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: NLRP3 INFLAMMASOME ACTIVATION MEDIATES SLEEP DEPRIVATION-INDUCED PYROPTOSIS IN MICE
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors investigated the mechanisms underlying sleep deprivation-(SD) induced neuroinflammation which have not been completely elucidated yet. For that purpose, microglia activation was investigated in a mouse model of SD and the stimulation of the NLRP3/caspase 1 inflammasome pathway was determined. Noteworthy, the authors show that SD-induced the NLRP3 inflammasome components and the release of cytokines typical of pyroptosis. Also, SD triggered an overexpression of GSDMD, a pore-forming protein associated to pyroptosis.
It would be interesting to determine whether pyroptosis occurs in microglia and whether different cell death programs induced by SD could occur in distinct cell types.

1. The work will benefit if the authors describe in more detail the structure of the NLRP3 inflammasome. In the introduction p. 5 lines 66-68 there is a mention to the NACHT domain of the platform protein NLRP3 but other protein domains in both NLRP3 and ASC and procaspase-1 contribute to the assembly of the NLRP3 inflammasome.

2. The results support that SD may induce NLRP3 inflammasome activation. Within this context I suggest the inclusion of the article “The NLRP3 inflammasome modulates sleep and NREM sleep delta power induced by spontaneous wakefulness, sleep deprivation and lipopolysaccharide” doi:10.1016/j.bbi.2017.01.012 in the references of the manuscript.

3. The number of animals used in each study should be stated in the legend of figures 2-4 as it is in figure 1.

4. In the Introduction, p. 5 line 73 please substitute P20 and P10 subunits by p20 and p10 subunits.

5. I suggest the Methods subsection of Hematoxylin and Eosin (HE) Staining should appear before the Immunohistochemistry (IHC) subsection since the description that the hippocampus CA1 region was immersed in 4% paraformaldehyde and then embedded in paraffin is indicated in the subsection of Hematoxylin and Eosin (HE) Staining.

6. The bar graphics regarding in vivo studies would be more informative if substituted by a type of graph were we could see the distribution of the values for each animal, such as an aligned dot plot.

7. The manuscript needs a thorough English Language review.
Experimental design: 1. The post-hoc test associated to the one-way analysis of variance (ANOVA) that was used in the statistical analysis of the results should be indicated in the methods section. Moreover, since Both ANOVA and Student’s t test assume data normality the authors should indicate whether the results pass a normality test (D’Agostino & Pearson omnibus normality test or the Shapiro-Wilk normality test). Otherwise the statistical analyses should be performed using suitable non-parametric tests like the Kruskall-Wallis test or the Mann-Whitney test.

2. It should be clarified in the manuscript whether the ELISA kits used for the detection of cytokines (figure 3), namely IL-1β and IL-18 detect the mature form of the cytokine or its pro-form. This is relevant regarding the conclusion that SD-induced NLRP3/caspase-1 pathway activation.
Validity of the findings: 1. Results section, page 9: The authors evaluate the effect of SD in neurodegeneration through the analysis of nuclear pyknosis and the TUNEL assay (Figure 1). It was observed that SD-induced nuclear pyknosis and apoptosis of neuronal cells. The SD protocol comprises a period of 6 hours sleep deprivation sustained for the 1-5 weeks.
1. 1. Which was the basis to select a period of 24 h as a recovery interval at the end of the 5th week of continued SD? Please indicate the rational in the manuscript.
1.2 A 24 h recovery period at the end of the SD protocol may inhibit SD-induced apoptosis in neuronal post-mitotic cells as concluded by the authors. Please give an explanation for this observation in the manuscript as well as for the decrease in the number of microglia.
1.3 The authors indicate that SD induces pyroptosis which is a cell death program distinct from apoptosis. Therefore the authors should better clarify with adequate markers whether SD-induced cell death regards apoptosis, pyroptosis or both mechanisms. Also, it would be interesting to understand whether SD-induced cell death in different cell types could be mediated by different mechanisms.

2. The authors investigated whether SD promoted the activation of NLRP3/caspase 1 pathway. As shown in Figure 2A, the NLRP3, Caspase-1, ASC as well as caspase-4 mRNA levels were significantly increased in SD group. Caspase-4 mediates non-canonical activation of the NLRP3 inflammasome (as shown in the article “Caspase-4 mediates non-canonical activation of the NLRP3 inflammasome in human myeloid cells” doi: 10.1002/eji.201545523) and GSDMD is a substrate of caspase-4. It would be interesting to discuss the possible contribution of caspase-4 for SD-induced pyroptosis.

3. In Figure 3 E regarding IHC for GSDMD level in mouse hippocampus CA1 region it would be helpful to indicate with an arrow the immunostaining for GSDMD. Indeed I am not sure whether GSDMD staining corresponds to the purple or the strong brown staining observed in the mice submitted to SD. Also, the color on the representative control image is very different even from the experimental condition SD+recovery, which according to the results is expected to be similar to control.

4. In the discussion, page 11 line 216, it is stated that SD induced the pyroptosis of neuronal cell. However the markers of pyroptosis were analysed in tissue homogenates which does not allow for the conclusion that pyroptosis occurred in neuronal cells. The only pyroptosis marker analysed by IHC was GSDMD (figure 3). The study of co-localization with a neuronal and/or microglia marker would be important to clarify this issue.
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: NLRP3 INFLAMMASOME ACTIVATION MEDIATES SLEEP DEPRIVATION-INDUCED PYROPTOSIS IN MICE
Review round: 1
Reviewer: 2

Output:
Basic reporting: 1.	Using a mouse model of sleep deprivation (SD) the authors showed SD-induced NLRP3 activation and pyroptosis. Although an interesting research topic, this is not novel. Recent published studies already demonstrated  that SD activates NLRP3 inflammasome activity in mouse neurons and astrocytes (Niznikiewicz et al., 2017; Smith et al., 2019; Zielinski et al., 2020) highlighting the potential of NLRP3 as a therapeutic target (Smith et al., 2019). These research works should be discussed. In this work, the authors suggest p38 MAPK signaling pathway mediates SD-induced activation of NLRP3 and pyroptosis. The authors should give more relevance to this result and better explore this mechanism in order to increase the novelty and the relevance of their work.

2.	The English language should be revised and improved throughout the article.  There are some incomplete sentences.
Experimental design: 3.	How the brains were fixed? Post-fixation or brain perfusion? This is not clearly mentioned in the methods section.

4.	To perform the statistical analysis the authors used one-way ANOVA and Student’s t-test. Do the results follow the normality?
Validity of the findings: 5.	I would suggest blocking the p38 MAPK signaling pathway with SB203580 in order to confirm the involvement of this pathway in SD-induced pyroptosis and NLRP activation and increase the validity of their findings.
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: NLRP3 INFLAMMASOME ACTIVATION MEDIATES SLEEP DEPRIVATION-INDUCED PYROPTOSIS IN MICE
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors clearly improved the manuscript, but I would suggest to carefully revise the English writting. There are still meaningless phrases.
For exemple: To further confirmed the p38 and ERK1/2 pathway in the SD induced NLRP3 and pyroptosis activation, the SD mice treated with or without the p38 and ERK pathway inhibitors,  SB203580 and U0126, and the NLRP3 and pyroptosis activation was analyzed by Western blot.Should it be "the SD mice were treated with or without the p38 ..."?
Experimental design: The authors improved the manuscript.
Validity of the findings: The authors improved the manuscript.
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: HIGH EXPRESSION OF PIMREG PREDICTS POOR SURVIVAL OUTCOMES AND IS CORRELATED WITH IMMUNE INFILTRATES IN LUNG ADENOCARCINOMA
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors described PIMREG as a novel prognosis biomarker for LUAD patients.
The article is well written, the analysis seems to be well performed. There are some points to clarify concerning the text.

1. Please, re-written lines 32-33. It was not shown that PIMREG decreases immune cells, only a negative correlation was found.

2. In the Discussion section, line 167, it is written that "PIMREG could be used as a biomarker for the DIAGNOSIS of LUAD". Please re-write this sentence,
as data did not show PIMREG for diagnosis, but prognosis.

3. Please re-write the sentence in line 197. The word "perfect" seems not to be the better choice to describe a biomarker.

4. Please re-write the sentence in line 200. It is not clear macrophage role.

5. Discussion section lines 206-208. The negative correlation of immune cells and PIMREG expression. It is not clear why "PIMREG may enhance tumor activity of lung adenocarcinoma through these immune cells". Please clarify.
Experimental design: Major points

1. Only the TCGA-LUAD cohort was evaluated in the study. It is a good practice to confirm the biomarker validity in other cohorts, as many as possible.
For LUAD, there are other cohorts with RNA-Seq that can be found at the GDC data portal or GEO.

2. The cut-off methodology was not described in the Methods section (median, best cut-off?). Please describe the method in detail because it is of vital importance for results reproduction.

3. Although the authors found a negative correlation between PIMREG and immune cells, the coefficient rho was small
(never more than 0.5 ou -0.5). The correlations are not strong, even with a significant p-value which was influenced by the high number of patients in this study. These results should be taken more cautiously and another cohort should be
used for validation.

4. Concerning cell culture and quantitative RT-PCR, the authors used b2-actin as an internal control. Did they test another housekeeping gene?

