Population Genetics of the Greater Grison (Galictis vittata; Mustelidae, Carnivora) and How Changes Since Miocene–Pliocene Have Shaped the Phylogenetic Relationships Between Grison Species and Other Neotropical Mustelid Genera

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

REVIEW: Population Genetics of the Greater Grison

This manuscript provides a standard but very extensive population genetic analysis of the Greater Grison based on mitochondrial DNA. While the effort is appreciated, the analysis feels excessive for the relatively small dataset and close relationships among samples. Many of the statistics add little insight.

Concatenating all loci into a single super-gene and using simpler K2 distances with bootstrap support would have been sufficient and more appropriate. Reporting values to four decimal places overstates the precision of the data, and the use of primary colors in the figures gives an amateur impression.

The species description and background is very detailed, a taxonomical review

Overall, it is a solid contribution to an understudied species, though somewhat over-engineered.

Four digits on many values are excessive and imply precission where there is none above two digits after the decimal.

Advice:

• don't use primary colors for your graphics, it looks amateur.

• AVOID phrases like: However, as we will see later, inferences using molecular

In all, a solid somewhat awkwardly over the top analysis and contribution.

Recommendation: ACCEPT with minor revisions

Author Response

Dear Editor,

In this document we respond to all the reviewers' comments, incorporating many of them into the new version of the manuscript. The new parts added to the text appear in red. Below, we answer in detail each of the questions or modifications requested by each of the three evaluators.

Reviewer 1:

1-Reviewer 1 said: “This manuscript provides a standard but very extensive population genetic analysis of the Greater Grison based on mitochondrial DNA. While the effort is appreciated, the analysis feels excessive for the relatively small dataset and close relationships among samples. Many of the statistics add little insight.”

Answer: We thank this reviewer for their comments. Regarding the number of statistics used, these are necessary, precisely because of the small sample size. With such a small sample size, some results could be highly spurious. Using multiple statistics allows us to analyze whether the results of some statistics are spurious, or random, or whether all the statistics, regardless of sample size, show the same pattern of results, thus lending reliability to the results obtained.

2-Reviewer 2 said: “Concatenating all loci into a single super-gene and using simpler K2 distances with bootstrap support would have been sufficient and more appropriate. Reporting values to four decimal places overstates the precision of the data, and the use of primary colors in the figures gives an amateur impression”.

Answer: If the sample size were identical for each gene analyzed, reviewer 1's idea would be perfect. The problem is that for G. vittata, the sample size is very heterogeneous for each marker analyzed (from 13 to 4 specimens, and they are not always the same). Similarly, the otter species analyzed were different for each mitochondrial marker used. This makes it impossible to create a single phylogenetic tree for the three databases used simultaneously. The reviewer's advice was followed, and all statistical values ​​have been reduced to two decimal places. The use of primary colors in certain figures has been modified following the reviewer's recommendations.

3-Reviewer 3: “The species description and background is very detailed, a taxonomical review

Overall, it is a solid contribution to an understudied species, though somewhat over-engineered.”

Answer: As the reviewer states, this work, in addition to presenting the first genetic data for G. vittata and L. patagonicus, is a review of the taxonomy and systematics of these neotropical mustelids due to the scarce knowledge of these Neotropical grisons.

Reviewer 4: “Four digits on many values are excessive and imply precission where there is none above two digits after the decimal. Advice:

• don't use primary colors for your graphics, it looks amateur.
• AVOID phrases like: However, as we will see later, inferences using molecular.
• Recommendation: ACCEPT with minor revisions

Answer: As mentioned previously, the four digits of all estimated values ​​were replaced with two digits, as requested by this reviewer. Similarly, primary colors were not used in some figures, and the words indicated by the evaluator were removed. We sincerely appreciate the evaluator's input.

Reviewer 2 Report

Comments and Suggestions for Authors

Title: Population Genetics of the Greater Grison (Galictis vittata; Mustelidae, Carnivora) and How Changes Since Miocene-Pliocene Shaped the Phylogenetic Relationships Between Grison Species and Other Neotropical Mustelid Genera

Authors: Manuel Ruiz-García and Juan Esteban Fula-Rodríguez 

Reviewed by Roger A Powell

GENERAL  COMMENTS

Title – The title is way too long.  Seems to me that this would suffice: “Phylogenetic Relationships Among Grison Species and Other Neotropical Mustelids”.

Simple Summary – This summary is too long to be a simple summary.  Readers want a short summary, preferably something less than half this length.

