Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CONTINUED NUCLEIC ACID TESTS FOR SARS-COV-2 FOLLOWING DISCHARGE OF PATIENTS WITH COVID-19 IN LU’AN, CHINA
Review round: 1
Reviewer: 3

Output:
Basic reporting: The article by Lyu et al., reports about the re-occurrence of positive COVID-19 tests as demonstrated by RT-PCR in 17 COVID-19 patients who had otherwise recovered and discharged from the hospital. They confirmed the same by conducting RT-PCR based testing on collect sputum, throat swab, a fecal or anal swab, and urine samples on the 7th and the 14th day following discharge. The study was conducted in a small cohort of patients n=67 in thee Lu'an Municipal Center for Disease Control and Prevention, Lu'an, China. The genes assayed for the tests were ORF and N for SARS-CoV-2.
The introduction calls for better details. Given that COVID-19 is a global pandemic, the current manuscript is very China-centric and I suggest adding a few international reports to lend more credibility to their hypothesis and conclusions. Specific details are as below
o	L 43-45: Being a global pandemic the authors must include the WHO patient discharge guidelines and guidelines issued by other major nations to aid the larger global audience understand the importance of the work done.
o	L 47- 51: Being a global pandemic I recommend the authors improve the information especially from outside of China at to add further rationale for their study. Since the authors explicitly mention that there have been previous reports of RT-PCR positivity in discharged patients a couple of lines on the knowledge gap being filled by the present study would enhance the read.
The language needs to be more lucid and well punctuated to avoid ambiguity. The authors may get the manuscript rewritten in proper English. There are places where the statements and observations do not match such as:
o	L69-70: What do authors mean by the statement “Unrecovered positive patients were those who remained negative from discharge to the end of study observation”? I think this line should be changed to “Recovered negative patients were those who remained negative from discharge to the end of study observation.”
o	L80: “A total of 17 (25.4%) out of the confirmed cases were recovered and were positive by RT-PCR” Does that mean the remaining cohort were still unrecovered and still suffering from the disease? Why were they discharged then?
Experimental design: The authors detected viral nucleic acids using RT-PCR based methodology in sputum, throat swab, fecal or anal swab, and urine samples of discharged patients.
o	L58: “confirmed COVID-19 cases in Lu'an Municipality, China, who met the discharge criteria” For a wider international audience it should be properly explained what the current discharge criterion at this point rather than putting it in the discussion and the clinical, laboratory, and radiological characteristics of the patients during hospitalization who tested positive following discharge can be put in the supplement for a clear understanding of the cases.
o	L61-62: Many cases of reinfection are being reported around the world a clarification on how did the authors rule out reinfection would have helped.
o	L 65-66: The authors have not mentioned the validity of the assay used. Multiple assays have been designed to detect COVID-19 and most assays have a limit of detection defined. So the authors should mention the RT-PCR test kit manufacturer assay details such as the limit of detection and Ct value cut off for a positive test. Further, any RT-PCR assay has its limit of detection clearly defined, please state the same in the methods they have not stated the sensitivity of the test? Does the Ct value of 40 count as positive or simple background noise?
o	A proper RT-PCR test is defined by the presence of controls. I thank you for providing the raw data, however the raw data supplied did not have any data regarding the controls and amplification curves of the tests. A true positive is always defined by a sigmoidal amplification curve.
o	L75-76: the alpha value of p <0.05 was considered statistically significant, the authors should explore using a stringent statistic p <0.01 for arriving at conclusions.
Validity of the findings: o	L 57-58: The cohort size of the study is very small (n=67) and only 17 were found to be having a positive RT-PCR test post-remission of all symptoms and only 1 had all samples which tested positive. For a study having implications as drastic as redesigning discharge criteria, the cohort size is too small for an effective conclusion. The authors should increase the cohort size before publishing their results.
o	L110-111: Infectivity of the virus from the low viral load is yet not proved the study would have been better presented if data about the infection spreading from discharged patients would have been presented their argument and thus would have corroborated the findings and conclusions drawn. Multiple papers in the recent past have used cell line based testing to validate the presence of the infectious virus in the biological sample (e.g. Wölfel R, Corman VM, Guggemos W, et al. Virological assessment of hospitalized patients with COVID‐2019. Nature. 2020; 581:465‐469; Xiao F, Sun J, Xu Y, et al. Infectious SARS‐CoV‐2 in Feces of Patient with Severe COVID‐19. Emerg Infect Dis J. 2020; 26:1920‐1922).  Wölfel et aI. 2020 did not find an infectious virus in the stool sample despite high concentrations of virus RNA, while urine samples never yielded a virus. I strongly suggest the authors conduct cell line-based studies before arriving at the conclusion they have arrived at based solely on RT-PCR testing

o	L119-121: The mere possibility of viral transmission is insufficient to change discharge criterion and evidence of transmission is this regard would have greatly helped. This indicates a larger cohort would have helped in strengthening the conclusions drawn.
Additional comments: These are trying times for the scientific community in general and the biomedical scientists specifically. I want to extend my best wishes to the authors for having reported this study, however, a few points need to be kept in mind before acceptance:
1.	The article in general needs to be rewritten IN PROPER LUCID ENGLISH LANGUAGE TO AVOID AMBIGUITY.
2.	China-centric parameters used but a better approach would be to also USE WHO GUIDELINES while reporting patient data for wider implications and understanding of the findings.
3.	For the RT-PCR, authors should mention the RT-PCR test kit assay details such as LIMIT OF DETECTION AND CT VALUE CUT OFF FOR A POSITIVE TEST.
4.	The detection of viral nucleic acid does not necessarily imply that viable infectious virions are present or that the virus can spread. Also, the absence of secondary infections indicates that there was no spread. Therefore the conclusions drawn are risky, and mere extrapolation of the findings and do not truly represent the findings. CELL LINE BASED STUDY TO DETECT ACTUAL INFECTIVE VIRIONS IS A MUST BEFORE THE MANUSCRIPT IS ACCEPTED FOR PUBLICATION with the present conclusions.
5.	It is difficult to work with COVID-19 samples and patients but cohort size may be expanded for better results.
Therefore I DO NOT RECOMMEND the publication IN ITS PRESENT FORM and suggest authors look into the suggestions made. The study was fairly executed but is far off when it comes to the conclusions drawn. There are some major queries as outlined above that the authors should address before the publication of this article


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CONTINUED NUCLEIC ACID TESTS FOR SARS-COV-2 FOLLOWING DISCHARGE OF PATIENTS WITH COVID-19 IN LU’AN, CHINA
Review round: 2
Reviewer: 1

Output:
Basic reporting: Although the manuscript has substantially improved, there are still some issued that should be addressed by the authors for its publication.

1. Authors mention on their rebuttal letter that the language of their manuscript has been refined and polished by using the English Language Editing service available from PEER J Publishing. I find that difficult to believe as there are still numerous spelling and grammar mistakes. I would emphasize that the manuscript would benefit from review by a native English speaker to assist with the sentence construction and spelling. I am highlighting some suggestions on the reviewed manuscript.
Experimental design: 2. In 2009, Bustin et al., proposed a number of guidelines targeting the reliability of results to help ensure the integrity of the scientific literature, promote consistency between laboratories, and increase experimental transparency. They mention that the nomenclature describing the fractional PCR cycle used for quantification is inconsistent, with threshold cycle (Ct), crossing point (Cp), and take-off point (TOP) currently used in the literature. These terms all refer to the same value from the real-time instrument and were coined by competing manufacturers of real-time instruments for reasons of product differentiation, not scientific accuracy, or clarity. Then, the authors proposed the use of quantification cycle (Cq), according to the RDML (Real-Time PCR Data Markup Language) data standard. These guidelines are highly used by groups with high quality standards on their qPCR assays. Thus, I would like to encourage the authors to adjust their manuscript to these guidelines.
Validity of the findings: 3. From my perspective, it is not clear for authors what a discussion means. The purpose of the discussion is to interpret and describe the significance of the findings in light of what was already known about the research problem being investigated, and to explain any novel insights about the problem after the authors taken the findings into consideration. Thus, the discussion implies contrasting the obtained results with those reported by others. And this is the main flaw of this manuscript. The lack of a deep discussion. I would like to see the manuscript discussion go deeper by comparing their results with that of other research groups. In its current form, authors do not compare their results against other studies (for example, authors may compare their findings with those of Weerahandi et al., 2021 and Lin et al., 2021). Authors also should emphasize the relevance of their study to reinforce preventive measures to control the transmissibility of the disease.

4. Finally, and once again, the conclusions are too shallow and simplistic. The conclusion should include summarizing the paper and focus possible future studies in broader perspectives of the paper. Authors should describe how does this study has contributed to knowledge gaps (which, due to the novelty of this pandemics are abundant).