5. Are there evidence for PIMREG at the protein level for LUAD patients? Could it be used as a biomarker? Is there any correlation between protein and mRNA level?
Validity of the findings: All data are available for reproduction. The conclusions are partially supported by the results because a validation cohort must be used.
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: HIGH EXPRESSION OF PIMREG PREDICTS POOR SURVIVAL OUTCOMES AND IS CORRELATED WITH IMMUNE INFILTRATES IN LUNG ADENOCARCINOMA
Review round: 1
Reviewer: 2

Output:
Basic reporting: a.	The language in the article is generally very clear and simple; a minor grammar check may be needed throughout the manuscript.
b.	The article has good mentions of some previous work. However, the introduction needs to be enhanced significantly to include a more detailed review of the current understanding involving the role of PIMREG in cancers as well as background information on PIMREG itself.
c.	Figure 4, Figure 6 font is too small. Overall resolution seems to be low, which makes it difficult to read figures effectively. Table formatting can be improved to enhance readability. Figure 1C acronyms need to be displayed and spelled out at least in SI.
d.	Results relevant to the presented hypothesis are appropriately included.
Experimental design: a.	The article is well within the scope of the journal. The article clearly defines the overall research goal. It would be helpful to more clearly define the specific research questions earlier on in the manuscript. The experiments are proposed in attempt to achieve the general goal of the research.
b.	In general, the investigations are carried out rigorously.  A brief mention of the ethical guidelines from human patient database use would be helpful to include in the methods.
c.	The materials and methods section provides a good overview of the analyses performed including the various statistical tests.
Validity of the findings: a.	In general, the data presented in the manuscript represents a solid base for the conclusions. Statistical analyses are robust and used appropriately.
b.	The conclusions from the studies are concisely stated. Expanding conclusions to include discussions of study limitations would be helpful.
Additional comments: 1.	Line 101 “There is a significant relationship between the PIMREG expression with the clinicopathological parameters such as T stage, N stage, Pathologic stage, gender, smoke, tumor status, number pack years smoked and age.” This statement is too general and does not seem to be backed by or referenced to the data. Such unreferenced claims need to be checked throughout the introduction and discussion.
2.	Fig 8, more details on the expression measurement should be provided (e.g. fold change, relative levels, etc.)
3.	More details on the choice of ADH1C primer in mRNA expression experiments are needed. Definitions needed for that gene and why it is used to detect PIMREG


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: HIGH EXPRESSION OF PIMREG PREDICTS POOR SURVIVAL OUTCOMES AND IS CORRELATED WITH IMMUNE INFILTRATES IN LUNG ADENOCARCINOMA
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors made changes and improve manuscript quality. PIMREG increased expression on other cohorts corroborates with the manuscript data.
Also, protein expression seems to agree with mRNA data, although only a representative picture was depicted.
There are a few misspelling sentences, mainly in the Methods section to be adjusted.
In the Discussion section, the authors claimed "Our study also demonstrated that PIMREG was correlated with immune infiltration
in LUAD, which might be the mechanism by which PIMREG affects the prognosis of LUAD."
Then, the authors rewrote the discussion in the last paragraph contradicting this sentence. Please, adjust the text.
Experimental design: The experimental design is suitable for the manuscript.
Validity of the findings: The manuscript is clear and the results sustain the conclusions.
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: HIGH EXPRESSION OF PIMREG PREDICTS POOR SURVIVAL OUTCOMES AND IS CORRELATED WITH IMMUNE INFILTRATES IN LUNG ADENOCARCINOMA
Review round: 3
Reviewer: 1

Output:
Basic reporting: The article was revised adequately.
Experimental design: The experimental design was evaluated in the previous review steps.
Validity of the findings: The results are clear.
Additional comments: The article is adequate and no further revision is needed.


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: SERUM ALBUMIN AND ALBUMINURIA PREDICT THE PROGRESSION OF CHRONIC KIDNEY DISEASE IN PATIENTS WITH NEWLY DIAGNOSED TYPE 2 DIABETES: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 1

Output:
Basic reporting: Authors demonstrated that low serum albumin and high microalbuminuria levels are strongly associated with CKD progression in diabetic patients. The paper is written in a good english and is conform to professional standards. The literature references are sufficient and fits with the field knowledge.
Experimental design: no comment
Validity of the findings: I think the paper is not so original as the relationship between initial damage from exposition to hyperglycemia, the occurrence of microalbuminuria and progression to CKD is already well demonstrated in literature. These paper confirms these findings.
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: SERUM ALBUMIN AND ALBUMINURIA PREDICT THE PROGRESSION OF CHRONIC KIDNEY DISEASE IN PATIENTS WITH NEWLY DIAGNOSED TYPE 2 DIABETES: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 2

Output:
Basic reporting: Major comments:
It is my impression that the terms renal function decline and CKD progression are used interchangeably throughout the manuscript.This is quite tricky and confusing as a decline in eGFR does not necessarily translate into progression in CKD.Also not all patients had CKD at baseline.Authors need to clarify and amend accordingly.Also the phrase ‘’kidney progression’’ in lines 27,31,38,111 is not correct and should be replaced.
Minor comments :
1.In line 82 replace word creatine with creatinine
2.In line 98 authors need to substitute phrase ‘’..mean age of progression and non-progression’’ with ‘’mean age of progressors and non-progressors’’
3.In line 156 authors need to supplement the word type in front of ‘’..2 diabetes’’.
Experimental design: Major comments:
1.There is a clear need to study natural course of T2DM and its complications in various ethnic populations and discover biomarkers that help predict development of these complications so as to be able to intervene early.There is also a paucity of data on the above with regards to exclusively newly diagnosed patients with T2DM.Authors of the present study are trying to address these issues by studying the change in renal function of a  relatively large cohort of Chinese people with newly diagnosed  T2DM.Although the follow up duration is not long a 30% change of eGFR from baseline allows for the study to capture early changes in renal function that could help predict adverse renal outcomes eg progression to CKD/ESRD.
2.The authors state that the primary endpoint of the study is a more than 30% decline in eGFR.This has been shown to consist a valid endpoint in the study of renal function decline in diabetes and the general population (Levey et al Am J Kidney Dis 2016).Since serum creatinine levels are subject to significant biological and preanalytic variability the authors should confirm whether this level of decline is sustained or is it just a one-off measurement at the end of follow up?
3.What is the proportion of people in the study on RAAS blockade given the known renoprotective effect of these agents(Lewis et al N Eng J Med 2001)?
4.What is the proportion of people with diabetic retinopathy (DR) in the study population?DR often coexists with kidney disease in diabetes(Afkarian et al JAMA 2016) and could also serve as a predictor of renal function decline in this population potentially even at an early stage(although it is most commonly seen with long-standing disease).
Validity of the findings: Major comments :
1.Have the authors looked into the diagnostics of multivariable logistic regression analysis model so as to confirm that their model is robust and generalizable?
Minor comments :
1.I would prefer the term multivariable compared to multivariate regression analysis.
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: SERUM ALBUMIN AND ALBUMINURIA PREDICT THE PROGRESSION OF CHRONIC KIDNEY DISEASE IN PATIENTS WITH NEWLY DIAGNOSED TYPE 2 DIABETES: A RETROSPECTIVE STUDY
Review round: 1
Reviewer: 3

Output:
Basic reporting: English revision is needed
Experimental design: Retrospective design is a relevant limit
Validity of the findings: The novelty is limited
Additional comments: The manuscript by Jian Zhu et al. describes a vaste retrospective study conducted on newly diagnosed type 2 mellitus diabetes patients. The study aimed to investigate risk factors associated with CKD progression. Authors found macroalbuminuria and low serum albumin as factors associated with kidney function decline.
The following issues can be raised:
1. The result on microalbuminuria does not add any new knowledge , since this is a widely accepted dogma in nephrology. Conversely, some novelty can be found in the association between low albuminemia and CKD progression. Low albuminemia could reflect in an impairment of nutritional status or in a blood diluition. Both conditions would be however present in aneddotic cases and are generally associated with AKI, that represents an exclusion criterion in this study. The underlaying pathogenic mechanisms still remain to be explained.

2. Authors missed to explain how their results would impact on clinical practice.

3. The English language quality deserve a revision by a English native speaker (eg  the expression wisely used by the authors 'kidney progression' sounds inappropriate, giving the idea of an improvement rather than a worsening. Authors might substitute it with'kidney failure progression'...)


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: SERUM ALBUMIN AND ALBUMINURIA PREDICT THE PROGRESSION OF CHRONIC KIDNEY DISEASE IN PATIENTS WITH NEWLY DIAGNOSED TYPE 2 DIABETES: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 1

Output:
Basic reporting: Language of the manuscript has improved.It reads better now.
Again there seems to be a misconception regarding microalbuminuria.There is no substance microalbumin as such but microalbuminuria as a defined level of excretion of albumin in the urine as compared to normo- and macroalbuminuria.Authors should address this in the manuscript.
Experimental design: My comments on experimental design have been addressed.
Validity of the findings: Further analyses as outlined in my review have been performed.Comments sufficiently addressed.
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: SERUM ALBUMIN AND ALBUMINURIA PREDICT THE PROGRESSION OF CHRONIC KIDNEY DISEASE IN PATIENTS WITH NEWLY DIAGNOSED TYPE 2 DIABETES: A RETROSPECTIVE STUDY
Review round: 2
Reviewer: 2

Output:
Basic reporting: Overall the manuscript is well presented and English language quality has been improved
Experimental design: Study design is appropriate and statistical analysis was complete
Validity of the findings: Novelty remains a bit limited, however the authors better underlined the utility of their findings and the current version appears more convincing.
Additional comments: The current version of the manuscript has been improved by the authors in accomplishment with my suggesions and comments.