Abstract – Similar problem as with the Simple Summary: this Abstract is too long.  I packs so much detail that readers can not keep track of much of it and, instead, get lost and forget what they read.  I recommend an Abstract no longer than 250 words.

line 13 – "Carnivore” is confusing because it can mean "member of the Carnivora" or "meat eater".  Paleontologist have a solution: Use "carnivore" for a meat eater and use "carnivoran" for an animal that is of a species that is a member of the Carnivora.  I have adopted this convention.  I recommend that you do, too.

Providing range maps for the species would be nice.

     I have trouble with your Introduction and Discussion.  You have material in your Discussion that I think belongs in your Introduction.
     A research paper tells a story that progresses through time, with a past, present and future.  The Introduction synthesizes the past and brings the reader to an author’s hypotheses and objectives.  The hypotheses or objectives direct the reader to research design and Methods, which are in the recent past.  The Methods lead to Results, which are the present.  Finally, the Discussion shows what an author’s results mean for future research.  There we have the story: past, present, future.
     An Introduction should synthesize the pertinent literature, leading readers logically through past research and past developments to the hypotheses or objectives of an author’s research.  The Introduction should start from a broad perspective and then narrow to the author’s research topic.  A good rule of thumb is that EVERY publication pertinent to an author’s research should be introduced in the Introduction.  This rule means that no new literature should be introduced in the Discussion.
     In contrast to the Introduction, which comes first and brings the reader through the past to the present (the Introduction synthesizes past work), the Discussion comes last and looks to the future.  A classic Discussion reiterates briefly the highlights of the Results and then explores what those new Results mean for future research.  The importance of the Results should be obvious from the Introduction, which synthesized ALL the pertinent, past research and which directed the readers to the author’s hypotheses or objectives.  The Results make clear which hypotheses (all presented in the Introduction) can be rejected and which can be refined for future research.  The Discussion should dwell on what the Results mean for future research.  What NEW hypotheses have been generated?  In a good Discussion, an author does not present hypotheses generated from the literature that might explain the Results; the Introduction should already have developed all the pertinent hypotheses that have come from past research and should have used those hypotheses to lead readers to the hypotheses tested or ranked in the Results.  And the Discussion should not present results from past research that are similar to or contrasting to your results because those results should have been handled in your Introduction.  New literature can be cited if the Results point towards completely new, unanticipated directions for future research.
     Limit the Discussion to topics related directly to the Results and to future research.  Avoid tangential topics.  The new ideas related directly to the Results, and the potential for future research related directly to the Results, will be exciting enough that tangential topics need not be addressed.  For most manuscripts and papers, another rule of thumb is that the appropriate length for a Discussion section is about ¼ or less of the text (from Title through Discussion).  Your Discussion is half of the text of your manuscript.  I got lost wading through it all.  Cut it down to ⅓ of its present length.

SPECIFIC  COMMENTS

I have suggested some revisions in wording here and there in the text.  I have highlighted those places.

Lines 79-80 – This statement is too extreme.  Yes, taxonomy is important but it is not all important.  For decades, fishers were included in the genus Martes though we understand now that they have their own genus, Pekania, and are not as closely related to the martens as are wolverines.  Despite this major mistake of taxonomy, no federal, state or provincial agencies had any trouble managing fishers because of the misguided taxonomy.  Including fishers in Pekania did not affect their conservation status or management.

Line 83 – Throughout your ms – “however” – Strunk & White, in “Elements of Style”, recommended against starting sentences and independent clauses with “however” followed by a comma when it means “nevertheless” or “nonetheless”.  Starting sentences with “However” followed by a comma has become common in biology (though not in other disciplines) but, nonetheless, Strunk & White’s rule still has merit for 2 reasons.  1) At the beginning of a sentence (or independent clause) and without a comma, “However” means “No matter how”.  “However often I get caught in the rain, I still do not learn to bring my rain gear.”  A reader can mis-anticipate the sentence to come when the sentence starts with “However” meaning “Nevertheless”.  2) “However,” (with the comma) can be a harsh jolt for a reader and, far too often, the sentence following “however” does not make clear exactly what from the previous sentence is to be compared to something in the following sentence.  When I find “However,” at the beginning of a sentence in my own writing, I use it as a red flag for a sentence that can be improved.  Try replacing “However” with “Nevertheless”, “Nonetheless”, “In contrast” or some other wording.  Try putting “however” into the interior of the sentence.  Try leaving “however” out of the sentence entirely.  I bet you will find that you can improve the sentence.  Try it.  Use the Search & Destroy capabilities of Word (TrackChanges) to find every “However,” and revise the sentences.  You frequently start sentences with “However,” but I have not marked those places.

