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: UNIVERSALITY VS EXPERIENCE: A CROSS-CULTURAL PILOT STUDY ON THE CONSONANCE EFFECT IN MUSIC AT DIFFERENT ALTITUDES
Review round: 1
Reviewer: 1

Output:
Basic reporting: The language is generally clear, though there are various passage where the paper could benefit from some careful proofreading. Examples:
•	Abstract: ‘consisting in’ should be ‘consisting of’
•	L 66.: ‘infants prefer listen’ should be ‘infants prefer to listen’
•	L 125: ‘(see(Prete et al., 2015))’ should be ‘(see Prete et al., 2015)’
•	Throughout the manuscript, the authors use the word ‘exposition’ when really they mean ‘exposure’. These two words have different meanings.

The article is structured appropriately and has appropriate figures and tables. It is generally self-contained with relevant results to the hypotheses.

I’m happy to see that the authors have shared their data. Ideally, though, this data would include stimulus-level responses, rather than just participant-level condition means. It would be a shame if this information is no longer available.

The theoretical background to consonance perception is not explained well. In particular, the description of how “harmony” may contribute to perceptual consonance is very unclear. It would be better to use the term “harmonicity”, because “harmony” has a special meaning in Western music theory that is different to how the authors use the word. It may also be worth mentioning the potential role of interference between partials (or roughness, or beating) in consonance perception. For a theoretical review, see the recent article “Simultaneous consonance in music perception and composition” by Harrison and Pearce.

To the extent that consonance perception is culturally determined, we would expect the consonance judgements of the Nepalese to reflect the distributions of pitch content in their musical style. However, the authors do not provide any account of the pitch structure of Nepalese music, or explain the way in which it deviates from Western music. This kind of detail is essential background for understanding the nature of consonance perception in Nepalese listeners, especially if the authors subscribe to a cultural account of consonance perception.

It is useful that the authors thought of hypoxia as a potential moderator of consonance judgements in this study. It would be sufficient to present this as a control analysis rather than a primary research question. However, given that the current paper considers hypoxia to be a primary research question, there needs to be more theoretical motivation in the introduction for why hypoxia might influence consonance judgements. It is currently not clear to me why one would anticipate that hypoxia would affect the relative consonance of musical chords. This discussion could be grounded in the three theories of consonance (harmonicity, interference, and culture) discussed earlier.

There are a couple of other historic cross-cultural studies of consonance perception that should be mentioned in the introduction:

> Butler,  J.  W.,  &  Daston,  P.  G.  (1968). Musical  consonance  as  musical preference: A cross-cultural study. The Journal of General Psychology, 79, 129-142.

> Maher, T. F. (1976). “Need for resolution” ratings for harmonic musical intervals:  A  comparison  between Indians and Canadians. Journal of Cross-Cultural Psychology, 7, 259-276.

The bar plots should be adjusted such that the lower tails of the error bars are visible.

The mediation analyses are a nice touch, but in other senses the statistical analyses and plots deserve to be extended. It would be useful to have a plot showing consonance judgements split by stimulus and by participant group, as in McDermott et al. (2016). This would help the reader to better understand the nature of the participants’ consonance judgements. It would be useful to replace the bar plots with figures that include raw data points, such that the reader can better understand the variability in the sample. It would also be useful to modify these was plots so that they reflect the repeated measures structure of the design, for example by plotting the difference between consonant and dissonant scores rather than plotting consonant and dissonant scores separately.
Experimental design: The research falls within the Aims and Scope of the journal. The research question is well defined and meaningful, and it is stated how the research fills an identified gap.

The methods are generally described in good detail. However, it is important for the reader to understand what chords were used in the stimuli. It is not sufficient to refer to another paper for this information, it should rather be included in the Methods.

One explanation for the lack of consonance perception in the highlanders could be that they did not understand the pleasantness judgement task. How did you ensure that the participants understood the task? Ideally there would be a control experiment where the participants demonstrated the ability to give reliable pleasantness judgements for a different kind of stimulus. Without such a control it is difficult to be confident in the present findings.
Validity of the findings: The participant groups in the study have sizes of six (Italians), six (Nepalese), five (lowland Sherpa), and five (highland Sherpa). As noted by the authors, it is harder to achieve large sample sizes with cross-cultural participant groups. Recruitment difficulties notwithstanding, it is very difficult to be confident in group difference findings when the groups number as low as five participants each, especially since the paper includes analyses at the individual-differences level. In particular, in the mediation analyses we see non-significant effects with nonetheless large point estimates, indicating that the statistical power is too low to identify effect sizes to any reliability. For comparison, the McDermott et al. (2016) study cited by the authors used sample groups of 64 (Study 1) and 50 (Study 2) Tsimane participants. In this sense the present study resembles more of a pilot than a final experiment. Indeed, the authors describe the study as a ‘pilot study’ in the title, but don’t mention the word ‘pilot’ elsewhere in the manuscript, and don’t hold back from describing themselves as being “quite confident about the validity of our conclusion.” If this paper really were documentation of a pilot study, then it would be useful to hear much more about what the authors learned from the piloting experience, and how they would conduct an improved study in the future. As it is, it is unclear that the paper represents an appropriate ‘unit of publication’ for PeerJ.
Additional comments: I reviewed an earlier version of the manuscript for another journal, and made many of the same comments provided above. While the authors did respond to some of the minor points I made in that previous review (e.g. adding ‘pilot’ to the title, fixing some typos, improving some unidiomatic English), they seem not to have responded substantively to any of the major comments. As a result, much of the above text is duplicated from my original review. I hope that this time the authors invest the time to improve the manuscript with the feedback from myself and the other 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: UNIVERSALITY VS EXPERIENCE: A CROSS-CULTURAL PILOT STUDY ON THE CONSONANCE EFFECT IN MUSIC AT DIFFERENT ALTITUDES
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors investigated the “consonance effect”, consisting in higher pleasantness judgments for consonant compared to dissonant chords – according to the Western definition of music in Italian and Himalayan participants. They were asked to express pleasantness judgments for consonant and dissonant chords at 1450 m and at 4750 m of altitude, with the aim to evaluate the effect of hypoxia on this task. Moreover, they considered the total mood disturbance as potential mediating factor.
Although I did some works on music, I do not consider myself expert of it, but I have more experience on cognitive processes, exercise and mood also in extreme conditions, so my feedback will be more on these aspects.
In my opinion it is interesting what the authors found in terms of TMD. I expected that between the first and second measurements there would be an increase (and not a decrease) of the total mood disturbance, because of the fatigue/stress/physical efforts (Di Corrado et al., 2014; Dupuy et al., 2014; Murgia et al., 2016). However, I agree with the interpretation of the authors that the concern before climbing vs. the gratification of after it might be an important factor, determining the results they observed.
Although there would be room for a large debate, the study by Shukitt-Hale & Lieberman (1996) – cited in the discussion of the present manuscript – indicates that mood states were adversely affected by duration and level of altitude, but with a main role of physical exertion. In this regard, the study by Murgia et al. (2016) indicates that physical exertion alone (after a multi-stage cycling race, thus independently from the altitude hypoxia) can significantly increase the TMD. Thus, it is possible to speculate that the participants of this study were not particularly affected by physical exertion and that hypoxia per se does not necessarily increase the mood disturbance.
I suggest authors to further work on this part of the discussion, including the observations above.

Minor points
There are some typos/errors to fix (e.g. “evidence” is an uncountable word and its plural is not allowed). I recommend an English proofreading before resubmission.

References
Di Corrado, D., Agostini, T., Bonifazi, M., & Perciavalle, V. (2014). Changes in mood states and salivary cortisol levels following two months of training in elite female water polo players. Molecular Medicine Reports, 9(6), 2441–2446.
Dupuy, O., Lussier, M., Fraser, S., Bherer, L., Audiffren, M., & Bosquet, L. (2014). Effect of overreaching on cognitive performance and related cardiac autonomic control. Scandinavian Journal of Medicine & Science in Sports, 24(1), 234–242. https://doi.org/10.1111/j.1600-0838.2012.01465.x
Murgia, M., Forzini, F., Filho, E., DI Fronso, S., Sors, F., Bertollo, M., & Agostini, T. (2016). How do mood states change in a multi-stage cycling competition? Comparing high and low performers. The Journal of Sports Medicine and Physical Fitness, 56(3), 336–342.
Experimental design: The research question are well defined, relevant & meaningful. The investigation has been conducted rigorously. Methods have be described with sufficient information to be reproducible by another investigator.
Validity of the findings: The data are robust, statistically sound, and controlled. As stated in the Basic reporting, authors should extend the discuss of TMD results.
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: UNIVERSALITY VS EXPERIENCE: A CROSS-CULTURAL PILOT STUDY ON THE CONSONANCE EFFECT IN MUSIC AT DIFFERENT ALTITUDES
Review round: 1
Reviewer: 3

Output:
Basic reporting: ********
For easier-to-read formatting of this review text, please use the Word document
********
1.	BASIC REPORTING
1.1.	Clear and unambiguous, professional English used throughout.
There is redundancy in how the task is described, in the Stimuli And Procedure section:
<STARTQUOTE> participants were instructed that very brief audio clips would be presented one at time, and that they will be required to express a pleasantness judgment
..
The participant was asked to listen to each stimulus and to express a pleasantness judgment
<ENDQUOTE>

At line 126, the same information seems to be repeated unnecessarily, within the span of a few lines:
<STARTQUOTE> an Italian sample was tested both at lower and higher altitudes with the aim to evaluate the possible effect of hypoxia at high altitude. Moreover, a mood questionnaire was also administered to this sample, in order to investigate the effect of hypoxia also on this variable. The second sample was Nepalese, with no or little exposition to Western culture and music. As for the Italian group, also this sample was tested both at lower and higher altitude
<ENDQUOTE>

Line 74:
<STARTQUOTE> the comparison among music preferences expressed by individuals belonging to very different cultures and living in countries very far apart from each other can help in shedding light on this topic
<ENDQUOTE>
This should probably not be a hard logical "and" here: I don’t think that countries need to be "very far apart" from one another to manifest widely different musical cultures/preferences that would be relevant in such a study. This is the case for instance with remote indigenous tribes of the Amazon investigated in previous studies (and perhaps also with the Nepalese communities investigated here, and described in the next passage), that despite being – on the map – relatively close to one another (and close also even to Western-permeated communities of S. America, e.g. to urban Bolivia) are nonetheless very different in their musical traditions.