However, I would like to suggest enclosure of  'Quality of life worsening' within ESRD outcomes in the introductory paragraph (please refer to and cite: D'Onofrio G. et al. Quality of life, clinical outcome, personality and coping in chronic hemodialysis patients. Ren Fail. 2017 Nov;39(1):45-53. doi: 10.1080/0886022X.2016.1244077)


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: WEIGHT-DEPENDENT SUSCEPTIBILITY OF TILAPIA TO TILAPIA LAKE VIRUS INFECTION
Review round: 1
Reviewer: 1

Output:
Basic reporting: -
Experimental design: -
Validity of the findings: -
Additional comments: PeerJ Reviewing Manuscript 59569v1: Weight-dependent susceptibility of tilapia to tilapia lake virus infection
This work demonstrated weight dependent susceptibility of tilapia to TiLV. Three different fish weights were used in injection test while 2 different weights were for cohabitation. The results suggested the smallest size (5 g) was the most susceptible to TiLV.
Comments:
1.	The main concern of the manuscript is the histopathological readings. Most of the descriptions are incorrect. In Fig 3, there was no sign of syncytial formation, cytoplasmic inclusion bodies, occasional congestion and hemorrhaging in the liver. No melanomacrophage centers (MMC) and pyknosis and karyorrhexis in the spleen. No hemorrhaging and congestion in the brain. No disruption of the gastric glands, hemorrhage in the lamina propria, and vacuolation in the intestines. Thus, the histopathological scoring in Table 1 is questionable making data interpretation regarding weight dependent susceptibility weaken/invalid.
2.	Cumulative mortality in Fig. 1 gave an impression that the experiment was performed at the same time. It might be better to present data as independent set up. Additionally, mortality from replicate tanks should be presented (instead of a combine total fish of 60) and statistical analysis can then be done.
3.	Information on fish age at 5, 25, and 65 g should be indicated.
4.	More data of other fish that are age-dependent susceptible to viral infection should be discussed. Any case of age-independent? Spring viremia of carp virus?
5.	In cohabitation, how to tell which fish were inducers or naïve?
6.	Lines 197-199, it was confusing what was the data for 5 and 25 g fish at 7 and 14 days.
7.	Sample at day 0 collected for qPCR should be in the method section. Were fish from control (no virus challenge) subjected to qPCR?
8.	A 28-day experiment for cohabitation experiment should be indicated in the method section.
9.	Lines 101-103 “For viral quantification, the samples were collected at 7, 14, and 22 days post-infection (dpi). For sample collection, the fish were euthanized using an overdose of an eugenol solution (3 mL/L) for 5 min.” should be read as “For viral quantification, the samples were collected at 7, 14, and 22 days post-infection (dpi). The fish were euthanized using an overdose of an eugenol solution (3 mL/L) for 5 min prior to fish necropsy and sample collection”
10.	TiLV quantification in Fig. 2 was assayed from livers of three fish but why more than three dots shown in the graph.


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: WEIGHT-DEPENDENT SUSCEPTIBILITY OF TILAPIA TO TILAPIA LAKE VIRUS INFECTION
Review round: 1
Reviewer: 2

Output:
Basic reporting: Good writing article, with interesting fundamental knowledge on the effects of fish weight/size to TiLV infection.
Experimental design: Good experimental design. However, some comments for improvement as stated below and in attached file.
Validity of the findings: Good data validity due to good experimental design. However, some comments for improvement as stated below and in attached file.
Additional comments: Fulfill the criteria of basic reporting, experimental design and validity of the findings. However, some comments for improvement as stated below and in attached file.


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: WEIGHT-DEPENDENT SUSCEPTIBILITY OF TILAPIA TO TILAPIA LAKE VIRUS INFECTION
Review round: 2
Reviewer: 1

Output:
Basic reporting: -
Experimental design: -
Validity of the findings: -
Additional comments: The revised manuscript has been satisfactorily improved. Just only one point; In the abstract, ± sign should be for SE values in the parentheses.


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: WEIGHT-DEPENDENT SUSCEPTIBILITY OF TILAPIA TO TILAPIA LAKE VIRUS INFECTION
Review round: 2
Reviewer: 2

Output:
Basic reporting: Please refer to the attached file
Experimental design: Please refer to the attached file
Validity of the findings: Please refer to the attached file
Additional comments: Overall, significant improvements have been made by the authors. However, minor additional info as follows should be made too:

In the abstract and discussion sections:
This study only did on 5, 25 and 65 g of fish. What happens if size bigger than that - until harvest size? Bigger size causes more economic losses, compared to smaller size - even smaller size more susceptible in this study. This might be the limitation of the study. Please indicate here and in the discussion.

In the discussion section:
Please scientifically discuss why smaller size is more susceptible compared to bigger size? Any scientific explanation? This should be stressed in the discussion, rather than just comparing with previous studies.

Please refer to the attached file.


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: CHILDHOOD MALTREATMENT AND SUICIDAL IDEATION IN CHINESE CHILDREN AND ADOLESCENTS: THE MEDIATION OF RESILIENCE
Review round: 1
Reviewer: 1

Output:
Basic reporting: None
Experimental design: None
Validity of the findings: None
Additional comments: This is a population-based cross-sectional survey, which aimed to estimate the possible associations between childhood maltreatment and suicidal ideation in a large group of Chinese adolescents. Though this is a well-organized manuscript, I have some comments for the authors for their consideration.
1. The manuscript mentioned that the purpose of this study is to explore the possible mediation. However, I found you examined the moderation in the result section. Why not refer to the moderating effect in the introduction?
2. The manuscript mentioned that “47 subjects refused to participate”. Authors should provide the reasons.
3. Authors should provide the references about “The Resilience Scale for Chinese Adolescents (RSCA)”.
4. The manuscript mentioned “Data were analyzed as previously described in Xiao et al., 2019.” Some questions for this sentence were as follow.
1. Please use “analyzed” to replace “analysised”.
2. What the kind of data was analyzed in Xiao et al., 2019?
3. I checked the articles from Xiao et al., 2019. However, the population in Xiao et al., 2019 focus on the “left-behind” children and adolescents. So, does the population in this study mainly included the “left-behind” children and adolescents?
5. Table 1 should provide P values for univariate analysis between SI and non-SI.
6. Line 184. Authors used the median of RSCA (94) to dichotomize study participants. Please add related references.


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: CHILDHOOD MALTREATMENT AND SUICIDAL IDEATION IN CHINESE CHILDREN AND ADOLESCENTS: THE MEDIATION OF RESILIENCE
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comments
Experimental design: no comments
Validity of the findings: Query # 1: In table 2, there were some significant variables such as ‘Grade’, ‘Study style’ and ‘Marital status of the parents’ for SI . The authors should list these positive results in the section of Results.

Query # 2: In Figure 3, the authors should make clear the path coefficient ‘-0.441’ between ‘Resilience’ and ‘SI’. As figure 3 were results of a group of path analysis, I’m not sure the source of the coefficient ‘-0.441’.
Additional comments: The study was well organized and presented in a proper way. The study was a meaning effort to explore relationship among resilience, childhood maltreatment and suicidal ideation in a Chinese adolescent sample. Further longitudinal and some larger sample of studies are merited to explore validity of results of the study.


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: CHILDHOOD MALTREATMENT AND SUICIDAL IDEATION IN CHINESE CHILDREN AND ADOLESCENTS: THE MEDIATION OF RESILIENCE
Review round: 2
Reviewer: 1

Output:
Basic reporting: None
Experimental design: None
Validity of the findings: None
Additional comments: I am satisfied with your revision. The minor problems still need to be addressed.
(1) For easy identification, please add a comma to thousands of numbers in the whole manuscript. For example, Please use "3,146" in the line 27.
(2) Please keep a consistent format between Line 195-197 and Line 204.
(3)Please add the footnote for OR and CI in the table 2 and Figure 2.


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: CHILDHOOD MALTREATMENT AND SUICIDAL IDEATION IN CHINESE CHILDREN AND ADOLESCENTS: THE MEDIATION OF RESILIENCE
Review round: 2
Reviewer: 2

Output:
Basic reporting: in line 202, 'aOR' should be a misspelling. Please check it.
Experimental design: no comments.
Validity of the findings: no comments.
Additional comments: The study was presented in a more proper way after its first revision.


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: CHILDHOOD MALTREATMENT AND SUICIDAL IDEATION IN CHINESE CHILDREN AND ADOLESCENTS: THE MEDIATION OF RESILIENCE
Review round: 3
Reviewer: 1

Output:
Basic reporting: None
Experimental design: None
Validity of the findings: None
Additional comments: None


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: SHAPE FAMILIARITY MODULATES PREFERENCE FOR CURVATURE IN DRAWINGS OF COMMON-USE OBJECTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: All standards listed here are met.
Some minor points in formulation are marked in the attached PDF.
Please provide an accurate definition of what you mean by "drawings".
Experimental design: All standards listed here are met.
See some minor points in the attached PDF.
Please describe whay you did not evaluate the familarity of objects depicted.
Please describe whay the aspect of three dimensionality was not assessed as an image feature in the drawings.
Validity of the findings: All standards listed here are met.
Please provide effect sizes for the results where appropriate.
Additional comments: The manuscript is well written and provides an insight into the research field.
The research was carried out and described accurately.