Line 94 – Fishers are in Pekania (here and elsewhere).  The yellow-throated and Nilgiri martens may be in Charronia.

Line 133 – Use the comparative (lower, higher) only when making a comparison.  “Lower”than what?  Lower than higher elevations?  That is ttrite.  You do not make a comparison here so just use “low”.  

Lines 217-218 – Altitude is distance above sea level in the air, while elevation is distance above sea level on the ground.  Planes fly at altitudes.  Grisons walk at elevations.

Line 313 – Table 1 does not include any references to bones but does include teeth.  This problem needs fixing or explanation.

Line 487 – Should “p =” be “π =” ?

Line 882 – This statement is too strong, as it was in the Introduction.  Species range does need to be known but taxonomy is not so important.  For decades, fishers and American martens were managed successfully in Minnesota, in New York, in Ontario, and in other states and provinces even though fishers were managed as Martes pennanti and not as Pekania pennanti.  Having the taxonomy wrong did not make any difference for management and conservation.

Comments for author File: Comments.pdf

Comments on the Quality of English Language

See my comments.

Author Response

Reviewer 2:

1-Reviewer 2 said: “Title – The title is way too long.  Seems to me that this would suffice: “Phylogenetic Relationships Among Grison Species and Other Neotropical Mustelids”.

Answer: We are keeping the title as is for two main reasons. First, the manuscript analyzes two different aspects: 1-the genetic structure of one species (Galictis vittata) and 2-the phylogeny of the three Neotropical Ictonychinae species, along with that of other genera of Neotropical mustelids. The title proposed by this reviewer only encompasses the second part of the research but not the first. The second reason is that the title of the special issue to which this article is addressed emphasizes the changes that occurred from the Miocene to the Pliocene and how these changes affected fauna worldwide. Therefore, we believe it is appropriate to align the article's title with the title of the special issue.

2-Reviewer 2 said: “Simple Summary – This summary is too long to be a simple summary.  Readers want a short summary, preferably something less than half this length. Abstract – Similar problem as with the Simple Summary: this Abstract is too long.  I packs so much detail that readers can not keep track of much of it and, instead, get lost and forget what they read.  I recommend an Abstract no longer than 250 words.

 Answer: Following this reviewer's recommendation, the Summary has been shortened, and the Abstract has also been shortened to 350 words, which is slightly more than what this reviewer requested but is within the 500-word limit that the journal potentially accepts.

3-Reviewer 2 said:” line 13 – "Carnivore” is confusing because it can mean "member of the Carnivora" or "meat eater".  Paleontologist have a solution: Use "carnivore" for a meat eater and use "carnivoran" for an animal that is of a species that is a member of the Carnivora.  I have adopted this convention.  I recommend that you do, too”.

Answer: Indeed, we followed this reviewer's advice and used the word Carnivoran throughout the text (reviewer 3 also made the same observation). Therefore, we followed the advice of both.

4-Reviewer 2 said: “Providing range maps for the species would be nice”.

Answer: The reviewer's idea is interesting, but we believe it would further lengthen the already lengthy manuscript. Furthermore, the text explains in great detail all the current knowledge regarding the geographic distribution of the three species analyzed.

5-Reviewer 2 said: “I have trouble with your Introduction and Discussion.  You have material in your Discussion that I think belongs in your Introduction.
     A research paper tells a story that progresses through time, with a past, present and future.  The Introduction synthesizes the past and brings the reader to an author’s hypotheses and objectives.  The hypotheses or objectives direct the reader to research design and Methods, which are in the recent past.  The Methods lead to Results, which are the present.  Finally, the Discussion shows what an author’s results mean for future research.  There we have the story: past, present, future.

Answer: Following this reviewer's advice, we have moved several sections of the Discussion to the Introduction, especially those concerning the morphological differences between the three species considered. Regarding this reviewer's opinion that the Introduction represents the past, the Results represent the present, and the Discussion represents the future, I would like to comment. I completely agree with this reviewer that the Introduction represents the past and the Results represent the present. I wholeheartedly agree. However, I disagree as much regarding the Discussion. Indeed, the Discussion should address the future (contributing new ideas, new hypotheses, etc), but by comparing what was known in the past with what has just been discovered in the present. Therefore, the Discussion will always include this contrast between past and present ideas, allowing us to project a future. Consequently, not everything from the past needs to be confined to the Introduction. The past plus the present will shape the future, and this should be reflected in the Discussion.