Finally, the "Aim" section goes into quite a lot of detail about the design and experimental manipulations, detail that is probably better suited (and that is indeed repeated) in the Methods section. The full design is not clear from the "Aim" section, which for instance does not specify whether the Nepalese sample were native of – and were tested at – low or high altitude; this is only clarified later in the Methods.
1.1.1	~Persistent confusions of terminology
Despite the subtle and complex relationship between the notions of 'pleasantness' and 'consonance' having been explained earlier, and despite this relationship being crucial for this paper, the former term seems to be used inter-changeably with the latter on multiple instances, alongside other several inaccurate usages of these and adjacent terms. I list some of those instances below.

<STARTQUOTE> Some evidences suggested music pleasantness to be a universal principle, independent from the cultural environment: for instance, a preference for consonance was described even in different species,(..)
<ENDQUOTE>
I think this leads to confusion and to a re-ambiguation (instead of a disambiguation) of terms. A subjective feature of music (pleasantness) cannot possibly constitute a universal principle. Furthermore, this wording also seems to ignore the fact that musical pleasantness can in fact arise out of multiple musical features, some not even in the pitch domain, e.g. groove (Witek et al., 2014).

This same confusion continues at line 69:
<STARTQUOTE>  (..) suggesting that music pleasantness might not be universal, developing instead with experience and listening habits, namely on culture (McDermott et al., 2016).
<ENDQUOTE>
The cited McDermott paper is about whether or not consonance preference – not musical pleasantness preference – is universal. Again, since these are notions central to the manuscript, I think the effort would be worth making to more clearly use each term in the right context.

The set of inter-changeably used terms includes not just 'pleasantness' and 'consonance' but also the general term 'music preference'. For instance, in the first paragraph of the Discussion, it is stated that
<STARTQUOTE> This first result would exclude the possibility that music preference is innate and biologically determined, because if this hypothesis were true we should have found a similar preference in all the samples tested.
<ENDQUOTE>
Again, I see no reason why consonance preference (which is the only type of preference tested here) would be expressed in such generic terms here, as "music preference". On the other hand, it is warranted to describe musical preference in general (with all of its facets) later in the conclusions, as the authors indeed do:
<STARTQUOTE> we could consider these results as a further evidence in favor of the crucial role of music exposition in music preference
<ENDQUOTE>

Cf line 111:
<STARTQUOTE> there is still a lack of results regarding the effect of altitude hypoxia on music pleasantness.
<ENDQUOTE>
Presumably it is the effect on preferences related to musical pleasantness that is intended here!

Cf line 355:
<STARTQUOTE> our results showed that neither pleasantness, nor consonance-dissonance difference, were influenced by altitude
<ENDQUOTE>
Once again, this is very confusing: isn't the consonance-dissonance difference computed in terms of pleasantness ratings? Then why list this as two findings when it is really just one?

1.2.	Literature references, sufficient field background/context provided.
Adequate.

1.3.	Professional article structure, figures, tables. Raw data shared.
Could the authors please also include the trial-by-trial data in the raw XLS file, and not just the subject-wise averages? Many thanks.

Figures:
•	Fig1
o	the figure caption reads "Measurements at low altitude at altitude and high altitude (ML, MA and MAH)". The Methods section only explains two of those: "lower and higher altitudes (respectively ML and MHA)". Presumably MA stands, then, perhaps for middle altitude (?!), however this abbreviation system does not seem to make sense, and is not consistent.
o	the x axis contains MHA instead of MAH as in the caption
o	it's not explained in the caption or elsewhere what the gray data points represent (e.g. corresponding to the "KJN base")
o	the stick figure icon seems to go with triangular data points to indicate measurements in the middle of the time series plot, but not also at the beginning (second data point), where the data point is circular
•	Figs 4 and 5
o	the values on the y axis are probably better displayed in 0.5 rather than 0.3 increments

1.4.	Self-contained with relevant results to hypotheses.
Cf line 54:
<STARTQUOTE> if harmony is the base for the consonance effect, how are consonant and dissonant chords evaluated in those cultures in which harmony is not present?
<ENDQUOTE>
Again, terms are being conflated here. If 'harmony' is to be used, then it would be worth giving at least a short clarification of what this term really taken to mean, in Western music. The sensory dissonance/consonance phenomenon of focus in this study is merely the 'vertical' dimension of harmony (i.e. concerning isolated chord presentations). However, a hierarchically-higher 'horizontal' dimension also exists (not of interest here), where the key notions of voice-leading, counterpoint and diatonic function are brought into the equation. The apparent 'paradox' of people liking dissonance then is simply – and elegantly – explained in this wider framework of harmony, as the simultaneous existence of a purely-negative perceptual connotation, stemming from the vertical/acoustical dimension (essentially: beating and roughness); with a further dimension provided by the horizontal dimension of harmony. This dimension (essentially: Western music conventions) is able to use this dissonance (in addition to other mechanisms) to create and control musical tension.
Relevant references from the music cognition literature, that may well be already known to the authors, should they wish to make this acknowledgement, are (Huron, 2001; Krumhansl, 1990; Terhardt, 1984; Tramo et al., 2001); and perhaps (Aldwell et al., 2010) on the music-theoretical side.

Also, see section 3.2.1 below.
Experimental design: 2.	EXPERIMENTAL DESIGN
2.1.	Original primary research within Aims and Scope of the journal.
Adequate.
2.2.	Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap.
2.2.1	~Hypotheses
<STARTQUOTE> Concerning altitude, due to the absence of previous evidence in this regard, we did not have specific expectations on the effect of hypoxia on music pleasantness evaluation
<ENDQUOTE>
Although too little may be known about the effects of altitude – namely high-altitude-induced hypoxia – on musical preferences to formulate a hypothesis, this factor was nonetheless of sufficient prior interest to investigate and manipulate, thus it would be good if this reasoning were more deeply thought-through. After all, this is presumably what this study aims to bring, over and above similar previous ones such as (McDermott et al., 2016) - see section 4.1 below. For instance, perhaps a link can be made to what prompted this factor to even be considered, as hinted at line 109:
<STARTQUOTE> It is well known that altitude hypoxia negatively affects some aspects of physiology and, specifically, sensory systems (Cingi, Erkan & Rettinger, 2010; Jha, 2012; Ruffini et al., 2015); however, to our knowledge, there is still a lack of results regarding the effect of altitude hypoxia on music pleasantness
<ENDQUOTE>

Although obviously no good in terms of formulating a hypothesis at this stage (which would be mere HARKing), perhaps a slightly more extended discussion starting from what is known about the effect of hypoxia on sensory systems in general would be warranted (perhaps from the wider " Environmentally-modulated metabolic.." project). Currently, this is only briefly touched upon in the Discussion:
<STARTQUOTE> Even if we did not have specific hypotheses in this regard, we also considered altitude hypoxia as a possible influencing variable, because of the well-known affecting role of such condition in sensory systems and cognitive performance (Cingi et al., 2010; Jha, 2012; Limmer & Platen, 2018; Ruffini et al., 2015), but our results showed that (..).
<ENDQUOTE>
2.3.	Rigorous investigation performed to a high technical & ethical standard.
Unfortunately, there are multiple issues here - see subsections below.
2.3.1	~Design/factors
Among the limitations typical of field studies in extreme environments, the authors cite: the difficulty in testing participants belonging to remote cultures, testing having been done at different altitudes, and the absence of control tasks. It would however be good to acknowledge that a further limitation of the study (whether one typical of such field studies, or simply a limitation of this specific study) is that the full factorial design was not tested, for instance that the low-altitude Sherpas were not also tested at high altitude (and vice-versa).

In the same vein, something that also strikes the reader as an arbitrary element of "missing data", is the fact that only the Italian group was measured on the Profile Of Mood State (POMS), with subsequent analyses of the effect of altitude on TMD; whereas it seems to have made sense equally to collect this data also for the other two native-low-altitude groups (She_L and Nep_L - see table below)! As suggested elsewhere, perhaps this can also be explained as a limitation typical of expedition/trekking studies.

The (incomplete) mixed design employed might be more clearly laid out in a table that clarifies the relevant conditions, manipulations and factor. This table might look something like:
	Western music familiar
	yes	no
Altitude	L	She_L
Ita_L	Nep_L
	H	Ita_L!	Nep_L!
She_H
where:
- _L or _H denotes that the group's native altitude is low or high respectively;
- "!" refers to testing done in conditions of altitude different to any one group's native altitude.