The definition of the drawings and the categorizations in hand-made, computer-made, shaded, not shaded is of course not exhaustive. Relevant seems to me the aspect of three-dimensionality, which was not or too little considered. Aspects of contrast in the drawing, which also has an effect on liking, could also have been taken into consideration.


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: SHAPE FAMILIARITY MODULATES PREFERENCE FOR CURVATURE IN DRAWINGS OF COMMON-USE OBJECTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: I think the direction of reasoning is opposite, because participants did the preference (or 2AFC) task first followed by the familiarity task. Therefore, participants’ familiarity judgment might be contingent on the preference (or 2AFC selection) made before the familiarity judgements. Therefore, the authors cannot conclude that the familiarity judgements modulate preference for curvature between tasks. Instead, preference judgments could modulate familiarity for objects. If the order of testing does not matter, then the same pattern of the results should be obtained when running the experimental tasks in the reversed order.
Additional comments: Why the authors collected 49 participants in the two experiments?

The authors note that the participants of the present study were treated in compliance with Declaration of Helsinki. If so, the descriptions related to the preregistration should be reported, because research registration is required in a publicly accessible database before recruitment of the first subject (#35).


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: SHAPE FAMILIARITY MODULATES PREFERENCE FOR CURVATURE IN DRAWINGS OF COMMON-USE OBJECTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: Mostly sound and clear, background well defined, references well chosen. See 'comments for the authors' for details.
Experimental design: Relevant, clear and sound. See 'comments for the authors' for details.
Validity of the findings: The data are provided, clear and consistent. However, the statistical analysis is unclear and potentially erroneous. This is my main concern. Please see 'comments for the authors' for details and suggestions for improvement.
Additional comments: In this study, the authors explored the interaction between familiarity and preference for curvature. The rationale of the authors is as follows. Preference for curvature has been consistently found in a wide range of experiments using different types of stimuli. However, curved objects are prominent in nature. We are therefore more familiar with curved than sharp-angled objects. As we tend to prefer visual features we are more familiar with, it is therefore possible that preference for curvature is not driven by curvature in itself, as a purely geometrical characteristic, but by the familiarity conveyed by curved objects. Hence, to understand better what drives preference for curvature, we need to spell out the interaction between familiarity and curvature. The research question raised by the manuscript is well delineated, relevant and meaningful.

While I find the experimental paradigm chosen by the authors to assess the interaction between familiarity and preference for curvature sound, I have a major concern, which I think should be tackled before publication.

Statistics -
My main concern is about the statistical analysis. Using mixed effects models is well-suited for analysing the type of data provided by the current experiment, which is a good point for the manuscript. However, I find the description of the mixed modelling used in both experiments incomplete, unclear, and, unless I am misunderstood, incorrect in Experiment 2. This obviously is an important concern as all the conclusions of the study are driven by the statistical analysis. To reassure the authors, I would like to say that solving this point may only be a matter of making the statistical inference explicit and correcting the model in Experiment 2. All the choices for the statistical models are justified stating ‘The maximal model that converged included participant and stimulus as random effects, as well as random slopes within participant’ (l. 266 and throughout). I am not sure I follow the procedure for model selection. The strength of mixed effects models is to allow one to draw general conclusions by controlling the idiosyncrasies of individual observers or stimuli. In the study, ‘participant’ and ‘stimulus’ should be considered as random effects whatever the choice for all the other sides of the modelling, or potential convergence. Choosing whether the intercept, slope, or both should be random can indeed be exploratory, but not whether ‘participant’ and ‘stimulus’ are random effects. What kind of convergence do the authors refer to for choosing the model? Maximal in which sense? A standard procedure for choosing a model is to compare different models in terms of information theoretic criteria (AIC, BIC or others). If the authors have followed a less conventional but sound method they should make it explicit in the manuscript. A plus would be to publish the code used to carry the statistical analysis alongside the data. Note that this concern applies to all the choices for the final statistical model in the paper.
Besides, my understanding is that the statistical model used in Experiment 2 is similar to the one used in Experiment 1. If it is not the case, this should be stated clearly. While fitting a linear mixed effects model is adapted to the data in Experiment 1 (modelExpt1 <- lmer( Liking ~ …) , this is not adapted for Experiment 2. In the second experiment, the outcome is binary (‘curved_preference_choice’ = 0, 1 or ‘sharp_preference_choice’ = 1, 0 to use the name of the variable in the spreadsheet). The inference process should therefore use a generalized mixed effects model and a binomial law (modelExpt2 <- glmer(curved_preference_choice~ …, family = binomial, …)). The remaining part of the statistical inference can then be similar to the one in Experiment 1.
Another point that could improve the quality of the analysis is to report p-values with a greater precision. Two significative numbers (in which case a true p = 0.001 should read p = 0.0010) would be a minimum.
In the last part of the analysis of Experiment 2, I don’t think that introducing another way to estimate the means in the different conditions adds anything. Mixed effects models provide much more information than this.
As a minor note, I think that the p-value for ‘Contour’ in Supplementary Table S1 should read < .001 ***, not > .001 ***.
Finally, as I have appreciated the authors’ good practice to add confidence intervals for the means, I would like to mention that adding confidence intervals for the Spearman-rank correlation is also important, even if not common yet (I am therefore not saying that this should be done here, that’s just a suggestion). A simple bootstrap procedure to do so can be easily implemented in R. I have used the papers and tutorials linked to this https://psyarxiv.com/h8ft7/to learn how.

Paradigm and methods -
The experimental paradigm chosen by the authors to assess the interaction between familiarity and preference for curvature is sound. A potential limitation to the conclusions is that the study uses self-reported familiarity. However, the authors clearly identify this limitation. Besides, I agree with the authors that repeated exposure to novel, unknown objects may rub out any effect of preference for curvature. I only have minor points to comment. I list them below.
l.173: The database includes almost 800 stimuli. What was the criterion to choose the 90 that were chosen?
l.180: If I am correct, the frame was 600 x 600 pixels and the stimuli were 300 x 300 pixels? I find the last part of the sentence hard to follow.
l.189, 196 (but also in Experiment 2): The task had 8 practice trials. With which stimuli, 8 from the set of 90, or other stimuli of the database? This is important as showing twice some of the stimuli may bias familiarity judgements.
l.188: ‘Each stimulus was presented on the centre of the screen until the participant responded on the sliding bar using the mouse.’ As written an inattentive reader may understand that the stimulus disappears once the mouse is moved. Consider changing for ‘Each stimulus was presented on the centre of the screen until the participant ***had*** responded on the sliding bar using the mouse.’
l.342: ‘In the 2AFC task, each pair of drawings was presented on the screen until participants responded as in previous studies (Munar et al., 2015; Corradi et al., 2018)’. Please insert comma after ‘responded’, otherwise the sentence implies that participants had to give the same answers as the ones that were given in previous studies to move on.
l.360 and following: the task in Experiment 2 is not explicit. We are told the words the instructions avoided but not what the instructions were. The paragraph ‘procedure’ for Experiment 2 should be as clear and explicit as the corresponding paragraph for Experiment 1, otherwise we don’t know what the participants were asked to do.

Writing -
The manuscript is generally clear even if some errors should be corrected and some explanation may be improved.
The main problem is in the statement of the conclusion given on lines 599-600. I f I am not misunderstood, when both objects are judged as equally familiar there is no clear preference in Experiment 2. The general conclusion as stated here, and in the abstract, is therefore abusive.
I list several other points below, following the order of the manuscript, not in order of importance.
Abstract: ‘unconventional participants’ is a bit odd; could you rephrase?
l.45: ‘using art-related stimuli’ -> ‘using non-representational art-related stimuli’ as art-related stimuli can be representational.
l.47: what do you mean by ‘shared preferences’?
l.133: ‘In this study, we examined preference for contour using drawings of common-use objects in two experiments’. Better ‘In this study, we examined preference for contour in two experiments using drawings of common-use objects’.
l.154: ‘or only model it’. The meaning of this part of the sentence, and hence of the full sentence, is not clear.
l.257: performed -> considered.
l.264: ‘as the influence of Contour’ -> ‘as assessing the influence of contour’.
l.281: ‘was our main objective’. As for line 264, should be rephrased.
l.321 and following: this interim discussion may be improved by explaining that the data suggest and interaction between familiarity and curvature to shape preference.
l.356: Both naïve and naïve are correct. The text uses both. Please choose one.
l.500: ‘Experiment 2, using the same stimuli and different participants, consisted of a 2AFC task simulating approach/avoidance responses’. Probably better: Experiment 2 used the same stimuli and different participants, and consisted of a 2AFC task simulating approach/avoidance responses’.
l.510: ‘Similarly, Corradi et al. (2018) found that the effect of preference for curvature decreased as the presentation time increased using the same task and stimuli.’ I suggest: ‘Similarly, and using the same task and stimuli, Corradi et al. (2018) found that the effect of preference for curvature decreased as the presentation time increased.’
I don’t understand the lines 576 to 584. In particular, the authors use ‘endorse’ with a meaning I am not familiar with.
l.602: ‘These findings support that…’ -> ‘These findings support the idea that…’.
l.608: ‘endorse’.
Misc.: several instances of ‘On another hand’ -> ‘On the other hand’.