References
Bustin, S.A., Benes, V., Garson, J.A., Hellemans, J., Huggett, J., Kubista, M., Mueller, R., Nolan, T., Pfaffl, M.W., Shipley, G.L., Vandesompele, J., Wittwer, C.T. 2009. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 55(4):611-622.
Lin, P., Chen, W., Huang, H., Lin, Y., Cai, M., Lin, D., Cai, H., Su, Z., Zhuang, X., Yu, X. 2021. Delayed discharge is associated with higher complement C3 levels and a longer nucleic acid-negative conversion time in patients with COVID-19. Sci Rep. 11(1):1233.
Weerahandi, H., Hochman, K.A., Simon, E. et al. 2021. Post-Discharge Health Status and Symptoms in Patients with Severe COVID-19. J. Gen. Int. Med. 1-8.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CONTINUED NUCLEIC ACID TESTS FOR SARS-COV-2 FOLLOWING DISCHARGE OF PATIENTS WITH COVID-19 IN LU’AN, CHINA
Review round: 3
Reviewer: 1

Output:
Basic reporting: Dear Authors,

You have made significant improvements to your manuscript in terms of the English language. Also, you have added the paper (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7276353/#r2) as a reference as I previously have suggested to you. It is great to read the improved version of the manuscript.
Experimental design: Dear Authors,
I think that it improves your manuscript the fact that you stated the deficiencies and limitations of your study. For example, I have suggested that you should conduct viral cell culture experiments, but you do not have access to a biosafety level 3 laboratory. This type of information is very important for future studies or publications.
Validity of the findings: No comment
Additional comments: Dear Authors,
You have improved your manuscript and followed the suggestions and recommendations that I have previously made. This type of work is relevant to the current and ongoing COVID-19 global pandemic. This reviewer understands the hard work and effort that went into improving your manuscript. Discharge protocols for COVID-19 patients, science-based, can save lives and valuable resources (e.g., PPE, ICU beds, and oxygen) during this pandemic.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: POTENCY OF BISRESORCINOL FROM HELICIOPSIS TERMINALIS ON SKIN AGING: IN VITRO BIOACTIVITIES AND MOLECULAR INTERACTIONS
Review round: 1
Reviewer: 1

Output:
Basic reporting: All terms and materials suitable for the subject have been provided.
Experimental design: A suitable experimental design was used to approach the experimental purpose.
Validity of the findings: It provides basic data that can be used as a cosmetic material for bisresorcinol.
Additional comments: This study confirmed the biological effects of bisresorcinol on tyrosinase, collagenase, and elastase in vitro. In addition, the interaction of bisresorcinol at the molecular level was confirmed using a docking technique.

1.	The inhibition mechanism for various enzymes in vitro will not be linear regulation as shown in Figure 2. Since enzymes have a tertiary or quaternary structure, the mode of action with a substrate or an inhibitor proceeds by hyperbolic or sigmoidal reaction. How can linear regulation come about?
2.	The raw data provided only conducted enzyme inhibition experiments on three concentrations, and how did Figure 2 show more than three concentrations?
3.	What is the yield of bisresorcinol extracted from Heliciopsis terminalis, and what is the purity of bisresorcinol used in the experiment? Please refer to this information in the paper or provide it as a supplementary material.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: POTENCY OF BISRESORCINOL FROM HELICIOPSIS TERMINALIS ON SKIN AGING: IN VITRO BIOACTIVITIES AND MOLECULAR INTERACTIONS
Review round: 1
Reviewer: 2

Output:
Basic reporting: BASIC REPORTING
The title, abstract, introduction, methods, results and discussion are appropriate for the content of the text. Furthermore, the article is well constructed, the experiments are well conducted, and analysis is well performed. The figures are relevant, high quality, well labelled and described.
Experimental design: EXPERIMENTAL DESIGN
The experimental design is original and the research is within the scope of the journal. Research question is well defined, relevant and meaningful. The methods are highly technical, ethical and logistical.
Validity of the findings: VALIDITY OF THE FINDINGS
All underlying data have been provided in detail. The findings are meaningful. The conclusions are well stated and relevant to the research questions.
Additional comments: This paper demonstrated the function of bisresorcinol purified from H. terminalis in skin aging. The authors investigate its inhibitory activity in vitro on collagenase, elastase, and tyrosinase using a series of spectroscopic techniques. The molecular interaction analysis was further evaluated using molecular dynamic simulation studies to decipher the binding sites of test compounds to collagenase, elastase and tyrosinase. In short, the authors demonstrate that the bisresorcinol purified from H. terminalis might be useful for inclusion in cosmetic products as an aging-enzyme antagonist.

Editorial Criteria
BASIC REPORTING
The title, abstract, introduction, methods, results and discussion are appropriate for the content of the text. Furthermore, the article is well constructed, the experiments are well conducted, and analysis is well performed. The figures are relevant, high quality, well labelled and described.
EXPERIMENTAL DESIGN
The experimental design is original and the research is within the scope of the journal. Research question is well defined, relevant and meaningful. The methods are highly technical, ethical and logistical.
VALIDITY OF THE FINDINGS
All underlying data have been provided in detail. The findings are meaningful. The conclusions are well stated and relevant to the research questions.

Overall, I think this paper is novel and will be of interest to others in the community of skin anti-aging strategies research. The statistical part is valid and makes sense. The authors make it comprehensive by investigating the molecular mechanisms behind its anti skin aging function in a qualitative manner. It can be assumed that the in vitro assays performed with the collagenase, elastase, tyrosinase and respected synthetic peptides are reliable tool to get first indications of the capability of bisresorcinol from Heliciopsis terminalis on skin aging. In general, the work is convincing except some major and minor comments below:

Major Comments:

I’m wondering if the cells used in culture were routinely subjected to mycoplasma testing or not? If so, please add sententense like “only cells that were negative for mycoplasma were used for experiments” in the Methods.

It’s better to add “all the chemicals used in this study were of analytical grade” or similar idea in the Chemicals and Instruments part.

Minor Comments:
The authors didn’t mention what tools were used to perform molecular interaction visualization for the selected receptors and ligand.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: POTENCY OF BISRESORCINOL FROM HELICIOPSIS TERMINALIS ON SKIN AGING: IN VITRO BIOACTIVITIES AND MOLECULAR INTERACTIONS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The title, abstract, introduction, methods, results and discussion are appropriate for the content of the text. Furthermore, the article is well constructed, the experiments are well conducted, and analysis is well performed. The figures are relevant, high quality, well labelled and described.
Experimental design: The experimental design is original and the research is within the scope of the journal. Research question is well defined, relevant and meaningful. The methods are highly technical, ethical and logistical.
Validity of the findings: All underlying data have been provided in detail. The findings are meaningful. The conclusions are well stated and relevant to the research questions.
Additional comments: This paper demonstrated the function of bisresorcinol purified from H. terminalis in skin aging. The authors investigate its inhibitory activity in vitro on collagenase, elastase, and tyrosinase using a series of spectroscopic techniques. The molecular interaction analysis was further evaluated using molecular dynamic simulation studies to decipher the binding sites of test compounds to collagenase, elastase and tyrosinase. In short, the authors demonstrate that the bisresorcinol purified from H. terminalis might be useful for inclusion in cosmetic products as an aging-enzyme antagonist.

Overall, I think the corrections look good to me. All my comments have been answered in a proper manner. Please find the original comments and feedbacks below:

Editorial Criteria
BASIC REPORTING
The title, abstract, introduction, methods, results and discussion are appropriate for the content of the text. Furthermore, the article is well constructed, the experiments are well conducted, and analysis is well performed. The figures are relevant, high quality, well labelled and described.
EXPERIMENTAL DESIGN
The experimental design is original and the research is within the scope of the journal. Research question is well defined, relevant and meaningful. The methods are highly technical, ethical and logistical.
VALIDITY OF THE FINDINGS
All underlying data have been provided in detail. The findings are meaningful. The conclusions are well stated and relevant to the research questions.

Overall, I think this paper is novel and will be of interest to others in the community of skin anti-aging strategies research. The statistical part is valid and makes sense. The authors make it comprehensive by investigating the molecular mechanisms behind its anti skin aging function in a qualitative manner. It can be assumed that the in vitro assays performed with the collagenase, elastase, tyrosinase and respected synthetic peptides are reliable tools to get first indications of the capability of bisresorcinol from Heliciopsis terminalis on skin aging. In general, the work is convincing except some major and minor comments below:

Major Comments:

I’m wondering if the cells used in culture were routinely subjected to mycoplasma testing or not? If so, please add sententense like “only cells that were negative for mycoplasma were used for experiments” in the Methods.

Response:
Thanks for your suggestion. The word we used “in vitro'' could be misunderstood, because cell culture was not performed in this study. The mentioned “in-vitro” experiments meant enzyme-inhibitory assay only. To avoid such misunderstanding, we have added the sentence “In vitro experiments that consisted of enzyme-inhibitory assays on collagenase, elastase and tyrosinase for the bisresorcinol and known inhibitors were carried out as follows.” Under the subtopic “Biological Activity Assays'' of the topic “Methods”

Feedback:
Thanks for the clarification. That makes sense to me.

It’s better to add “all the chemicals used in this study were of analytical grade” or similar idea in the Chemicals and Instruments part.

Response:
Thanks for the suggestion. We put the sentence “All the chemicals used in this study were of analytical grade.” in “Chemicals and Instruments” part.

Feedback:
It looks good to me. Thanks!

