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: YAK RUMEN MICROBIAL DIVERSITY AT DIFFERENT FORAGE GROWTH STAGES OF AN ALPINE MEADOW ON THE QINGHAI-TIBET PLATEAU
Review round: 2
Reviewer: 2

Output:
Basic reporting: The manuscript presents an improvement in relation to the first version. However, there are some parts that it is necessary to change. In general; methodology, results and discussion were improved; however, the introduction has some grammar mistakes and it is essential to develop in detail the importance of the study and its new contribution. There are some names of phyla and expression ‘et al’ that are not written in cursive, authors must change them.
The abstract should be re-organized. It is necessary to write the importance of the study before the overall
Line 23 In this study, the diversity…  this sentence is methodology should be after the overall.
Line 26-28 This work investigated the rumen bacteria… this sentence is the overall and should be introduced in line 23, after the sentence that finishes in 3000- 5000.
Line 33 The word strongly is not necessary for the description.
Line 34 Bacteroidetes and Firmicutes must be in cursive script.
Line 36-37 Authors did not try to separate samples. It is better to write that taxonomic groups did not present differences regarding gender.
Line 38-39 Re-organize the sentence like this; The rumen microbiota was enriched with functional potentials that were related to ABC transporters, the Two-component system, Aminoacyl-tRNA biosynthesis, and metabolism of Purine, Pyrimidine, Starch and sucrose metabolism.

Line 46 QTP is enough, this abbreviation was mentioned in the abstract.

Line 48 the harsh environment of the QTP, with its low oxygen… the harsh environment of the QTP that possesses low oxygen…

Line 49 Natural alpine meadows in the QTP have qualities… Natural alpine meadows in the QTP also have qualities

Line 51 Change: The growth period of grass is approximately 100 to 150 d per year, to The growth period of grass is approximately from 100 to 150 d per year.

Line 70 Previous studies have shown…

Line 71 and seasons all impact on ruminal microbes…

Line 73-74 (Xue et al.,2017) reported… in this sentence, authors need to write only the year in parentheses. Xue et al. (2017) reported.

Line 81 In this paragraph, the authors could mention some taxonomic groups detected in yak rumen.

Line 88 High-throughput sequencing technology has provided microbial…

Line 89 Change the word and for as well as.
Line 90-92 This is a short description of the research, but it is necessary to write the importance of this study and its contribution.
Experimental design: Authors improved considerably the methodology, however, there are some mistakes that it is necessary to change.

Line 96 besides this project is… besides this project was
Line 99 Change word granted to granting.
Line 102 sample plot is 3265 m… sample plot was 3265 m.
               The sample plot has… The sample plot had
Line 103 Delete comma
Line 103-106 Was the place´s information monitored by the authors? or was information provided by a special office? If it was provided for other office or research group, authors should mention it.
Line 108- 109 These ten yaks without supplementary feeding during the experimental stage were allowed access to water … These ten yaks, without supplementary feeding during the experimental stage, were allowed access to water.
Line 113-119 If authors monitored these percentages, these values they would be in results as a supplementary table. In the case that authors receive the information of another group, they should mention it after the description of the forage.
Line 125 to investigated… to investigate.
Line 130-132 Change: This method has been used in previous studies (Sun et al., 2018; Xue et al., 2017) and (Ramos-Morales et al., 2014) have shown that stomach tube and rumen cannulation methods do not result in changes of rumen microbiota, to ´This method has been used in previous studies (Ramos-Morales et al., 2014; Xue et al., 2017; Sun et al., 2018), and they have shown that the stomach tube and rumen cannulation methods do not have any influence on the results´.
References must be organized from the older to the younger.
Line 136 Delete word ´were´.
Line 143 If authors used stool samples, it means that they studied gut microbiome. The manuscript is focused on rumen microorganisms, authors should delete or change this word.
Line 151 If you use a sequencer that was in your laboratory, this statement is right. If you take the service of a company, you should mention the place where the samples were sequenced.
Line 154 Version of software is enough, it is not necessary to write the web page.
Line 163- 164 In this case, reference is adequate, it is not necessary to write the web page.
Line 171 Mentioning the web page is right in this case.
Line 177 web page was mentioned before, it is not necessary to mention it again.
Validity of the findings: Description of the results was improved, there are some mistakes that it is necessary to change.

Line 192 The CP content was highest… The CP content was the highest
Line 199 Authors should write a short conclusion of this table.
Line 205 Authors described 65,92% of total OTUs, there are three zones that were not mentioned; 1599, 1297 and 1038 correspond to 34.08% of OTUs. Authors should add information about them.
Line 210 with highest number… with the highest number
Line 211- 212 with highest number… with the highest number
Line 214- 217 I suppose these values are the average of the three values detected in samples, authors should mention it in the paragraph.
Line 222 Firmicutes in cursive script.
Line 224 Proteobacteria in cursive script.
Line 228 Firmicutes in cursive script.
Line 269 Change word furthermore for another, such as in addition or moreover, authors used this word at the beginning of the paragraph.
Line 278 The sentence: female could not be separated from male samples should be improved, authors described a process, it is better to write that it was not detected differences in PcoA between female and male samples.
Line 294 The word but is informal, it is better to use the word, however.
Line 319 Which is the difference between low and high-quality diet? Is it about different kind of nutrients, food or plants?
Line 325- 328 This information could be in the introduction. In this part, it would be appropriated to write results obtained in this study are according to previous studies.
Line 330 Authors should emphasise the importance of obtaining similar abundance in different studies since this indicates the relevance of both phyla in the degradation of the plant cell wall.
 The study mentioned, was it about yak or another animal?
Line 331 Bacteroidetes in cursive script.
Line 332 Firmicutes in cursive script.
Line 334 The relative abundance of Firmicutes in the WGY group was lower
335 than in REGY and GY; this sentence describes a result. It should not be considered in this part of the manuscript.
Line 340 You should write: Xue et al. (2005) and Ley et al. (2006) showed...
Line 342 Delete the word ´but´, the word ´and´ is better in this case.
Line 348- 350 If authors detected phylum Synergistetes, this sentence should be related to the results of the study. However, if the authors did not detect this phylum, they should discuss why they did not detect it. In addition, anaerobic environments possess several phyla that authors could consider in the discussion since they could be in these animals.
Linea 353 Where were these families found? Yaks? Humans?
Line 356 change to: de Menezes et al. (2011).
Line 360 Delete the dot after the word In.
Line 362 The pathways to degrade plant cell wall have been broadly studied in Bacteroidetes during the last years. Authors could improve the discussion using this information.
Line 369 Research by (Long et al., 1999) … Research by Long et al. (1999)
Line 401 (Bolnick et al. 2014) have shown… Bolnick et al. (2014) have shown
Line 402 Rats are not a ruminant animal. Perhaps the authors wanted to describe other microorganisms in rat’s bodies.
Line 423-430 This sentence is very long and difficult to understand. Authors should write their ideas newly.
Line 442 This study reports the effects of on rumen microbial diversity… this study reports the effect of the forage on rumen microbial diversity
Line 446 that the gender of the animal had less impact on yak ruminal microorganisms… this effect is not related to the gender of the yaks.
Line 446-449 Authors should also consider the study of archaea in conclusions.
Additional comments: No 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: NURHACHIUS LUEI, A NEW ISTIODACTYLID PTEROSAUR (PTEROSAURIA, PTERODACTYLOIDEA) FROM THE EARLY CRETACEOUS JIUFOTANG FORMATION OF CHAOYANG CITY, LIAONING PROVINCE (CHINA) AND COMMENTS ON THE ISTIODACTYLIDAE
Review round: 1
Reviewer: 1

Output:
Basic reporting: Clear, unambiguous, professional English language used throughout - Professional article structure, figures, tables.

The English language is not always clear and unambiguous in this manuscript. I tried to correct it when I could (see attached doc file), but it is still in need of a linguistic review by an English speaking paleontologist. The language is professional, but sometimes the terminology is not the appropriate one (see for example Edmund, 1969 for the terminology relative to the dentition and related structures). EDMUND A. G. (1969) - Dentition. In GANS, C. & PARSONS, T. S. (eds.), Biology of the Reptilia, 1:117-200, Academic Press, London and New York.
Figures show some flaws and their number should be increased (see below).

Intro & background to show context. - Literature references, sufficient field background/context provided

The Introduction gives sufficient field background. However, it is rather chaotic, sometimes redundant and with parts that can be eliminated. I restructured it in the revised text that is attached as doc file.
A small part of the Introduction was moved into the Material and Methods section, because the latter lacked any reference to the Materials.

Literature well referenced & relevant.

The authors cite a lot of papers. Some citations are unnecessary and should be eliminated (e.g., that of Seeley, 1901 at line 343). It is surprising that more recent papers about the age of the Jehol Group formations are not mentioned in the Introduction (e.g, Chang et al., 2009).

Structure conforms to PeerJ standards, discipline norm, or improved for clarity.

Apparently, the authors followed the general PeerJ standards about manuscript structure, but did not care about details of PeerJ standards (e.g., how to write a reference list, how to write figure captions, where reporting grants etc.). This is annoying for a reviewer.
In the References, papers must be listed according to the author names in alphabetic order.
The reference list is written without following any standard or rule and contain a lot of mistakes.

Is the Beipiao Pterosaur Museum a public repository? Most journals do not accept papers about fossils that are deposited in private collections or similar institutions, mainly if they are holotypes.

Figures are relevant, high quality, well labelled & described. - Professional article structure, figures, tables.

The figures must be improved, mainly those showing fossils. A general problem is the poor quality of the drawings that does not allow recognizing many important details reported in the text. Below, there is a brief list of suggested improvements.

