PONE-D-24-04047Campylobacter jejuni and Campylobacter coli, determinants and antimicrobial resistance pattern among under-five children with diarrhea in Amhara National Regional state, Northwest Ethiopia.PLOS ONE

Dear Dr. Worku,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

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Eric Ogola, BVM,MPH,CASMI Fellow

Academic Editor

PLOS ONE

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: No

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: No

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Line 45: Among the Campylobacter isolates, C. jejuni were commonly isolate (73.3%).......

Kindly add 95%CI to all proportions in the manuscript either in the text or the tables. This provide a better expression of the proportion uncertainty.

Reviewer #2: This is a very important manuscript presenting the results of a study that assessed the prevalence of Campylobacter spp among children aged under-five years with diarrhoea. This study has many strengths including:

1- It is addressing a zoonotic disease that is opportunistic in nature

2- The study used advanced assay techniques (MALDI-TOF MS) to identify the organisms and included antibiotic susceptibility testing.

3- The sample size is fairly large

However, the manuscript has some important weaknesses that should be addressed before publication as summarised below:

- Language - there are numerous grammatical errors in the manuscript that are distracting from the important message. The authors could seek professional language editing services to improve the quality of this manuscript.

- The abstract methods do not include some crucial information about how the study was conducted and only focusses on laboratory methods.

Introduction - although the authors mention that they used an advanced laboratory technique (MALDI-TOF MS) to identify the organisms, they do not mention what is the advantage of this method over traditional methods.

Methods - source population - the two sentences in this section are very similar and repetitive.

Sampling technique - it is not very clear how the children were selected and what case definitions of diarrhoea were applied.

Data analysis - the cut off for including variable in a multivariate model is given as 0.25. This threshold is very high, the authors should consider repeating this analysis using a lower threshold.

Results - There are two Table 4s in this section. One appears after Table 6. The abbreviations used in the second Table 4 should be spelt out in a footnote below the table.

Discussion - the authors should revise the first paragraph in this section to summarise the main findings in the study.

The authors note that the prevalence of campylobacter spp was similar to that found in other studies in Ethiopia but higher than other countries such as Kenya. Could this be attributed to the different laboratory techniques? In general, this section focusses so much on comparing study findings with previous study at the expense of explaining the implications of their findings for public health.

conclusions - the following statement is not fully backed up by data presented in this manuscript:

"Culture and sensitivity test should be carried out prior to medication of diarrheal patients"

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Reviewer #1: No

Reviewer #2: No

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PONE-D-24-04047R1Campylobacter jejuni and Campylobacter coli infection, determinants and antimicrobial resistance pattern among under-five children with diarrhea in Amhara National Regional state, Northwest Ethiopia.PLOS ONE

Dear Dr. Worku,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Please submit your revised manuscript by Jun 07 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:

• A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Tebelay Dilnessa, MSc

Academic Editor

PLOS ONE

Journal Requirements:

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

Additional Editor Comments:

• The title, ‘Campylobacter jejuni and Campylobacter coli infection, determinants and antimicrobial resistance pattern among under-five children with diarrhea in Amhara National Regional state, Northwest Ethiopia’. The word ‘resistance’ better be replaced by ‘susceptibility’. Then modify the objective and description of tables accordingly.
• The affiliation better which was ‘⁵Animal Health Institute, Sebeta, Ethiopia’ should be linked to the author (s) or removed.
• The email addresses of authors were not needed here in the main manuscript as it appears in the system except the corresponding author.
• Lines 39 &40: ‘Suspected colonies were analyzed using matrix-assisted laser desorption ionization-time of flight mass spectrometry to confirm the species.’ It is better written as, ‘The suspected colonies were analyzed using matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) to confirm the species.’
• Lines: 41&42: ‘Logistic regression was used to analyze the associated factors.’  It was better written as, ‘Logistic regression was used to analyze the associated factors with P-value <0.05’.
• Lines 43-45: ‘Among the 428 samples, 30 (7.0%) (CI: 4.5-9.3%) were culture-positive for Campylobacter species. The prevalence of C. jejuni and C. coli among under-five children was 5.1% (CI: 3.0-44 7.0%) and 1.9% (CI: 0.7-3.3%)’. Here the sign (%) from the CI part should be removed because by default it has already known. Similarly, the number of isolates with the total better written together with the percentages. This works for the whole abstract and result part of the paper. For example, for the above better written as, “Among the 428 samples, 30 (7.0%) (CI: 4.5-9.3) were culture-positive for Campylobacter species. The prevalence of C. jejuni and C. coli among under-five children was 22/428 (5.1%) (CI: 3.0-44 7.0) and 8/428 (1.9%) (CI: 0.7-3.3)’.
• Line 64: ‘………health infrastructure is prevalent. [1].’ Better written as, ‘………health infrastructure is prevalent [1].’
• Line 88: ‘……… in animal husbandries. [12,13].’ Better written as, ‘,… in animal husbandries [12,13].’
• Similarly, line 94: ‘……………..and tetracycline. [14–16].’  Better written as, ‘…….. and tetracycline [14–16].
• Line 96 &97: ‘Binary and multivariable logistic analysis was used to identify associated risk factors. A P value less than 0.05 at a 95% confidence interval was statistically significant’. Better written as ‘Bivariable and multivariable logistic regression analysis was used to identify associated risk factors. A P-value < 0.05 at a 95% confidence interval was considered as statistically significant’.
• In the data analysis part, it is better you add something about the interpretation of ‘MALDI-TOF MS assay.  
• Line 100: ‘MALDI-TOF MS provides a rapid…….’ It is better written as, ‘Matrix-assisted laser desorption ionization-time of flight mass spectrometry provides a rapid…….’ As the start of a sentence will not be acronyms/abbrevations.
• Line 100: ‘…….Campylobacter species identification.’ It is better written, ‘……….Campylobacter species identification.’ Similarly, the binomial nomenclature should be followed throughout the document.
• Lines 103&104: ‘………pattern of Campylobacter jejuni and C. coli among under-five children with diarrhea.’ It is better written as, ‘………pattern of C. jejuni and C. coli among under-five children with diarrhea.
• Lines 116, 121, 123, 124, 137 and 128: ‘Bahr Dar’ should be corrected as ‘Bahir Dar’
• Line 199: ‘Data quality’ better written as ‘Data quality assurance’
• Lines 232- 235: Table 1 of the last column ‘Campylobacter infection (%, CI)’, the confidence interval (CI) was not much important and better be removed. Similar cenario existed in Tables 2 &3.
• Line 243: ……Campylobacter species, better written as ……..Campylobacter species,
• Line 259: The subheading ‘Potential risk factors and their relationship with Campylobacter enteritis’ It is better written as, ‘Potential associated factors and their relationship with Campylobacter enteritis’
• Line 272: Table 5 should be supplemented with P- value of COR and P-value of AOR.
• In the result part, you said nothing about how may culture positives confirmed by MALDI-TOF MS assay as positive and how many also became negative.
• The presentation of associated factors better be presented as, for example lines 276-278: ‘Consequently, children residing in peri-urban areas and having contact with domestic animals were 4.91(1.92-12.54) and 5.03(2.18-11.62) times more likely to contract Campylobacter gastroenteritis.’ It is better presented as, ‘Consequently, children residing in peri-urban areas and having contact with domestic animals were 4.91 (95%CI: 1.92-12.54, P=?) and 5.03 (95%CI: 2.18-11.62, P=?) times more likely to contract Campylobacter gastroenteritis.’ Sometimes we can also write as (AOR: ?, 95%CI: ? P=?). This works for all associated factors analysis.
• Lines 425-434: Authors contribution should be removed as the system creates automatically.
• Beyond the comparison, the discussion part requires scientific explanation and reasoning.
• It is better also you follow the manuscript writing protocol for PloS One, especially font size, font type, reference list writing, table and figure preparation and whether figures submitted within the main manuscript or not.
• Proof readig of the whole manucript is needed.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #3: (No Response)