Minor Comments:
The authors didn’t mention what tools were used to perform molecular interaction visualization for the selected receptors and ligand.
	Response:
Thanks for the suggestion. In this study, we have used two packages to perform molecular interaction visualization, which were BIOVIA Discovery Studio software and Visual Molecular Dynamics (VMD). We have added the sentence at the end of the method section, with a track change, as followed “Molecular interaction between the compound and a protein receptor was analyzed and visualized using BIOVIA Discovery Studio software (BIOVIA, 2020). The structure of the protein binding site compound was visualized using Visual Molecular Dynamics (VMD) package (Humphrey, Dalke & Schulten, 1996)” to respond to your comment. The additional reference for VMD was also added in the reference section.
Feedback:
That is very clear. Thanks for the clarification. And thanks for adding the citations.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EVALUATING ACTIVE VERSUS PASSIVE SOURCES OF HUMAN BRUCELLOSIS IN JINING CITY, CHINA
Review round: 1
Reviewer: 1

Output:
Basic reporting: English needs to be improved.
Experimental design: no comment.
Validity of the findings: no comment.
Additional comments: This is an interesting paper that provides useful novel information on performance of two detection modes for human brucellosis and the factors associated with human brucellosis in Jining City, China and got the conclusion that active detection mode is better for early diagnosis of HB and therefore proposed three measures including strengthening public education in high-risk area and improving diagnosis in both hospitals and CDCs. The findings are interesting and of significance to prevention and control of HB. Thus it can be considered for publication in this journal after some revision.
Some general comments:
1.Although the language is generally readable, there are many mistakes in spelling and grammar and wrong use. Thus proofreading/editing by a native English speaker is recommended. Some examples are given as follows:
L70 “mortality” should be “morbidity”;
L86-87: “due to the positive cases were not initiative”
2. Please check and ensure the citation of refs is correct. For example Line 91, text mentioned 2018 edition, but the ref 17 is actually 2012 edition.
3. “the positive detection rate” isn’t a typical epidemiological term, it is better to be replaced by percentage of test positive or test prevalence.
4. The test prevalence (or the positive detection rate used in this manuscript) should be displayed with 95% CI.
5. In conclusion, the proposed three measures are not conclusions, thus should be moved into the Discussion section.

Some specific comments:
Abstract
1.	Line 27, you should use the term of “high-risk” throughout the manuscript, do not add space inside the phrase.
2.	Line 28-29, the sentence of “Data were analyzed that experienced the greatest number of cases during the period under observation” is difficult to understand. Please rewrite this sentence.
3.	Line 35, 37, regarding P values, the should be in italics and small letter.

Introduction
4.	Line 54-55, here should put “contaminated ” before “raw meat and dairy products”.
5.	Line 70, “mortality” should be replaced by “morbidity” because brucellosis rarely results in death in humans.

Materials and Methods
6.	Line 86-87, “due to the positive cases ” is suggested to revise “since these cases” .
7.	Line 91, the reference biography for the diagnosis criteria for brucellosis in human should be updated to 2018 edition.
8.	Line 92-96, please use the correct ref about the 2018 edition.
9.	Line 99-103, it is suggested to integrate clinical signs and laboratory procedures into the case definition.
10.	Line 110-110, Odds ratio (OR) should be added into the section of data analysis.
11.	Line 112, regarding multivariate logistic regression model, please describe how to evaluate the prediction accuracy of the model. Besides, it is suggested to include population source as a covariate to determine whether the active source is more likely to detect HB infection than the passive source.

Results
12.	Line 131, clinical manifestations should be written in the plural.
13.	Line 132, P values should be in italics.
14.	Line 139, with the lower levels of education
15.	Line 157-168, the section of factors associated with the risk of HB could be improved with more details after reanalysis.

Discussion
16.	Line 172-173, please refer to latest publications in China.
17.	Line 194-196, “The participants from hospitals had more similar clinical symptoms and signs but with less contact history of sheep or cow, which may reveal a higher misdiagnosis rate of HB based on the clinical manifestation of HB.” The points are confusing and the sentence has grammar mistakes.
18.	Line 197-198, The points are confusing and the sentence has grammar mistakes.
19.	Line 220, the first letter of Brucella should be capitalized, and the word should be italicized.
20.	Line 223-225, “For passive detection, strengthen the monitoring on the population with a clear history of occupational exposure, especially the co-presenting of hyperhidrosis.” – this sentence is grammatically flawed.
21.	Line 241-242, “Third, we did not perform the culture of the Brucellosis to give a gold standard result, which may have a certain bias on the detection rate.” –“Brucellosis” should be replaced with “Brucella spp.”; “detection rate” should be “prevalence”.

Acknowledgment
22.	Line 259-260, the font size is not consistent.

Table 1
23.	The statistical analysis methods on different types of variables should be described in the footnote.

Figure 1
24.	The first letter of Brucella should be capitalized, and the word should be italicized.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EVALUATING ACTIVE VERSUS PASSIVE SOURCES OF HUMAN BRUCELLOSIS IN JINING CITY, CHINA
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: The grouping method is not scientific
Validity of the findings: I don't think the results are consistent with reality.
Additional comments: There were more clinical symptoms in the active detection group than in the passive detection group, and the positive rate of brucellosis was higher than that in the passive detection group. I think these are not consistent with reality. In addition, the method section states that the diagnostic criteria are 2018 (Line 91), while the reference is 2012?


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ACCURACY OF TRIGGERING RECEPTOR EXPRESSED ON MYELOID CELLS 1 IN DIAGNOSIS AND PROGNOSIS OF ACUTE MYOCARDIAL INFARCTION: A PROSPECTIVE COHORT STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: 1. I recommend changing the title of the study be like:
"Accuracy of Triggering Receptor Expressed on Myeloid Cells 1 in Diagnosis and Prognosis of Acute Myocardial Infarction: Retrospective Cohort Study"
2. The paper requires language editing.
3. Introduction:
	Line number 45, using the "MACEs" abbreviation is not acceptable, the abbreviation should be defined previously.
	Line number 46, using the "MI" abbreviation is not acceptable, the abbreviation should be defined previously.
	The gab of knowledge is not clearly outlined in the introduction; the authors should explain the gab clearly.
Experimental design: 4. I suggest the method section be structured based on Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Statement: https://www.strobe-statement.org/index.php?id=available-checklists
5. From line number 77 to 81, the distribution of patient and healthy persons in groups "GSE66360, GSE61144, and GSE60993" is not clearly identified.
6. Actually which statistical software is used SPSS or R studio? I'm confused that authors say in the abstract they used R software and in the Statistical analysis part of the Method Authors say they used SPSS.
7. Authors should regulate any online tools they used in the method section clearly.
Validity of the findings: 8. Table 2, 3, and 4: the authors should define all the abbreviations mentioned in any of the tables below them.
9. Table 1, 2, 3, and 4: the authors should define all the units of measurement.
10. Providing more interpretation of results in the discussion section with more evidence in comparing your results with the previous studies.
11. Providing of the limitations, strength, and recommendations sections.
12. I suggest the result section be structured based on Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Statement: https://www.strobe-statement.org/index.php?id=available-checklists
Additional comments: The paper requires language editing.
Small sample size.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ACCURACY OF TRIGGERING RECEPTOR EXPRESSED ON MYELOID CELLS 1 IN DIAGNOSIS AND PROGNOSIS OF ACUTE MYOCARDIAL INFARCTION: A PROSPECTIVE COHORT STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Authors of the paper named “Diagnosis and prognosis value of Triggering Receptor Expressed on Myeloid Cells 1 in patients with acute myocardial infarction” aimed to examine whether this protein can be used as a novel biomarker of AMI. Using the bioinformatics, authors have examine differentially expressed genes between healthy control individuals and AMI patients. TREM-1 was singled out as a significantly upregulated gene in AMI patients. To confirm results obtained analysing GEO, authors have measured TREM-1 levels in plasma samples of 147 individuals (35 healthy controls and 112 AMI patients), obtained 4h after admission. Plasma levels of TREM-1 were significantly upregulated in AMI patients. Upregulation of TREM-1 was also corelated with the severity of CAD as the triple vessel group had noticeably higher levels than single vessel patients. At the same time hyperglycaemia seemed    not to have a significant impact on TREM-1 expression. Finally, Cox regression analysis of the patients involved singled out TREM-1 next to Hb and IVS as a factor that strongly affects 6-month outcome in AMI patients.
Experimental design: General comment:
Scientific question is well formulated. It is methodically and logically well formulated paper, easy to follow and with a clear message. The experimental design is suitable to address the raised question.
Validity of the findings: Impression of this reviewer is that some of the conclusions are not fully supported with the data
Additional comments: Major comment:
Authors have concluded that TREM-1 can be indeed used as biomarker. As it is written in the text of the manuscript, TREM-1 is expressed on myeloid cells (neutrophils, monocytes, macrophages). These cells are the main responders to the cardiac ischaemic injury. Number of invading neutrophils and monocytes during the inflammatory phase directly corelates with the tissue damage. From the presented data, it seems that that TREM-1 is actually a marker of neutrophil/monocyte recruitment. Any state that elevates number of these cells will also elevate levels of plasma TREM-1. Therefore, can we really say that TREM-1 is an AMI biomarker??? It would be helpful if the authors would show TREM-1 levels in the later time points.
Additionally, it is not clear why authors have chose TREM-1 at the same time ignoring the S100A12 when this gene was also strongly upregulated in AMI patients?
This reviewer finds a bit strange that other S100 proteins didn’t come up in bioinformatics data, namely S100A8 and S100A9. Can you comment on this?
Minor comment:
1.	Is TREM-1 corelated with neutrophils and monocytes numbers in the blood?
2.	Can you please provide NLR (neutrophil-lymphocyte ratio) upon admission and how this number corelates with TREM-1 plasma levels?
3.	Introduction, line 66 “still remains further” please change into “still requires further”
4.	Introduction, line 69, please rephrase the sentence.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ACCURACY OF TRIGGERING RECEPTOR EXPRESSED ON MYELOID CELLS 1 IN DIAGNOSIS AND PROGNOSIS OF ACUTE MYOCARDIAL INFARCTION: A PROSPECTIVE COHORT STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: 'no comment'
Experimental design: 'no comment'
Validity of the findings: 'no comment'
Additional comments: The article is acceptable for me.
Thanks for the respectful following of the reviewers' comments.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SIMILAR SEED DISPERSAL SYSTEMS BY LOCAL FRUGIVOROUS BIRDS IN NATIVE AND ALIEN PLANT SPECIES IN A COASTAL SEAWALL FOREST
Review round: 1
Reviewer: 1