 Figure 1. Many skeletal elements (e.g., the premaxilla, squamosal, quadratojugal, odontoid, the lower temporal fenestra, the cervical vertebrae 6 and 7, etc.) that are mentioned in the text are not indicated in figure 1. All identified bones and structures that are mentioned in the text must be indicated in this fundamental figure. Teeth and tooth positions must be indicated and numbered.
The boundaries of quadrate, quadratojugal, jugal and squamosal, that between lacrimal and jugal and those among the cervical vertebrae (mainly cervicals 6 and 7) must be drawn.
Portions of the skeletal elements that are represented only by the print of the bone (e.g., the ceratobranchial I) must be marked by dashed lines.
The beginning and the end of the prid must be clearly identifiable; mark it with arrows. The same should be done for the "dentary sulcus" in the mandibular symphysis.
According to PeerJ rules, the two parts of the figure must be indicated as A and B.

Figure 2. "Fossil position" can be eliminated in the figure. It is sufficient to write in the caption that the star indicates the site where the fossil was found.

Figure 3. The resolution of this figure is very low in the pdf I received. It is 137 kb, well below the 900 dpi that is the minimum size for a PeerJ figure. I do not understand how this could be possible. When I submitted to PeerJ, the system did not allow me to attach figures below 900 dpi: I had to enlarge the figure (without any real reason to do it).
Anyway, it would be wise to mark the nodes mentioned in the text and report the names of the relative clades (in the drawing or in the caption).
I also suggest the authors to run a Bremer test to see how 'resistant' the nodes are, reporting the corresponding values in this figure.
The number of this figure must be changed, because it is first mentioned in the text after figure 4.

Figure 4. Fig. 4A is redundant respect to Fig. 1 and must be eliminated. As in Figure 1, all bones and structures that are mentioned in the text must be indicated in the drawings. Tooth positions must be all numbered. The quality of the drawings must be improved.

Figure 5. The quality of the drawings must be improved (e.g., alveolar margins and crown-'root' boundary must be drawn) and the bones and structures that are mentioned in the text and are important for the distinction of the two taxa must be indicated.

Figure 6. There are no letters (A, B and C) in the three photographs.

Further figures must be added, showing clearly all the diagnostic features of the new species in comparison to the other species of the genus Nurhachius.

The numbering of the figures does not follow the order of citation in the text (see, for example, the first citation of Fig. 3 at line 303 and that of Fig. 4 at line 248).

The captions of the figures are not written following the rules given by PeerJ. They must be carefully rewritten following the rules. The captions with my revision are in the doc file of the text.

Raw data supplied (see PeerJ policy).

Yes.

Self-contained with relevant results to hypotheses.

Yes, but see other comments.
Experimental design: Original primary research within Scope of the journal.

Yes.

Research question well defined, relevant & meaningful. It is stated how the research fills an identified knowledge gap.

The paper is the description of a new pterosaur species and contain a phylogenetic analysis that gives original results in the definition of the Istiodactylidae and closer taxa. This is clearly defined, although some linguistic flaws, redundancy and the occasional use of inappropriate terminology make difficult the reading and interpretation.
More comments are in the doc file (which I had to transform into a pdf to upload it here; thus, read "doc file" as pdf, below) with my revision of the text.

Rigorous investigation performed to a high technical & ethical standard.

Yes, but the definition of the characters that are diagnostic of the new taxon must be better supported and explained. This is also the case of the revised diagnosis of Nurhachius.
Furthermore, the authors seem to confuse "diagnosis" and the "description of a node in a phylogenetic analysis", which are two things that are conceptually different.
More points and suggestions are in the doc file with my revision of the text.

Methods described with sufficient detail & information to replicate.

In general, yes. However, it is unclear which of the mentioned specimens have been studied personally by the authors and which data have been essentially taken from the literature.
Validity of the findings: Impact and novelty not assessed.

Novelty and importance of the described specimen are assessed.

Data is robust, statistically sound, & controlled. All underlying data have been provided

Data are sometimes ambiguous. Definition of the characters that are diagnostic of the new taxon as well as the revised diagnosis must be better supported and explained.
More comments are in the doc file with my revision of the text and general comments below.

Conclusions are well stated, linked to original research question & limited to supporting results.

See points above and comments in the doc file with my revision of the text.
Additional comments: 4. GENERAL COMMENTS
The authors use the term "stem-group istiodactylids" throughout the text. This term is incorrect for at least two reasons:
FIRST: The terms "stem" and "crown" should be used only for taxa that have living representants. I know that they are often incorrectly used otherwise, but see: Jefferies, R.P.S. (1979). The origin of chordates – a methodological essay. In M.R. House (ed.). The origin of major invertebrate groups. London ; New York: Academic Press for The Systematics Association. pp. 443–447.
Pterosaurs are totally extinct, thus "stem" and "crown" cannot be used for them.
SECOND: Even if the use of the term "stem" were correct, Haopterus gracilis, Hongshanopterus lacustris and Archaeoistiodactylus linglongtaensis would not be "stem-group istiodactylids" but the stem-taxa of the clade Archaeoistiodactylus linglongtaensis + Haopterus gracilis + Hongshanopterus lacustris + Istiodactlylidae. Anyway, that would be a purported clade, because Archaeoistiodactylus linglongtaensis was shown to be even outside the Pterodactyloidea.

It is better to mention Haopterus gracilis, Hongshanopterus lacustris and Archaeoistiodactylus linglongtaensis as "taxa reported as close to the Istiodactylidae" here. Haopterus gracilis and  Hongshanopterus lacustris are "taxa closer to the Istiodactylidae according to the phylogenetic hypothesis produced by the authors and Hongshanopterus lacustris is the sister-taxon of the Istiodactylidae.

As it is reported below in the text, Istiodactylidae are defined by Andres et al. (2014) as "the least inclusive clade containing Istiodactylus latidens Seeley and Nurhachius ignaciobritoi. It should be specified at the beginning of the paper.

Lines 96-99 - No, this is not necessarily the case. It depends upon the local sedimentation rates and the presence or not of hiatuses within the sections. In fact, both formations have dramatically different thicknesses in different localities. Since the thicknesses of the formations are locally very variable, comparisons should be done only within a same local stratigraphic section, anyway.

Line 104 - This section is called Material and Methods because you must list here the materials that are the subject of your study. You did not list anything.

Line 105 -  You must report here which specimens you studied personally and which other were coded or described only based on the literature.

Line 143: Why the specific name is N. luei and not N. lüi? You should explain it.

Line 146: Where is this Museum? If it is in the town of Beipiao, you must write: Beipiao Pterosaur Museum,  Beipiao, Liaoning Province, China. Is this museum a public istitution?

Line 151: This should essentially be a differential diagnosis: a list of features distinguishing N. luei from L. ignaciobritoi. Is it?  Why do you define it "a combination of features"?

Lines 168-169: I cannot see this feature in figure 1. Please draw better it and indicate with arrows where it begins and where it ends.

I suggest to read Edmund (1969) as for the terminology to use when dealing with dentition and related structures. EDMUND A. G. (1969) - Dentition. In GANS, C. & PARSONS, T. S. (eds.), Biology of the Reptilia, 1: 117-200, Academic Press, London and New York.

Nurhachius igniaciobritoi -  Longchengpterus zhaoi
The holotype of Longchengpterus zhaoi should coded separately from the holotype of Nurhachius igniaciobritoi in the phylogenetic analysis to test whether the two specimens fall as sister taxa or not. This would be pivotal is assessing whether the two specimens belong to a same species.
Supporting as much as possible with unambiguous evidence the belonging of the holotype of Longchengpterus zhaoi to Nurhachius igniaciobritoi is fundamental in the subsequent comparison with the specimen here described (BPMC-0204) and its attribution to a distinct species.

DIAGNOSTIC FEATURES OF NURHACHIUS (lines 132-138)
Lines 132-138: A diagnosis and the description of a node in a phylogenetic analysis are two things that are conceptually different. A phylogenetic analysis gives a phylogenetic hypothesis based on the data (always partial) reported in the data matrix and does not include all the known taxa. A phylogenetic hypothesis is destined to change by definition. The diagnosis is a taxonomic tool to unambiguously distinguish the taxon from all other similar taxa and must be as stable and definitive as allowed by the fossil record.
Rewrite the diagnosis listing the combination of characters that is diagnostic of Nurhachius. This means a combination of features that are all shared by N. igniaciobritoi and N. luei and that does not occur in any other pterosaur as such a combination. It should be reported when the feature is unknown in one or more of the taxa belonging to the Istiodactylidae (i.e., it is ambiguous).

Comments on characters
<<Slight dorsal deflection of the palate*>>

It is not the whole palate, it is the palatal surface of the tip of the snout, if I understood correctly.

<<Lower temporal fenestra slit-like>>

You call otherwise this fenestra in the text. Can you reliably identify the shape of this opening in N. igniaciobritoi? Based on your Fig. 4, it seems not the case.

<<dentary mandibular symphysis with gradual taper of the lateral margins>>

In which view? If you mean "in dorsoventral view", I think that this is the plesiomorphic condition within the Pterosauria.

<<triangular, laterally labiolingually compressed teeth tooth crowns lacking carinae>>

Labiolingually compressed tooth crowns is a synapomorphy of Hongshanopterus lacustris+Istiodactylidae. Thus, the diagnostic feature is only " tooth crowns lacking carinae".

<< anterior mesialmost teeth relatively longer than others>>

"Relatively longer" is a rather ambiguous definition. Anyway, this feature is plesiomorphic within the Pterosauria.

<< crowns with both labial and mesial slight concavities*>>

I do not understand this feature. In the description of the teeth of N. luei, the authors write "The lingual surface of the crown is concave with a well-marked longitudinal depression", which is not the same. The authors must clarify this point and consider the possibility that depressions could be caused by the collapse of the pulp cavity.