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: SHAPE FAMILIARITY MODULATES PREFERENCE FOR CURVATURE IN DRAWINGS OF COMMON-USE OBJECTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: Thank you for answering all my questions and adressig all my concerns.


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: SHAPE FAMILIARITY MODULATES PREFERENCE FOR CURVATURE IN DRAWINGS OF COMMON-USE OBJECTS
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors successfully responded to my previous comment.


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: SHAPE FAMILIARITY MODULATES PREFERENCE FOR CURVATURE IN DRAWINGS OF COMMON-USE OBJECTS
Review round: 2
Reviewer: 3

Output:
Basic reporting: I find that all the changes made by the authors have improved the manuscript and respond well to all the queries I had during the first phase of review. Both experiments are now clearly explained and fully reproducible.
Experimental design: No comment.
Validity of the findings: All the changes regarding the statistical inference and report have been satisfactorily implemented. The statistical modelling is now clear, coherent, and explicit.
Additional comments: I am happy with all the improvements made on the manuscript. They solve satisfactorily all the issues I found in the first submission. I therefore would like to congratulate the authors on this submission. I may have missed it, but I haven’t found any link or reference to the code used for the statistical inference in the manuscript. I think that it would be useful for the reader to have access to the osf entry created for this work. Please find below a short list of minor concerns in the updated manuscript.

Line 118: ‘e.g.’ instead of ‘i.e.’ in ‘for more realistic (i.e., photographs) or more abstract stimuli (i.e., irregular polygons).’?

Line 218: ‘Lastly, the apparent position of the objects in relation to the viewer was frontal view in 24 pairs, and ¾ view in 21 pairs.’ -> Lastly, the apparent position of the objects in relation to the viewer corresponded to a frontal view in 24 pairs, and to a ¾ view in 21 pairs.

Line 221: ‘Out of the computer-made drawings, 2 pairs were shaded, 13 pairs were not shaded, 8 pairs were in a ¾ view, and 7 pairs were in a frontal view.’ -> I think that ‘in ¾ view’ and ‘in frontal view’ are more correct, please check.

Line 333: ‘The familiarity selection task showed that the curved shapes were selected as the most familiar ones with an average of .49,…‘ ‘In a proportion’ or similar instead of ‘average’?

Line 491: same as line 333.

Line 507: please spell out ‘OR’ the first time it is used.

Line 562: ‘with 1499 samples suggested for a test at the 0.5 and 0.1 level’. Do you mean the 0.05 and 0.01 levels.

Lines 665 and 667: ‘In particular, Pepperell (2012) indicated that he uses a curvilinear perspective, instead of a linear perspective, because it is closer to the phenomenology of the perceiver.’ and ‘Straight lines in nature can appear curved in perception, a phenomenon that artists have shown (Pepperell & Haertel, 2014).’

I am struggling a bit with the thread here. Isn’t the statement more general than the one given in the first sentence (‘he uses’), and could both sentences be merged? For example, something such as ‘In particular, Pepperell (Pepperell (2012),  Pepperell & Haertel, 2014) has shown that artists use curvilinear perspective instead of linear perspective because it is closer to the phenomenology of perception.’


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: SHORT-CHAIN SOLUBLE POLYPHOSPHATE FERTILIZERS INCREASED SOIL P AVAILABILITY AND MOBILITY BY REDUCING P FIXATION IN TWO CONTRASTING CALCAREOUS SOILS
Review round: 1
Reviewer: 1

Output:
Basic reporting: -	Overall, the text is well written, yet a few point of attention are:
o	Line 62: parenthesis before DCPD
o	Line 75: “Some studies showed short-chain (n<20) soluble polyphosphates (poly-P) superior over ortho-P fertilizers in increasing crop productivity”, should be written as “polyphosphates (poly-P) are superior over ortho-P fertilizers” or “polyphosphates (poly-P) to be superior over ortho-P fertilizers”
o	Line 76: “This may largely ascribe to its slow release characteristic” should be written as “This may largely be ascribed to its slow release characteristic”
o	Line 89: a “iii)” should be added here fore the last point
o	Line 212: “repeated P application significantly promoted P vertical migrant” should be written as e.g. ““repeated P application significantly promoted vertical P migrantion”
o	Line 140: “phosphorous”; Line 157, Line 255: “phosphorus”. Try to be consistent with the use of the P abbreviation.
o	Line 284: “their interact with soil particles in a wide-range of soil conditions.” Interact should be changed to interaction.
-	A lot of relevant literature has been cited in the introduction, yet the authors should revise the statements that were made (see also comments on validity of the findings).
-	Figures are clear and relevant.
-	Line 116: The method used to apply the single fertilizer application is not explained clearly. “The amount of P fertilizer was uniformly mixed with the upper 20 mm of soil” but also the authors write that “the P solutions were sprayed on the upper soil surface of the soil cylinder”.
-	Line 287: “In summary, our study showed that ammonium-polyphosphate application significantly increased soil phosphorus mobility and availability”. Sentence should be revised: the phosphorus mobility and availability increased compared to what?
-	The raw data are presented very clearly.
Experimental design: -	Line 133: “all cylinders were placed into a -80℃ freezer for 12 hours”. Why was opted for this freezing method? Freezing of the column is generally not required to sample the soil in the column along with the column depth. Can the freezing affect the P speciation, e.g. by concentrating the solution P concentration during the freezing step, or by slightly changing the pore structure? Has this method been used before in comparable work?
-	Soil cylinder experiment: No pH measurement was performed on the obtained soil samples from the column, although acid-base effects are expected with the use of the reported P fertilizer compounds. The obtained soil pH can also strongly affect to the availability and mobility of the fertilizer P and the soil P.
-	An important issue that was not addressed sufficiently in this work, either in the introduction or in the experiments, is the application form of the fertilizers. For example, the MAP fertilizer was dissolved prior to application, while most literature studies cited in the manuscript report on fertilizer trials with granular MAP, which is also the most commonly used form of this fertilizer. Why did the authors opt to dissolve the MAP prior to application?
Validity of the findings: -	Figure 2: in the loamy soil, it seems like there is a data point missing for the single APP treatment at 10 mm? How does this affect the modeling, and the statement made on line 207, as for example the single application of MAP at 10 mm depth could have a quite comparable value for Olsen P?
-	A main concern on the interpretation of the results of this study refers again to the application form of the fertilizer MAP, and more attention should be given to this in the discussion. Also, the authors refer to literature findings in the discussion, that clearly relate to a granular MAP application (e.g. Hedley & McLaughlin, 2005 and by Lombi et al., 2004), while no granular fertilizers were tested in this study. Therefore, I believe the interpretation of the results of this study should be done more carefully with respect to not only the P fertilizer source but also the application form (dissolved MAP in this study, granular MAP in many literature studies).
-	In line with the previous comment, the authors should revise the literature statements that are made in the manuscript, because the corresponding citations do not always confirm the statements made in the manuscript. In that sense, the discussion definitely still needs to be improved.
-	The study investigates both P availability and P mobility from different P fertilizer sources and application methods. The introduction focusses on the efficiency (PUE) of polyphosphate fertilizer relative to conventional orthophosphate fertilizers. The matter of P mobility (P leaching) is, however, not addressed. A high P availability is obviously a positive aspect for fertilizer efficiency; however, a high mobility of P in the soil is not always beneficial for the PUE. Naturally, from a chemical perspective, there is a link between P availability and P mobility, but also this is not discussed. Could the authors elaborate more on the importance of both P availability and P mobility of the different P fertilizer sources?
Additional comments: -	Line 281: “Therefore, future study should combine using edge-cutting technologies such as atomic force microscopy (AMF), Raman spectrometer and Fourier transform infrared spectrometer (FTIR) and X-ray absorption near edge structure (XANES) to provide in situ direct evidence regarding the behaviors of polyphosphate fertilizer and their interact with soil particles in a wide-range of soil conditions.”. This statement is too broad. What information can still provide additional knowledge on the behavior or P and poly-P in calcareous soils? What samples are interesting to be analyzed with which techniques?


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: SHORT-CHAIN SOLUBLE POLYPHOSPHATE FERTILIZERS INCREASED SOIL P AVAILABILITY AND MOBILITY BY REDUCING P FIXATION IN TWO CONTRASTING CALCAREOUS SOILS
Review round: 1
Reviewer: 2

Output:
Basic reporting: English Editing required
Title did not express the body of article I suggest to revise it.
Article structure figures and tables need improvement
Lack self contained with relevant results to hypothesis
Experimental design: The author is talking about mobility of P in Soil But I did not see hydraulic properties of the soil in text. Please provide the Hydraulic properties of Soils after experimental Set Up. The soil column develop with disturbed soil which ultimately affect the Hydraulic properties of soils. So, the mobility of P with out soil Hydraulic properties is under question?
2ndly, the solubility of the different P sources is missing in text. Please provide the solubility rate.
Methods need improvement for replication of experiment.
There is need to correlate the identified knowledge gap with findings
Validity of the findings: The detail about the statistical tests they performed on the data before doing the ANOVA since it is a parametric test, normality and homogeneity of variance should be verified.
Additional comments: This study is of great importance. The Introduction section is well written with recent references. I appreciate the fact that the authors described the methodology used in detail in the study. It helps to understand the results section thoroughly. However, I have some concerns about the different parts of the manuscript. Moreover, the paper lacks many major details. I suggest a major revision to address a few issues. If the authors address carefully the comments, I’ll recommend publication of the manuscript in the journal. Please, do include more implications of the results of the study in the conclusion and abstract sections.