Output:
Basic reporting: In the present manuscript, Liu et al. compared the bird-seed dispersal of an alien (Phytolacca americana) and a native plant species (Cayratia japonica), two species that produce fleshy fruits with similar morphological traits and on the same season. These fruits were promptly (i.e., as soon as they mature) consumed by 11 bird species. The study concludes that the alien species has no constraints in its seed dispersal by native frugivorous birds, which promotes its establishment and expansion through disturbed microhabitats (e.g., forest clearings) at the study area.
•	I find it an interesting study; however, it needs a thorough revision in the English language so that the text can be clearly understood. I suggest corrections for some examples in the section “General comments”.
•	The introduction seems to be overall well structured, but I think that there are sections that need a bit of more work, especially between lines 63-75. There are poorly constructed sentences here (lines 63-64 and 74-75) that renders their message hard to convey to the reader.
•	The manuscript’s structure conforms to PeerJ standards. The raw data and the permits for the field work were supplied.
•	There is a mismatch in the number of figures cited in the main text and at the end of the manuscript (4 vs. 7).
•	There is also some missing information regarding the meaning of the values presented in both tables and in the barplots (please see my comments in the section “General comments” for more details).
Experimental design: •	This research lies within the scope of the PeerJ journal.
•	The research question is present at the end of the introduction section. However, it is buried within an extremely long sentence (lines 85-90), which makes it to be somewhat difficult to understand. I suggest the authors to rewrite this long sentence in a clearer manner and split it into smaller sentences.
•	In the methodology section, there are multiple missing details regarding the experimental design that must be provided (lines 118, 120-125, 130-131).
Validity of the findings: •	The results of the study seem reasonably interpreted and the conclusions related to the research question. However, it is difficult to critically assess the results without a more detailed description of the sampling design (see my comments in the next section).
Additional comments: Line 26: delete the word “much”. It has a subjective meaning.

Line 27: “…which were attracting the visitation of local birds.”. I don’t see this statement anywhere in the results and discussion sections. Moreover, it was not tested. Please, delete it or develop/justify this idea in the manuscript.

Line 30: At the beginning of the sentence, replace “11” by “Eleven”.

Line 35: Replace “larger amount of the fruits” by “higher number of fruits”.

Line 43: Replace “Seed dispersal was” by “Seed dispersal is”. Here and throughout the manuscript, especially in the introduction section and in the description of the study site, the authors should use the present tense when presenting facts.

Line 63-64: It is not easy to understand what the authors are trying to say in this sentence.

Lines 85-90: The sentence is too long. Consider splitting it.

Line 86: Why did you assess the seed dispersal of only one native species (i.e., Cayratia japonica)? I read in lines 113-114 that there were more fleshy-fruited plant species at the study site. Was it because they were scarce? Because they produce fruits that are morphologically different to those of Phytolacca americana and you only wanted to test fruits with similar morphological traits?

Line 110: It is enough to state only the months when migratory birds pass through the study site. Adding also the season is redundant, so you may delete it.

Line 118: You inspected plants every 50 metres in transects or randomly? How many plants of each species did you sampled?

Line 119: This is the first mention to these variables: “fruit characteristics”, “fruit height”, and “microhabitat”. Therefore, you must state what they stand for. For example, what is the “fruit height” variable? Maximum fruit height on each plant? Average fruit height on each plant? Height of each measured fruit?

Line 120: How many fruits per individual plant were measured?

Line 121: How was the plant density measured? Within random plots? Which area?

Lines 123-124: The number of ripe and missing fruits were assessed in the whole plant or sampled in specific branches? And it was assessed in how many individual plants?

Line 125: What was the sampling design to find those seedlings? Was it within plots around selected individual plants? If that was the case, what was the plot area?

Lines 130-131: How many individual focal plants were sampled?

Line 136: I suggest replacing “feeding frequency number” by “feeding visits” or other straightforward term.

Line 137: Based on Table 2, I presume that the “feeding duration” is the number of seconds that each bird spends on each individual plant. Therefore, I suggest changing the text segment “feeding duration per time” to “feeding duration (in seconds)”.

Line 153: What kind of generalized linear model? The authors should detail what kind of model was used.

Lines 155-156: Apart from “first stopping distance”, the meaning of these variables is not very clear (see my comments above). What is the “feeding amount per time”?

Line 175: Replace “As time going on” by another phrasing. For example, “As the time progresses”.

Line 180: It is insufficient to present only the p values. Please, provide the coefficient estimate and standard error, statistical test, degrees of freedom, and p- value. These parameters should also be presented separately for the two models (one for the alien and another for the native plant species).

Line 182: Replace “11” by “Eleven”.

Lines 194-195: Change “was the species whose first stopping distance longest after” to “was the species with the farthest fist stopping distance after”.

Lines 196-197: Please, present the results of the Mann-Whitney test in the same way it is in line 171. Additionally, for consistency, both the t-value and degrees of freedom should be reported for all t-tests.

Line 202: Replace “300” by “Three-hundred”.

Line 205: Delete “much”. It is a subjective term. Alternatively, you may consider replacing it by “significantly”.

Line 210: Move this sentence to the methods section.

Lines 216-217: Since you found significant differences in the ANOVA tests, why not use post-hoc tests to assess which treatment(s) is/ are leading to such differences? The authors state that seed ingestion by birds promoted seed germination, which is supported by the results. But this benefit might not be significantly different from the hand-cleaned treatment.

Line 249: Replace “7” by “Seven”.

Line 253: There is a mistake in the way these references are presented.

Lines 262-263: The authors found no “(…) obvious “preference” in the selection and utilization of the seeds (…)”. It seems to me that this lack of selection was found because all bird species were analysed as a whole, which might mask the variability in the response of each species. When I look at the data provided in Table 2, I see that most bird species display substantial differences in “feeding frequency” and/or “feeding time” between plant species. This suggests that, although most birds visit both plant species, they still have “preferences” toward either the alien or the native plant. Of course, a formal analysis would be needed to test this assertion.

Line 302: “itis” to “it is”.

Line 303: “nativeone” to “native one”.

Lines 303-305: Cumbersome sentence. Consider rewriting it.

Table 1: What do these values represent? Are they mean ± standard deviation?

Table 2: Are these values representing means? What is the sample size? A value representing the variability in measurements (i.e., standard deviation) should also be included.

Figures: The number of figures cited in the text do not match those found at the end of the manuscript (4 vs. 7 figures, respectively). This is because the plots belonging to the same figure are separated in distinct figures, which is odd.

Figures 4 and 5 (at the end of the manuscript): What do the error bars represent? This must be indicated in the figure caption.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SIMILAR SEED DISPERSAL SYSTEMS BY LOCAL FRUGIVOROUS BIRDS IN NATIVE AND ALIEN PLANT SPECIES IN A COASTAL SEAWALL FOREST
Review round: 1
Reviewer: 2

Output:
Basic reporting: I am attaching the original .pdf document with the corresponding annotations and suggestions that make up most of my review.

The question addressed in this work is interesting and important for me to answer when it comes to understanding how are the dispersal processes by frugivorous animals of invasive species in community and facilitation contexts. In general terms, the work is self-contained and both the data collected and the corresponding analyzes are necessary to address the question posed.

I believe that the level of English must be significantly improved for publication, there are many sentences that must be rewritten and many others shortened.

The general structure seems correct to me, although it is necessary to review the redundant information and delve more deeply into some points of greater interest and scope. Tables and figures are correct, in the .pdf notes I suggest several modifications that I think can improve the figures and tables qualitatively. There is a serious error in the code of the figures that is carried over almost the entire manuscript!