DIAGNOSTIC FEATURES OF THE NEW SPECIES (lines 151-156 and 379-388)

<<The quadrate is inclined at 150° instead of the 160° of N. igniaciobritoi .>>

It is a difference of only 10°. Is it really meaningful? Can you exclude it is caused by crushing or slight disarticulation? How is the slope variability within a sample of pterodactyloids belonging to a same species (e.g., Pterodactylus antiquus and Pteranodon longiceps)? Is it higher than 10°, thus the variability you observed in the three Nurhachius specimens may be intraspecific?

<< The medial process of the pterygoid, is broad and plate-like, whereas it is reduced in N. igniaciobritoi >>

Show it in a figure. Can you exclude that it is reduced in N. igniaciobritoi because it is broken?

<<The dentary median dorsal sulcus of the mandibular symphysis extends until the first pair of dentary teeth, whereas it reaches the sixth pair of dentary teeth in N. igniaciobritoi>>

Show it in a figure. Can you exclude that the difference is of taphonomic origin?

<< The odontoid (pseudotooth) lacks a lateral foramen, whereas the foramen is present in the referred specimen of N. igniaciobritoi (see Martill, 2014, fig. 7C-D).>>

That foramen is on the left lateral side in N. igniaciobritoi. Can you see the left lateral side of the odontoid in BPMC-0204? You do not figure this important feature, thus I cannot know it. Show it in a figure.

<<The odontoid has a smooth blunt occlusal surface, whereas the surface is sharp in N. igniaciobritoi >>

Can you see the occlusal surface of the odontoid in BPMC-0204? The occlusal surface should be the apex, if the odontoid is dorsally directed. You do not figure this important feature (which is not indicated in Figure 1), thus I cannot know it. I suppose it should be blunt (which is the opposite of sharp), but it is necessary to add a photograph of the structure to show it.

<<The odontoid is dorsally directed, whereas it is slightly anterodorsally directed in N. igniaciobritoi (but see Martill, 2014, p. 57, right column, lines 21-23)>>

Again, you must show it in a figure. You must prove that there is a substantial difference between the direction of the structure in N. igniaciobritoi and in BPMC-0204.

<The ceratobranchial I of the hyoid apparatus accounts for 60% of the mandibular length, whereas it accounts for 60% of the mandibular length in N. igniaciobritoi.

Actually, this does not appear to be a diagnostic feature in the Discussion section, because both species result to have the same relative length (cf. lines 382 and 387). Is it a typo?

<<12 tooth positions in each side of the upper jaw; 11 tooth positions in each side of the lower jaw.>>

This is not reported as a diagnostic feature in the Discussion section. Comparison with tooth count of  N. igniaciobritoi is not discussed.
Consider that tooth count can depend upon the growth stage of the individual and is often intraspecifically variable (see Edmund, 1969). Does the tooth count variability within the sample of a pterodactyloid pterosaur represented by an adequate number of specimens support your statement? A difference of 2-3 tooth positions could be not significant.

<< The mandibular teeth extend distally beyond the symphysis, whereas they are confined to the symphysis in N. igniaciobritoi>>

Does the variability in the distalmost teeth positions within the sample of a pterodactyloid pterosaur represented by an adequate number of specimens support your statement?

Lines 389-90: <<both specimens of Nurhachius ignaciobritoi come from the upper part of the Jiufotang Formation>>.

This must be demonstrated.

REFERENCES
The reference lists is not written following the rules given by PeerJ. It is a chaotic list of citation written following different styles and full of mistakes. I tried to correct part of the mistakes but I did not do all the job. The authors must rewrite it in a correct way.

Lines 64-77 - It surprising that the references regarding the dating  of the Jehol Group units are quite old and more recent papers (like Chang et al., 2009) are not mentioned.

Line 64 - Gradstein et al. (2004) is an old and partly obsolete reference. The International Chronostratigraphic Chart of ICS (IUGS) is updated twice per year. See www.stratigraphy.org

-------------------------------------------------------------------------------------------------------------------------------------
CUSTOM CHECKS

New Peerj species policies
- The name of the new taxon was not mentioned in the title.
- The new species was probably entered in Zoobank because it has a LISD, but I cannot check whether it is correct or not, of course (searching for it by Google does not give any result).

New species checks
Do you agree that it is a new species?
I agree that it may be a new species, but the authors must support their statements in a more clear and exhaustive way. This is explained in the General Comments and in the doc file with my revision of the text. Diagnostic features must also be explained by detailed figures. The differences with the other taxon/taxa must be clearly shown to the reader.

The authors affirm that "both specimens of Nurhachius ignaciobritoi come from the upper part of the Jiufotang Formation". This is an important information, because the two purported species would result to have lived in two distinct intervals of the geological time that are separated by some million years (as underlined in the Introduction). However, Authors do not give any evidence or reference to support their statement. The stratigraphic provenance of the two specimens of N. ignaciobritoi must be demonstrated.

Is it correctly described e.g. meets ICZN standard?
I discussed this in the doc file with my revision of the text. The new species is correctly described, but the emended diagnosis of Nurhachius must be reconsidered. A diagnosis of a taxon and the description of a node in a phylogenetic analysis are two things that are conceptually different.

Raw data check - Review the raw data.
If you consider the data matrix of the cladistic analysis as "raw data", it is impossible to review all the characters, character states and character state codings in only 10 day. I did not do it. Anyway, the data matrix was not produced ex novo for this paper, but was taken from a previous one. Therefore, fact that the terminology used in the character and character state definitions is sometimes inappropriate, is not a fault of the authors of the manuscript under review. As for the codings of the newly added taxa, their correctness is a personal responsibility of the authors. I never saw those specimens personally, therefore I cannot judge the correctness of the codings.

Running the data matrix gave the same results as that obtained by the authors.

Image check - Check that figures and images have not been inappropriately manipulated.
The figures do not seem to have been manipulated, but they contain some mistakes and can be substantially improved (also graphically).


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: NURHACHIUS LUEI, A NEW ISTIODACTYLID PTEROSAUR (PTEROSAURIA, PTERODACTYLOIDEA) FROM THE EARLY CRETACEOUS JIUFOTANG FORMATION OF CHAOYANG CITY, LIAONING PROVINCE (CHINA) AND COMMENTS ON THE ISTIODACTYLIDAE
Review round: 1
Reviewer: 2

Output:
Basic reporting: Despite the fact that I' not a native English speaker myself, I was able to detect several grammatical errors and stylistic issues. The text seems to be written by more than one author with completely discrepant styles, and some excerpts are rather confusing (see the annotated PFD file I provided). Stylistic standardization will improve clarity and is extremely recommended in this case. I also recommend a careful review by a native English speaker prior to publication.

The literature references are adequate and the authors provide an up-to-date background for the accessed issues.

Raw data was provided by the authors. Figures are competent (with the exception of Figure 3, which came to me in a exceptionally poor quality). They are, however, insufficient to support the conclusions drawn out by the authors, as some key features of the new specimen lack proper illustration.

The study is self-contained, all the results being relevant to the hypotheses.
Experimental design: This is an original research, based on a specimen never before adequately studied and illustrated. The primary research question (whether or not the new specimen represents a new taxon) is relevant and well defined. The author hypothesis was properly tested by anatomical comparisons and a robust phylogenetic analysis. Methods are properly described and the provided raw data enables replication.
Validity of the findings: The author hypothesis was duly tested with the use of adequate methodology, making the results sound, so that I was convinced of their main statements. Some relevant information lacks proper illustration (see the annotated PDF file).
Additional comments: This fine MS describes a new Chinese istiodactylid based on a wonderfully-preserved specimen, adding information on this curious pterodactyloid clade. The MS has several merits justifying its publication in PeerJ, not the least of them being the resolution of the phylogenetic position of the enigmatic taxon Archaeoistiodactylus linglongtaensis.

In spite of that, I have several concerns that must be accessed by the authors before a final version is ready for publication. All of them are detailed in the annotated .pdf file I provide together with this report.


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: NURHACHIUS LUEI, A NEW ISTIODACTYLID PTEROSAUR (PTEROSAURIA, PTERODACTYLOIDEA) FROM THE EARLY CRETACEOUS JIUFOTANG FORMATION OF CHAOYANG CITY, LIAONING PROVINCE (CHINA) AND COMMENTS ON THE ISTIODACTYLIDAE
Review round: 1
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This manuscript describes a unique specimen that is of great value to pterosaur specialists. It has been enjoyable to read it and review the phylogenetic analysis, with which I agree. The revision of the Istiodactylidae lineage was necessary as the authors detailed, but even those taxa close related with the istiodactylids following the phylogenetic definition established by Wang et al. (2005).

My primary concern is on phylogenetic concept of “stem group”, which should be avoided. The concept “stem group” is strictly related with “crown group”, which was introduced for classifying living organisms and their extinct relatives (Jefferies, 1979; Willmann, 2003). Thus, a stem group is used for those extinct relatives more closely related to the crown group than to any other extant organisms, but excluding the crown group (i.e., the most recent common ancestor of the living lineage and its descendants). Due to Istiodactylidae is an extinct lineage, the proposed term “stem-group istiodactylids” makes no sense. It could be substituted by a statement as “lanceodontians close to istiodactylids than anhanguerians”, but I really encourage the authors to propose a taxonomic definition of this lineage, either from Haopterus or even all the lineage from Lonchodraco+Ikrandraco.

Also, please take into account italics and punctuation marks (see below).