6-Reviever 2 said: “Limit the Discussion to topics related directly to the Results and to future research.  Avoid tangential topics.  The new ideas related directly to the Results, and the potential for future research related directly to the Results, will be exciting enough that tangential topics need not be addressed.  For most manuscripts and papers, another rule of thumb is that the appropriate length for a Discussion section is about ¼ or less of the text (from Title through Discussion).  Your Discussion is half of the text of your manuscript.  I got lost wading through it all.  Cut it down to ⅓ of its present length”.

Answer: We have shortened the Discussion. Some paragraphs have been moved to the Introduction, and others have been removed from the manuscript. The text is now 48 pages long. Of these 48 pages, 15 are Discussion (31.25%), which is practically the 1/3 proportion that this reviewer suggests the Discussion should have in relation to the manuscript text.

7-Reviewer 2 said: “Lines 79-80 – This statement is too extreme.  Yes, taxonomy is important but it is not all important.  For decades, fishers were included in the genus Martes though we understand now that they have their own genus, Pekania, and are not as closely related to the martens as are wolverines.  Despite this major mistake of taxonomy, no federal, state or provincial agencies had any trouble managing fishers because of the misguided taxonomy.  Including fishers in Pekania did not affect their conservation status or management”. Line 882 – This statement is too strong, as it was in the Introduction.  Species range does need to be known but taxonomy is not so important.  For decades, fishers and American martens were managed successfully in Minnesota, in New York, in Ontario, and in other states and provinces even though fishers were managed as Martes pennanti and not as Pekania pennanti.  Having the taxonomy wrong did not make any difference for management and conservation.

Answer: Following this reviewer's comment, we have removed several phrases from the introduction and discussion that refer to the importance of taxonomy for biological conservation.

8-Reviewer 2 said: “Line 83 – Throughout your ms – “however” – Strunk & White, in “Elements of Style”, recommended against starting sentences and independent clauses with “however” followed by a comma when it means “nevertheless” or “nonetheless”.  Starting sentences with “However” followed by a comma has become common in biology (though not in other disciplines) but, nonetheless, Strunk & White’s rule still has merit for 2 reasons.  1) At the beginning of a sentence (or independent clause) and without a comma, “However” means “No matter how”.  “However often I get caught in the rain, I still do not learn to bring my rain gear.”  A reader can mis-anticipate the sentence to come when the sentence starts with “However” meaning “Nevertheless”.  2) “However,” (with the comma) can be a harsh jolt for a reader and, far too often, the sentence following “however” does not make clear exactly what from the previous sentence is to be compared to something in the following sentence.  When I find “However,” at the beginning of a sentence in my own writing, I use it as a red flag for a sentence that can be improved.  Try replacing “However” with “Nevertheless”, “Nonetheless”, “In contrast” or some other wording.  Try putting “however” into the interior of the sentence.  Try leaving “however” out of the sentence entirely.  I bet you will find that you can improve the sentence.  Try it.  Use the Search & Destroy capabilities of Word (TrackChanges) to find every “However,” and revise the sentences.  You frequently start sentences with “However,” but I have not marked those places.

Answer: We fully agree with the reviewer. All instances of "However" in the text at the beginning of sentences have been replaced with other words.

9-Reviewer 2 said: “Fishers are in Pekania (here and elsewhere).  The yellow-throated and Nilgiri martens may be in Charronia”.

Answer: We have included these genera in the description of genera within the mustelid subfamilies.

10- Reviewer 2 said: “Line 133 – Use the comparative (lower, higher) only when making a comparison.  “Lower”than what?  Lower than higher elevations?  That is ttrite.  You do not make a comparison here so just use “low”.  Lines 217-218 – Altitude is distance above sea level in the air, while elevation is distance above sea level on the ground.  Planes fly at altitudes.  Grisons walk at elevations.

Answer: Following this reviewer's advice, we have corrected those two aspects in the text.

11-Reviewer 2 said: “Line 313 – Table 1 does not include any references to bones but does include teeth.  This problem needs fixing or explanation.”

Answer: Table 1 now explains that these samples were fragments of mandibles that included teeth from animals that had been found run over.

12- Reviewer 2 said: Line 487 – Should “p =” be “π =” ?

Answer: Indeed, some Greek letters were misaligned in the text. The actual symbol is p.