Clarifying the design in this way would more clearly highlight which comparisons were within subjects (i.e. effect of altitude-change from low to high) vs between subjects (i.e. effect of low vs high native altitude), even if the effect of the Western factor (unlike Altitude) is in fact tested not via factorial ANOVA but via mediation analysis.

The providedTable 1 does not clarify these essential design parameters. Also, this table is, to my eyes, not necessarily suitable for the main article text, since the full raw-data table (with extra data on ratings) is available, and since variables such as "foreign language spoken" are not at all essential to the study. This Table also has some missing data (e.g. in the Y/Mus column).

I find it odd that, although She_L and She_H are two different groups, the authors chose to describe their overall sample as consisting of three "groups": Italians, Nepalese and Sherpa, with 2 "subgroups" in the latter group, on native altitude grounds. Perhaps clarify that both of the two 'subgroups' and the two 'groups' are in fact on the same hierarchical level, and together form the levels of your Ethnicity factor (as used in some of the ANOVAs).

Cf line 130:
<STARTQUOTE> As for the Italian group, also this sample was tested both at lower and higher altitude, in order to evaluate the possible interaction between culture and hypoxia on the consonance effect.
<ENDQUOTE>
I think it would be good to clarify whether hypoxia is a temporary 'acute' state (that low-altitude-native populations are susceptible to), or a 'chronic' state (typical of high-altitude-native populations).
As I understand (e.g. from here), hypoxia refers by definition to the former, i.e. the effect of high altitude upon an individual NOT habituated to high altitudes. If that’s true, then comparing the performance of a low-altitude-native group at low and at high altitude is not qualitatively the same as comparing a low- vs a high-altitude-native group, as hypoxia (as construed above) only affects the low-altitude natives. Thus, simply distinguishing the statistical nature of each comparison (Altitude being a within- and between subjects factor, respectively), I feel it is a problem that the NativeAltitude factor (yes/no) is nowhere modelled in this.

A further unrelated (and major) problem is that, according to the XLS file provided, the SherpaLow and SherpaHigh groups seem to not be homogeneous in terms of being familiar/not with W. music (one participant who gave the opposite response, in each group); and the same for the Nepalese group.
Despite this, the "familiarity with Western music" factor seems to be defined and tested categorically, along precisely the lines of group membership, as if this variable was homogeneous within each group. This is even more of a problem given the very low sample size (N=5 for She_L).
Thus, the two categorical factors are haplessly confounded, and only the first Mediation Analysis, which uses the binary variable of familiarity with Western music, with no group attribution is reliable! However, that analysis seems to not be considered central by the authors (Fig3 only shows the second Mediation Analysis), and instead the main conclusion that consonance preference is cultural, is primarily based on the ANOVAs, where the Western factor is, as I explain, imperfectly coterminous with group membership as used as a factor.

Related, while the difficulty of finding participants in remote groups can perhaps explain the non-homogeneity (in terms of familiarity with W. music) in the Sherpa group, perhaps the authors can explain why this was also the case in the Nepalese group, where presumably (?) those difficulties did not apply.
2.3.2	~Analyses
<STARTQUOTE> Then, by means of two mixed repeated-measures ANOVAs, Ethnicity (Italian, Nepalese, Sherpa) was considered as a between-subjects factor, and Chord (consonant, dissonant) was used as a within-subject factor. Pleasantness evaluations recorded at the lower altitude were used as the dependent variable in the first ANOVA, and those recorded at the higher altitude were used in the second ANOVA.
<ENDQUOTE>
This ANOVA would be described as a mixed-model ANOVA, as opposed to a "mixed repeated-measures" ANOVA, as it has both a within- and a between-subjects factor.

<STARTQUOTE> In all ANOVAs post-hoc tests were carried out by means of Duncan test.
<ENDQUOTE>
Does this multiple-comparisons-correction measure also correct for the fact that – due to the incompleteness of the native altitude X tested altitude X ethnicity X Western music familiarity design matrix (see above) – you have had to use four separate ANOVAs? If not, I would suggest to mention this as a statistical limitation, in the same place where the low number of subjects is mentioned; and explain perhaps that both were consequences of the difficulty of getting (enough) subjects in each cell of the design.
The low sample size limitation might also be linked/qualified in terms of the word "pilot" that appears only in the title but nowhere discussed/qualified in the paper.
2.3.3	~Stimuli
It would be good if the 24 chords were made a part of the Supplementary Materials – in both music notation as well as waveform/MP3 format. Otherwise, it is very difficult for readers to imagine what those chords sounded like, in order to judge the amount of dissonance in each for themselves. Prete et al. 2015's SM does not seem to hold this information either.

Also, the number of unique chord types (that is, disregarding transpositions) used as exemplars here across the consonant and dissonant categories, was N=6. This makes for little variability within each category, inasmuch as this variability is used for inference (generalisation). This is at least the case in comparison to the variability found in other similar studies, i.e. behavioural studies, based on single-chord presentations (chords of 3 notes or more), with definitions of dissonance not based on harmonicity.
Such studies used stimulus sets with e.g. N=15 chords (Lahdelma & Eerola, 2016), N=28 chords (Park et al., 2011), N=80 chords (Popescu et al., 2019), etc. Outliers in this sense can be found in the literature, e.g. N=3 stimuli (Foo et al., 2016), although these are generally fMRI studies, with the corresponding limitations.
Still, this low variability should, in my opinion, be acknowledged as a limitation, and perhaps be again linked to the limitations that existed upon the total testing time.
2.4.	Methods described with sufficient detail & information to replicate.
Adequate.
Validity of the findings: 3.	VALIDITY OF THE FINDINGS
3.1.	Meaningful replication encouraged where rationale & benefit to literature is clearly stated.
The hypoxia hypothesis not being clearly formulated (see section 3.2.1) is particularly problematic since the study of the effect of hypoxia seems to be this study's main addition, above and beyond McDermott 2016, as indeed stated in the Discussion at line 384:
<STARTQUOTE> Compared to [McDermott's (2016)]  findings, the present study adds the evidence of null effects of other demographic information on music preference, as well as it shows a null effect of hypobaric hypoxia on the pleasantness of music.
<ENDQUOTE>

3.2.	All underlying data have been provided; they are robust, statistically sound, & controlled.
<STARTQUOTE> the difference of pleasantness evaluation between consonant and dissonant chords was also calculated, obtaining an overall measure in which negative scores corresponded to unpleasant judgments, positive scores corresponded to pleasant judgments and a score of 0 corresponded to a neutral judgment (neither pleasant nor unpleasant).
<ENDQUOTE>
I find this description utterly confusing, and not at all matching with the fact that each pleasantness judgment was rated on a 4 point-Likert scale. I expected to read that negative scores corresponded to a dissonance preference, and positive to a consonance preference, but this is not what this text states!
Furthermore, this description confusingly suggests that the consonant/dissonant manipulation was done within-chord (manipulations upon a single starting/baseline chord), whereas that is not how the Stimuli section describes them – these were just different chords. Thus, I don’t see how average pleasantness ratings that are equal between Cons and Diss chords amount to a "neutral judgement", when at most this can be described as equal preferences (e.g. the subject likes both Cons and Diss chords).
3.3.	Conclusions are well stated, linked to original research question & limited to supporting results.
The statistics are reported thoroughly and completely. However, as per my observations for the incompleteness of the factorial design, several aspects of the four ANOVAs strike me as odd:
- The first two ANOVAs use partly overlapping groups (Ita, Nep), whereas the Sherpa groups are different between the first and second ANOVAs. The statistics do not "know" (i.e. model) this, and so the ANOVA assumptions regarding the variance within each population are not met. Thus, I do not think this kind of analysis allows drawing results that pool from across this dataset, but instead imposes they be treated as independent results.
- Fig 4 is meant to show the 3-way ANOVA (with altitude included as a factor), but only the Ethnicity and Chord factors are shown in the bar plot. Data should also be broken down by Altitude, even if there is no sign. effect of it.
- The result of the first two ANOVAs are nowhere plotted; perhaps these could at least go into the SupplMat?
- You say:
<STARTQUOTE> Furthermore, consonant chords were judged as more pleasant by lowlanders than by highlanders
<ENDQUOTE>
But I thought the relevant DV was meant to be the difference, upon which what you call the "consonance effect" is based; thus, reporting this effect on the raw Consonance DV in isolation seems to me rather opportunistic, or at best a posthoc side-track.

Also, the second mediation analysis (Fig 3) used both recorded variables (Consonance, Dissonance) as well as their difference, as dependent variables; whereas the significant effect in the first mediation analysis is reported only for the Difference measure – although presumably once again all three were tried as DVs?...

The first paragraph of the Discussion states
<STARTQUOTE> the effect was completely absent in highlander Sherpas, the subsample who declared to listen to music less frequently and, importantly, not to listen to Western music. This latter result is the most important of the present study, because it shows that music exposition is the key factor determining the consonance effect:
<ENDQUOTE>
Whereas in most of the paper, control for enculturation as a possible cause of consonance preference is done by the variable of exposure to Western music, now the authors unexpectedly also throw in the variable of amount of music (of any kind, not just Western) listened to, the "Music/day (hours)" variable in the XLS.
The reasoning behind mixing notions like this is to me, again, incomprehensible: although it can be assumed that any non-Western music remaining in those listening habits is probably not consonance-heavy, there is no assurance of this at all. But even if this assumption is correct, then surely even frequent listening of this type of music should still lead to an absent consonance effect, right? And if this argument is correct, then likewise low values of music listening (as the highlander Sherpas report) should not be taken as proof that consonance preference is culturally-based; only their reported low familiarity with Western music should be taken as such proof - but there, we have the issue presented above with non-homogeneity in the She_H and Nep groups in terms of this variable.