I have noted a few additional issues:
Line 36 – use italics in the Latin phrase “et al.” and delete comma after “Lü”
Line 37 – I recommend to write “Istiodactylus sinensis” instead of “I. sinensis”
Lines 38-39 – avoid to use the term “stem-group istiodactylids”
Line 57-59 – use italics in the Latin phrase “et al.”
Line 65 – the word “old” should be deleted
Line 87 – “and also the bird Jeholornis” should be added a reference
Line 109 – if you used the last version of TNT, you should referred as TNT 1.5 (Goloboff & Catalano, 2016)
Line 167 – “2019” instead of “2016”
Lines 440-441 – “Haopterus was recovered as a basal ornithocheiraean” following Andres et al. (2014) definition (i.e., the least inclusive clade containing Ornithocheirus simus and Anhanguera blittersdorffi) this is not true. In Holgado et al. (2019), Haopterus was recovered close but out of Ornithocheirae. A proposal of how it should be rewritten: Haopterus was recovered close to anhanguerians than istiodactylids.

Additional references
Goloboff, P. A., Catalano, S. A. (2016). TNT version 1.5, including a full implementation of phylogenetic morphometrics. Cladistics 32(3), 221–238.
Jefferies, R.P.S. (1979). The Origin of Chordates: a methodological essay. Academic Press for the Systematics Association, London. pp. 443–447.
Willmann, R. (2003). From Haeckel to Hennig: the early development of phylogenetics in German-speaking Europe. Cladistics 19, 449–479.


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: NURHACHIUS LUEI, A NEW ISTIODACTYLID PTEROSAUR (PTEROSAURIA, PTERODACTYLOIDEA) FROM THE EARLY CRETACEOUS JIUFOTANG FORMATION OF CHAOYANG CITY, LIAONING PROVINCE (CHINA) AND COMMENTS ON THE ISTIODACTYLIDAE
Review round: 2
Reviewer: 1

Output:
Basic reporting: This is a rereview of a major revision. The authors have corrected the points, etc. pointed out by previous reviewers. I have noted minor errors and other points that could be corrected, but ms. is publishable with minor revision.

See my detailed comments and marked up ms. in the attached pdf.
Experimental design: OK
Validity of the findings: OK
Additional comments: See my detailed comments and marked up ms. in the attached pdf.


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: THE SECRET LIFE OF DEEP-SEA SHRIMPS: ECOLOGICAL AND EVOLUTIONARY CLUES FROM THE LARVAL DESCRIPTION OF SYSTELLASPIS DEBILIS (CARIDEA: OPLOPHORIDAE)
Review round: 1
Reviewer: 1

Output:
Basic reporting: The language should be improved throughout to ensure that the reader can clearly understand. The authors need to be more specific in details and avoid using vague or descriptive words.

Some words are missing from the sentences such as “to” and “the”.

The flow of the introduction and discussion seems disjointed and hard to follow.
The author needs to clearly indicate what the papers main aims are? Introductions have the main aim of showing the reader the problem and the proposed solution.

I would start with the problem: no or limited verified published information on larval development for Oplophorid shrimps other than plankton samples.

Solution: Describe the development from reared larvae. Why is this important: this can give information about the species ecology and evolutionary development. Add some background information surrounding larval development strategies.

The authors should give the reader the information of why was the investigation was carried out and why this species was specifically analysed?

I feel there is lost opportunity within the article to give valuable information on the specifics of the larviculture process. The egg dimensions and number hatching is reported in the methods rather than the results. No survival information is provided. The growth / size data could be provided in a table.

The article structure seems to be good with standard sections.

The figures are clear and presented well. However, the scale bars could be made clearer.

No raw data was attached. The table presented provides data on the number of broodstock, number of eggs and number of larvae. No data on the survival or growth for the larvae is provided.

Some citations do not follow PeerJ guidelines. Some references are not cited within the manuscript.

Specific Details:
Line 25: I recommend replacing “outburst” with “attention”.
Line 33-34: Seems like a stand-alone statement. Could you explain or go onto explain why this is important?
Line 35: The use of the Gurney as a reference throughout the manuscript doesn’t seem correct as not all the authors are being cited. See appropriate peerJ referencing styles. i.e. “Gurney & Lebour (1941) described the…”.
Line 42: Please re-phase. You could use “an” instead.
Line 44 – 46: This sentence is hard to follow. Please re-phase and be clear on your main point that the quantity of egg yolk is in some regards correlated to the larval feeding strategy, i.e. either lecithotrophic or planktotrophic. A simple explanation of what theses two strategies are should be provided and how this relates to the grouping of the species / split of the genera.
Line 47: A leading sentence is needed here to flow from larval feeding strategies to larval morphology. The main point is also missed: that within the supergroup there is a wide variety of zoeal development stages before metamorphosis.
Do you mean “Only a small quantity of the species, within the Oplophoroidea superfamily have the larval developmental stages described from samples which were collected from plankton tows. These were …”. This paragraph needs clarity in how it leads to the next paragraph. Can you explain why Systellaspis debilis was focused upon?
Line 60-62: Please re-phrase. I find that sentence hard to understand, between which two stages? The authors should clearly state that there is confusion as to whether there is another stage before metamorphosis.
Line 67- 69: The finding that larvae are full lecithotrophic should be moved to within the results section and not shown within the introduction unless there is background knowledge corresponding to this?
Line 74: Please give the full name before using acronyms. i.e. Instituto Português do Mar e da Atmosfera (IPMA).
Line 80: Was there any cleaning of the eggs prior to transfer? Can you explain in the methods how you measured the eggs, for example longest width x length.
Line 80: The table should be presented within the results.
Line 82-83: “hatching that in some cases occurred almost one and a half month after collection” Should be in the results section. Could say instead along the lines of ”for a maximum of .. days?”.
Line 84: please be more specific as to what the larvae were transferred to. A 30 ml glass beaker? Information on the larval conditions should be indicated. For example, Was the conditions similar to that of the eggs - kept at a salinity of 35± 1, a temperature of 18+ 1 ºC, and a photoperiod of 12 h light: 12 h dark? Please clarify. Was the water replaced with autoclaved or filtered SW. If it was filtered than more information should be given.
 Line 86: Can you give information on the Artemia sp. and how it was prepared, for e.g.  similar to the methods used by ….
Line 88-90: Could be more succinct. “When the larvae reached the decapodid stage they were fixed in 4% borax buffered formaldehyde along with the exuviae for later morphological analysis.”
Line 98: “is referred to”
Line 115: An average should be accompanied with SD or SE.
Line 139 and throughout: I would avoid the word “very” or “little” as it seems too descriptive or ambiguous.
Line 166 and throughout: Fig 3A & 3A1
Line 433: I’m not sure what you mean by sequence? DNA sequence / larvae stages / development? Please clarify.
Line 433-434: Please re-phase. . I would try to avoid the use of his, ours and we. Rather specifically refer to each. For example “However, Gurney & Lebour, (1941) indicates that the second zoea stage as … whereas the results from this study indicate that ….
Line 433-437: Please make this sentence easier to understand.
 Line 442 & throughout: I would avoid using “are in fact”. Please re-phase.
Line 448: I would replace “now presented” with “in this study”.
Line 454: “when he says” – please clarify.
Line 454-456: This needs to be in a separate paragraph and expanded upon with the paragraph below.
Line 456 & 458 & throughout: “by the present”
Line 465: give example of some of the decapods that show developmental stage plasticity.
Line 465-466: please re-phase. For e.g. Gurney … indicated that the differences observed
Line 467: Please clarify what you mean by “individual variation in the degree of development”
Line 468: Please re-phase. Be more specific, for e.g. “The plasticity shown in the number of larval stages is more common…”
Line 469-470: Can you explain why the proposal of the species being full lecithotrophic would increase the possibility of plasticity in the number of larval stages?
Line 474 - 477: “a larve récemment éclose” I do not understand this. Is this comparative forth stage or decapodid? Please re-phase and indicate more focus to the reader what it is you trying to show.
Line 480: Remove Fig 1.
Line 480-481: This should be in a new paragraph. There needs to be a clear flow through the discussion with a sentence introduction to each new paragraph building on the focus of the discussion. For example. “We propose that S. debilis is a full lecithotrophic species due to the findings within this study showing complete metamorphosis without the addition of food. Being lecithotrophic has been described as a probable evolutionary consequence of the habitat where this species inhabits (REF).“
Line 485: please re-phase sentence starting with “in that situations” as it is unclear.
Line 497: The sentence is a little confusing as you have said that S. debilis is full lecithotrophic. I think it would be beneficial to the reader to introduce the different energetic aspects of larval development within the introduction or the beginning of the discussion, highlighting the difference between planktotrophy, full, primary and secondary lecithotrophy. I would give further examples of similar species (if known) of each. In addition, do the authors assume that feeding will commence again at the onset of metamorphosis as direct observation of the development of the mouth parts?
Line 498: I am unsure of the use of the word "characters". Should this be characteristics? Or simply use "the larval morphology between … and  .. are similar".
Lines: 498-499: Are you comparing all the species in two sets of genus? Should this not be Systllaspis spp. etc?
Line 500: Unsure of the word “series”. Should this be species?
Line 502: I think the readers may find it confusing when referring to “the author”. I think this should be changed to be specific. Which author are you referring to?
Line 512: general readers may not be familiar with the term “derived character”. This paragraph needs more of an introduction. It is unclear what the focus is for the reader and why the discussion has moved onto evolutionary development?
Line 514: the reference to the Fig 2, clade 1 is confusing. This should either be shown within the manuscript itself or the phylogenetic analyses clearly described to the reader.
Line 519: Please do not use the words actually and quite, and be specific. I am unsure what was observed? Please re-phrase this sentence.
Line 527-539: Similar to the earlier comment for line 497. The term “nutritional vulnerability” needs to be clearly explained and introduced earlier in the discussion.  This may be a good paragraph to build upon to introduce the different energetic aspects of larval development.
Line 540-554: This paragraph could be made more succinct. It is unclear what you are trying to agree with or dispute? Please make this clearer and tie it in with the rest of the discussion.
Line 544: “more superficial depths” please be specific.
Line 546: there is a full stop between described and demonstrate. Please re-phrase the sentence.
Line 547: you refer to evidence but have not  made clear or provided supporting results. It should be identified as speculation that the larvae reside within the mesopelagic zone and the argument given.
Line 551-554: Seems like stand-out sentences that need more explanation in relation to the studies aims and its implications, or it should be removed.
Line 571: I could not find a citation for this reference within the manuscript.
Line 614: I could not find a citation for this reference within the manuscript.
Figures: Really nice figures and clear drawings. Can the scale bars be relocated to avoid confusion and made clearer?
Within Figure 2 caption: C’ should be C1.
Within Figure 2 caption: Should A1 be included in the 0.05 mm scale? Please check all correspond to the correct scale.
Experimental design: The research question was not well stated nor was the potential implications of such research indicated and should be provided clearly and succinctly.
The morphological definitions and figures are of a high standard and are clear and precise.
More information is needed within the methods detailing the larval culture conditions to allow it to be reproducible by another investigator.
I think the manuscript would benefit with more information such as, egg development, hatch rate compared between years, larval survival data of each stage.
Yolk quantity or yolk nutritional quality using biochemical analysis would have been beneficial to the study.