Reviewer 3 Report

Comments and Suggestions for Authors

This paper describes new genetic data for the genera Galictis and Lyncodon, which have previously only been reported either rarely or not at all.  This study provides new data on evolutionary relationships and history, and population genetics (including  genetic diversity and genetic heterogeneity between populations) based  on three mitochondrial genes.  Overall the paper is well presented and written.

However, this kind of study is unusual these days when whole genome sequencing is becoming the norm and it would be good to understand why this was not done.

I have made numerous  minor comments and corrections on the paper attached.

One particular pint is that by including G. cuja and Lyncodon in the same genus Lyncodon, you would make that genus paraphyletic.  The authors' options are to keep the same taxonomic arrangement or include all three species in Galictis.

My main comment is that the paper is too long and it gets confusing trying to keep up with the vast array or results and discussion. I strongly recommend splitting it up into three papers as follows;

1. Phylogeography  of G. vittata
2. Population genetics of Galicitis/Lyncodon
3. Taxonomy and evolutionary history of Galictis/Lyncodon

Comments for author File: Comments.pdf

Author Response

Reviewer 3

1-Reviewer 3 said: “This paper describes new genetic data for the genera Galictis and Lyncodon, which have previously only been reported either rarely or not at all.  This study provides new data on evolutionary relationships and history, and population genetics (including genetic diversity and genetic heterogeneity between populations) based on three mitochondrial genes.  Overall, the paper is well presented and written.”

Answer: We are immensely grateful for this reviewer's comment regarding the general idea of ​​the manuscript.

2-Reviewer 3 said: “However, this kind of study is unusual these days when whole genome sequencing is becoming the norm and it would be good to understand why this was not done”.

Answer: This is a very interesting question. Indeed, nowadays, if sufficient financial resources are available, the ideal would be to analyze complete mitogenomes or even metagenomic data (WGS, SNPs, etc.). However, what happens if you only have two hairs from a specimen, a 0.5 cm² piece of skin, a 3-gram fragment of bone, or a single tooth? It is very difficult to obtain high-quality DNA in sufficient quantity to generate results for complete genomes. Furthermore, one needs financial resources to carry out such large-scale analyses. The interesting thing about this study is that, despite not having sufficiently large samples to extract high-quality DNA in large quantities, with sequences of small fragments of mitochondrial DNA well-balanced in terms of their evolutionary rates per million years, we obtained results regarding the phylogeny of Mustelids that are practically indistinguishable from those obtained in studies that analyzed more than 20,000 base pairs. This means that even with very small and very low-quality samples with small fragments of some genes, results can be obtained that are really important for reconstructing the evolutionary history of species, whose samples are very difficult to obtain.

3- Reviewer 3 said: “I have made numerous minor comments and corrections on the paper attached”.

Answer: All the corrections this reviewer made to the text have been incorporated into this new version of the manuscript. In fact, all the information in the results section regarding the divergence times between the different taxa analyzed has been moved to a new table, as requested by this reviewer.

4-Reviewer 3 said: “One particular point is that by including G. cuja and Lyncodon in the same genus Lyncodon, you would make that genus paraphyletic.  The authors' options are to keep the same taxonomic arrangement or include all three species in Galictis.

Answer: Indeed, as this reviewer points out, that is one of the important findings of this manuscript. Potentially, there could be different solutions to resolve the possible paraphyly of the genus Galictis.

5- Reviewer 3 said: “My main comment is that the paper is too long and it gets confusing trying to keep up with the vast array or results and discussion. I strongly recommend splitting it up into three papers as follows;

1. Phylogeography  of G. vittata
2. Population genetics of Galicitis/Lyncodon
3. Taxonomy and evolutionary history of Galictis/Lyncodon”

Answer: This version of the manuscript has been shortened by about three pages compared to the previous version, especially the Discussion. The reviewer's suggestion is interesting and would allow us to publish three articles instead of one. However, we believe that the number of results obtained is not large enough to justify dividing them into three separate articles. These are initial results that have shown very interesting patterns, but they are still preliminary. With much larger sample sizes analyzed, it might be possible to divide the manuscript into three separate articles, but we believe we have not yet generated enough results to do so. Likewise, the structure of the results and discussion addresses precisely the three themes mentioned by this reviewer, whose contributions we deeply appreciate.

The two authors emphatically thank the three reviewers because their comments helped improve the present version of the manuscript.

Reviewer 4 Report

Comments and Suggestions for Authors

Comments are arranged by numbers of referred manuscript lines in ascending order, not by the level of perceived importance.