<STARTQUOTE> In the present study we also explored the possible effect of some demographic features on the consonance effect, such as age, schooling and playing music: the results revealed that none of these variables influences music pleasantness, stressing that the exposure to a particular kind of music is the only variable responsible for the consonance effect.
<ENDQUOTE>
Since not ALL demographic variables were tested, but only a select few, I think the conclusion needs to be toned down, by qualifying the term "only", e.g. to say that exposure is likely the most salient predictor.

<STARTQUOTE> Only the mood was influenced by hypoxia, with the Italian sample revealing lower mood disturbance scores (index of tension, depression, anger, fatigue and confusion) at higher compared to lower altitudes. We speculate that this difference could be due to the fact that once climbed the mountain, the more positive mood can be due to a kind of gratification compared to the possible concern before the climb. This result agrees with the findings of Karinen and Tuomisto, who reported that during a prolonged expedition in Everest mountain well-motivated participants were capable to maintain a good mood state (Karinen & Tuomisto, 2017); on the contrary, Stavrou and colleagues, who reported that hypoxia was capable to negatively affect the mood, exacerbating the effect of bed rest and ambulatory confinement (Stavrou et al., 2018). Other evidences linked mood and altitude hypoxia, affirming that mood states were adversely affected by duration and level of altitude, but with a main role of physical exertion (Shukitt-Hale & Lieberman, 1996). Heinrich and colleagues recently affirmed that acclimatization plays a big role in cognitive and mood alterations during a sojourn at altitude (Heinrich et al., 2019). To be noted that none of our participants suffered remarkable altitude sickness symptoms. Thus, from these findings and from our results, we support the idea that hypoxia may increase the mood disturbance during non-rewarding conditions or in association with altitude sickness symptoms, but that hypoxia per se, or hypoxia during motivating conditions does not promote mood disturbances.
<ENDQUOTE>
I fail to see how this unexpected – and extended – discussion of the effects of altitude on mood is relevant in the Discussion of a paper on the universality of consonance preference, especially when the argument does not come back to this focal topic.

<STARTQUOTE> the fact that our findings strictly resemble those recently described by McDermott and colleagues allows us to be quite confident about the validity of our conclusion
<ENDQUOTE>
The conclusion of the present study indeed rest heavily on McDermott's 2016 study, in supporting the conclusion that consonance preference is cultural and not biological. To note, however, is that said McDermott study has been heavily criticised methodologically, e.g. by Bowling et al. (2017), Zatorre (2016) and others. This might or might not convince the authors to reconsider the degree to which their results (themselves qualified, for what it's worth, by the criticisms in this review), together with the McDermott result, really constitute strong evidence for the stated hypothesis. Might a more ample/general framework be more suited, e.g. Cross' proposal of music as a “biocultural phenomenon” (2003), at least until the literature has some more definitive results on the biology/culture question?

<STARTQUOTE> in a cross-cultural study with Western and African (Cameroon) participants, Fritz and colleagues (Fritz et al., 2009) found no group differences in judging both emotional content (happy, sad, fearful) and pleasantness of Western and African music, suggesting a biological basis of music processing, independent from the culture.
<ENDQUOTE>
It seems to me the authors only incompletely acknowledge the arguments in favour of the opposite view. Stated as above, the reader might remain unconvinced as to the side taken by the authors. This is because the Fritz et al. study actually concluded that "sensory dissonance [seems to] universally influence the perceived pleasantness of music". Thus, if this study is cited as proof for a biological basis of music pleasantness, it seems that, for authors to retain a conviction in the cultural basis hypothesis, some further discussion is necessary!
Additional comments: 4.	GENERAL COMMENTS
********
For easier-to-read formatting of this review text, please use the Word document
********
In general, I found the premise of the study justified. The highlighted need for cross-cultural studies, and the attempt to accrue more evidence to further address the nature-nurture question in music cognition (with regards to consonance preference, and perhaps more generally), are all valid. As a behavioural study on consonance preference across cultures, what this study – part of a wider empirical project to study aspects of human physiology across various conditions – is certainly a very welcome addition to the literature.

However, as you will read in the sections below, I am afraid I also found the manuscript unsatisfactory in several crucial respects, to do with the methodology, hypotheses, clarity of writing, and coherence of reasoning. I do think that at least the most substantive of my comments would have to somehow be addressed before considering publication, which I hope that the Authors will consider doing (and hope that the Editor will allow the revision). My recommendation is essentially "accept conditional upon satisfactory major revisions".

I welcome further discussion and would be happy to review a revised version of the manuscript. If the paper is eventually accepted (which I hope it is, assuming satisfactory revisions), then I would very much hope and encourage the authors to agree to publish the review history, in order to continue (what I regard to be) the good policy of open peer review practiced by PeerJ (and other journals).

I have largely structured my comments under the suggested headings of the PeerJ review template. I have included some subheadings of my own to those, their names prefixed with a ~. When my comments pertain to a particular section of the text, I am quoting that section using
<STARTQUOTE> text formatted as such.
<ENDQUOTE>
I have tried to place comments inside the relevant subheadings as explained, rather than structuring them in terms of major/minor comments – hopefully it will be clear (also from this heading structure) which comments relate to 'deep structure' and which to 'form'. Please also note the annotations made in the PDF version of the manuscript, which contain predominantly minor/surface comments (e.g. suggestions re clarity of phrasing/wording etc.), that the authors may wish to also take into account.

*******************
5.	REFERENCES
Aldwell, E., Schachter, C., & Cadwallader, A. (2010). Harmony and Voice Leading. Cengage Learning.
Bowling, D. L., Hoeschele, M., Kamraan, Z. G., & Fitch, W. T. (2017). The Nature and Nurture of Musical Consonance. Music Perception, 35(1), 118–121.
Cingi, C., Erkan, A. N., & Rettinger, G. (2010). Ear, nose, and throat effects of high altitude. European Archives of Oto-Rhino-Laryngology: Official Journal of the European Federation of Oto-Rhino-Laryngological Societies (EUFOS): Affiliated with the German Society for Oto-Rhino-Laryngology - Head and Neck Surgery, 267(3), 467–471. https://doi.org/10.1007/s00405-009-1016-6
Cross, I. (2003). Music as a Biocultural Phenomenon. Annals of the New York Academy of Sciences, 999(1), 106–111. https://doi.org/10.1196/annals.1284.010
Foo, F., King-Stephens, D., Weber, P., Laxer, K., Parvizi, J., & Knight, R. T. (2016). Differential Processing of Consonance and Dissonance within the Human Superior Temporal Gyrus. Frontiers in Human Neuroscience, 10. https://doi.org/10.3389/fnhum.2016.00154
Huron, D. (2001). Tone and Voice: A Derivation of the Rules of Voice-Leading from Perceptual Principles. Music Perception, 19(1), 1–64. https://doi.org/10.1525/mp.2001.19.1.1
Jha, K. N. (2012). High Altitude and the Eye. Asia-Pacific Journal of Ophthalmology (Philadelphia, Pa.), 1(3), 166–169. https://doi.org/10.1097/APO.0b013e318253004e
Krumhansl, C. L. (1990). Cognitive foundations of musical pitch. Oxford University Press, USA.
Lahdelma, I., & Eerola, T. (2016). Mild Dissonance Preferred Over Consonance in Single Chord Perception. I-Perception, 7(3), 2041669516655812. https://doi.org/10.1177/2041669516655812
Limmer, M., & Platen, P. (2018). The influence of hypoxia and prolonged exercise on attentional performance at high and extreme altitudes: A pilot study. PLOS ONE, 13(10), e0205285. https://doi.org/10.1371/journal.pone.0205285
McDermott, J. H., Schultz, A. F., Undurraga, E. A., & Godoy, R. A. (2016). Indifference to dissonance in native Amazonians reveals cultural variation in music perception. Nature, 535(7613), 547–550. https://doi.org/10.1038/nature18635
Park, J. Y., Park, H., Kim, J., & Park, H.-J. (2011). Consonant chords stimulate higher EEG gamma activity than dissonant chords. Neuroscience Letters, 488(1), 101–105. https://doi.org/10.1016/j.neulet.2010.11.011
Popescu, T., Neuser, M. P., Neuwirth, M., Bravo, F., Mende, W., Boneh, O., Moss, F. C., & Rohrmeier, M. (2019). The pleasantness of sensory dissonance is mediated by musical style and expertise. Scientific Reports, 9(1), 1070. https://doi.org/10.1038/s41598-018-35873-8
Ruffini, R., Di Giulio, C., Verratti, V., Pokorski, M., Fanò-Illic, G., & Mazzatenta, A. (2015). Adaptation of olfactory threshold at high altitude. Advances in Experimental Medicine and Biology, 837, 19–22. https://doi.org/10.1007/5584_2014_70
Terhardt, E. (1984). The Concept of Musical Consonance: A Link Between Music and Psychoacoustics. Music Perception, 1(3). http://search.proquest.com/docview/1300644337/citation/33847C4B99D341EAPQ/1
Tramo, M. J., Cariani, P. A., Delgutte, B., & Braida, L. D. (2001). Neurobiological foundations for the theory of harmony in western tonal music. Annals of the New York Academy of Sciences, 930(1), 92–116.
Witek, M. A. G., Clarke, E. F., Wallentin, M., Kringelbach, M. L., & Vuust, P. (2014). Syncopation, Body-Movement and Pleasure in Groove Music. PLoS ONE, 9(4), e94446. https://doi.org/10.1371/journal.pone.0094446
Zatorre, R. (2016). Human perception: Amazon music. Nature, 535(7613), 496–497. https://doi.org/10.1038/nature18913