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: SHORT-CHAIN SOLUBLE POLYPHOSPHATE FERTILIZERS INCREASED SOIL P AVAILABILITY AND MOBILITY BY REDUCING P FIXATION IN TWO CONTRASTING CALCAREOUS SOILS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The new title for the paper suggested by another reviewer is, in my opinion, definitely not an improvement. With this new title, it is not clear what the reference situation is for the ‘increased’ availability and mobility of short-chain soluble polyphosphate fertilizers, while in the previous title this was well stated (“Short-chain soluble polyphosphate outperforms orthophosphate fertilizers…”). I believe it is important to somehow mention in the title that this study compares polymeric phosphate fertilizer with orthophosphate fertilizer.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The issues I addressed in the previous version of the manuscript were sufficiently addressed in the revised version, I thank the authors for the changes that were made.


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: GEOGRAPHIC DISTRIBUTION MODELING AND TAXONOMY OF STEPHADISCUS LYRATUS (COTHOUNY IN GOULD, 1846) (CHAROPIDAE) REVEAL POTENTIAL DISTRIBUTIONAL AREAS OF THE SPECIES ALONG THE PATAGONIAN FORESTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The authors need to improve their English in some parts (see specific comments). It remains to improve the information in the introduction, includes more examples of how rare the study of micro-snails; provide microhabitat data and its importance for the persistence of snails in similar latitudes, temperate ecosystems, or neighboring countries such as Chile. PeerJ standards are fine. Figures can be better, especially Figures 1 and 5. Raw data is fine.
Experimental design: The authors should improve some aspects of the transects and provide more details to ensure replicability. The niche distribution model is well explained, but you should provide examples of similar work with smaller numbers of samples for reliable models. The questions could be more precise.
Validity of the findings: The new antecedents for this species are relevant and must be considered and validated, but improvements are required. The new morphological data of the shell and the internal parts are quite good. You need yes or yes to improve the microhabitat part. Improve the delivery of information in the niche model.
Additional comments: Thank you for giving me the opportunity to review the work of the authors Cuezzo, Medina & Nieto. The work consists of showing new morphological evidence of both the ultrastructure of the shell and the internal parts of Stephadiscus lyratus, a forest micro-snail. Furthermore, due to the scarce information on these snails, the authors provide data on microhabitats, a potential model of an ecological niche, and the representation of this species of micro snail in the system of protected areas of the states of Chile and Argentina. The work is a good contribution from the taxonomic and ecological point of view. I agree that the authors are making an important contribution to a species of which almost nothing is known. However, better work needs to be done. In the current form, the authors need to improve the background in the introduction, clarify their objectives and some methods. In addition, the results present necessary information that must be removed since it does not contribute to the development of the proposed objectives. The authors are not conducting a systematic review of the genus Stephadiscus, but rather providing new morphological and ecological data, which I repeat is tremendously valuable. In the discussion, the authors focus more on their results, but they speculate on some points and that should be corrected.

Introduction
Line 2. Remove this: “Stephadiscus Hylton Scott, 1981 is a genus of the diverse…” Star with “Charopidae is…”
Line 6. Remove: “was the researcher that most..” and after described insert “ many”
Line 8: please specify what you mean by anatomical (shell morphology, internal parts morphology?).
Line 9: this phrase “Also, adequate fieldwork to estimate the current distributional range of genera and species has not been done” is an affirmative sentence. Are you sure that any other researchers made this task before?
Line 11. Insert here: Stephadiscus, Hylton Scott, 1981, included…; delete “initially”
line 17. Add “Currently, the distribution of Stephadiscus…”
Lines 15-17. “The taxonomic position of Stephadiscus striatus Hylton Scott, 1981 from northeast Argentina and Venezuela, will have to be reconsidered since it seems to belong to Punctidae (Miquel & Barker, 2009)” please rephrase. It can read some rare this phrase.
Line 18. Change: “The distribution of the Stephadiscus” to “The Stephadiscus distribution is restricted to…”
Lines 21-24. This phrase is some unconnected from the main text “Biogeographically, southern South America belongs to the Andean region, proposed among others by Morrone (2018), who divided this region into three subregions, the Subantarctic, Central Chilean and Patagonian. The Andean region has a closer relationship to the Austral region in the Austral kingdom (Morrone, 2018)” I suggest change the sense and combine phrases, for example, after “The distribution of the Stephadiscus is restricted to Patagonia at both sides of the Andes from S 36° towards the southernmost portion of the continent, including Malvinas islands and southern archipelagos (Miquel & Barker, 2009; Miquel & Araya, 2013) and belong to the Andean region following Morrone (2018). This region was divided into three region: Subantarctic, central Chile and Patagonian (Morrone 2018?)
Line 25. Delete from this line this: “designated as the genus type species” and incorporated in brackets in line 13 after lyratus
Line 28. This have some cite or is only a personal observations of the authors?
Line 37. Please provide other cites. There is a wide literature about modelling
Line 38. Delete “the” species…
Line 39. Delete, in this sense and rephrase please.
General comment: is important for this work reveal that modelling niche for land snails is an important tool, especially to know your distribution/potential distribution. In current form, the introduction needs more information. I suggest read, and incorporate these articles:
1.	Weaver, K. F., Anderson, T., & Guralnick, R. (2006). Combining phylogenetic and ecological niche modeling approaches to determine distribution and historical biogeography of Black Hills mountain snails (Oreohelicidae). Diversity and Distributions, 12(6), 756-766.
2.	Beltramino, A. A., Vogler, R. E., Gregoric, D. E. G., & Rumi, A. (2015). Impact of climate change on the distribution of a giant land snail from South America: predicting future trends for setting conservation priorities on native malacofauna. Climatic Change, 131(4), 621-633.
3.	Mumladze, L. (2014). Sympatry without co-occurrence: exploring the pattern of distribution of two Helix species in Georgia using an ecological niche modelling approach. Journal of Molluscan Studies, 80(3), 249-255.
General comment: the authors show a second aim: “second, to characterize the microhabitat where it is found”. However, nothing about microhabitat is said in the introduction. Microhabitat can influence distribution and abundance of land snail by environmental temperature or food supply and is essential for the occurrence of them. Also, obviously, can influence the potential distribution using modelling. As the ecology of land snail in south America is scarce, I suggest taking account a new paragraph with articles in similar latitudes or ecosystems as:
1.	Barahona-Segovia, R. M., Riveros-Díaz, A. L., Zaror, S., Catalán, R., & Araya, J. F. (2019). Shelter, ecophysiology and conservation status of Plectostylus araucanus (Pulmonata: Bothriembryontidae) in the fragmented Maulino Forest, central Chile. Revista Mexicana De Biodiversidad, 90, 1-11. (For Chile)
2.	Gómez, P., Espinoza, S., Hahn, S., Valenzuela, M., & Ormazábal, Y. (2020). Forest variables associated to the occurrence of the endemic pulmonate Macrocyclis peruvianus (black snail) from the Maule Region of central Chile. Caldasia, 42(2), 306-312. (For Chile)
3.	Murray, J., Johnson, M. S., & Clarke, B. (1982). Microhabitat differences among genetically similar species of Partula. Evolution, 316-325. (For Micronesia)
4.	Müller, J., Strätz, C., & Hothorn, T. (2005). Habitat factors for land snails in European beech forests with a special focus on coarse woody debris. European Journal of Forest Research, 124(3), 233-242. (For Germany)

M&M
Lines 63-65. Cite? I suggest Smith-Ramírez, C. (2004). The Chilean coastal range: a vanishing center of biodiversity and endemism in South American temperate rainforests. Biodiversity & Conservation, 13(2), 373-393.
Line 70. Jungle? This is wrong. Valdivian evergreen forest is a rainfall forest but not a jungle. Remove this please.
Line 78. Please provide cites.
Line 95: how meters have you linear transect?
Lines 96-99. How many roots, trunks or stones survey per transect? What is the sampling effort per site? How many persons per site? More details are necessary to ensure replicability.
Lines 128-129. Delete “as part of the 60”
Line 141. Are 24 points a good number? When I work in modeling, the minimum required is 30 or 40 records for the most reliable distribution model. Please provide other articles when after spatial autocorrelation delete many occurrences.
Line 171: full occurrences are 60, 37 or 24? Please specify. If are 24, please remember to give more examples with a low number of occurrences. If are 60, why use full dataset if you are applied a spatial correlation?
Line 171. Final models or final model? Please be consistent through the text
Line 75. Define G region here

