Decoding Anotylus (Thomson 1859) Beetle Diversity: DNA and External Morphology Match in Área de Conservaciόn Guanacaste, Costa Rica

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

It is overall a very nice study and I enjoyed reading the manuscript. I see, however, one major issue and some smaller problems, which I list below:

- It is surprising that the authors did not attempt to make any species delimitation analysis. In my opinion, this is a must in such a study (major issue)

- I strongly argue that more than one outgroup should be added to the analyses to increase the robustness of the estimations. 

- When measuring the body length, the usual problem is the extent of the abdominal segments' retraction. Depending on this, the length can vary and is not a good estimator. I rather suggest measuring the length of the forebody (head+pronotum+elytra) to ensure comparability between specimens.

- Why the supports' values on the tree are given with two decimal numbers? Or is the first number indicating support from one analysis and the second from the other analysis? If so, it is not clear and confusing for the reader.

- Please, take the number of currently described species of Staphylinidae from the Catalogue of Life where it is most updated

- line 119: remove double l in "Oxytellinae"

After implementing these changes, I strongly recommend the publication of the manuscript.

Comments on the Quality of English Language

There are some stylistic errors (e.g. missing commas), which makes reading slightly uncomfortable, but it just requires minor changes.

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

This is an interesting and well thought-out piece of work.  I wonder if any of the recognised species has been described and personally I consider that this should be researched and if appropriate the species should be described and named.

Author Response

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Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

General comments:

This manuscript seems not well written in editorial and grammatic terms. And as a taxonomist, I do not think the morphological part of this study has much value to science. The species delimitation in Anotylus as well as many other oxyteline or largely staphylinid genera is based on aedeagal or sexual features in combination with other qualitative features. The morpho traits used in this study are commonly present in the descriptions of new species, but they are in most cases not at all diagnostic. So I will not be surprised to see the overlapping pattern in Figure 3. And the authors did not discuss the assembly of Anotylus along with the evaluation, which is very disappointing. A great deal of models or hypotheses have been proposed to explain the elevational variation of species assembly, like the Mid-Domain Effect, Area Effect, climate factors, etc. The authors should test those hypotheses and elucidate the change of species assembly. That being said, I advocate the acceptance of publication after a major revision. I think the authors should be encouraged (to improve their study and modify the manuscript) rather than be rejected, as few such studies can be found in the rove beetle community.

Specific comments:

Page3 lines 105-106: Actually most of staphylinid or oxyteline genera do not have "singular morphological character unique" to themselves. When you read the Ref [56], you must find this. And in "However, [57] suggested the genus...", who did that? There are a lot of editorial errors in the text, esp. in References, the authors should check the text carefully.

Page 4 lines 189-190: Did you perform the ML phylogenetic analysis in MEGA 11 or another programme? And why bootstrap with 7500 replicates? Although it is plausible, but most commonly it is set like 1000 or 2000 or like wise.

Pages 4-5 lines 196-206: How did you estimate the area of elytra and eyes? Elytra are irregular polygons or even not polygons (they may have curved margins). And eyes are convex surfaces. And the authors found the elytral area explains much more specific level variation than other variables, but the elytral areas may vary with the body size, did you calibrate such variables according to the individual bodysize?

Page 5 lines 220-233: I do not like such interpolation to do with missing values. It is okay to use that, but when you calculate the degree of freedom, please do not count the filled missing values in. They exist simply for making the calculation executable, they did not have any information about the variation of the sample. Authors usually overlooked that and made the degree of freedom overestimated and thus inflate the statistical significance.

Page 6 lines 240-244: Since I cannot find the supplementary files (Table S3), I can hardly make sure the use of ANOVA (and post-hoc comparisons) is correct. Very popular issue here is using simple ANOVA in lieu of phylogenetic ANOVA, as species here is not statistically independent (see your phylogeny). I hope the authors noticed that and use it in right way.

Page 9 lines 310-311: "Neotropical Anotylus. As has been done", it should be a comma rather than a full stop here.

Ref 55: The surnames and given names of the authors of this article were confused. And the name of the publication is wrong. It is Zootaxa not "English". Similar errors are also found in Ref 85.

References: when citing the name of a journal, the authors should use either the full name or short name (abbreviation), not mix in a same article. And if you use the full name of PNAS with "of the United States of America", you must keep using this form across the whole article.

Comments on the Quality of English Language

I notice the authors are all working with the universities from English-speaking countries. That is why I do not understand the quality of this MS looks not that good, just regarding to the writing. The authors may need to put more effort or time into the editing work.

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

I appreciate the changes that the authors provided. I think, however, that my major complaint - lack of the species delimitation analysis - is not satisfied. While I acknowledge that it was a project with limited scope, running one more analysis would not be difficult. I do not say it has to be translated to the formal species description, it is more for confirming the phylogenetic results, which are not the best for detecting the lineages at this taxonomic level. I explicitly think about e.g. ASAP or ABGD analyses. After including this, I think the manuscript will be ready for publication. 

Author Response

Reviewer #1

I appreciate the changes that the authors provided. I think, however, that my major complaint - lack of the species delimitation analysis - is not satisfied. While I acknowledge that it was a project with limited scope, running one more analysis would not be difficult. I do not say it has to be translated to the formal species description, it is more for confirming the phylogenetic results, which are not the best for detecting the lineages at this taxonomic level. I explicitly think about e.g. ASAP or ABGD analyses. After including this, I think the manuscript will be ready for publication. 

 

Response:

Thank you for your thoughtful comments in each round.  There are many algorithms that detect putative species based on sequence variation.  Indeed, the detection of sequences sufficiently divergent to be useful as species hypotheses is what the BIN algorithm in BOLD accomplishes (an analysis specific to this gene region and this taxonomic level).  Following the reviewer’s suggestion, we have added the following sections:

Line 180: “We compared one other method of species delimitation via genetic distance (e.g. Assemble Species by Automatic Partitioning (ASAP) [78]) with the BIN-derived species hypotheses.”

Line 269: “The best ASAP estimate of species via sequence divergence (asap-score of 2.5) estimated 14 species within the 18 BINs by lumping together the four species pairs with less than 5 % inter-specific sequence divergence. As these four species pairs are characterised by sympatric elevational distributions (Table 1) and marked (if not always significant) PC1 scores of morphometric differentiation (Table S3) we follow the BIN divisions as the first step in this integrative taxonomy. An ASAP partition that agreed with BIN divisions was one of the ten best partitions (asap-score 7.5).”

Reviewer 3 Report

Comments and Suggestions for Authors

Ref 55: the name of the first author is "Lü" not "Lu". 

Ref 60: there is a domain link string inserted in that reference ("<br class="br"/>"), please remove it.

Please do not take the MDPI style as a scapegoat. Formatted data often makes mistakes, but it is the author's responsibility to check and correct them.

For me, no more issues. But there may be more minor errors that I have not found. Anyway, I am delighted to see this manuscript finally be published with Diversity. 

Comments on the Quality of English Language

I am not native speaker. I did not find any more English issues in terms of grammar and can understand the main text, but I am not so sure of its "quality". 

Author Response

Reviewer #3:

Ref 55: the name of the first author is "Lü" not "Lu". 

Ref 60: there is a domain link string inserted in that reference ("<br class="br"/>"), please remove it.

 

Response:

Thank you for your thoughtful comments in each round.  Citation 55 and Citation 60 have been corrected.