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: UNIVERSALITY VS EXPERIENCE: A CROSS-CULTURAL PILOT STUDY ON THE CONSONANCE EFFECT IN MUSIC AT DIFFERENT ALTITUDES
Review round: 2
Reviewer: 1

Output:
Basic reporting: This has been significantly improved since the last submission.
Experimental design: The methodological reporting has been improved, with useful information about the musical stimuli. One note is that the authors included the chord <C, E, A flat> in the 'consonant' stimulus set, but in Western music this chord (termed the augmented triad) is generally considered dissonant and inharmonic. This is not a huge problem for the validity of the finding, but in future studies the authors might consider instead using the consonance-dissonance stimuli of Josh McDermott's group, which fulfil a similar function to these stimuli but are more consistent with consonance judgements in Western music.
Validity of the findings: The authors have improved their commentary on their findings, making a better job of restricting their conclusions according to the methods employed and the results obtained.
Additional comments: The paper has been improved a lot, and now the main weakness is in the Introduction. Still, as commented by me and by Tudor Popescu, the theoretical account of consonance is somewhat incoherent. The paper really needs to provide a clear definition of consonance, and it needs to explain whether consonance is being treated here as being terminologically equivalent with pleasantness, as in the operationalisation of Josh McDermott's papers. I dislike the term 'consonance effect' - I have not seen it before literature, and it is problematic because it implies that the only valid form of consonance is the consonance system of Western music.

The definition of harmonicity is likewise incoherent. The authors write this:

> A harmonic chord (a set of notes played together) sounds as pleasant if it is consonant, and consonance is defined by the specific intervals among notes (Bowling & Purves, 2015). According to this view, in fact, consonance is associated with pleasantness, and dissonance with unpleasantness (the so-called “consonance effect”; Prete et al., 2015, 2019).

Here, it just seems like you're equating harmonicity with consonance and pleasantness. This is not useful. Here is how I think the terminology should be defined (not in these specific words, of course):

Consonance - a concept in Western music theory concerning how notes may be combined into chords. Consonance is associated with pleasantness, stability, and relaxation.

Pleasantness - this is how you operationalise consonance in the present study, consistent with many previous studies (e.g. Bowling et al., McDermott et al.) While pleasantness is only a part of the music-theoretic notion of consonance, it is the easiest to study with nonmusicians and nonWesterners.

Harmonicity - this is when the pitches in the chord are related by simple integer ratios of frequencies. It is a potential contributor to pleasantness and consonance, with this effect being possibly moderated by cultural exposure.

Interference - this is when the spectral components in a chord are close enough to elicit masking and beating. It is a potential contributor to pleasantness and consonance, with this effect being possibly moderated by cultural exposure.

I hope this is clear?

Minor comments:

l. 60: 'deepen' should be 'deeper'

l. 104-106: unclear, please clarify

l. 124: not sure what you mean by lacking modulation frequency

l. 125-126: the definition of tonal hierarchy is not very accurate, please revise (you can have a tonal hierarchy without notes deviating from the scale)

l. 252: pc should be PC

l. 411: complains should be complaints


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: UNIVERSALITY VS EXPERIENCE: A CROSS-CULTURAL PILOT STUDY ON THE CONSONANCE EFFECT IN MUSIC AT DIFFERENT ALTITUDES
Review round: 2
Reviewer: 2

Output:
Basic reporting: Fine.
Experimental design: Fine.
Validity of the findings: Fine.
Additional comments: The authors have satisfactorily fulfilled all my requests.


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: UNIVERSALITY VS EXPERIENCE: A CROSS-CULTURAL PILOT STUDY ON THE CONSONANCE EFFECT IN MUSIC AT DIFFERENT ALTITUDES
Review round: 2
Reviewer: 3

Output:
Basic reporting: Adequate.
Experimental design: Adequate.
Validity of the findings: Adequate.
Additional comments: I congratulate the authors for having thoroughly addressed my concerns, comments and suggestions. The text has been much improved and clarified, and so have the figures. I regard this revision as being suitable for publication.
As a small remaining point, I would still suggest to include the music notation (score) for the 24 chords, if available; I did not see this in the Supplemental Materials. If this is not available, the fact that at least the audio files are now included is certainly still helpful.


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: INTEGRATING POPULATION GENETIC STRUCTURE, MICROBIOME, AND PATHOGENS PRESENCE DATA IN DERMACENTOR VARIABILIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: Some word spelling, grammar and writing format should be checked throughout the manuscript. Specific points:
Line 387, “(Dergousoff and Chilton, 2011)” should be changed to “Dergousoff & Chilton, 2011” according to the requirement of the Journal.
Line 421, “Norther” should be changed to “Northern”
Line 265 and 285, please confirm the writing form.
Please unify the format of the titles at all levels, and carefully double checked the references of the manuscript.
Experimental design: no comment
Validity of the findings: no comment
Additional comments: Lado et al. studied the intergration of population genetic structure, microbiome, and pathogens presence data for Dermacentor variabilis. This research is of great significance for tick-borne diseases. And the following is my comments.

1. In this study, the reason for choosing Dermacentor variabilis and a brief introduction of this species should be given in the part of “Introduction”.

2. From the figure 1, the number and distribution of samples are relatively fewer. Are the selected sampling localities representative?

3. Whether need to consider the influence of sex and developmental stages in the present study. And why the authors choose to detect the Rickettsiales, not other pathogens?

4. Line 91-95, “In amphibians, Griffiths et al. (2018) showed that at the landscape level, the microbiome was not structured according to host population structure even though the genetic distance among hosts was still significantly correlated to microbial community dissimilarity when controlling for geographic distance.” This sentence is not very clear, please rewrite it.

5. Line 400-402, “Thus, the addition of microbiome, pathogen presence, host sex, and geographic location led to conclusions that are, in general, consistent with host genetics”. Host information or references were not mentioned in this part, and how is this conclusion drawn?


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: INTEGRATING POPULATION GENETIC STRUCTURE, MICROBIOME, AND PATHOGENS PRESENCE DATA IN DERMACENTOR VARIABILIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: There is no scientific question in this article. The authors should clearly define their objectives and their scientific question.
I strongly advice the authors to increase the number ot tick samples or focus their study on the tick microbiome analysis.
Experimental design: There is no scientific question in this article.
The number of samples is too low. Authors cannot reasonably perform population genetic and pathogen analysis on only 64 ticks collected in 12 different sates (tick pathogen analysis regularly performed on only one tick).
Validity of the findings: The number of samples is too low to perform this type of studies.
No negative controls for the tick microbiome analysis.
Additional comments: In this study, authors generated microbiome and pathogen presence data for Dermacentor variabilis, and integrated those data sets with population genetic data, and metadata for the same individual tick specimens. The total number of ticks collected in 12 states was 64 ticks. Clustering and multivariate statistical methods were used to combine, analyze, and visualize data sets. As mentioned by the authors, interpretation of the results is challenging, likely due to the low levels of genetic diversity and the high abundance of a few taxa in the microbiome, and to my opinion the too low number of samples. From the tick microbiome analysis, they found that Francisella was dominant in almost all ticks, regardless of geography or sex. Nevertheless, results showed that, overall, ticks from different geographic regions differ in their microbiome composition. Additionally, DNA of Rickettsia rhipicephali, R. montanensis, R. bellii, and Anaplasma spp., was detected in D. variabilis specimens. Finally, authors summarize their study only saying that « this is the first study that successfully generated microbiome, population genetics, and pathogen presence data from the same individual ticks, and that attempted to combine the different lines of evidence. »

Authors are right saying that interdisciplinary approaches are essential and required to improve our understanding of TBP ecology. The integration of these different approaches is the key to improve our understanding. However, I did not find this data integration in the paper and the authors do absoultely not answer and hypothesize to this potential link between tick population genetic, tick-borne pathogens and microbiomes. The objectives are unclear. No scientific questions emerge from this study. The number of samples is too low to perform this type of integrated studies and obtain accurate data for the analysis of tick genetic population and pathogen prevalences. At no time authors discuss about these limitations. In my opinion, this study cannot be considered for publication and I strongly advice the authors to increase the number ot tick samples or focus their study on the tick microbiome analysis.

Major :
- There is no scientific question in this article.
What is the scientific question in this study ? The authors mentioned L.36-38. « This is the first study that successfully generated microbiome, population genetics, and pathogen presence data from the same individual ticks, and that attempted to combine the different lines of evidence ». Combining different approaches in a study is not a scientific question. What is the goal of this study ? For each approach, authors have descriptive results but no links between these different results appear in the study. Authors are completly right when they say that « we still do not fully understand how host genetics and environmental factors interact to shape the microbiome of organisms, or how pathogenic microorganisms affect the microbiome and vice versa. They are still right when they say that « the integration of different lines of evidence may be the key to improve our understanding of TBDs ecology. ». These data and their combined analysis are crucial to keep improving our knowledge on TBD ecology. With such a context in the introduction, I expected that this study would provide new results and hypothesis. After reading the entire article, I did not find any accurate and relevant information about that.
The authors should clearly define their objectives and their scientific question.