I would have thought that one of the major questions of the research was the comparison between the two treatments /years, i.e. fed and un-fed? Factors such as survival / growth / microbiology (bacteria levels) to base the proposal of the larvae being lecithotrophic are not shown?
Was the feeding behaviour specifically observed for as mentioned within the text? If so this should be included in the methods. However, it would be hard to observe feeding behavior with live Artemia especially within the first developmental stages without the use of a microscope. What was the size / stage / nutrition of the Artemia as this can have an effect of the way larvae feed, at least for carnivorous larvae - for e.g. see Pochelon et al., 2009. Feeding Ability of early zoeal stages of the norway Lobster Nephrops norvegicus? Biol. Bull. 2016:335-343.
Was any quantification made on feeding rates i.e. the amount of feed provided compared to the amount of feed recovered to be certain that the larvae did not feed on the food provided?

Specific Details
Line 77: Is there any more information on the broodstock such as, what depth were they captured, what size, what moult period were they in?
Line 97: Similar to your earlier work and could be used as a reference: Bartilotti, Catia & Salabert, Joana & Dos Santos, Antonina. (2016). Complete larval development of Thor amboinensis (De Man, 1888) Decapoda: Thoridae) described from laboratory-reared material and identified by DNA barcoding. Zootaxa. 4066. 399. 10.11646/zootaxa.4066.4.3.
Line 110: Please clarify development until metamorphosis.
Validity of the findings: A fantastic achievement to culture species to metamorphosis for the first-time, identifying the species to be full lecithotrophic is a significant improvement to the knowledge gap for this species.

The discussion needs to have a clearer flow for the reader to follow and clearer focus on results related to the aims of the study.

The discussion seems disjointed with several topics discussed. The discussion starts by comparing the morphology in this species with that of other authors. The authors need to make the readers aware of the need to replicate of detailing the developmental stages of this species and the comparison with known descriptions from Gurney and Lebour (1941). The results indicate that are some differences and then indicate why this is important.

The issues of development stage plasticity needs to clearly augmented and the implications of this. More information indicating why this may occur should be included for example, different locations, depth, nutritional value within the yolk? The authors give no recommendation on further investigation, such as genetic analysis.

The proposal of the species being full lecithotrophic needs to fully proposed and augmented rather than stated as such. The proposal of the species being full lecithotrophic increasing the possibility of plasticity in the number of larval stages is not well explained or backed by any literature.

The discussion then jumps to the proposal that classifying larvae as lecithotrophic be based primarily on the size of the eggs is not well introduced to the reader and should be improved. Does this have any relation to the original aims of the paper?

The discussion then compares morphology of Systellaspis and Oplophorus, which I feel would be better suited to earlier in the discussion when comparing to Gurney and Lebour, (1941).

The proposal to study the evolutionary correlation between the two developmental pattern extremes in the mesopelagic environment between species is good but needs a clearer explanation and introduction.

The discussion then jumps back to the yolk reserves and the nutritional quality, although no specific details of nutrition were looked at within this study.

I feel from line 540 the discussion is not focused. The proposal that the species live in deeper layers of the water column needs a clearer explanation and linked with supporting results and/or literature.
The discussion needs to give the reader some implications of the findings and would benefit with a summary of conclusions in relation to the major findings of this paper.
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: THE SECRET LIFE OF DEEP-SEA SHRIMPS: ECOLOGICAL AND EVOLUTIONARY CLUES FROM THE LARVAL DESCRIPTION OF SYSTELLASPIS DEBILIS (CARIDEA: OPLOPHORIDAE)
Review round: 1
Reviewer: 2

Output:
Basic reporting: I think this is a very nice piece of work. I have really enjoyed reading it.
This is a well-structured and well-written manuscript, authors did a great job. I think that is a detailed and complete study in a taxonomic group with few validated larval data, therefore this work provides relevant information.
I am not a native English speaker, therefore I have not tried to correct language, although I think it is correct.
I have only some minor comments, questions, and suggestions for helping to improve it.
Experimental design: Materials & Methods.
I would like to see some details about how eggs were conserved alive during cruises and transported to the laboratory. I mean, all larvae hatched at lab facilities? or in some cases some larvae were obtained onboard and later transfer to the laboratory on land? Also, I see that from females 1, 4, 8 and 9 you did not get any larvae, and in the case of female 10, just one from 11 eggs. They were not viable eggs? Or dead by….? I mean surely you get eggs in different degree of development, are there any relationship between eggs hatched and eggs not hatched with time of development? I see some eggs hatched after one month and a half of culture, so they were collected in early phase of development.

Respect to larvae hatched, I cannot see any comment about swimming behavior. These zoeae were active swimmers?  In the figures of general view of zoeae I see short natatory setae, sound like not well swimmers.

In the description:
Line 96. Be more precise, “…long plumose setae on the exopods of maxillipeds and on…”, insert "of maxillipeds"
Line 110. Yolk was present in all stages, even decapodid, but what about first juvenile? According to amount of yolk in decapodid I guess that is still present in juvenile, and you obtained at least two juveniles (did you check this feature?) . In the case of Dugastella valentina, another caridean with abbreviated lecithotrophic development, yolk was observed until third juvenile stage (see Rodriguez & Cuesta, 2011).

Line 144. In the scaphognathite you describe 9-11 marginal plumose setae but in fig. 2F I see also a distal long  stout process. I suggest to describe it.
Line 188. In the scaphognathite you describe 11-14 marginal plumose setae. But in fig. 3F I see only 10 marginal plumose setae and still a distal long stout setose process that you drawn perfectly differentiate of the rest of setae of this appendage. I suggest to describe it correctly.

In the setation of endopods of maxillipeds you describe all setae as plumose. I am not sure (based on your illustrations) that all of them are plumose, maybe some are simple setae? They look really small and without setules.

Lines 205-208. I agree that uropods are absent, but visible under integument in telson. Anyway, remember that uropods are appendages of sixth pleomere. I suggest to replace the sentence in telson to uropod description. As Uropods (Fig. 3K): absent, but already visible under telson integument.

Lines 232 and 287. The same that I comment above respect to distal long stout setose process still present in scaphognathite. It is just disappearing in the decapodid (fig. 6F).
Line 254.I cannot see in fig.4J that first pereiopod endopod is 5-segmented. I see lightly segmentation is marked, but not clear segments differentiated. And, of course this segmentation is more difficult to see in pereiopods 2-5.

Respect to zoea IV, you say in your description that pereiopods are all functional. What about maxillipeds are they functional as natatory appendages?? I mean, the pleopods are still not functional. How these larvae (zoea IV) move? “walking”? swimming?
Validity of the findings: I think that conclusions are based on data obtained and are well supported. I have only some comments for take into account in the Discussion section.

Line 443. Gurney describe the zoea I as having uropods “under skin”, you say that in the present work are not visible, but you could add “until zoea II”, where they are visible (fig. 3K).

Lines 483-487, and 519-523. I agree with citations here included, González-Gordillo et al. (2010) and Kattner et al. (2003), but they refer to brachyurans and lithodids, I suggest to check Rodriguez & Cuesta (2011), and references herein,  where you can find examples of caridean (closer taxa) with direct, abbreviated and extended development (in this case evolutionary selection is not the same for all freshwater species). Also, you can find caridean larval descriptions (closer to Oplophoridae than brachyuran and lithodids) and see similar reduction of setation of mouthparts due to lecithotrophy adaptation. Maybe you can find more arguments to include in your discussion about larval lecithotrophy, mouthparts not functional, etc.

Rodríguez A & Cuesta JA (2011) Morphology of larval and first juvenile stages of the kangaroo shrimp Dugastella valentina (Crustacea, Decapoda, Caridea), a freshwater atyid with abbreviated development and parental care. Zootaxa, 2867: 43-58.
Additional comments: As mentioned above, I think this is a good work, therefore I have only comment some questions for helping you to improve it.

These are my last comments:
References
I check the manuscript and I cannot find the citation to dos Santos (1999) and Zariquiey-Alvarez (1968), I suggest to cite where appropriated or remove these references.

Figure captions
The figure captions are included twice, first from lines 617-66, and second from lines 668-717.
In figure 7, lines 662-664, the term “epipodite” is not appropriated used.