Results
Lines 201-220. Your work is not a systematic review, therefore these phrases are unnecessary here. Please remove.
Line 223. Delete body
Line 224. Add “body” black…
Lines 224-260. Why the authors combine taxonomy with ecological modeling? This link in the introduction is missing or at least, poorly developed.
Line 264. Stephadiscus lyratus was mainly found living on or under the bark of fallen trees or under humid logs in contact with the ground. How is percentage per microhabitat? The authors realize transect and on each, search actively land snails per microhabitat type, therefore, authors can obtain a percentage of microhabitat use.
Line 267. Rephrase
Line 269. Temperature buffer? Please clarify
Lines 275-334. This is completely unconnected with the aims proposed by the authors. In a taxonomical article this information must be placed in the introduction in many cases, for example, “S. lyratus has the largest shell diameter (DM= 5.5 mm) among all species classified in Stephadiscus. It is a very conspicuous species regarding its shell coloration and sculpture”, can include in the present work in introduction section. Again, I think that this a hybrid work with Taxonomy and modeling niche and this link must be clear. Remove this lines of your present work
Line 340. Met? Also, show the AICc value for best fitted model and compare it with other models.
Lines 342-345. What is the values of AICc weight for BIO variables?
Line 367. Which IUCN category are II, Ib respectively? Please rephrase

Discussion
Line 378. Deleting the information of lines 275-334, you must place this authority names in this line-
Line 380. Always, after a continuum point, you must provide the full name of the species. Review through the text.
Lines 382-383. Please rephrase.
Line 398. To the area in Chile? Replace for “to Chile”
Lines 411-415. “From the seven environmental variables used in the EGD analysis of S. lyratus, the main constraining variables are the temperature (Isothermality, Minimum Temperature of Coldest Month, and Seasonality) and Annual Precipitation. Thus, temperature and precipitation were the most important abiotic factors to explain S. lyratus occurrence”
What do you hope will happen to the micro snail with climate change? How will the abundance or distribution of the micro snail vary with potential changes in humidity and temperature? Please include a short text about it.
Lines 415-420. Sentence too long, please separate into two phrases. In addition, use this example in the introduction to show the scarce sources of information in this taxonomic group.
Lines 423-427. This part begins with the importance of microhabitats for S. lyratus, but in the introduction, the authors not present anything of this structural key element. Moreover, the description is very qualitative and the authors not present percentages, proportions or other metrics to reveal the importance of fallen barks, for example, we find 60% of all individuals under fallen barks vs 12% of individuals in logs in contact with humid soil. This type of information is key to understand future conservation status if the aim of this part is that.
Lines 428-430. Provides cites. I suggest:
Régnier, C., Achaz, G., Lambert, A., Cowie, R. H., Bouchet, P., & Fontaine, B. (2015). Mass extinction in poorly known taxa. Proceeding of Natural Academy of Sciences, 112, 7761–7766.
Régnier, C., Fontaine, B., & Bouchet, P. (2009). Not knowing, not recording, not listing: numerous unnoticed mollusk extinctions. Conservation Biology, 23, 1214–1221.
Line 448. Change “evaluation” for “assess”
Line 458. Small size?
Lines 468-471. Provide cites
Lines 471. In line 455, the authors said that “Empirical data on the life history of small land snail species in their natural habitat are hardly available worldwide” but, in the 471-473, the authors said “The combination of the ectothermic physiological traits of this species, low dispersal capacity, probable low fecundity producing only few eggs, and its narrow habitat requirement (forest specialist) turns S. lyratus into a vulnerable species”. The authors realized experimental thermal or supercooling experiment in S lyratus? Do the authors present information on dispersal capacity or habitat requirements? I understand that is probable, but, the information is not available and therefore, you can suggest that this species could be vulnerable, but not use affirmative sentence.
If you want to declare S. lyratus as vulnerable species must apply an IUCN Red List, for example, B criterion, the most rapid assessment for this species and see if it meet the thresholds
Lines 457-471. The authors explain largely physiological traits that could be the cause for S. lyratus to be considered a vulnerable species, but, is too long explanation for a final speculation. Reduce importantly this and be concise.

Conclusions
Line 497. Anatomical parts is internal parts? Review through the text
Point 5 of conclusion is not supported and is only speculation without experimental or natural dataset and therefore I suggest remove it. Conclusions must be of your results.
In addition, one aim is to characterize the microhabitat, but the authors nothing concludes about this. Please replace the point 5 with the current information by microhabitat conclusion

Figures
Figure 1. The green color of Patagonian forest is to clear
Figure 5. Describe what mean VTF, MSF, etc… the legend must be sufficient to understand all aspect of the figure


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: GEOGRAPHIC DISTRIBUTION MODELING AND TAXONOMY OF STEPHADISCUS LYRATUS (COTHOUNY IN GOULD, 1846) (CHAROPIDAE) REVEAL POTENTIAL DISTRIBUTIONAL AREAS OF THE SPECIES ALONG THE PATAGONIAN FORESTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: All my comments are in the General comments section.
Experimental design: All my comments are in the General comments section.
Validity of the findings: All my comments are in the General comments section.
Additional comments: General comments
In this contribution, the authors present new data on the morphology and distribution of the micro snail Stephadiscus lyratus in the Patagonian Forests. Also, they developed a niche model to identify potential distribution areas in neighboring subecoregions. Their results showed new morphological diagnostic traits for the species and that suitable climates exist in the Valdivian Temperate Forests subecoregion.

I am not an expert in snails, so there is nothing that I can comment on the taxonomic findings. Still, I realize that these contribute to the biology and taxonomy of the group due to the authors' careful morphological analyses that, as they mentioned in the manuscript, are not common for these invertebrates.

I will focus on the distributional data and analysis. I think that the niche modeling was technically well conducted. The authors were strict in the methodology and presented all the information necessary to replicate the analysis. My concerns, however, are in the motivation of this analysis and the interpretation of their results.

First, If the idea was to test the hypothesis that this snail has a broader geographic distribution than the one proposed by Scott (restricted to the Subpolar Magellanic Forest), I think that the work is incomplete. This is because with the niche model you cannot test this hypothesis; actually, the potential distribution obtained with the niche model is a hypothesis itself, so you need field data from the region where the model indicated that there are suitable conditions for the species to test the initial hypothesis.

Second, although the modeling procedure was technically adequate, its use was overly simplistic. The authors used it only to describe the potential distribution of the species and the variables that were more informative according to maxent, without any attempt to understand why those variables were important and the implications of this information in the light of conservation, climate change and other topics that the authors present in the Discussion. In other words, this contribution is mostly descriptive but very little analytic. I strongly suggest that the authors make an effort to make a stronger link between the morphology that they studied here, the physiology known for the species and the distribution, to present more robust support for their inferences.

Specific comments
L34. New paragraph for Modern methodologies…

L36. Eliminate and/spatial. ENMs use only the environmental data obtained from the locations.

L43-45. I understand what you mean, but it is not correctly expressed. It is not by identifying scenopoetic variables related to the species that makes it possible to identify areas where other populations inhabit. It is the combination of environmental conditions in the relevant scenopoetic variables that really matter.

L54. There is no mention of collecting permits.

L61. We followed…

L73. Not vegetation, but plants

L158. Actually, several of your environmental variables are correlated, but not at the level of your threshold, which, by the way, is pretty high.

L275. I think this section, while interesting, is overly long. I suggest sending it all or at least most of it to Supplementary Information as it does not make part of the findings of the study. Also, it should not go in the Results section, but in the Discussion.

L411. New paragraph for From the seven…

L414-415. You only used temperature and precipitation information to build the models, so this sentence is tautological. Here it would be useful to discuss why such environmental variables were the important ones for the snail distribution. In other words, what limits the distribution of the species and why.

L.475-492. This part of the Discussion regarding the possible effects of climate change in the snail can be improved with actual data specific for the region. Not only future projections but also if there is something that the authors can say about how the climate has changed in Magellanic Subpolar Forests in the last decades and how these changes affect the species' physiology in light of your findings. For example, regarding the environmental variables that you found important for the distribution of the species. Otherwise, it is just speculation

L501-505. This conclusion is definitively misleading. The fact that the niche model indicated that there are suitable climatic conditions for the snail in the Valdivian Temperate Forest does not mean that the species is there, and thus that your initial hypothesis is not supported. You obtained a distributional hypothesis with your modeling analysis, not a test of your initial one; you cannot test one hypothesis with another hypothesis. Moreover, your fieldwork found no evidence of the species' presence in the area of the Valdivian Temperate Forest where you sampled. As you mentioned, your results may guide future fieldwork in the areas where the model indicates potentially suitable areas, but until then, you cannot accept or reject your initial hypothesis.

Figure 5. It would be helpful to present in the map the occurrence and sampling sites (where you did not find the species). Also, you can include the protected areas that you used in the analysis.

Table 1. I think that If you are interested in an analysis of habitat preference, it is better to do it with the occurrences, not with the potential distribution map.


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: GEOGRAPHIC DISTRIBUTION MODELING AND TAXONOMY OF STEPHADISCUS LYRATUS (COTHOUNY IN GOULD, 1846) (CHAROPIDAE) REVEAL POTENTIAL DISTRIBUTIONAL AREAS OF THE SPECIES ALONG THE PATAGONIAN FORESTS
Review round: 1
Reviewer: 3

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The manuscript is adequate in content, methodology and results.


