Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Dear Authors,

After reviewing the manuscript entitled "Prevalence, Antimicrobial resistance profiles, and risk factors Analysis of Campylobacter spp, from Dogs in Kelantan, Malaysia."

I see that the article is very important from a veterinary point of view.

This article study the prevalence and the antimicrobial resistance and the risk factors of one of the leading cause of gastroenteritis (Campylobacter) in dogs in a region of Malaysia.

Kindly find the list of my comments regarding this work:

01- In the Abstract, the term "Abstract" is duplicated kindly remove one copy of it.

02- In the Abstract, you do not highlights on the results concerning the risk factors for Campylobacter infection in dogs.

03- In the list of Keywords, you are kindly invited to add the term "Malaysia".

04- In the Introduction, The first paragraph is very long, I suggest to dived it into two paragraphs.

05- In the Introduction, In line 48, you are invited to put this reference when you talked about the presence of Campylobacter in dogs.

xxx Fecal prevalence of Campylobacter spp. in house dogs in Lebanon: A pilot study.

xxx Prevalence of Campylobacter spp. in a cross-sectional study of dogs attending veterinary practices in the UK and risk indicators associated with shedding.

06- In the Introduction, Lines 48-52, when you talked about the transmission of Campylobacter from animals to human, you are kindly invited to use the following articles as references:

xxx Prevalence, antimicrobial resistance and risk factors for campylobacteriosis in Lebanon.

xxx Prevalence and Antimicrobial Resistance of Campylobacter Species and Associated Factors Among Under-Five Children with Diarrhea at Randomly Selected Public Health Facilities in Mekelle, Tigray, Ethiopia.

07- In the Introduction, Lines 60-62, why some information are in italic? Kindly put it in the regular form.

08- In the Introduction, Lines 64-66, why some information are in italic? Kindly put it in the regular form.

09- In the Materials and Methods, Paragraph "Bacterial Isolation and Identification", you mentioned the follow "... those positive for catalase and oxidase, negative for urease, and positive for Hippurate were presumptively identified as Campylobacter positive." Are you sure that all Campylobacter species are positive for Hippurate? Kindly verify this point!!!

10- In the Materials and Methods, Paragraph "Bacterial Isolation and Identification", you mentioned the follow "Phenotypically confirmed isolates were preserved in Müller–Hinton broth supplemented with 50% glycerol and stored at −80 °C for further analysis." How can you explain that Campylobacter isolates will survive in Müller–Hinton broth?!! It is an exigent bacteria and Müller–Hinton broth is a basic medium!!!

11- In the Materials and Methods, Paragraph "Antimicrobial susceptibility testing", you mentioned the follow "Antimicrobial susceptibility testing (AST) for Campylobacter isolates was performed using the Kirby-Bauer disk diffusion method on Mueller-Hinton agar" How can you explain the use of Mueller-Hinton agar for the culture of Campylobacter species? You are invited to replace (Mueller-Hinton agar) by (Enriches Mueller-Hinton agar).

12- In the line 191, you are kindly invited to replace "Cj0414" by "cj0414".

13- In the line 192, you are kindly invited to replace "gyrB" by "gyrB".

14- In the line 207, an information is missed "The antimicrobial resistance profiles of Campylobacter helveticus and Campylobacter were broadly similar", I think that you should replace it by "The antimicrobial resistance profiles of Campylobacter helveticus and Campylobacter jejuni were broadly similar".

15- In Table 1, Why there is no information regarding to the sensitivity of Campylobacter jejuni towards Kanamycin and Chlorophenicol?

16- In the line 225, you should remove the number "3.4" keep "3.3".

17- In the Whole manuscript, you should put the name of bacterial species in italic. Example Lines 229 and 232 Campylobacter spp. are not in italic.

18- In the line 252, this section should have the number 2.4 instead of 2.5.

19- You are invited to align the information in your tables.

20- Why in you study, you was limited to data regarding Campylobacter jejuni and Campylobacter helveticus??

21-  In the Discussion, It is good to mention the impact of Campylobacter infection on human health, noting its transmission from Animals to Humans (mainly trough chicken "Broilers") and its impact on human life mainly on children, you can use the following articles as references for these points:

xxx Prevalence of Campylobacter spp. in broilers in North Lebanon.

xxx Prevalence, Laboratory Findings and Clinical Characteristics of Campylobacteriosis Agents among Hospitalized Children with Acute Gastroenteritis in Lebanon.