- The number of samples is too low to perform this type of studies.
Authors cannot reasonably perform population genetic and pathogen analysis on only 64 ticks collected in 12 different sates (in three states, only one tick was collected). 8 states have a number of collected specimen < 5. This is unbelivably low to (i) have an accurate estimation of pathogen prevalences, (ii) identify and compare tick microbiome composition and diversity and (iii) perform robust and significant statistical analyses. For three of these 8 states, authors collected only one tick ; One (in North Dakota) was positive for Rickettsia montanensis… so authors concluded that the prevalence was 100%. This conclusion does not make sens. Authors need to have more ticks to estimate the pathogen prevalences (with standard deviations…).
Performing this type of studies and integrating the different approaches needs to collect and analyze much more ticks to have a more accurate estimation of both pathogen prevalences and population genetic. 64 ticks in 12 states (almost 50% only in Ohio) is too low.
The authors mentioned in the abstract : « Interpretation of the results is challenging, likely due to the low levels of genetic diversity and the high abundance of a few taxa in the microbiome ». I guess the low number of samples is probably another reason.

- Negative controls
L177-194 : Concerning the tick microbiome analysis, authors did not mention if they performed negative controls ? Several negative controls have to be done at each step of the process (crushing, extraction, amplification…). These controls have then to be sequenced to exclude contaminant sequences from the final dataset. In low biomass samples, (i.e. individual tick samples), these contaminants may represent a large part of the obtained sequences, and thus generate considerable errors in downstream analyses and in the result interpretation. Please provide detailed information about the controls you performed in this study.

Minor :
L.42-111 : The introduction part is too long. The part from L.69-79 is, in my opinion, not essential. Please shorten the introduction and get straight to the point stating clearly the objectives of this study.

L. 117 : 64 adults : Please provide information about the number of males and females, their localization. Authors cana dd these information in the Table 1.

L.198-199 : What do you mean by « the dimensions of the data were therefore substantially reduced. »

L.260-264 : What about R2 (R) ? Did you merge your sequences ? With your primers, you should obtain amplicons around 400-420bp. You mentioned 251bp. I do not understand these results.

L.263-264 : « the average number of sequences per tick specimen included in downstream analyses was fairly low, ranging from a few hundred to ~15,500 ». The number of sequences was strongly different between samples. Did the authors homogeneize this number of sequences to obtein an equal number in each sample ? This step is essential to compare samples. Please add this information in the M&M part.

L.272 : Rickettsiella is also a tick endosymbiont. Please improve this sentence with some other newer references.

L.297-299 : Please provide the stress value R of the NMDS. An NMDS ordination with a stress value around or above 0.2 is deemed suspect and a stress value approaching 0.3 indicates that the ordination is arbitrary. Stress values equal to or below 0.1 are considered fair, while values equal to or below 0.05 indicate good fit.


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: INTEGRATING POPULATION GENETIC STRUCTURE, MICROBIOME, AND PATHOGENS PRESENCE DATA IN DERMACENTOR VARIABILIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: Some writing format should be checked throughout the manuscript. Take some lines for example: Line 117, 286, 306, 311.
Besides, there are so many mistakes in the writing of the references in the MS. For example: Line 482, “12(268)” should be changed to “12: 268”
Line 582, “PLoS ONE 8” should be changed to “PLoS ONE 8:e61217”.
Line 617, “Genome Biology and Evolution 7” should be changed to “Genome Biology and Evolution 7(3):831–838”.
Line 627, “1-19” should be changed to “e0155559”.
More, I won’t list the other mistakes in the part of “References”, please check carefully according to the Journal Guideline.
Experimental design: no comment
Validity of the findings: no comment
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: INTEGRATING POPULATION GENETIC STRUCTURE, MICROBIOME, AND PATHOGENS PRESENCE DATA IN DERMACENTOR VARIABILIS
Review round: 2
Reviewer: 2

Output:
Basic reporting: the authors’ have not given raw data, i.e. how many total sequences were there, how many sequences per sample before filtering, how many sequences before and after quality control (QC), mean +/- and OTU +.- per sample. This is very necessary information. Information about sequences detected in negative controls are also crucial. Please provide in the supplemental material an Excel file with the OTU tables (before releasing the OTUs detected in controls and after releasing these OTUs).
Do the authors submitted their sequences to GenBank ? please provide the Accession Numbers !
Experimental design: I understood the authors considered ticks per « regions » (East, West and North). If I am right (based on the size of circles on the figure 1), the authors have considered the « western cluster » with only four specimens (this is an hypothesis because all these information are missing in the ms, please provide the number of specimens per region). I reiterate my comment : for me, the number of samples is unbelivably low to (i) have an accurate estimation of pathogen prevalences, (ii) identify and compare tick microbiome composition and diversity and (iii) perform robust and significant statistical analyses. Estimation of pathogen prevalences need standard deviations, please provide these information in the table 2.
 A part of the author answer is : « As for the low prevalence, unfortunately it is very low. And this would not be fixed by increasing the number of ticks by a hundred… this is the reality ». Your answer means that you can identify and generalize the tick microbiome and the prevalence of tick-borne pathogens in the West coast analyzing only four ticks. I am sorry, I am not convinced by this answer at all. All the epidemiologists around the world try to get the higher number of specimens to better estimate the prevalence of pathogens and the authors assure that increasing the number of ticks is not necessary and only few specimens is needed. If you really think that this answer is correct and relevant, please write very rapidly an opinion paper on this : this future paper should interest all the epidemiologists.
Validity of the findings: The authors mention in their answer that the reads did not merge well. Could they include this information in the MS and try to explain this result? In my opinion, this could suggest that the quality of the sequencing and/or the preparation of the amplicon library were not optimal. Could the authors provide the sequencing quality controls? What was the range (number of bp) the authors used for merging reads ?
Concerning my remark on the lack of negative controls, the authors only mentioned they used two negative controls. Because these negative controls are crucial for the next steps of the sequence analyses, we need much more information about these controls… What kind of controls ? At which steps these controls have been performed ? please provide details on these controls.
Additional comments: I thank the authors for their answers. I am sorry but I have still some remarks/questions :
I understood the authors considered ticks per « regions » (East, West and North). If I am right (based on the size of circles on the figure 1), the authors have considered the « western cluster » with only four specimens (this is an hypothesis because all these information are missing in the ms, please provide the number of specimens per region). I reiterate my comment : for me, the number of samples is unbelivably low to (i) have an accurate estimation of pathogen prevalences, (ii) identify and compare tick microbiome composition and diversity and (iii) perform robust and significant statistical analyses. Estimation of pathogen prevalences need standard deviations, please provide these information in the table 2.  A part of the author answer is : « As for the low prevalence, unfortunately it is very low. And this would not be fixed by increasing the number of ticks by a hundred… this is the reality ». Your answer means that you can identify and generalize the tick microbiome and the prevalence of tick-borne pathogens in the West coast analyzing only four ticks. I am sorry, I am not convinced by this answer at all. All the epidemiologists around the world try to get the higher number of specimens to better estimate the prevalence of pathogens and the authors assure that increasing the number of ticks is not necessary and only few specimens is needed. If you really think that this answer is correct and relevant, please write very rapidly an opinion paper on this : this future paper should interest all the epidemiologists.
The authors mention in their answer that the reads did not merge well. Could they include this information in the MS and try to explain this result? In my opinion, this could suggest that the quality of the sequencing and/or the preparation of the amplicon library were not optimal. Could the authors provide the sequencing quality controls? What was the range (number of bp) the authors used for merging reads ?
Concerning my remark on the lack of negative controls, the authors only mentioned they used two negative controls. Because these negative controls are crucial for the next steps of the sequence analyses, we need much more information about these controls… What kind of controls ? At which steps these controls have been performed ? please provide details on these controls.
Finally, the authors’ have not given raw data, i.e. how many total sequences were there, how many sequences per sample before filtering, how many sequences before and after quality control (QC), mean +/- and OTU +.- per sample. This is very necessary information. Information about sequences detected in negative controls are also crucial. Please provide in the supplemental material an Excel file with the OTU tables (before releasing the OTUs detected in controls and after releasing these OTUs).
Do the authors submitted their sequences to GenBank ? please provide the Accession Numbers !


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: INTEGRATING POPULATION GENETIC STRUCTURE, MICROBIOME, AND PATHOGENS PRESENCE DATA IN DERMACENTOR VARIABILIS
Review round: 3
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
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: RELIABILITY AND VALIDITY OF AN EQUANIMITY QUESTIONNAIRE: THE TWO-FACTOR EQUANIMITY SCALE (EQUA-S)
Review round: 1
Reviewer: 1

Output:
Basic reporting: This is a timely and important research topic and I commend the authors for there pursuit of this publication.

The writing is clear and relevant and the discussion around craving / attachment is significant.

There are however a number of clear oversights in this field. Firstly there has been some significant published work in the field of equanimity. Firstly an operational definition specifically tailored for Western psychology was published by Weber (2019) in clinical psychology forum. This is very important given the two facing nature of equanimity. Both 'inner' and 'outer' equanimity are clearly defined and it would seem an oversight to not include these at the forefront of discussion under the western implementation. I would expect both these definitions to be included as they formulate a timely and pertinent aspect of equanimity going forward.

