especially important considering that animals are commonly colonized by a wide diversity of microorganisms, including potentially pathogenic species.”

Response:
We sincerely thank the reviewer for these encouraging comments and for recognizing the relevance of infection prevention and microbial contamination in veterinary clinical environments.

Comment:
“The findings highlight a pressing need to strengthen infection control practices in veterinary settings, particularly in resource-limited environments. From a One Health perspective, the persistence of microorganisms on surgical instruments may facilitate indirect transmission between animals, humans, and contaminated surfaces. Environmental contamination plays a well-documented role in the transmission of healthcare-associated infections, with estimates suggesting that 20–40% of such infections may result from cross-contamination via surfaces or equipment.”

Response:
We appreciate the reviewer’s valuable observation. In response, the Discussion section was revised to better emphasize the importance of infection prevention and instrument reprocessing practices in veterinary healthcare settings, particularly under resource-limited conditions. The One Health perspective was also clarified and more directly linked to the findings of the present study.

Comment:
“Consistent with previous reports, microbial persistence on medical and surgical instruments is especially evident in settings where low-level disinfectants are used. In the present study, contamination was consistently observed in clinics employing low-level disinfection protocols, whereas no contamination was detected in the clinic using a high-level disinfectant. These findings underscore the importance of appropriate disinfection strategies in reducing microbial burden and limiting transmission risks.”

Response:
We thank the reviewer for this important observation. In the revised manuscript, statements regarding disinfectant performance were carefully moderated to avoid causal overinterpretation because of the limited sample size and observational exploratory design of the study. The revised text now emphasizes that residual contamination was more commonly observed in clinics reporting the use of low-level disinfectants, whereas lower contamination frequencies were observed in clinics implementing high-level disinfectant protocols or combined reprocessing strategies. Additional caution regarding interpretation of these findings was also incorporated into the Discussion and Conclusions sections.

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Dear Authors,

After Carefully reviewing the revised version of your Brief Report entitled "Residual Bacterial Contamination of Surgical Instruments After Chemical Disinfection in Veterinary Clinics in Veracruz, Mexico".

I see that the paper is more suitable for publication in its present form.

Best Regards,

Author Response

We sincerely thank the reviewer for the positive evaluation of our revised manuscript and for considering the paper suitable for publication in its present form.

We greatly appreciate the reviewer’s time, constructive feedback, and valuable comments throughout the review process, which significantly helped improve the quality and clarity of the manuscript.

Best regards,
The Authors

Reviewer 2 Report

Comments and Suggestions for Authors

I would like to thank the authors for the revised version of the manuscript. I think the paper has been clearly improved compared with the previous version. The exploratory nature of the study is now better stated, and the conclusions are more cautious and more consistent with the limited sample size and observational design. The topic is relevant because infection control and instrument reprocessing in small veterinary clinics remain important issues, especially in settings where sterilisation may not always be available. The detection of residual bacterial contamination after routine chemical disinfection provides useful preliminary information and may encourage better infection-prevention practices in veterinary clinical environments.

However, I suggest a few minor improvements before publication. First, the authors should provide the exact p-value for Fisher’s exact test instead of only reporting “p > 0.05”. This would improve transparency, especially because the sample size is small. Second, Table 1 should be reformatted because some words are split awkwardly and the table is difficult to read in its current form. Third, the authors may add one clear sentence in the discussion stating that bacterial recovery after disinfection does not necessarily prove infection risk or transmission risk, since bacterial load, virulence factors, and antimicrobial susceptibility were not assessed.

I also suggest a final English and formatting revision. The manuscript is generally understandable, but some sentences could be polished to improve clarity and flow. In addition, the page numbering and layout in the reference section should be checked before publication. Overall, I think this revised manuscript is scientifically improved and can be considered for publication after minor revisions.

 

Comments on the Quality of English Language

English is generally understandable, but it would benefit from a careful final language revision to improve clarity, flow, and readability. I think the scientific message is clear overall; however, some sentences are long and could be simplified, especially in the introduction, methods, and Discussion sections.

I also suggest checking minor grammatical issues, consistency of terminology, and formatting problems in the table and reference section. These revisions would make the manuscript easier to read and would improve the professional presentation of the study.

Author Response

Reviewer Comments:

We sincerely thank the reviewer for the careful evaluation of our revised manuscript and for the positive and constructive comments. We appreciate the recognition of the improvements made in the revised version and the acknowledgment of the relevance of the study.

1. Exact p-value for Fisher’s exact test
   Response: We appreciate this suggestion. The exact p-value obtained from Fisher’s exact test has now been included in the Results section to improve transparency of the statistical analysis.
2. Table 1 formatting
   Response: Thank you for this observation. Table 1 has been reformatted to improve readability and avoid awkward word splitting and layout issues.
3. Clarification regarding infection/transmission risk
   Response: We agree with the reviewer. A clarifying statement has been added to the Discussion section indicating that bacterial recovery after routine disinfection does not necessarily demonstrate a direct infection or transmission risk, since bacterial load, virulence factors, and antimicrobial susceptibility were not evaluated in this study.
4. English language and formatting revision
   Response: The manuscript has undergone an additional English language and formatting revision to improve clarity, readability, grammatical consistency, and overall presentation. The reference section formatting and page layout were also carefully checked and corrected where necessary.

We thank the reviewer again for the valuable comments and positive assessment of our work.