1. Contrary to lines 21, 34–35, 39, 282, 311, 346, 352, 428–429, 431, 433, 494, 505, 508, 603, 611–612, 671, 711–712, 860, 867, 871, 924, 978, 985, 1202, 1207, 1702, 1709, 1711, and elsewhere, D-loop is neither a gene nor a gene part.

2. Referring to Galictis vittata and Lyncodon patagonicus as “virtually unstudied at the molecular level” (line 23) is inadequate and appears to reflect bibliographic incompleteness of this manuscript, rather than the actual situation. While these species are certainly not among best molecularly studied, they have been better studied at the molecular level than many other mustelids, carnivorans, or mammals. This is demonstrated by the following list of articles reporting results of molecular studies of Galictis vittata and/or Lyncodon patagonicus (these are articles that just come to my mind, so the list is certainly incomplete): Fulton & Strobeck 2006, Koepfli et al. 2008, Wolsan & Sato 2010, Sato et al. 2012, Bornholdt et al. 2013, which are cited in this manuscript (but note that despite citation elsewehere in this manuscript, 2 of these articles are omitted in the relevant paragraph from line 225 to 247; and Systematic Biology 52 (2003): 571–593 (https://doi.org/10.1080/10635150390235368), Science 307 (2005): 81 (https://doi.org/10.1126/science.1102832), Molecular Phylogenetics and Evolution 52 (2009): 632–642 (https://doi.org/10.1016/j.ympev.2009.05.036), Molecular Phylogenetics and Evolution 54 (2010): 726–745 (https://doi.org/10.1016/j.ympev.2009.10.033), BMC Biology 10 (2012): 12 (https://doi.org/10.1186/1741-7007-10-12), Methods in Ecology and Evolution 8 (2017): 1276–1283 (https://doi.org/10.1111/2041-210X.12729), PLoS One 16 (2021): e0240770 (https://doi.org/10.1371/journal.pone.0240770), Sato & Wolsan 2022 (Molecular systematics of the caniform Carnivora and its implications for conservation; pp. 41–59 in E. Do Linh San et al., eds., Small Carnivores: Evolution, Ecology, Behaviour, and Conservation; Wiley-Blackwell, Hoboken; https://doi.org/10.1002/9781118943274.ch2), Chemical Senses 47 (2022): bjac033 (https://doi.org/10.1093/chemse/bjac033), Chemical Senses 50 (2025): bjaf045 (https://doi.org/10.1093/chemse/bjaf045), which are entirely omitted from this manuscript. These omitted articles are relevant to this manuscript and should be referred to where appropriate. It is of note that from among the omitted articles, the PLoS One 16 (2021), Chemical Senses 47 (2022), and Chemical Senses 50 (2025) articles are of particular importance given that the first (in PLoS One) publishes complete mt genomes of Galictis vittata, and the two others (in Chemical Senses) report novel and important molecular results from Lyncodon patagonicus.

3. Contrary to line 74, Mustelinae contains only 2 extant genera, Mustela and Neogale (Neovison is a junior synonym of Neogale), and Ictonychinae contains 6, rather than 5, extant genera (Poecilictis needs to be added; for rationale, see African Zoology 61 [2026]: 55–62, https://doi.org/10.1080/15627020.2026.2638562).

4. Line 77: “Those same studies” here is misleading because what is written following this phrase (lines 77–100) is heavily biased toward the phylogenetic hypothesis of Koepfli et al. 2008, whereas the more widely and better supported hypothesis that places Ictonychinae sister to Lutrinae (see Fulton & Strobeck 2006, Wolsan & Sato 2010, Sato et al. 2012, all cited in this manuscript; and Nyakatura & Bininda-Emonds 2012—https://doi.org/10.1186/1741-7007-10-12 and Hassanin et al. 2021—https://doi.org/10.1371/journal.pone.0240770, which are omitted from this manuscript) is ignored. Instead this biased treatment, inform the reader about the two alternative hypotheses and provide the adequate references.

5. Contrary to line 85, libyca is of Poecilictis, rather than Ictonyx (see, e.g., Hayder et al. 2026—https://doi.org/10.1080/15627020.2026.2638562).

6. Lines 90–91: Replace “The most outstanding” by “Two”.

7. Lines 98–100: Correct “patagonian” to “Patagonian” (line 98), add the name of the New World clade (Lyncodontini), replace “ictonychins” (which refers to Ictonychinae, rather then Ictonychini) by Ictonychini, and end the sentence with the proper reference (Sato et al. 2012).