Secondly, there is the Equanimity Barriers Scale that is regarded as the first scale to be published around equanimity. In an obvious narrow field of enquiry as you quite rightly highlighted, it would be remiss to not include a scale that specifically mentions the term equanimity in it. This also would seem to compliment the development of a scale to coincide with hedonic and neutral tone. This should really be considered as a scale in the methodology given the expected convergent validity. This is published under current psychology in an impact factor journal and so should be included at least in the discussion as a key aspect of secular implementation of equanimity.

Thirdly, the well established concept of 'de-centering' should be given some attention - this is possible the closes psychological component related to equanimity and so in any paper that considers equanimity as a construct should distinguish it from related constructs of non-judgement, acceptance and de-centering. The only way for a narrative that combines both Buddhist and secular mindfulness together and potentially makes it more robust is a narrative that unifies constructs/ motivations and ethical underpinning. And does not lean towards fundamentalist thinking of 'this should be the mindfulness way, this should be the Buddhist way' - what you are doing is attempting to join mindfulness more closely with the Buddhist concept of equanimity so it has to be adapted to suit the western audience. Only by complimenting both and finding unification and commonality is this narrative going to be taken seriously. Otherwise it will fall to a rejection type debate.

I commend the overall paper but for me it reads a little - this is mindfulness - this is Buddhist - but you know what is a better approach is 'look why is the arrival of equanimity pertinant to western science? To do this an oversight of an already existing western two facing definition (and that is the important aspect  - as it is this that connects self to other - from inner compassion to outer compassion ) No other defintion attempts to do this - they all point to the vague nature of balanced mind, even tempered etc. This is not entirely useful to fully operationalising this construct. Then again - in western science barriers are a huge aspect of equanimity given the relative individual differences that we all have. Include these aspects and re-situate the discussion to have some more purpose the paper becomes more refined.
Experimental design: This is of a good quality and is clear with good intention.

The research questions are meaningful and rigorous investigation has certainly been implemented.

The methods are good however I do feel the oversight of the equanimity barriers scale is an issue. This should be considered.
Validity of the findings: The data appears to be highly useful and meaningful to a generalised population.

The conclusions should also link up with the re-defined introduction and tied in with were we are at with western psychology - what we need to do going forward - further studies using all existing equanimity scales in order to differentiate equanimity from what currently exists. Otherwise we are left in the same place - people say why introduce this Buddhist concept - 'de-centering' does it for you - why introduce it when acceptance and non-judgement are the same. This is a serious consideration to what you are trying to achieve.
Additional comments: Please continue this good work. I totally value this attempt and some minor work should make this more refined and useful for taking equanimity onto the next stage of research.


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: RELIABILITY AND VALIDITY OF AN EQUANIMITY QUESTIONNAIRE: THE TWO-FACTOR EQUANIMITY SCALE (EQUA-S)
Review round: 1
Reviewer: 2

Output:
Basic reporting: -	The language used throughout this paper (grammar and spelling) would benefit considerably from editing by a native speaker of English. Please see the annotated PDF for several suggestions. Please note that this is not an exhaustive list and there may be more edits required. Also, the formatting of the references needs to be checked and fixed. If APA is being used, it needs to be checked that capitalization is correctly applied.
-	The introduction and background are well-structured but lack some information that would better situate the reader within the literature on equanimity and mindfulness. For example, an explanation of the difference or relationship between equanimity and mindfulness would provide greater context for some of the research cited. Similarly, it is recommended to expand more on the other current equanimity scales and their drawbacks to make a stronger case for the scale. The subscales proposed in the scale also need more explanation and better support. Please see the annotated PDF for specific comments. The research that has been referenced in the manuscript thus far, however, is well done and relevant.
-	The structure of the manuscript conforms to PeerJ standards and discipline norms. It is also presented clearly.
-	All the tables presented are relevant and of good quality. I would recommend checking that the titles comply with the applied formatting style. Please check that the titles and tables appear on the same page. A comment on adding information about the relationship between the outcome variables and meditation practice (in the demographic information) was also added to the annotated PDF.
-	The raw data are available online and the link is provided.
Experimental design: -	With regards to the research being within the scope of the journal, the study falls within the social sciences, if I am not mistaken. The journal states that “PeerJ does not publish in the Physical Sciences, the Mathematical Sciences, the Social Sciences, or the Humanities (except where articles in those areas have clear applicability to the core areas of Biological, Environmental, Medical or Health sciences).” I am unsure of how this impacts this submission.
-	The research question is well defined, relevant, and meaningful. The authors make a significant contribution to the field of equanimity and mindfulness research.
-	The investigation is rigorous and opens avenues for many other research opportunities. It appears that the research was conducted using high ethical standards.
-	Overall, the methods have been described with enough detail. Once the language has been edited, it will enhance the clarity of this section for better comprehensibility.
Validity of the findings: -	All underlying data relevant to the investigation seem to have been provided and seem robust, statistically sound, and controlled.
-	Speculations are identified as such.
-	Conclusions are well-stated and linked to original research question and future research directions.
Additional comments: The manuscript appears to meet the journal’s article requirements and the authors seen to have adhered to the ethical approval statement. No identifiable information was found in the data files. The experiment contributed to impelling this field of research forward and seems to have been conducted ethically.

This was a very interesting paper to review. The authors make a very important contribution to the field. I look forward to reading their revised manuscript and future research.


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: RELIABILITY AND VALIDITY OF AN EQUANIMITY QUESTIONNAIRE: THE TWO-FACTOR EQUANIMITY SCALE (EQUA-S)
Review round: 1
Reviewer: 3

Output:
Basic reporting: This article is extremely clear with good professional English used. The references were good. The data was clear and well presented. The hypothesis was clearly stated with a good discussion of the outcome.
Experimental design: No comment. A good well designed study.
Validity of the findings: No comment.
Additional comments: I really enjoyed this paper and I think that the fact that the sample participants were non meditating adds to the very real need for the concept of equanimity and the impact it could have on mental health both generally and as part of interventions.

I wish them all luck in their next 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: MODIFYING EFFECTS OF LEAF LITTER EXTRACTS FROM INVASIVE VERSUS NATIVE TREE SPECIES ON COPPER-INDUCED RESPONSES IN LEMNA MINOR
Review round: 1
Reviewer: 1

Output:
Basic reporting: Literature references, sufficient field background / context provided, but
check the literature list.
Not in reference list: González-Guerrero et al., 2016; Palmgren and Nissen, 2011; Sinclair and Krämer, 2012; Khellaf and Zerdaoui, 2009, Madhusudhan et al., 2009.

Check the literature cited carefully. There are 19 items in the references, which are missing from the text.

Drawings are legible except Fig. 5 in which the line markings are hardly visible.
Experimental design: Original primary research within Aims and Scope of the journal.
The manuscript refers to global trends in environmental protection and exploring the possibilities of limiting the toxicity of heavy metals (in this case copper) by extracts from leaf mulch from trees of various species. The authors aptly used Lemna minor, which is a recognized natural indicator of toxicity in the world. In addition, a large number of physiological studies deserve recognition, which show the reactions of plants (Lemna minor) to copper and the possibility of reducing its toxic effect by extracts from the leaves of the studied tree species.
Research question well defined, relevant & meaningful. It is stated how research fills an identified knowledge gap.
Rigorous investigation performed to a high technical & ethical standard.
Methods described with sufficient detail & information to replicate.
Validity of the findings: All underlying data have been provided; they are robust, statistically sound, & controlled.
Conclusions are well stated, linked to original research question & limited to supporting results.
Additional comments: Line 1-2 Change the title to a more understandable one
Line 15-44 In Abstract, the research background, the purpose of the work, methods, results without providing unnecessary details and conclusions should be more demonstrated. The current form has a lot of details that take up a lot of space and make understanding difficult
Line 99 - should be supplemented with cyanobacterial toxicity  (Literature)
Line 18-29 The data presented should be described in a more understandable way
In the Results chapter, when describing the response of individual parameters of physiological activity, it would be beneficial to insert at the beginning a general sentence about the effect of treatment on this parameter. Then you can document it with detailed results just as the authors did.
The discussion is written properly.
Drawings are legible except Fig. 5 in which the line markings are hardly visible.
Not in reference list: González-Guerrero et al., 2016; Palmgren and Nissen, 2011; Sinclair and Krämer, 2012; Khellaf and Zerdaoui, 2009, Madhusudhan et al., 2009.

Check the literature cited carefully. There are 19 items in the references, which are missing from the text.

Line 477-482 - The text is repeated in 483-488.

The manuscript should be linguistically corrected, since many sentences are too long, making it difficult to understand the text.
The results obtained are interesting, bring a lot of hitherto unknown information and therefore, after applying the proposed corrections, they should be published.