Table 1. I miss one date in this table, the size of the females. It would be nice to relate female size and number of eggs. If authors missed these data, maybe some comment in the Materials and Methods would be welcome. For example, the number of eggs was related with females´size or after fished the dead females could bear less eggs (loss during the process). I am curious about differences between females with just 2-3 eggs and other with 12 eggs.


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: THE SECRET LIFE OF DEEP-SEA SHRIMPS: ECOLOGICAL AND EVOLUTIONARY CLUES FROM THE LARVAL DESCRIPTION OF SYSTELLASPIS DEBILIS (CARIDEA: OPLOPHORIDAE)
Review round: 1
Reviewer: 3

Output:
Basic reporting: Overall: There are small grammatical and formatting errors throughout the manuscript.  Please read carefully.

Many parts are well-written but there are areas of the manuscript where the English needs improvement.  This is mostly the result of incorrect word choice that makes the manuscript hard to read in some areas.  Please consult with a native English speaker to help with this part.

In some parts the text needs to be expanded.

The drawings are very nice!
Experimental design: There is some missing information on the experimental design.  For example, I am confused by the addition of the 2010 study.  What is the purpose of mentioning this study?  Are results part of this manuscript?

Also, It is unclear “how many” individuals were examined for each stage.  Please provide this information for each stage in the descriptions.

Other concerns are listed in the "general comments" below.
Validity of the findings: Overall fine, suggestions provided in general comments.
Additional comments: Overall: There are small grammatical and formatting errors throughout the manuscript.  Please read carefully.

Many parts are well-written but there are areas of the manuscript where the English needs improvement.  This is mostly the result of incorrect word choice that makes the manuscript hard to read in some areas.  Please consult with a native English speaker to help with this part.

Title:  I would consider shortening the title.  It is a bit long winded for the description of the study.

Introduction:

Line 25: “recent outburst” is not the right use of the term here.  Consider recent “attention”

The Wong et al. paper did not resurrect the family Acanthephyridae so please add the reference for this on line 33.

Overall the introduction is well written and important basic life history knowledge for mesopelagic shrimp.  There are some additions I would like to see:

1.	The introduction is very centered on oplophorid larvae, but what is missing is an overview on our state of knowledge for mesopelagic decapod shrimp as a whole.  It would be great for the authors to “justify” why this work is important for deep sea biodiversity.  This is not described in the paper much at all.
2.	It was unclear how the present study differs from what is already known from Gurney and Lebour 1941.  It needs to be emphasized how the present study enhances our knowledge on Systellaspis debilis and how it provides “better” or “more” knowledge then what is already in print.  This is also not expanded well.

With these additions, the introduction will be a very nice contribution.

Methods:

The authors state in 2010 the shrimp did not eat Artemia.  Is there are referenced study for this?  If so, please add it.

I am curious about the lack of feeding.  Is it possible that the larval development is “affected” by not eating?  Has there been any studies that look at the effect of this in the literature?  Some justification needs to be given as to how not eating should NOT affect the larval rearing process.  One could assume as these shrimp are deep sea species, food would would be scarce anyways, but I would like to see some expansion here.

Results

I am very confused on why the authors are discussing the 2010 study.  Is that part of this study?  If so, this needs to be developed.  It is unclear how the 2010 study fits into this study.

Also, the authors begin to make conclusion in the results section and this needs to be moving to the discussion.  The results are to just report the results and not “discuss” the findings yet.

It is unclear “how many” individuals were examined for each stage.  Please provide this information for each stage in the descriptions.

Discussion

I do not understand this sentence:
We agree with the author when he says that probably there is
455 some variation in the number of stages of the same species, since the characters presented for the
456 telson agree well with those described by present study for this larval stage.

Do the authors think there are possible more than 4 stages?  Please revise to clarify.

The discussion on the two developmental patterns is very interesting and a valuable addition to the manuscript.

I would like to see the discussion dived into 2 sections: 1 that discusses the morphological characters and variability between studies and 1 that discusses lecithotrophy and the relation to evolutionary history.

The author do a very nice job comparing their findings with past literature on morphological differences between studies.  It becomes a bit confusing what the final “conclusions” are concerning the variability between studies.  At the end of section 1, I would like to see a conclusive paragraph that summarizes the major findings.  For example, is it possible that the authors did not capture all the life stages?  Or are they confident they described the full series and the other studies are incorrect?  I was confused by all the different comparisons being made. The summary paragraph can summarize the findings of this study along with the others in a concise story.

Line 544: What are superficial depths?  Acanthephyra spp. are hard to generalize as they display a WIDE distributional range with some species “stylorostratis” not migrating at all (Gulf of Mexico depth min of 750m) and others being shallow.

Line 544-547: the sentence is not complete.  Please revise and describe the larval behavior.

I really appreciated the drawings in the manuscript.  They look fantastic and I highly commend the authors for drawing the photophore pattern!!!!!


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: THE SECRET LIFE OF DEEP-SEA SHRIMPS: ECOLOGICAL AND EVOLUTIONARY CLUES FROM THE LARVAL DESCRIPTION OF SYSTELLASPIS DEBILIS (CARIDEA: OPLOPHORIDAE)
Review round: 2
Reviewer: 1

Output:
Basic reporting: I have read carefully this revised version and compare it with the previous one. Also I have read the rebuttal letter by the authors. I think this is a clear improved version. I have no main corrections or comments.
Therefore, I think this manuscript version can be acepted in the current form for publishing.
Experimental design: It have been improved following some suggestions by referees.
Validity of the findings: The new discussion is improved with new data, comments and comparison with new citations. Findings as better supported now.
Additional comments: I think the revised version is clearly improved and deserve to be published.
I have found only a very minor thing to correct. In line 584 I think that references must be cited by year of publication, so "Dos Santos (1999), Torres et al. (2014) and Pochelon et al. (2017)", I do not understand the current order (based on...?).


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: 13C VALUES OF GLYCOLYTIC AMINO ACIDS AS INDICATORS OF CARBOHYDRATE UTILIZATION IN CARNIVOROUS FISH
Review round: 1
Reviewer: 1

Output:
Basic reporting: This paper provides a good data for effect of dietary carbohydrate on the d13 C values of amino acids in Atlantic salmon using compound specific stable carbon isotope analysis. In this study, Atlantic salmon were reared either on a control diet with high carbohydrate (corn starch) or on an experimental diet with low carbohydrate (red seaweed, Palmaria). The control diet was a mixture of fish and pea protein (58%), lipids (20%), carbohydrate: wheat gluten (9%) and corn starch (18.7%).  The experimental diet added a low carbohydrate ingredient, Palmaria in all the same composition without corn starch. Authors found that there is a wide difference in d13 values of glycolytic amino acids (Ser, Gly and Ala) from diet to body tissues. They suggested that glycolytic amino acids are associated with dietary carbohydrates.

Although the paper contains potentially interesting information, my main concern with this paper is that authors need to clarify the composition of main three dietary nutrients (proteins, lipids, carbohydrates) within high carbohydrate diet (Control) and low carbohydrate diet (Experiment). Furthermore, Authors need to clarify the d13 C values of each component. For example, although both corn starch and wheat gluten are carbohydrate, d13C values of corn starch are approximately -9 to 11‰ and wheat gluten is around -25 to -30‰. Average of both mixture is around -20‰. Palmaris is composed of 22% protein and no carbohydrate and d13 values of Palmaris is -20‰. Both control and palmaris added are -20‰ but there is difference in each component. Furthermore, there is other influence of lipids on both Control and Experiment. Authors need to explain how d13C values of lipids are consistent both two diets and then argue that d13C values of glycolytic amino acids came from Palmaria carbohydrate, not from Palmaria lipids and fish oil.

Other recommending on this paper is that it is much better to do more focusing on effect of carbohydrate on glycolytic amino acid syntheses, even though all data came from amino acids. This paper is trying to explain three nutrients (amino acids, carbohydrate and lipids) on one experiment and it is quite confusing. So I suggest it needs to rewrite on each paragraph focusing on the influence of carbohydrate on d13C values of amino acids. Several NEAAs also are strongly related to lipids when there is less amount of proteins in diets. Authors need to explain more why Ser, Gly and Ala was significantly affected by the isotopic values of energy components (carbohydrates) of diet. Alanine is formed by the addition of an amino group to pyruvate, a precursor of glucose. Glycine is synthesized from serine, which is synthesized from 3-phosphoglycerate.

Additionally, authors tried to put all NEAA and EAA data on Figures, but I suggest that all EAA results (no difference) should be sent back to supplementary information and put only the NEAA data on figures to make clear on the main finding.
Experimental design: Experimental design on this paper is based on feeding experiment to test the effect of seaweed carbohydrate on amino acids on fish. Authors compared Atlantic salmon with and without low carbohydrate seaweed Palmaria. All data were well presented and results are interesting for scientific community. However, authors need to do more works on the composition of each diet (control and seaweed inclusion) based on the macronutrients (protein, lipid, carbohydrate) and require more explanation on carbohydrate and glycolytic amino acids.
Validity of the findings: All underlying data were provided well and Conclusions are well stated.
Additional comments: Detailed comments

Line 98-100: Authors mentioned Atlantic salmon and metabolic fate of plant carbohydrate. Plant has protein, lipid and carbohydrate. I think authors need to explain why plant carbohydrate is important in carnivorous fish instead of plant protein or plant lipid. Authors mention Palmaria is  low carbohydrate and high protein and it is confusing.

Line 224-225: Salmon muscle contains a lot of lipids compared with other species. Without extracting lipids from salmon muscle tissues, d13C values of amino acids would be biased due to lipid effect. Authors need to explain why the lipid correction was used on muscle tissues and how it is correct. There is increased demand for dietary lipid as an energy source when no carbohydrate in Palmaria. And the sentence “to affect on d15N values” is not relevant and focusing on d13C values and please delete.