8. Line 187: Note that Sato et al. 2012 (cited in this manuscript) and Chem. Senses 47 (2022): bjac033 (https://doi.org/10.1093/chemse/bjac033) and Chem. Senses 50 (2025): bjaf045 (https://doi.org/10.1093/chemse/bjaf045), both not cited, provide exact localities for Lyncodon patagonicus.

9. Line 188: Correct “neotropical” to “Neotropical”.

10. Contrary to line 225, there are only 2 extant grison species (Galictis vittata and Galictis cuja). Lyncodon patagonicus is not a grison.

11. The paragraph from line 225 to 247 needs a serious revision to incorporate the relevant articles listed in Comment 2 above. Note that this paragraph lacks reference not only to the articles omitted from this manuscript (Comment 2), but also to two important molecular studies that are cited elsewhere in this manuscript, Fulton & Strobeck 2006 and Wolsan & Sato 2010. To compare these studies’ datasets to those discussed in this paragraph, the former study is based on 79 carnivoran species, with 20 genera and 36 species of mustelids (including Galictis vittata), and 5 nuclear markers totalling 2974 bp, while the latter study is based on a very large dataset of 52 musteloid species, with 22 genera and 43 species of mustelids (including Galictis vittata), and 25 nuclear and 29 mitochondrial markers totalling 27,965 bp.

12. Lines 266—271: Revise this to make it clear to the reader that the problem with obtaining DNA from the 3 Galictis and Lyncodon species, which resulted with a very small DNA sampling in this study, refers specifically to this study’s sampling, and not to the overall available sampling. Note that other authors managed to sequence multiple nuclear genes and much longer mitochondrial sequences from these species (see articles listed in Comment 2).

13. Lines 273 and 289: The relevant references need to be added (see Comments 2 and 11).

14. Line 301: While this manuscript uses some GenBank sequences (generated by Bornholdt et al. 2013), one can wonder why other Galictis and Lyncodon sequences published by other authors and available in GenBank are not used despite the fact the DNA sampling used in this study is very small.

15. Lines 310–313 (Table 1): Voucher information (reference numbers, collections) for the sequences obtained in this study needs to be added. Providing here GenBank accession numbers for these sequences would also be useful.

16. Lines 849–850: Correct “ichthyonychin” to “ictonychine”.

17. Line 859: Correct “Mustelid” to “mustelid”.

18. Lines 859–863 (Table 8): Add divergence time estimates from Sato et al. 2012.

19. Lines 865–930: Note that comparing solely divergence time posterior means without to consider 95% posterior intervals can be misleading and should be avoided in a serious comparison. Therefore, revise this text to avoid the subjective and unjustified overestimations of similarity with Koepfli et al.’s 2008 temporal estimates and dissimilarity with those of Sato et al. 2012.

20. Lines 957–963: The sampling of this study may be the largest with respect to the number of extant Lyncodontini individuals (Lyncodontini is the name of the Galictis–Lyncodon clade; use it), but is very small (as compared to previous studies) with respect to DNA. Make this clear and revise the last sentence (lines 959–963) to avoid the striking overestimation of this study’s importance for the “taxonomic, systematic and evolutionary knowledge of the extant Neotropical Ictonychinae species”. Given the actual current knowledge in these areas (exemplified by the results of studies that are omitted in this manuscript; see Comment 2), the very small DNA sampling, and the fact that this sampling is only of mtDNA, this study predicts rather a limited contribution to the knowledge of supraspecific relations in the extant Lyncodontini). However, given a relatively large sampling of Galictis vittata individuals and the focus on mtDNA, this study’s results are important at the infraspecific level. Instead of putting importance where it does not belong, focus on the real populational importance of this study (this actually refers to the whole manuscript).

21. Contrary to lines 1118–1120, this study is not “the largest genetic analysis” of Galictis vittata to date because previous studies examined much more DNA data of this species (only regarding mtDNA, Hassanin et al. 2021—https://doi.org/10.1371/journal.pone.0240770 examined the whole mitochondrial genomes of this species).

22. Lines 1185–1188: This is a speculation. This and the following taxonomic conclusions (lines 1191–1198) are not justifed and as such should be dropped.

23. Lines 1210–1211: Use either “two studies” or “three most comprehensive studies” and also discuss Wolsan & Sato 2010 because Koepfli et al. 2008 is less comprehensive than Wolsan & Sato 2010. However, it would be more useful and ethical if all relevant publications (some of those listed in Comment 2) were referred to in support of the discussed alternative hypotheses.