In the discussion, I miss a deeper thinking on the interpretation of the results obtained. Address issues such as the effect of this loss of dispersal events for native species caused by “disperser theft” caused by an invasive species. Also, some examples with other mutualistic or antagonistic interactions (for example, it is common to study the effects of invasive species in pollination or parasitism systems). This type of cases could be useful to understand the role that similarity-dissimilarity of traits has in the adaptation of invasive species in a context of biological interactions.
Experimental design: As in the previous section, specific suggestions and recommendations can be found in the attached document.

I have liked reading the experimental design, although there are several points that should be explained more specifically. It is an ambitious design as it covers quite a few important points in a dispersion system, which is highly valuable.

Practically all the samples are poorly explained in the text (crop size estimations, seedling count, methodology of focal observations, selection and justification of study sites, etc.). I believe that it is necessary to be more specific when explaining how each of these samplings has been carried out and why it has been in each specific way.
The number of fruits used to characterize both species seems little to me and it would be good to be able to measure more fruits and explain how they have been collected.

Clearly detailing the procedures for these types of measures significantly improves attributes such as replicability or repeatability in field studies. If it is a problem of space in the text, I suggest detailing these procedures in Supplementary Material.
Validity of the findings: NO COMENT
Additional comments: I would like to congratulate you. The question addressed seems interesting to me and the way to approach it is correct. I miss a clearer definition of the starting hypotheses and a more in-depth development of the discussion, paying attention to the consequences and natural processes that these results suggest.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SIMILAR SEED DISPERSAL SYSTEMS BY LOCAL FRUGIVOROUS BIRDS IN NATIVE AND ALIEN PLANT SPECIES IN A COASTAL SEAWALL FOREST
Review round: 1
Reviewer: 3

Output:
Basic reporting: The authors explore the hypothesis that the invasive plant Phytolacca americana being similar in fruiting to the native plant Caryatia japonica can take advantage of a pre-existing frugivorous assemblage and so have a successful population establishment.

The idea and objective of the study is very interesting and original. The objective of the study is clear and justified. Also, there is few literature on invasive plants’ seed dispersal, so this study is a valuable contribution.

However, there are important issues that need the attention of the authors to allow a better understanding.

English language needs to be thoroughly revised throughout the whole manuscript. The current state of the manuscript makes reading comprehension difficult and slow. Verbal tenses are often mistakenly used (e.g. the use of past for present events), and there are many parts of the text that need rephrasing for a better understanding. I have corrected the English in the abstract and introduction section (see pdf attached) to give an idea to the authors of the necessary corrections. I am not a native speaker so some of my corrections may not be appropriate. I suggest the authors ask for the help of an English native speaker, or someone with an advanced level of it.

The manuscript would also benefit from some additional references on the topic of invasive fleshy-fruited plant species dispersal and seed dispersal in general (e.g: Traveset, A., & Richardson, D. M. (2014). Mutualistic Interactions and Biological Invasions. Annual Review of Ecology, Evolution, and Systematics, 45(1), 89–113.)

The structure is clear, and the article is well self-contained. Figures and table need a revision, more explanation in their captions, as well as more information the material and methods section (see detailed comments in General comments).
Experimental design: The idea of the study is well defined, and it fills a knowledge gap on invasive plants seed dispersal.

The general hypothesis could be complemented with minor ones, to better support why the specific experiments have been settled. Also, some of the recorded data is not used in the discussion (such as bird feeding behaviour or stopping distance). I believe this data can be very useful to discuss the legitimacy of the different species as dispersers (see in General comments).

Materials and methods section needs more developing. It is not clear how many study sites there were. Also, some experiments are not clear or are not fully supported by a previous hypothesis (such as the seedling growth by microhabitat or the birds stopping distance).

Authors should also include the number of individual plants studied for each experiment. From the metadata I can guess 5 individual plants were marked for the calculation of removal rates, but it is not clear how many individuals were used to calculate bird visiting/feeding rates and how many plants were used to measure the fruits characteristics and fruit height. The choice of different statistical tests would also benefit from additional explanation.
Validity of the findings: All underlying data but plants density have been provided in the metadata. The use of some statistical tests could be better clarified, but all test values are duly reported.

I have one point regarding the conclusions and discussion. Seed dispersal is not explicitly measured in this study, so I advise authors to review all those parts in the manuscript where they talk about it. While several species are acting as dispersers (proven by the faeces you germinated), the bird identity of these faeces is not made explicit. Some of the frugivore species that you documented may be acting as pulp thieves (peckers) or seeds predators (e.g. Eophona migratoria; Shi, Y., Li, X., & Guo, Z. (2012). Tree fruits consumed by Eophona migratoria in winter. Journal of Biology, 29(3), 20-23).

Please be aware of this when taking conclusions or speculating.

In addition, I have some more specific comments that the authors should address. They are detailed in “General comments” sections by lines to facilitate the revision process.

Kind regards,

Elena Quintero
Additional comments: Abstract

Line 23: In the biological cycle of a plant, germination comes first to seedling grow. Therefore, I recommend stating before results on seed germination and latter results on seedling growth based on microhabitats.
Since differential seed deposition is not measured, I wouldn’t consider seedling establishment as a proxy. There are many survival factors that can differentiate proportions of seed deposition and seedling establishment.

Line 35: when you say “carried by birds” you only consider consumed/transported fruits right? I imagine you not accounting for birds pecking on fruits. Bird feeding behaviour is not very much discussed in the manuscript. I think you can use this behaviour data to discuss on the role of birds as legitimate disperser.

Line 36: Careful you did not measure seed dispersal per se, to do that you would need to prove that bird consumers actually act as dispersers (you only did this during the germination experiment with unidentified bird feces). Consider changing “seed dispersal” for “fruit consumption”.

Line 37: “preferred” is a bit confusing. I suggest you use other term as “grow more abundant in”.

Line 38: Start phrase with: “Our results support that...”. No need to use “could” in this phrase, if you have actually proved that seed germination is better with bird ingested seeds, you can speak safely.

Introduction

Most of this section is written in past tense, when events are presently occurring. Please check all verbal tenses throughout the text and change it to present when necessary.

Line 47-49: link both sentence by a comma instead of a point for better comprehension.

Line 50-51: this is a big claim, do you mean “one of the most important processes”? Seed dispersal while being very important process is not the only important one.

Line 55-57: Why does this process need to be rapid? Please provide a reference if leaving this phrase.

Line 57-60: link both sentence by a comma instead of a point for better comprehension.

Line 70-74: this is in contraposition to what you say in line 58, where it only mentions “local” birds. Maybe change to “frugivores present in the local area”.

Line 74-75: Please rephrase this sentence, since it is hard to follow.

Line 78: what do you mean by “artificial” trees?

Line 81: Perhaps you have confused Li et al 2019 reference, with this one:
Li, N., Yang, W., Fang, S., Li, X., Liu, Z., Leng, X., & An, S. (2017). Dispersal of invasive Phytolacca americana seeds by birds in an urban garden in China. Integrative Zoology, 12(1), 26-31. The reference you citate does not study Phytolacca americana.

Line 82: please change feed by consumed.

Line 85: please change dominate by dominant

Line 89: seedlings do not “select” microhabitat purposely, maybe choose a different term that applies better like “thrive”, “grow”,…

Materials & Methods

It may be interesting to mention somewhere that Cayratia japonica is a climbing plant, to better understand its seed dispersal and seedling growth.

Line 108-109: I suggest you write common names with small letters, and Latin names between parenthesis.

Line 119: What do you mean by fruit height? Is it the mean height at which the fruits are located in the plant? Please clarify.

Line 121-123: this is the first notice for the reader of a second study site. How many sites did you study? If there were two study areas (at least for some of the experiments), you should also include it in the materials & methods section.

Line 121-123: these results do not have supporting numbers anywhere or statistical significance. Can you please provide it?

Line 123-124: I don’t follow this experiment. How did you exactly measure the ripening rate? Did you always referenced ripen fruits to the original crop size or did you calculate the percentage based on the remaining fruits? I find confusing the biological meaning of this relationship.

Line 126: why did you only selected two microhabitats? Are these two the most important ones or clearly distinguished? Maybe you had a prior hypothesis based in these 2 microhabitats? Please explain the reader why you took this decision.

Line 130: How many observation hours did you do and how many plants did you observe?

Line 133: As I understood, the focal “subject” in this case is the plant, not the bird. I suggest you remove “focal” in this phrase to avoid confusion. Maybe you mean something like “birds that visited the plant were observed until they were out of sight”?

Line 134:  this information is not present in the results neither in the metadata. Behaviour is divided in swallowed or pecked. Did you use ‘dropped fruits’ or ‘taken in flight’ for some experiment? If so, please justify it.

Line 141: How many seeds did you planted for each experimental group? From the metadata I see you used 20 bowls with 5 seeds each, is that correct? Please include it.

Line 143: Do you have the identity of the bird feces? Were they all from the same bird species?

Line 153: Could you please justify why you used a GLM and not a Linear model? was it because the data followed a non-liner distribution?