Line 255-256: This sentence is not correct. D13C values of marine sourced proteins were higher than the terrestrial protein sources especially Gly is highest in marine proteins and it is used for detecting the consumption of marine food in human diets. Please correct.

Line 272-273: Authors need to explain why Ala is negative and Ser and Gly is positive in d13C values offset between fish and diets on Discussion part. It suggests there is different pathway on synthesis on both negative alanine and positive serine or glycine. Please focus on three glycolytic amino acids along with corn starch or non-starch carbohydrate.

Line 303-304: Authors need to mention this corn starch in Intro part. I suggest authors need to explain how corn starch is different from non-starch polysaccharides.

Table 1: This table should have percentage of protein, lipid, carbohydrate and include each carbon stable isotope values from Control and Experiment diet. Please add three nutrients and carbon stable isotope values of three nutrients. Especially, please add composition of carbohydrate in Palmira.

Figure 2: I suggest that only B is enough for showing the results and better to delete A or delete EAA data on Figure 2 and simplify the figure.

Figure 3: Figure 3 is very confusing and difficult to figure out the difference between control and Palmaria inclusion.  Please use only compound diet and Palmaria focusing on difference.


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: 13C VALUES OF GLYCOLYTIC AMINO ACIDS AS INDICATORS OF CARBOHYDRATE UTILIZATION IN CARNIVOROUS FISH
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: See 'General comments for the author'
Validity of the findings: See 'General comments for the author'
Additional comments: Dear Authors,

This letter constitutes my review of the manuscript titled “13C values of glycolytic amino acids as indicators of carbohydrate utilization in carnivorous fish” by Yiming V. Wang et al., for PeerJ.

In this study, the Authors investigated the effects of high- and low-digestible carbohydrates in diets to the carbon isotopic composition of amino acids in Atlantic salmon. I feel that the data reported are novel and may be valuable in that they impact how we can understand carbon isotopic composition of amino acids in organisms. However, I feel that the interpretation/explanation of the data observed is insufficient in this manuscript, because I have great concerns (which are described below) in several places in this manuscript. I therefore recommend ‘major revision’ to the handing editor of PeerJ. Sorry for negative comments, but I will be grateful if the Authors positively receive my comments, and if my comments are useful to enhance the value of the revised manuscript.

1.	From this manuscript, I did not understand why the cap-delta 13C (muscle vs. diet) values are significantly large or small among non-essential amino acids such as Ser, Gly, Ala, Glx, and Pro, but negligible for Asx. It will be better if the Authors can explain specific mechanism to make such variation in the cap-delta 13C values among non-essential amino acids.
2.	Also I did not understand why the variable delta-13C value are related to biosynthetic activity of non-essential amino acids. I think that abundance of non-essential amino acids (except for Alx) should be significantly increased from compared to that of essential amino acids, if the biosynthetic activity of non-essential amino acids is significantly large enough to change in the delta-13C value between salmon and diets for the amino acids. It will be better if the Authors show change in the molar balance (i.e., increase the of non-essential amino acid abundance, except for Alx) of amino acids from diets to salmon.
3.	I am very much doubtful of digestibility of corn starch in the Atlantic salmon. I think that the salmon have no chance to feed on any starch in wild environments, and therefore that they have little or no digestibility of corn starch. Moreover, in general, for human, we can easily digest cooked (e.g., boiled) starch, but hardly digest raw starch. It will be better if the Authors show digestibility of ‘raw’ corn starch for salmon that used in this study. Probably, reference literature(s) for line 303-304 is(are) required.
4.	In this manuscript, a little information is available for the controlled feeding experiments of the salmon. I think that, for example, initial size of the salmon before experiments and change in the biomass size (e.g., length, weight, etc.) during the experiments are at least required. These data will be very useful for readers of this manuscript, to recognize that the 14 weeks are enough in this experiments. Perhaps, it will be better if the Authors show the growth carves of salmon during the trials in this experiment.

Please do not hesitate to contact me if you have any questions about my review.

Sincerely yours,

July 6, 2019
Yoshito Chikaraishi
ychikaraishi@lowtem.hokudai.ac.jp


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: 13C VALUES OF GLYCOLYTIC AMINO ACIDS AS INDICATORS OF CARBOHYDRATE UTILIZATION IN CARNIVOROUS FISH
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I have four concerns on the previous manuscript, and I recognize that the Authors have made revision sufficiently against the review comments (1), (3), and (4), but not the comment (2) about change in the molar balance of amino acids from diets to salmon.

I appreciate that the Authors added a supplemental table (Table S5) for showing AA compositions for diets and salmon. However, my original concern is increasing when I saw them. The Authors have explained that the variable delta-13C value are related to biosynthetic activity of non-essential amino acids (except for Alx). If this explanation is true, I think that abundance of non-essential amino acids (except for Alx) should be significantly increased compared to that of essential amino acids, because the biosynthetic activity of non-essential amino acids is significantly large enough to change in the delta-13C value while the biosynthetic activity of essential amino acids is zero. However, in Table S5, I did not find such increase of non-essential amino acid concentration in the AA composition from diets to salmon. Thus I think that ‘the Authors’ explanation for the delta-13C value (i.e., large biosynthetic activity of non-essential amino acids)’ and ‘observation for the AA composition (i.e., little increase of non-essential amino acid concentration)’ may be contradictory to each other.
I think that this concern is not main point of this study, but I would like to see potential/possible explanation for this contradiction.

Sincerely yours,
August 10, 2019
Yoshito Chikaraishi
ychikaraishi@lowtem.hokudai.ac.jp


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: CHEMICAL CHARACTERISATION OF POTENTIAL PHEROMONES FROM THE SHOULDER GLAND OF THE NORTHERN YELLOW-SHOULDERED-BAT, STURNIRA PARVIDENS (PHYLLOSTOMIDAE: STENODERMATINAE)
Review round: 1
Reviewer: 1

Output:
Basic reporting: Overall, this manuscript contains interesting data that certainly improve our knowledge of chemical signals in bats. Although the language and tone are appropriate, the presentation lacks some detail and clarity.

Pheromones have been defined in a variety of ways and the authors clearly state their working definition, but given the semantic debate over this term, it may be best to avoid it altogether. Since the function of this chemical signal is still unknown, it may be better to use a more general term like semiochemical or chemical signal. Additionally, the description of the two categories of pheromonal effects (lines 49-55) is curious as it seems to conflate different classification schemes. Further, the definition of primer pheromones is inconsistent with previous definitions (see Wyatt 2010). Rather than try to classify semiochemicals, it might be more useful to give a general sense of diversity of functions. This issue of defining pheromones reappears in the discussion (e.g. lines 287-290). It may be true that sexual stimulation and attraction are the most common functions assigned to pheromones, but perhaps this is because the term pheromone is more often invoked for chemical signals involved in reproduction. There numerous examples of mammalian chemical signals that are not directly used for mate attraction and stimulation.

The introduction incorporates some relevant background, but the overall organization could be improved. For example, in the second paragraph the authors make a case for the importance of chemical signaling in bats (although citations are sparse), but the details of variation in vomeronasal organ and associated genes seems out of place. As the authors mention later, the absence of the VNO does not preclude the use of intra-specific chemical signals. The variation among bats is interesting, just not relevant at this point in the manuscript. If the authors insist on keeping this content in the introduction, it would be helpful first introduce the structural variation, then discussion the genetic variation. Wible and Bhatnagar (Journal of Mammalian Evolution, 1996, 3:285-314) described variation in VNO morphology before the more recent genetic methods.

I appreciate at the introduction concludes with a clear statement of their objective and brief statement of the methods used. This objective is clearly met with the data presented. With a bit of polishing this manuscript will be a great foundation for further study of chemical communication in Sturnira spp.

Detailed remarks:
line 27: Consider rewording “mostly highly developed” to “more developed”
line 81: “A number of species in the family Emballonuridae [have] sacs…”
line 86: This description of S. bilineata behavior seems to conflate salting with hover displays.
line 105: Consider replacing “unstudied” with “understudied” as the latter half of the paragraph shows the trait is not completely unstudied.
line 95: Much of the work on S. bilineata described here was done prior to Schneeberger et al 2016. Also, the chemical composition has been examined by Caspers et al (Chemical Signals in Vertebrates 11, 2008, pp. 151-160).
line 126: extra parenthesis in citation
line 150: ambiguous sentence
line 175: “We carried out [v]olatile…”
line 228: It should be Bray-Curtis dissimilarity
line 244 (and elsewhere): The phase ‘status groups’ is initially confusing. Define clearly.
line 315: Intended meaning of “specificity” is unclear. Consider rewording to emphasize sexual dimorphism.
line 334: sentence fragment
line 349: cumbersome and potential run-on. Consider splitting.
line 392: “…further work [is] needed to confirm…”
line 408: Bhatnagar and Meisami (1998) appears in the reference list but is not cited in the text.
Experimental design: The overall design of the study seems sound and the methods used to identify the individual compounds are rigorous, but several details are missing from the description of the methods. For example, during what months were the samples collected? If these bats are seasonal breeders, one might expect seasonal variation in such a sexually dimorphic secretion. Given that males are classified as either reproductive adults or sub-adults, it would be helpful to have more detail about the assessment of reproductive status. Are all adults assumed to be reproductive or was the size/state of the testes examined?

The description of where the samples were collected from the bats is unclear (line 149-151). At first I thought two samples were collected, one from the shoulder patch and one from the surrounding area, but later realized only a single sample type was analyzed. Line 257 refers to the shoulder gland fur, whereas most other instances refer to the fur surrounding the gland. This may be semantic nit-picking, but the location of the hair samples is critical to the study.