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: MODIFYING EFFECTS OF LEAF LITTER EXTRACTS FROM INVASIVE VERSUS NATIVE TREE SPECIES ON COPPER-INDUCED RESPONSES IN LEMNA MINOR
Review round: 1
Reviewer: 2

Output:
Basic reporting: The manuscript concerns the  exploring of the possibilities of reducing copper toxicity by leaf mulch extracts from tree two species using Lemna minor as an indicator of toxicity. The effect of litter extracts and copper as assessed on the basis of many parameters of physiological activity in Lemna minor. The work is interesting and brings many new elements.
The manuscript requires proofreading before publication.
The title should be shortened to make it easier to understand
In Abstract it contains a lot of details that take up a lot of space and make understanding difficult. Its content should articulate research background, the purpose of the work, methods, results and conclusions.
In the Results chapter, it would be beneficial to present an additional conclusion on the impact of treatments on individual assessment parameters. Currently, only detailed data is often provided on the basis of which the reader must guess the existing relationships
The drawings are legible except for Fig. 5 in which the line markings are hardly visible.
The captions under the drawings are too long.
Not all literature references cited in the text are in the reference list and vice versa
The manuscript should be linguistically corrected,
The results obtained bring a lot of new, previously unknown information from a scientific point of view and after the proposed correction of the text they should be published.
Experimental design: The research methodology is carried out correctly in accordance with international standards
Validity of the findings: The results obtained bring new information previously unknown in the literature. Noteworthy is the large number of physiological tests performed, which indicate Lemna minor's reactions to the treatments used
Additional comments: The manuscript may be published after the adjustment. Comments are given above.


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: MODIFYING EFFECTS OF LEAF LITTER EXTRACTS FROM INVASIVE VERSUS NATIVE TREE SPECIES ON COPPER-INDUCED RESPONSES IN LEMNA MINOR
Review round: 1
Reviewer: 3

Output:
Basic reporting: The English needs work; in many places, there are too many citations to support a single statement - it's not clear whether they each portray a different aspect of the statement or why so many are needed; structure is fine; figures are fine.
Experimental design: I think the basic question is whether DOC from invasive vs native species alter Cu toxicity to Lemna, and this would be additional reason to limit invasive species.  In that case, the experimental design is appropriate.
Validity of the findings: The effect of DOC or DOM on dissolved cationic metal toxicity to aquatic organisms, such as Lemna, is reasonably well-studied. DOM or DOC has been included in a number of regulatory Biotic Ligand Models for metals.  Research has also been conducted on how various different sources of DOC have different modifying effects on metal toxicity - relative to their affinity for the metal thus creating a metal-complex that is less or not bioavailable.  This might be in play in this study, not sure - but the authors should talk about this.  Generally, the very great detailed biochemistry assays are interesting, but the link to toxicity endpoints is less obvious.
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: MODIFYING EFFECTS OF LEAF LITTER EXTRACTS FROM INVASIVE VERSUS NATIVE TREE SPECIES ON COPPER-INDUCED RESPONSES IN LEMNA MINOR
Review round: 2
Reviewer: 1

Output:
Basic reporting: Enter 'no comment' if you have nothing to add.
Experimental design: Enter 'no comment' if you have nothing to add.
Validity of the findings: Enter 'no comment' if you have nothing to add.
Additional comments: Enter 'no comment' if you have nothing to add.


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: MODIFYING EFFECTS OF LEAF LITTER EXTRACTS FROM INVASIVE VERSUS NATIVE TREE SPECIES ON COPPER-INDUCED RESPONSES IN LEMNA MINOR
Review round: 2
Reviewer: 2

Output:
Basic reporting: No additional comment to the previous review
Experimental design: No additional comment to the previous review
Validity of the findings: The results obtained bring a lot of new, previously unknown information from a scientific point of view and in its current form it  should be published.
Additional comments: In the revised manuscript submitted for re-review, all my comments were taken into account. That is why I recommend it for publication.


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: SAFETY EVALUATION OF FOUR FABA BEAN EXTRACTS USED AS DIETARY SUPPLEMENTS IN GRASS CARP CULTURE BASED ON HEMATOLOGICAL INDICES, HEPATOPANCREATIC FUNCTION AND NUTRITIONAL CONDITION
Review round: 1
Reviewer: 1

Output:
Basic reporting: faba bean was fed to big grass carp for improve the muscle texture, the fish is named with crisp grass carp. the mn is helpful to understand the effects of faba bean on grass carp. the results is benifical to the crisp grass carp production.
Experimental design: why the experimental diets are very different protein content?
Validity of the findings: no comment.
Additional comments: 1.why the experimental diets are very different protein content? the protein content of FBA and FBW is very higher than control group, the results indicated FBW can improve the growth performance, we can conclude the reason is the higher dietary protein intake. why FBW protein is so higher? Can you supply the composition of FBW,FBA?
2.information of the commercial diet need to be supplied.
3.whteher the growth performance is same to the published paper(Effects of four faba bean extracts on growth parameters, textural quality, oxidative responses, and gut characteristics in grass carp,Aquaculture,2020,https://doi.org/10.1016/j.aquaculture.2019.734620)? but the resutes of growth is not consistent.you should give me a reasonable explanation.


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: SAFETY EVALUATION OF FOUR FABA BEAN EXTRACTS USED AS DIETARY SUPPLEMENTS IN GRASS CARP CULTURE BASED ON HEMATOLOGICAL INDICES, HEPATOPANCREATIC FUNCTION AND NUTRITIONAL CONDITION
Review round: 1
Reviewer: 2

Output:
Basic reporting: it is very difficult to read the data in Table 3 due to the format
Experimental design: no comment
Validity of the findings: no comment
Additional comments: In this manuscript, four FB extracts (water extract, alcohol extract, proteins, and residues) were mixed with commercial feed and fed to grass carp (Ctenopharyngodon idellus) using whole FB and commercial feed as controls and examined their beneficial and detrimental effects for the safety evaluation. It is a worthy work. However, there are some issues that need to be clarified.
Main concerns:

Line 90, preparation of faba bean extracts should be briefly introduced and their corresponding contents in different diets should be added.

Line 91, the author referred that 16 kg was used for each of the four experimental diets (each for 16 kg), it might be not very reasonable due to not enough diets to feed all sample fish. In the fish culture, grass carp with a body weight of 750 ± 50 g were randomly allocated into six groups with two tanks per group, and each tank was maintained at a density of eight individuals. Fish were fed for 100 days and the feed amount of each day was 3%-5% of fish weight. And on 50 d, three fish were randomly taken from each group. Therefore, for the diet of each group, it was 0.75*2*8*(3%-5%)*100－0.75*3*(3%-5%)*50≈43.5 kg at least.

Line 166-167, the author referred that four FB extract groups had markedly higher hepatopancreatic weight than the FB and control groups on 100 d (P < 0.05), it is not accurate as FBP extract groups (19.12±4.64ab) had not markedly higher hepatopancreatic weight than the FB groups (9.82±2.35b) and the control group (10.95±3.66b) on 100 d. And please give an explanation why the intestinal weight of the control group was significantly lower than those of other groups (P < 0.05) on 100 d, and why many of the hepatopancreas, Visceral fat and Intestinal weight in different groups on 100 d were reduced compared to 50 d, although a limitation of this study were given that the experimental time lasted for 100 d through fall and winter in the discussion.

Line 172-183, in the hematological analyses, it is very difficult to read the data in Table 3 due to the format, which needs to be improved. It is necessary to explain why these hematological parameters such as red blood cell (RBC), hematocrit (HCT) and hemoglobin (HGB) contents were chosen. Furthermore, many results in the hematological analyses were questionable. For example, the author referred that clear differences were observed on 100 d with small deviations, where FB and FB water extract groups showed lower counts of red blood cell (RBC), hematocrit (HCT) and hemoglobin (HGB) contents, some of which were statistically significant (P < 0.05), but in fact only FB water extract groups showed significant difference in RBC.

Line 205-206, in Fig 3, total cholesterol content (TC) levels were markedly low on 50 d with no significant differences between groups, but increased by 100 d, which is very large difference and change, a reasonable explanation should be provided.

Minor comments:

The location of Fig 5B and C was missing.
The format of references should be consistent, e.g. line 344 and 353 DOI did not need to change a line; line 389, DOI should not be bold.


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: SAFETY EVALUATION OF FOUR FABA BEAN EXTRACTS USED AS DIETARY SUPPLEMENTS IN GRASS CARP CULTURE BASED ON HEMATOLOGICAL INDICES, HEPATOPANCREATIC FUNCTION AND NUTRITIONAL CONDITION
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment。
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The revision of mn had been improved by the author， but i still had some question
1.about protein difference of experimental diets，whether FB water and alcohol extracts improved growth performance?  because the high protein,we cant make a conclusion in the abstract.
2.such as the new reference should be cited by the mn,
Xiaoxia Li, Shijun Chen, Jijia Sun et al. Partial substitution of soybean meal with faba bean meal in grass carp (Ctenopharyngodon idella) diets, and the effects on muscle fatty acid composition, flesh quality, and expression of myogenic regulatory factors. Journal of the World Aquaculture Society. 2019.
LianGan, Yan-zhiWang, Shi-junChen et al. Identification and characterization of long non-coding RNAs in muscle sclerosis of grass carp, Ctenopharyngodon idellus fed with faba bean meal. Aquaculture, 2020,516:734521.
Wei-HuaXu, Hong-HongGuo, Shi-JunChenet al. Transcriptome analysis revealed changes of multiple genes involved in muscle hardness in grass carp (Ctenopharyngodon idellus) fed with faba bean meal. Food chemistry,2020,  314:126205.
Proteomic and metabolomic basis for improved textural quality in crisp grass carp (Ctenopharyngodon idellus C.et V) fed with a natural dietary pro-oxidant.Food Chemistry ,2020,325:126906.