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #3: Partly

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3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #2: I Don't Know

Reviewer #3: Yes

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4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #3: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: No

Reviewer #3: No

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2: I have the following observations:

1. Two versions of the manuscript are presented which are very different. The first version is marked as "revised" and the second marked "Revised manuscript with changes with yellow". I assume the second one was supposed to be the revised manuscript highligting the changes that have been made. The authors need to thoroughly review their work before submission to avoid this type of confusion.

2. There are some grammatical or typographical errors in the manuscript. The article could benefit from thorough language editing. Below are some few examples:

- title - "antimicrobial resistance pattern" an "s" should be added to "pattern"

- Abstract - "children with under-five year age disproportionally affected with foodborne illness."

- "The prevalence of C. jejuni and C. coli among under-five children was 5.1% (CI: 3.0- 7.0%) and 1.9% (CI:0.7-3.3%). - "respectively" may be needed at the end of this statement.

- "The rest might be either refuse to participate or unable to get specimen"

- "Clinical characterization of under-five children" might sound better if written as "Clinical characteristics of children aged under-five years"

3. The details of how data was analysed - especially the bivariate and multivariate are lacking in both abstract and main manuscript. What was the outcome of interest? What measures of association were calculated? How were covariates progressed from bivariate to multivariate models? Since the data

4. Sampling of children - in some places the authors refer to "convenient sampling" yet in other places they refer to "consecutive sampling". They need to be consistent.

5. "The rest might be either refuse to participate or unable to get specimen" - the authors only talk of the 428 participants. This statement implies that there were "others" who are not quantified. It is also not clear what proportion of these "others" were excluded based on the two criteria mentioned. Were these "others" significantly different from the included participants?

6. "Data of the current study showed that 8 variables analyzed by Bivariate logistic regression had P value < 0.25 and considered for further analysis using multivariable logistic regression model.The Model was checked by Hosmer-Lemeshow goodness-of-fit." Should be refiened and moved to the data analysis section rather than results.

Reviewer #3: Basic questions to the Author Lanuage, Fragmented sentence, punctuation and similar questions should edited through out the manuscrpit as indicated in the comment.

Sampling and selection of participants should be clearly indicated

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: No

Reviewer #3: No

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[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Attachments

Attachment

Submitted filename: Campyello bacter.docx

Campylobacter jejuni and Campylobacter coli infection, determinants and antimicrobial resistance patterns among under-five children with diarrhea in Amhara National Regional State, Northwest Ethiopia

PONE-D-24-04047R2

Dear Dr. Worku,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Tebelay Dilnessa, MSc

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Some proof reading is required.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #3: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #2: Yes

Reviewer #3: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #3: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

Reviewer #3: No

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2: The authors have made a good effort at revising the manuscript, which now reads better. I have no further comments.

Reviewer #3: As it it is indicated above the the revision of manuscripit using standard English should be checked again before publication or acceptance of the document through out the whole document.

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: No

Reviewer #3: Yes: Demissie Assegu Fenta

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