22- In the Discussion, It is good to mention that during COVID-19 pandemic we remarkably see the increase use of non-specific antibiotic, which increase the antimicrobial resistance trend worldwide, you can refer to the following article concerning this point:

xxx Antibiotic misuse during the COVID-19 pandemic in Lebanon: a cross-sectional study

23- In the Discussion, It is importance to talk about the future strategies to combat antimicrobial resistance in bacteria, here I suggest to highlight on the impact of using new antimicrobial therapy such as the trojan horse strategy or targeting bacterial metallophores to fight bacterial resistance. You can use the following papers to explain this point:

xxx Siderophore–drug complexes: potential medicinal applications of the ‘Trojan horse’ strategy

xxx Towards new antibiotics classes targeting bacterial metallophores.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Title: Prevalence, Antimicrobial Resistance Profiles, and Risk Factors Analysis of Campylobacter spp. from Dogs in Kelantan, Malaysia

1. Abstract: The abstract does not mention any risk factor analysis, despite this being part of the study title. Please clarify whether such analysis was conducted and include a brief summary of it in the abstract if applicable.

2. Lines 60–66: Please clarify the reason for using italicized text in this section. If it is not standard nomenclature (e.g., scientific names), consider removing the italics for consistency and readability.

3. Line 100 – Sample Size Justification: The manuscript states that a total of 50 rectal swabs were collected from dogs. However, there is no explanation of how this sample size was determined. Please provide justification or a calculation to support the adequacy of this number. Also, considering only 14 Campylobacter isolates were obtained, the sample size may be insufficient to draw robust conclusions, particularly regarding antimicrobial resistance and risk factor analysis.

4. Risk Factor Analysis – Methods Section: There is no mention of risk factor analysis in the Methods section. If this was conducted, please provide a detailed description of the variables assessed, statistical tests used, and the criteria for significance. If not, the study title should be revised accordingly.

5. Figure 1: Please label the Y-axis to clarify what is being measured or represented in the figure.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The present study, "Prevalence, Antimicrobial resistance profiles, and risk factors Analysis of Campylobacter spp, from Dogs in Kelantan, Malaysia" studied the prevalence, evaluate the antimicrobial resistance 
profiles, and identify risk factors associated with infection in dogs from Kelantan, Malaysia. Overall, the work is significant in respect of the increasing AMR; however, has few concerns that need to be addressed before further processing of this manuscript.

1. line 60-62, 64-66: Why italic
2. Write why to study Campylobacter where many other pathogen cause AMR and could transmit to human. 
3. Write more background on the major issues on the AMR in dogs and how it impacts humans health. 
4. Cite previous studies that reports AMR transmission from dogs to human.
4. In the objective section write why it is important to study the prevalence as well as AMR in this population?
5. Write the primer sequences for 16s rRNA,gyrB gene and Cj0414 gene and their amplification conditions. 
6. Did you sequence these genes, if so please write the accession no.
7. Figure 1.: remove the chart title work or add a title.
8. 286-287: Add reference for "The occurrence of Campylobacter spp. in domestic animals, particularly companion animals such as dogs, holds significant public health relevance due to their established zoonotic potential"
9. Discuss more on Zoonotic Potential of Campylobacter.
10. Line 301-302: Write the reference for the previous studies.
11. line 310: write the author name [23] and do same throughout the manuscript.
12. Write how the present study findings helps in human health management.
13. Discuss the probable consequence of the high levels of antimicrobial resistance in veterinary and public health.
14. There are several grammatical errors. please correct
15. Write a paragraph in the discussion for limitation and recommendation (based on this research).

Comments on the Quality of English Language

Minor English language correction is required 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

Dear Authors,

After reviewing the revised version of the manuscript entitled "Prevalence, Antimicrobial resistance profiles, and risk factors Analysis of Campylobacter spp, from Dogs in Kelantan, Malaysia."

I just have one point regarding mentioning that Muller-Hinton medium used in the study was an Enriched-Muller-Hinton medium and not a normal Muller-Hinton medium.

Otherwise, I see that the article is good in its present form, thanks to the modifications you made,

Best Regards,

Author Response

Reviewer 1- Round #2

Comments and Suggestions for Authors

After reviewing the revised version of the manuscript entitled "Prevalence, Antimicrobial resistance profiles, and risk factors Analysis of Campylobacter spp, from Dogs in Kelantan, Malaysia."

I just have one point regarding mentioning that Muller-Hinton medium used in the study was an Enriched-Muller-Hinton medium and not a normal Muller-Hinton medium.

 

Otherwise, I see that the article is good in its present form, thanks to the modifications you made

Response: Thank you for your valuable feedback and for your positive comments on the revised manuscript.

Regarding your observation, I confirm that we used Enriched Muller-Hinton medium in the study, not the normal Muller-Hinton medium.

 

 

 

 

Reviewer 2 Report

Comments and Suggestions for Authors

Thanks for your intention to edit and improve your manuscript. 

Comments:

Figure 1: Adjust the Y-axis so that the tallest bar reaches 100.

Adjust the column widths in Table 5 to improve readability.

Author Response

Review comment 2 #round 2

Comments and Suggestions for Authors

Thanks for your intention to edit and improve your manuscript. 

Comments:

Figure 1: Adjust the Y-axis so that the tallest bar reaches 100.

Adjust the column widths in Table 5 to improve readability.

Response: Thank you for your comment and suggestion. We have amended the figure 1, and table 5 as suggested.