Line 156: depending on what did you use t-test or U-test? Please clarify

Results

Please check figure numbers and letters, they are mixed up (e.g. figure 1A and 1B = figure 1 and figure 2). Figure captions need more explanation, especially explain what axis represent and error bars.

Line 169: this is very interesting. Did any of the two plants had a longer fruiting period than the other?

Line 171-172: please provide numbers and statistical significance if you have measured this. You may also want to include this in table 1.

Line 175-176: Say explicitly Cayratia turns black and Phytolacca turns purple black for more clarity.

Line 177: no other animal was observed during your focal observations, but you did not monitored plants during the afternoon/night. Please state this only holds true for the time you monitored.

Line 180-181: I am not sure to understand this experiment. How do you make sure missing fruits are consumed by birds and not fallen?

Line 182, 202 and 249: when starting a sentence with a number it should be written it in letters, please adjust this.

Line 182-190: I suggest you shorten this section by stating only the names of those species feeding in one of the two plants. The rest (a total of 7) are common and previously mentioned in M&M and in fig. 3.

Line 194-195: why did you measure stopping distance? Was this a proxy for minimum dispersal distance. This is not explained in the introduction hypothesis.

Line 198: I suggest you change “feeding amount each time” by “number of fruits consumed per visit”.

Line 203: why seedlings are searched around mother trees? Weren’t they searched in microhabitats?

Line 210: move this sentence to Materials and methods section.

Line 212 and line 216: “controls” is confusing, I suggest you change it to “groups”, since the controls would be hand-cleaned seeds and intact fruits (not defecated seeds).

Discussion

Line 227, 288, 300 and 26: English correction: The term much similar in this article is often mistakenly used. “Much similar” seems to refer to the comparison of the 2 plants with another unknown system. I recommend you remove the “much” and leave just “similar” for all instances throughout the text.

Line 227: I recommend changing “indicates” by “suggests”, since you are speculating.

Line 236: can you provide a reference for the fact that Phytolacca contains more seeds? If you also calculated this, you should include it in the M&M and results sections.

Line 237: Seed dispersal effectiveness is not only based in consumption frequency but also in quality factors such as gut treatment, microhabitat deposition or seed predation that are not accounted in this study (see Schupp 1993 Vegetatio https://doi.org/10.1007/978-94-011-1749-4_2.). I advise something more restrained as: Phytolacca can use these traits as an advantage over Cayratia improving the effectiveness of its dispersion.

Line 244: This is very interesting! you can link it to plant colonization processes in forest gaps (especially worrying for invasive species). See: Burnham, K. M., & Lee, T. D. (2010). Canopy gaps facilitate establishment, growth, and reproduction of invasive Frangula alnus in a Tsuga canadensis dominated forest. Biological Invasions, 12(6), 1509-1520.
Or
Martins, S. V., Júnior, R. C., Rodrigues, R. R., & Gandolfi, S. (2004). Colonization of gaps produced by death of bamboo clumps in a semideciduous mesophytic forest in south-eastern Brazil. Plant Ecology, 172(1), 121-131.

Line 251-252: since you have recorded bird feeding behaviour, I suggest you use this information to discuss the potential of species as dispersers (i.e. the 4 species of peckers probably are not dispersing much), and the implications.

Line 261-262: these are already mentioned results and do not need to be repeated unless they are followed by an explanation.

Line 276-279: The seed germination Inhibition hypothesis you mention holds true for many fruits species, not only invasive ones (see: Cipollini, M. L., & Levey, D. J. (1997). Secondary Metabolites Of Fleshy Vertebrate‐Dispersed Fruits: Adaptive Hypotheses And Implications For Seed Dispersal. The American Naturalist, 150(3), 346–372. https://doi.org/10.1086/286069)

Line 284-285: Is not possible to generalize for all species. Some may be seed predators/pulp peckers. Since you have not identified the identity of the bird’s faeces, you should indicate so.

Conclusion

Line 301: I suggest you replace the world “system” by “mutualistic relation”

Line 302 and 303: need space between words “it is” and “nativeone”

Figures

For a better understanding of figure 1 I think it would help plotting fruit removal along *time* for both species, so it shows depletion rates. See for example figure 1 in Cullen, A. C., Farley, K. E., Pagano, S. S., Gallagher, F. J., & Holzapfel, C. (2020). All in the timing: How fruit nutritional content influences the timing of fruit consumption of two invasive shrubs. Plant Ecology, 221(10), 951–963. Yet, I am aware this may not be in the scope of the authors, so just consider it a suggestion.

Figure 3. the interaction links indicates the number of visits or the number fruits consumed? You indicate links are ‘relative’, do you mean relative to plants or to what?
Also, I propose you add a grey colour to the links, to facilitate interpretation, otherwise with too many black lines are a bit messy.

Figure 4. I suppose y-axis is a mean number since it has error bars. How is it calculated? (e.g. per site, per m2…)

Figure 6. y-axis shows *accumulated* seed germination


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SIMILAR SEED DISPERSAL SYSTEMS BY LOCAL FRUGIVOROUS BIRDS IN NATIVE AND ALIEN PLANT SPECIES IN A COASTAL SEAWALL FOREST
Review round: 2
Reviewer: 1

Output:
Basic reporting: The English language was considerably improved in the current version of the manuscript.  I would like to congratulate the authors for the good work. The figures are now corrected but there are some typos in the captions (for more details, please see my general comments below).
	Although the manuscript was notoriously improved (especially in grammar and the provided methodology), I still have a few comments that are detailed under “General comments”.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Lines 31-34: This sentence needs to be rewritten (at least lines 31-32; consider dividing it in two sentences) because it is hard to understand. I suggest the authors to rewrite to something such as “There were no significant differences in bird feeding frequency, feeding duration, and first stop distance between the alien and the native plant species. These similar results were shown across …”.

Lines 50-51: “Birds are the most common seed disperser in the world…” Unless you have a reference to support this statement, I suggest rephrasing to “Birds are among the most common seed dispersers worldwide”.

Line 51: Change “meditated” by “mediated”.

Line 62: “different seed dispersal systems” still seems a bit vague to me. Given the references presented afterwards, it seems to me that rephrasing to “Most alien plant species have different traits when compared to …” would fit better and have a more straightforward meaning.

Line 75: State that the native plant species also produces fleshy fruits.

Line 77: Because it starts a sentence, do not abbreviate the genus. Write “Phytolacca americana”.

Line 116: Do not use the word “trees” to refer to the individual plants because they are not trees. Instead, change here and where applicable in the manuscript (lines 119, 120, 124, 174, 175) to, e.g., “20 sampled individuals from each species” or “20 sampled focal plants from each species”.

Line 116: Write the units in which you measured fruit size.

Line 118: In that case, don’t you think that it would make more sense to rename the variable “fruit’s heigh” to “maximum fruit’s height”?

Lines 160-161: The use of post-hoc tests (and which one was used) after ANOVA should be stated here.

Line 162: Please, provide a reference for R software and state that it was used to make the plots.

Line 170: Because it is starting a sentence, the generic epithet in Phytolacca americana should not be abbreviated.

Lines 181-182: Please, provide the regression estimated parameters (i.e., the beta coefficient).

Line 191: For consistency, throughout the manuscript, always abbreviate the generic epithet of the scientific names except when the name is written for the first time or when it is starting a sentence.

Line 236: There is a typo here. Change “Americana” to “americana”.

Lines 255-256: Cumbersome sentence. For example, the part where it is mentioned that those are pecking birds should come at the beginning of the sentence, before saying that pecking may have damaged the seeds.

Line 256-258: Why were they considered seed dispersers? I’m assuming that it is because they ingest the whole fruit, but this information must be stated here.

Line 268: It would be easier to understand this sentence if the authors would replace “feed amounts” by, for example, “number of consumed fruits”.

Line 304: “in autumn in a coastal seawall forest”. This part of the sentence doesn’t add any important information here because the fruits of both plant species are similar irrespective of the season and habitat (right?). Therefore, I suggest the authors to delete this text segment.

Table 2: In the caption, the first sentence doesn’t look right. I suggest the authors to replace it by, e.g., “Bird feeding behaviour on the native and alien plant species.”

Table 2: As you did in the main text, I suggest the authors to replace the name of the variable “feeding amount” by another name that makes it easy to understand what was really measured here.

Figure 1: In the caption, there is a typo. Change “analysi” to “analysis” and delete “(A) Phytolacca americana”.

Figure 2: There is a mistake in the caption.

Figure 3: Replace “Accumulate” by “Cumulative”. Also delete “which planted averagely in 20 bowls.”.

Figure 4: Replace “30” by “Thirty” because it is starting a sentence.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SIMILAR SEED DISPERSAL SYSTEMS BY LOCAL FRUGIVOROUS BIRDS IN NATIVE AND ALIEN PLANT SPECIES IN A COASTAL SEAWALL FOREST
Review round: 2
Reviewer: 2

Output:
Basic reporting: The authors have substantially improved the quality of the manuscript. The improvement in writing and the level of English is evident. The organization and design of the figures and tables has also improved.