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: GEOGRAPHIC DISTRIBUTION MODELING AND TAXONOMY OF STEPHADISCUS LYRATUS (COTHOUNY IN GOULD, 1846) (CHAROPIDAE) REVEAL POTENTIAL DISTRIBUTIONAL AREAS OF THE SPECIES ALONG THE PATAGONIAN FORESTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: 1. English editing service is necessary through the text. I suggest an excellent tool for improving this problem in the review letter.
2. Literature reference provided sufficient background.
3. Article adequately build.
4. Self-contained with relevant results to hypotheses.
Experimental design: 1. Follow primary research, although some results still need to connect with the aims proposed.
2.  Research question well addressed.
3. High technical and ethical standards.
4. Methods well developed.
Validity of the findings: 1. For micro snails, is important to understand more about their ecology.
2. For modelling is well supported
3. Bullet point in the conclusion needs work and connected with the original aim.
4. Some speculation is provided, although not all well supported.
Additional comments: The manuscript improves your content, but English still needs review through the text. I suggest an excellent tool for non-native speakers called trinka.ai (https://www.trinka.ai/). This an efficient Grammarly and writing English corrector used specifically for non-native speakers and scientists in many parts.

Also, I do not agree with some parts of the answers given in the rebuttal letter in the remarks section, since those comparisons can be made in a dichotomous species key, thus making sense of the previously mentioned systematics for this species. The systematic section needs improving.

Other comments as follow:

Title
¿Why the taxonomy reveals potential distribution areas? I suggest ordering the words used in the title.

Abstract
English review is needed.

Introduction
L80. “(Couthouy in Gould, 1846)”.
When the authors insert taxonomical description by some author included in the work of a second taxonomist, please use italic for “in”. The correct form according to the Zoological code of nomenclature and ZooBank is (Couthouy in Gould, 1846)”. Correct through the text

L111-112. Add references for this affirmation.

L198. Delete “plus” and replace it with a ‘+’ symbol.

M & M
L214. What is the terminology used by the authors? This is key in the description of the internal anatomical parts. Please provide the most updated terminology used in malacology.

Results
L388. “Rainforest provide…” or temperate forest? be consistent. I think that the most adequate term is temperate.

L398-399. “but probably are associated…” This is speculative and I suggest removing it. However, the authors can discuss this briefly.

L411. After the dot place the complete name of the genus “… 1.5mm). Stephadiscus lyratus …”

L414. “Couthouy in Goul” in italic

L407-485. I do not agree with the author putting the comparisons with other Stephadiscus species and other genera in full text ¿Why don't the authors incorporate a dichotomous species key using whorls, shell, and other taxonomical traits and solve the systematics from this point of view? This resolves my doubts about why systematic and not a simple taxonomical ordination. In addition, I think that the synonyms, the type species, and syntype for your work do not add new information for the species because this already described. Also, I think is necessary to delete lines 328-341. A systematic review is not proposed in your aims, so, are necessary changes about this theme.

L499-503. Based on the proposals made by the second reviewer, the authors incorporate in lines 298-315 a full paragraph for the answer. However, I find it unnecessary, in results, to describe what the BIO3 means (499-503).

Discussion
L556-558. For this reason, the dichotomic species key is completely necessary. The differences with other genus or even, with similar species of the same genus might be resolved using this tool.

L559. Internal anatomical parts?

L587. Change to the format of reference citations in text.

L593. “… main constraining variables are (related with) the temperature…” many grammatical errors like this are present in the text. Please, use trinka.ai or a specialized English edition service.

L605. “Temperature Seasonality, and Mean Temperature” not use a capital letters.

L607. ¿Bell shaped or bell-shaped like the results? Be consistent.

L608-609. Not use a capital letters for BIO variable names.

L637. References?

L652. Add a dot at the end.

L663. References?

L667-668. Use format for references in the text.

L679-682. I think that this paragraph is unconnected from the conservation issues and is repeated literally from the lines 626-629. Please, authors must be careful with this.

L683-685. I still considered that this is speculation of the authors and not based on the information available. However, is true that land snails have low vagility and that they are vulnerable to habitat fragmentation, fires, or climatic change, but not, for this reason, the statement in the rebuttal letter “does not deserve any additional experimentation to sustain it” is true.

Conclusions
Bullet 5. The statement “The combination of the ectothermic physiological traits of this species, low dispersal capacity…” not supported by the current data. When authors say physiological trait, I imagine, e.g., rollover speed, supercooling or freezing point, or physiological performance at different controlled temperatures to permit the readers to understand which are the environmental limits for the species. These limits are not the environmental variables explored here because, as the authors mentioned in your manuscript, microhabitat buffer these environmental conditions, and therefore, the performance can change under these conditions. Also, the physiological traits can change along latitudinal gradient permitting several phenotypical expressions for the same environmental pressure.

Acknowledgments
Include editor and reviewers.


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: QUANTIFYING THE EFFECTS OF DELISTING WOLVES AFTER THE FIRST STATE BEGAN LETHAL MANAGEMENT
Review round: 1
Reviewer: 1

Output:
Basic reporting: This paper is clearly written with appropriate structure and citations.  The results are relevant to the hypotheses and the inferences are presented clearly.

This paper analyzes existing data on the size of the Wisconsin wolf population, reported offtake by human hunters, and rates of disappearance of radiocollared wolves.  These data are combined in a very simple population model to assess whether the policies implemented by wildlife management agencies and the state government after delisting from the ESA are likely to achieve the stated goal of keeping population size constant.  The authors use cumulative incidence functions fit to disappearance rates of  radiocollared wolves in periods with and without hunting to demonstrate that increased legal hunting is associated with a substantial increase in mortality rates due to other causes, including illegal killing.  Simple models that begin with the reported initial population size, allow it to grow at empirically measured rates (one high and one low) and subtracting out the number of legal and additional mortalities shows that the policy is likely to cause a substantial decrease in population size in a single year.

The strength of this paper is that the results just outlined are presented in a succinct and convincing manner.  I find that the data are used appropriately, the methods and assumptions are stated clearly, and the inferences are supported by the data.  One caveat is that the expected population sizes from these models are indeed expected, rather than demonstrated, but this could be viewed also as a strength, because the expectation (and its logic) is laid out proactively and can be tested.

The weakness of this paper is in the framing, which is often presented in language that I feel is likely to be inflammatory in a way that is not necessary, and indeed may interfere with productive evaluation of competing hypotheses about the mechanisms underlying the policies that are being evaluated.  Even in cases in which I personally agreed with a sentiment being expressed (implicitly or explicitly), I often felt it might be framed in a more even-handed manner.
Experimental design: The central question is well defined, and the methods used to assess the hypothesis were appropriate.  This work addresses an important knowledge gap, about the consequences for population dynamics of an abrupt change from full protection under the Endangered Species Act to high levels of legal recreational killing.
Validity of the findings: The underlying data are presented clearly and explained appropriately.  The analysis is clear and appropriate.  The conclusions are stated clearly and summarized appropriately in the Tables and Figures.
Additional comments: This paper analyzes existing data on the size of the Wisconsin wolf population, reported offtake by human hunters, and rates of disappearance of radiocollared wolves.  These data are combined in a very simple population model to assess whether the policies implemented by wildlife management agencies and the state government after delisting from the ESA are likely to achieve the stated goal of keeping population size constant.  The authors use cumulative incidence functions fit to disappearance rates of  radiocollared wolves in periods with and without hunting to demonstrate that increased legal hunting is associated with a substantial increase in mortality rates due to other causes, including illegal killing.  Simple models that begin with the reported initial population size, allow it to grow at empirically measured rates (one high and one low) and subtracting out the number of legal and additional mortalities shows that the policy is likely to cause a substantial decrease in population size in a single year.

The strength of this paper is that the results just outlined are presented in a succinct and convincing manner.  I find that the data are used appropriately, the methods and assumptions are stated clearly, and the inferences are supported by the data.  One caveat is that the expected population sizes from these models are indeed expected, rather than demonstrated, but this could be viewed also as a strength, because the expectation (and its logic) is laid out proactively and can be tested.
The weakness of this paper is in the framing, which is often presented in language that I feel is likely to be inflammatory in a way that is not necessary, and indeed may interfere with productive evaluation of competing hypotheses about the mechanisms underlying the policies that are being evaluated.  Even in cases in which I personally agreed with a sentiment being expressed (implicitly or explicitly), I often felt it might be framed in a more even-handed manner.

 For example line 280 criticizes “the vagaries of opaque electoral politics and patronage lavished upon interest groups” to conclude that “the history of political scapegoating of wolves will likely repeat itself”.  Line 247 states that “only a government that values evidence as a way to inform decisions would heed scientists. Although we have stressed the conservative estimation procedure we used, we fear decision-makers will not heed the cautions.”  In these passages, the intent of actions by others is being stated as though it was known, as are the values on which the actions were based, along with the likelihood of future actions.  I think it would probably be better to avoid statements about intentions and values in the context of the simple population data that were analyzed here.

I find the Methods and Results sections well written and without this problem, but I would suggest revision of the material outside lines 109-242 to allow the analysis and results to stand more simply on its merits.  I agree with the authors that there are many individuals with strongly held biases in this matter.  I think this analysis makes an important contribution to the assessment of policy actions taken rapidly after delisting to allow (or even promote) high levels of human caused mortality.  I am nonetheless concerned with the framing.