Given that some solvents can interact with plastics, what type of vials were used to store the samples? Where any blanks (same solvent and vial but without the hair sample) run as controls? Also, the authors note that the MCT solvent stabilizes the secretion, but it should still be reported approximately how long the samples were stored before analysis. On a similar note, how well does the solvent stabilize the secretion, if volatiles were able to be extracted from the headspace via SPME?

The chosen statistical methods are reasonable, but the large differences in dispersion between groups may impact the results (see Anderson and Walsh. Ecological Monographs, 83(4), 2013, pp. 557–574).

Some additional details will also allow the reader to better understand how the data were analyzed. For example, the authors report there were 15 major peaks, but is unclear what criteria were used to define peaks as major. Perhaps this distinction is obvious from the chromatograms, but a sample chromatogram was not provided. Similarly, it seems the relative areas are calculated relative to only the total area of the major peaks not the total area, both major and non-major; this should be made more clear. The software that was used to identify peaks and calculate the area should also be indicated.

It is also unclear to me why the authors have chosen to analyze both relative and absolute areas. It’s completely understandable that the size of the hair sample could be precisely controlled, and using relative areas is an obvious solution, so why also analyze absolute areas at all? From Table 1 it appears that males consistently had higher peak areas and this may be interesting in its own right. If the fur samples were approximately similar in size, but the adult males consistently had much stronger signals, that may suggest adult males have more active glands. This is qualitatively interesting, even if it can be rigorously quantified. When examining composition, it seems more appropriate to use relative abundances. It is unclear which set of responses (absolute or relative) was used for the ANOSIM. Additionally, the number of permutations used in both the ANOSIM and SIMPER procedures should be reported.

The authors clearly state their compliance with ethical guidelines and animal handling protocols.
Validity of the findings: The authors do a nice job of connecting their findings to other studies that found similar compounds in semiochemicals of other species as well as dietary components. This was a clear strength of the discussion. Overall the discussion of the findings is appropriate, with one exception. The authors claim that their findings “strongly suggest a specific role in male-female attraction” (lines 40 & 285). This is not necessarily true. The sexual dimorphism of the glandular product does suggest a role in mating, but it is just as likely used for male-male competition as it is for mate attraction. At this point, one also cannot rule out its role in species recognition.

On lines 36 and 281 the authors suggest that the NMDS was the analysis used to test for differences. This is incorrect. NMDS is helpful for visualization, but the statistical significance is derived from the multivariate analysis of similarity.

Additional comments:
line 238: The text implies that only 5 samples were collected, when it seems that 5 samples were collected from each age/sex class.
line 237- 238: This sentence implies that all 15 peaks differed significantly, but that is not true.
line 259: sample size numbers are confusing because they refer to numbers of peaks, not samples
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: CHEMICAL CHARACTERISATION OF POTENTIAL PHEROMONES FROM THE SHOULDER GLAND OF THE NORTHERN YELLOW-SHOULDERED-BAT, STURNIRA PARVIDENS (PHYLLOSTOMIDAE: STENODERMATINAE)
Review round: 1
Reviewer: 2

Output:
Basic reporting: see general comments
Experimental design: no comment
Validity of the findings: no comment
Additional comments: This is an interesting paper which reports a first chemical characterisation of secretions which may have semiochemical roles in the bat Sturnia parvidens. As the authors observe, the differences between the secretions of adult reproductive males, juvenile males, and females indicate that these differences may be important in courtship and even mate choice in adults. The potential roles of these semiochemicals will no doubt be the focus of future investigations.

However, the introduction perhaps over emphasises novelty (e.g. Line 57 “yet very little is known about the possible role of pheromonal communication in this group” and Line 63 “ little work on olfactory systems in bats “) even though novelty is not a criterion for publication in peerJ. The current ms is an interesting piece of work in its own right. The introduction could be constructively rewritten to help the reader place the paper in the context of the fairly extensive previous work on semiochemicals in bats, currently cited later in the Introduction and in the Discussion, bringing them early into the Introduction. None of the previous work detracts from the interest of the current paper which builds on the solid work of others.

More specific points follow.

1.	Line 49 ff Rewording vs signature mixtures/pheromones with priming effects to remove the confusion of the terms. The concepts are perhaps more clearly explained in my 2017 short review [Wyatt, TD (2017) Pheromones. Current Biology 27: R739–R743.]  “Many pheromones … have both immediate ‘releaser’ effects and longer lasting primer effects.“  and  “It is worth separating the concepts of pheromones and signature mixtures (it was a mistake in my 2003 book to combine them). It is precisely because individuals have different odour profiles that these can be learnt and used to distinguish different animals.”

The current text reads:  Line 49 ff  “There are two general categories of pheromonal effect: primer pheromones are fixed chemical signals that elicit a defined response in the recipient animal (behavioural or physiological), while signature mixtures are emitted signals which are learned by the conspecific receivers and often have complex and variable chemical profiles. These “individual mixtures” or “signature odours”, which act in social communication though learning, rather than eliciting a specific behavioural or endocrine response, may identify a specific individual or social group (Wyatt 2010, Dehnhard 2011).”

Suggest change to “  “There are two general categories of semiochemicals within a species: pheromones, chemical signals which elicit a stereotypical behavioural or physiological response in the recipient animal, while signature mixtures are odour cues which are learned by the conspecific receivers and often have complex and variable chemical profiles. These “individual mixtures” or “signature odours”, which act in social communication though learning may identify a specific individual or social group (Wyatt 2017, 2014, Dehnhard 2011).”

2.	Pheromones (in mammals) are not exclusively detected by the VNO (see Section 9.3.1 in Wyatt 2014 or Figure 4 in Wyatt 2017) [as noted in the ms Lines 127-8]. It would be worth also adding here, after “potential pheromones.” , “Pheromones in mammals are also detected by the main olfactory system (Wyatt 2014, 2017) so the lack of functioning VNO receptors does not rule out the possibility of pheromones.”

3.	Could the authors integrate the information about bat VNO in the paragraph starting Line 56 with the paragraph starting Line 116.   (in the later paragraph we learn that Sturnia parvidens does have a VNO and functional Trpc2.)

4.	Line 97 “did not undertake actual chemical characterisation of the wing-sac liquids” does not seem to match the cited paper [Schneeberger et al. (2016)] which reports in its methods:
“Wing sacs were wiped out with a piece of cotton of ∼5 mg that had previously been washed with dichloromethane (99.9%) to remove potential contaminating substances. We stored the samples in a Teflon-capped glass vial (2 ml RotilaboH, Karlsruhe, Germany) at -80◦C until analysis. After sample collection, we released all bats at the site of capture.
[…]
Chemical Analysis
All samples were prepared for analysis and analyzed at Bielefeld University. Prior to chemical bouquet extraction, we added 100 µl of dichloromethane (DCM) to each sample”

5.	Line 285:  “This strongly suggests a specific role in male–female attraction (rather than just species recognition”   . A sex pheromone can have both functions.

6.	Line 287 ff As there has been no behavioural or physiological investigations of the secretions yet, I would delete Lines 287-306 inclusively. Such speculation is not helpful to the reader.

7.	Line 308 Suggest delete the sentence starting “For example, in mixed”.

8.	Line 329 – description of P hastatus and its sexually dimorphic glands might be better in the introduction? Interesting.
Discussion

9.	Lines 337-376.  Delete the words comparing the molecules found in this species with those of unrelated species. Whether a molecule is a pheromone in another species is largely irrelevant (unless it’s a close relative).  Tells us nothing about the likely importance in this species.

10.	Line 393 – suggest change “odour signatures” to “secretions”

11.	Minor:  Wyatt 2010 missing from reference list


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: CHEMICAL CHARACTERISATION OF POTENTIAL PHEROMONES FROM THE SHOULDER GLAND OF THE NORTHERN YELLOW-SHOULDERED-BAT, STURNIRA PARVIDENS (PHYLLOSTOMIDAE: STENODERMATINAE)
Review round: 2
Reviewer: 1

Output:
Basic reporting: The revised manuscript is greatly improved and I look forward to seeing it published. The revised introduction provides adequate background and the discussion addresses how the results fit within the current state of knowledge.
Experimental design: The methods now provide sufficient detail to fully understand the procedure and replicate the results. Supplemental figure 1 is a nice addition, as it really highlights how distinct the adult males' profiles are, relative to females and juvenile males. It would be helpful to see a similar figure for the headspace analysis as well. Labeling all the peaks may not be feasible, given the number of compounds listed in table 2, but perhaps those that differ significantly can be indicated.

It is still unclear whether the reported ANOSIM results are based on absolute or relative peak areas.  Figure 2 contains four plots, but only two statistical results are included in the text. The methods seem to imply the relative abundances were used, but this should be stated more clearly.

In the methods, the authors state they use a SIMPER analysis to identify which compounds underlie the differences between the groups; however, the footnote of Table 2 states the significance was assessed by a one-way ANOVA.  Are the differences reported in the text based on the SIMPER analysis as suggested by the methods or one-way ANOVAs as suggested by the table footnote?
Validity of the findings: The revisions adequately address the initial over-emphasis on mate attraction.
Additional comments: Line 195: missing parenthesis
Line 307: replace "using non-metric multi-dimensional scaling" with "multivariate analyses"


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: CHEMICAL CHARACTERISATION OF POTENTIAL PHEROMONES FROM THE SHOULDER GLAND OF THE NORTHERN YELLOW-SHOULDERED-BAT, STURNIRA PARVIDENS (PHYLLOSTOMIDAE: STENODERMATINAE)
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors have made significant and helpful improvements to the ms in response to the reviewers' comments.  I particularly appreciated the detailed responses and the availability of the tracked-changes ms version which made it quick to assess revisions.