In the general comments section some suggestions and necessary changes are detailed, indicating the specific line in each case.
Experimental design: A detailed description of the ratios for ripen fruits and missing fruits is still missing.
Some other suggestions on experimental design are noted in the general comments section.
Validity of the findings: NO COMMENT
Additional comments: Lines 31 -34:  The wording of this sentence makes it difficult for me to understand, even though the result is simple and robust. It could perhaps be said: “Similar results were shown across alien and native species in bird foraging behavior (feeding frequency, feeding duration and first stop distance) indicating that a similar seed dispersal relationship had been established between local frugivorous birds and both plant species.

Lines 83-87: At the end of the introduction, perhaps before this paragraph of objectives, it might be good to briefly develop the hypothesis or starting point, what facilitation or competition processes can be found and what consequences they could have. I consider that an explanation of this would improve the quality of the manuscript as it would help the reader to understand why it may be necessary to compare these seed dispersal systems.

Line 109: Confusing. Perhaps “small number of other fleshy fruited plant species such as Lycium..”

Line 114: Change “..we randomly inspected..” for “ we randomly chose 20 focal trees from each target species along the coastal..”

Line 118: Fruit´s height is confusing for me, in Table 1 for Cayratia japonica the average Fruit height is 0.2 (m). How have you measured this trait? Did you measure the distance of the tallest fruits to the ground?

Lines 119-121: Despite the McDonnell reference, it is not clear to me how these ratios are calculated. If values of 100% are reached, I suppose that everything refers to an initial available cropsize. Is that correct? How are the missing fruits calculated? Is it a subtraction or have you based on signals in the Infructescence? These results are key to affirming that there is no limitation in the dispersal of any of the target species, so it is necessary to define well how they have been calculated.

Line 130: “20 focal stands”. In each focal stand could inhabit more than one individual plant. There are stands with both species co-ocurring or are species-specific stands?  It is confusing because the incongruence between "focal individual sampling" and "20 focal stands". Be more specific please.

Lines 179-180: Perhaps an example of how these values have been measured in the field, which was measured, and how these ratios have been calculated, can be useful to understand and suggest clarifications to improve the explanation of these results.

Line 212: The selection of the bar.plot of fig. 3 leads to a misunderstanding. What is being compared in the t-tests are the microhabitat preferences within each species. I think the best thing to do would be to represent the seedlings found in each microhabitat of one species next to each other, and in the other part of the graph the other species. Substitute the order of place (Forest gap and understory) for the species, and use the colors for the Microhabitats. It is only a suggestion and there are many possibilities, but what is being compared in the statistical test does not agree with the supporting figure.

Line 213: “We randomly..” In M&M section you explain: Seedlings were found within plots (r=25m, n=30) around the individual selected plants. Maybe just write: "We found 300 P. americana seedlings.."


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: SIMILAR SEED DISPERSAL SYSTEMS BY LOCAL FRUGIVOROUS BIRDS IN NATIVE AND ALIEN PLANT SPECIES IN A COASTAL SEAWALL FOREST
Review round: 2
Reviewer: 3

Output:
Basic reporting: English language has greatly improve in this new version. The manuscript has a good and clear structure as well as a clearly defined goal. Supporting literature is enough and most data is provided.
Experimental design: The research question is very well supported (i.e. the comparison of dispersal systems for the species). Authors could increase the justification and significance of the manuscript by briefly adding a general hypothesis at the end of the introduction (e.g. invasive plant taking advantage of an already established dispersal system of a native fruiting plant and the implications this may have).

Materials and methods are better explained in this new version. Yet I am still missing a few more details on the section “Fruit and seedling investigations”. It is not fully clear to me how authors performed some experiment (see general comments below).

Table 2 has greatly improved and the merging of figures 1 and 4 looks much better, allowing for comparisons between the native and the invasive species.

Overall, I congratulate the authors for the great improvement of the manuscript and after addressing the issues raised in here, I think it will be ready for publication.
Validity of the findings: No comment
Additional comments: Abstract

Line 17-18: Local birds do not disperse plants with that purpose (establish populations), that is rather the purpose of the plants.

Line 25: I suggest writing the words “types”, “colors” and “periods” in singular.

Lines 31-34: very long phrase, please consider splitting in two. Also significance values are not in context since you are not stating comparisons in the sentence.

Introduction

Line 55: there is a red dot, change to black font.

Lines 62-63: What you say here is in contraposition to what you say in lines 71-73. From what I understand from the text I suppose you mean that, based on the literature available to date, it appears that the dispersal systems of invasive plants differ from those of native plants in the area they colonize. However, I am not certain that this is what the authors mean. Please review these two sentences.

Line 82: change the second ‘and’ for a ‘by’.

Material and Methods

Line 109: add species after ‘fruiting’

Line 116: how many fruits per plant did you take?

Lines 117-118: The ripening period length refers to the dates you have done field work or to the natural phenology of both plants? Please clarify.

Lines 119-121: If you did this experiment as in McDonell et al 1984, please reference it for further support. However McDonell et al. used bagged branches as reference to calculate percentage, but I think you did not. Could you please explain with further detail how you calculated ‘ripe and missing fruits percentage’? To what total number is this calculation referred, to the number of ripen fruits present in the previous revision maybe? Or were the percentages visually estimated?

Line 121: to what does the r in the parenthesis refers to? radius of the plots? Please clarify in the text.

Lines 121-122: As I get from the supplementary data you provide, for the seedling microhabitat experiment you selected 15 plots per species, of which half corresponded to forest gap and the other half to understory, is this right? Please develop a bit more the experiment description.

Line 125: These 20 plots are new to the reader (as far as I understood), therefore please introduce them as such. When you say “of the 20 plots” seems confusing, as if they have been mentioned previously.

Lines 133-134: is this information 'number of fruits per visit' present in the table 1?

Line 139: this reference is out of context here, I suggest deleting it.

Results

Lines 168-169 and lines 172-174: since you only have 2 species, please indicate which has bigger fruits and which is higher.

Lines 174-175: Please revise the English in this sentence. This is the first notice you distinguish between plant’s age (not stated in M&Ms), also in the Table 1 tree density is grouped by species (not divided by age). Did you perform any statistical analysis? Supplementary data for tree density is not provided.

Lines 189-197: Please stick to common or latin names for the birds species, but do not use them interchangeably.

Line 210: Did you test differences in germination between species? In figure 3 seems that Caryatia seedlings germinate in a higher proportion.

Line 213: randomly found? If I understood correctly an active search for seedlings was done in 20 plots (so, not random).

Discussion

Lines 255-256: it is not possible to tell how much each bird dispersed (since this was not actually measured), please use conditional when speculating. Maybe these birds did not even disperse at all.

Lines 256-258: These numbers do not coincide with the previous statements, are you considering all species as potential dispersers (including peckers)? Either change the sentence or move before lines 255-256.

Line 261-263 and 272: choose between using latin or common names for birds.

Line 269:  P. americana showed significant higher frugivory, and this may give it an advantage over C. japonica, however but you can’t prove it actually had more effective dispersal. This depends on the different birds feeding on it (some frugivores may be more effective than others, or some may not even disperse) the differential contribution between bird species is key for determining dispersal effectiveness.

Line 290-292: germination difference between species could actually be tested with you data, did you do perform any analysis?

Line 309: I suggest replacing “this coastal..” by “Dongchuan coastal…“

Tables and figures

Table 2: feeding frequency and feeding amount need more explanation. It is not clear if feeding frequency refers to number of fruits per visit, or total number of visits. Then feeding amount is not clear if refers to number of plant individuals visited or number of birds visiting at the same time.

Figure 2: some error in the figure legend, one sentence is duplicated.
Link colours are now blue and black and looks messy. I suggested filling the link with a light grey color in order to differentiate it from the background.
Also if enough space I suggest adding birds latin name abbreviations above the rectangles, so it would facilitate visualisation.

Figure 3. Accumulate needs a “d” in the end


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CLINICAL, SEROLOGICAL AND EPIDEMIOLOGICAL FEATURES OF HEPATITIS A IN LEÓN, NICARAGUA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The manuscript clearly described prevalence, clinical features and risk factors of acute hepatitis A in  León, Nicaragua during 2006-2010.
Experimental design: In this study cases enrolled are symtomatic cases presented to healthcare facility by advertise, some of asymtomatic/mild cases might not be included.
Validity of the findings: This study reveal the informative epidemiologic data of acute hepatitis A in  León, Nicaragua during 2006-2010.
Additional comments: Reviewer's report: The manuscript entitled "Clinical, serological and epidemiological features of hepatitis A in León, Nicaragua
” This study described prevalence, clinical features and risk factors of acute
hepatitis A in  León, Nicaragua during 2006-2010. The majority of cases are young population.  In the basic knowledge, HAV infection is mostly asymtomatic among young children, In this study cases enrolled are symtomatic cases presented to healthcare facility some of asymtomatic/mild cases might not be included. This study reveal the informative epidemiologic data of acute hepatitis A in  León, Nicaragua during 2006-2010.

Reviewer comments
1.Result part: Authors should add table of demographic data/clinical data of enrolled cases, classified by age group, this would be informative table. And result in clinical part and  biochemical lab should be summarized in the table.

Quality of written English: Acceptable

Declaration of competing interests: I declare that I have no competing interests.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CLINICAL, SEROLOGICAL AND EPIDEMIOLOGICAL FEATURES OF HEPATITIS A IN LEÓN, NICARAGUA
Review round: 1
Reviewer: 2

Output:
Basic reporting: The subject of this study is interesting for the high endemicity of hepatitis A infection, however, the presentation of the data needs to be revised to make its readability.
Experimental design: The study design could provide a flow chart to make it clear.
Validity of the findings: Figure 1 is published, this could be confusing its originality.
The tables are suggested to revise, then clearly presented in the main text.
Finally, the presentation of the validity of the findings should be supported by measurements.
Additional comments: 1. In the abstract, the " the aim of the study " please clarify. Is the statement of the aim of the study consistent with the title or main text?
2. In the study conduct, it is suggested to add a time flow chart of baseline survey and intervention.
3. Please provide the inclusion and exclusion criteria clearly.
4. Were the participants excluded including vaccination or past infection?
5. In the Results, figure 1, Does it present in the previous study? （Mayorga et al., 2016）Did it provide new information in this study?
6. In the Results,
As we can see from the raw data, it is suggested to revise table 2 or the main text. Given the total number of participants and the number, measurements are inconsistent with Table 2, the readers hardly understand the result in the main text and Table 2.
As we can see in Table 2, the trend of seroprevalence is increasing with age, even the small sample size in each aged group. It may be compared between the measurement group and the symptomatic group.
7. In the Results,
As we can see from the raw data, it is suggested to revise table 1, given it provides very little information.
It may be compared between the measurement group and symptomatic group including clinical features and laboratory findings.
8. In the Results, ‘’ Logistic regression analyses”, it is suggested to exclude missing data (symptomatic group), even past infection cases, or subclinical cases. This could be more exact to predict the risk factors.
9. In the Discusses, the authors emphasized the high endemicity in this area, it is suggested to explain by using the data of measurement group or by aging.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CLINICAL, SEROLOGICAL AND EPIDEMIOLOGICAL FEATURES OF HEPATITIS A IN LEÓN, NICARAGUA
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The article entitled " Clinical, serological and epidemiological features of hepatitis A in León, Nicaragua’’ has been reviewed again. After the revision, the manuscript could be present as the clinical, serological, and epidemiological features of hepatitis A.
However, the author’s response to query No.1. in the “abstract”, the " the aim of the study " is not described more clearly. “To monitor and document endemicity and disease burden of acute hepatitis A” mean “The clinical manifestations and seroprevalence of HAV” have been reported in the previous study or in this study.  The “an ongoing vaccine effectiveness study” is ongoing in the future or now in this study. Most studies may have one objective, the introduction reported three objectives. I felt confused. The statement of the aim of the study in the abstract and introduction was suggested to compatible with the title.
The study flow chart, presentation of the data, and the tables have been revised.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CLOSTRIDIUM BUTYRICUM RELIEVE THE VISCERAL HYPERSENSITIVITY IN MICE INDUCED BY CITROBACTER RODENTIUM INFECTION WITH CHRONIC STRESS
Review round: 1
Reviewer: 1

Output:
Basic reporting: See my comments below
Experimental design: - Original primary research within Aims and Scope of the journal.
N/A, this is an editorial decision from PeerJ.

See my comments below for the rest
Validity of the findings: See my comments below
Additional comments: The focus of this manuscript is to test the therapeutic property of a probiotic in a model of post-infectious Irritable Bowel Syndrome. Two main features of IBS are addressed in this study 1/hypersensitivity and 2/microbiome changes.
This topic is highly relevant in Gastroenterology.  PI-IBS is a seductive hypothesis and quite well studied in both basic and clinical aspects.  Unanswered questions in the field relate to understand precisely the short- and long-term effect on the microbiome (not only in terms of composition), and why (and how) such changes in microbiome back to 'normal' has some impact on host physiology after the infectious event (e.g. hypersensitivity, hyperexcitability of enteric neurons, unbalance proteolytic balance at mucosal surface). These outstanding questions in the field of PI-IBS are not particularly well addressed in the present paper.
Interestingly, authors focused on Clostridium butyricum, which is not one of the first probiotic that comes into (my) mind. Changes in microbiota taxonomy are somewhat expected and not very original over what is published in the field. Behavioral score for hypersensitivity is valid, but definitely not state of the art.

Overall and specific comments:
Language: The manuscript reads well, the EN language is good and I found no obvious typos. This is appreciated. I would not request any additional reviewing on that aspect.
Ethics: This study uses animals, specifically mouse model. Ethical aspects seem to be in place, with protocols and SOPs authorized by the host institution.
Data availability: I have been able to access to all 96 replicates from this study in PRJNA698375 project. Labeling of samples is explicit enough if any deeper investigation is required. At this step of reviewing, I have consulted but not reprocessed raw data.
- Antibiotics are not therapies recommended for PI-IBS (except at the stage when there is actual infection). I do not understand the clinical rationale of including such group in this paper.
- It is unclear which type of statistical test is used in each Figure. The small paragraph in the Method section is not enough informative.
- It is unclear the bioinformatic pipeline that has been used to perform 16S analysis. In particular Figure 6 panel B.
- This graphical abstract of in vivo studies is appreciated. But I do not understand this comment from the graphic "C.butyricum/antibiotics gavage continued for 1 weeks"
- Figure 2: I am not convinced there is a significant difference (* P<0.05) between C.rodentium only versus C.rodentium+C. butyricum (orange versus magenta?). To me a 2-way ANOVA would have been more appropriated to statistically analyze this type of dataset. How can the authors explain the small difference in response to 0.1ml injection? This small difference is likely to impact the response to other volume distention.
- I always considered C.rodentium infection to have a huge impact in microbiota composition and overall decrease in richness of OTUs. This is expected because microbial niche is vastly dominated by such enteropathogen (although this is usually temporary). This is not the case here. How the authors can explain such difference? Additional references to support this finding would be appreciated.
- I would recommend author to balance the view of C. butyrium as a strictly beneficial microbes. It has been demonstrated elsewhere that such stain can be considered as a pathogen. This is quite important to consider for clinical relevance of using C. butyrium as a human 'drug'
- C. rodentium is rather a good model for attaching/effacing type of E. coli pathovar such as enteropathogenic E.coli (EPEC) but not so much for EHEC/STEC pathovars O157H7.
- line 98: are you sure 5-6 weeks-old B6 mice weight in average 13-15 g?
- Are the necessary steps have been taken to reduce cage effect at the beginning of the experiment? Such as mixing the bedding between all cages?
- Reduction of OTU: In the text, it could be informative to remind the reader how much OTU have been recovered at the basal timepoint.
- Please remove any non-informative details on taxon ordering such as 'norank_f_' ,'_NK4A136_group' or g_norank_f_norank_o'


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CLOSTRIDIUM BUTYRICUM RELIEVE THE VISCERAL HYPERSENSITIVITY IN MICE INDUCED BY CITROBACTER RODENTIUM INFECTION WITH CHRONIC STRESS
Review round: 1
Reviewer: 2

Output:
Basic reporting: English language needs editing.
Intro & background to show context. Literature well referenced & relevant.
Structure conforms to PeerJ standards, discipline norm, or improved for clarity.
Figures are relevant, high quality, well labelled & described.
Experimental design: Original primary research within Scope of the journal. Research question well defined, relevant & meaningful. It is stated how the research fills an identified knowledge gap.
Rigorous investigation performed to a high technical & ethical standard. Methods described with sufficient detail & information to replicate.
Validity of the findings: Conclusions are well stated, linked to original research question & limited to supporting results.
Additional comments: Regarding manuscript entitled “Clostridium butyricum relieve the visceral hypersensitivity in mice induced by Citrobacter rodentium infection with chronic stress”, the authors aimed to examine whether the intervention of C. butyricum or antibiotics could affect the etiology of visceral hypersensitivity.
The study is interesting and well designed. However, minor revision is needed before its acceptance.
Add the aim of the study in the abstract at the end of background subsection.
Add clear hypothesis at the end of introduction before the aim of the study.
Clarify the method used for statistical analysis in each parameter (Student’s t-tests or one-way ANOVA).
Language editing is important before its acceptance.
Please avoid using “we”


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CLOSTRIDIUM BUTYRICUM RELIEVE THE VISCERAL HYPERSENSITIVITY IN MICE INDUCED BY CITROBACTER RODENTIUM INFECTION WITH CHRONIC STRESS
Review round: 2
Reviewer: 1

Output:
Basic reporting: See my final report below
Experimental design: See my final report below
Validity of the findings: See my final report below
Additional comments: Authors have revised the manuscript well according to my initial comments, with modifications highlighted in the text. As a minor comment, I am not much convinced by the rebuttal for Figure 2 statistics. That being said I will let the scientific community to make their own judgment on this particular data and interpretation.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CLOSTRIDIUM BUTYRICUM RELIEVE THE VISCERAL HYPERSENSITIVITY IN MICE INDUCED BY CITROBACTER RODENTIUM INFECTION WITH CHRONIC STRESS
Review round: 2
Reviewer: 2

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: .