24. Line 1221: The late Oligocene to early Miocene European Plesictis is the oldest known mustelid (technically it is the oldest known stem mustelid), its oldest known species is Plesictis plesictis, and the oldest known occurence of this species dates to 24.7–23.3 million years ago (Ref. 144 plus Acta Palaeontologica Polonica 44 [1999]: 223–230 [http://www.app.pan.pl/archive/published/app44/app44-223.pdf], Zoological Science 20 [2003]: 243–264 [https://doi.org/10.2108/zsj.20.243], and Molecular Phylogenetics and Evolution 53 [2009]: 907–922 [https://doi.org/10.1016/j.ympev.2009.08.019]). North American fossils reported as “Plesictis” by Ref. 145 do not represent Plesictis.

25. Contrary to lines 1222–1223: The “32.4 to 30.9 mya in Asia” date is for the origin of Musteloidea, not Mustelidae, and Kollias & Fernandez-Moran (not solely Kolias) cited this date after Sato et al. 2012.

26. Contrary to line 1225, Promartes and Plesiogale are very distantly related musteloids, and neither is closely related to Guloninae.

27. Lines 1252–1263: Do not overestimate the differences that actually may not exist (see Comment 19), compare hypotheses rather than publications, and provide all relevant references in support of the discussed hypotheses (see Comment 4).

28. Lines 1291–1292: Results of Sato et al. 2012 are here omitted despite the fact that this study perfomed a more comprehensive and better supported biogeographic analysis than that of Koepfli et al. 2008. This and many other examples throughout this manuscript show that, for any reason, Koepfli et al. 2008 is favored over other relevant and often better supported studies, which helps neither this manuscript nor science. Revise this manuscript accordingly to remove this problem.

29. Line 1294: Add reference to the most recent relevant study of Baranogale (Annales Societatis Geologorum Poloniae 94 [2024]: 255–272, https://doi.org/10.14241/asgp.2024.09).

30. Lines 1304 ff.: Revise this using information from Journal of Systematic Palaeontology 22 (2024): 2348032 (https://doi.org/10.1080/14772019.2024.2348032) and references within.

31. Line 1374 ff.: Discuss the hypothesis of semiaquatic origin for Lyncodontini (see Chemical Senses 50 [2025]: bjaf045, https://doi.org/10.1093/chemse/bjaf045).

Author Response

Dear Editor of Animals,

We are sending you all the changes and modifications to the manuscript entitled "Population Genetics of the Greater Grison (Galictis vittata; Mustelidae, Carnivora) and How Changes Since Miocene-Pliocene Shaped the Phylogenetic Relationships Between Grison Species and Other Neotropical Mustelid Genera," following the recommendations of the fourth reviewer. Many of this reviewer's comments are invaluable in enriching the manuscript in several aspects.

1- Reviewer 4 stated: Contrary to lines 21, 34–35, 39, 282, 311, 346, 352, 428–429, 431, 433, 494, 505, 508, 603, 611–612, 671, 711–712, 860, 867, 871, 924, 978, 985, 1202, 1207, 1702, 1709, 1711, and elsewhere, D-loop is neither a gene nor a gene part.

Response: Following the reviewer's advice, the word "gene" has been replaced with the word "marker" whenever D-loop is mentioned.

2- Reviewer 4 said: Referring to Galictis vittata and Lyncodon patagonicus as “virtually unstudied at the molecular level” (line 23) is inadequate and appears to reflect bibliographic incompleteness of this manuscript, rather than the current situation. While these species are certainly not among the best molecularly studied, they have been better studied at the molecular level than many other mustelids, carnivorans, or mammals. This is demonstrated by the following list of articles reporting results of molecular studies of Galictis vittata and/or Lyncodon patagonicus (these are articles that just come to my mind, so the list is certainly incomplete): Fulton & Strobeck 2006, Koepfli et al. 2008, Wolsan & Sato 2010, Sato et al. 2012, Bornholdt et al. 2013, which are cited in this manuscript (but note that despite citation else we here in this manuscript, 2 of these articles are omitted in the relevant paragraph from line 225 to 247; and Systematic Biology 52 (2003): 571–593 (https://doi.org/10.1080/10635150390235368), Science 307 (2005): 81 (https://doi.org/10.1126/science.1102832), Molecular Phylogenetics and Evolution 52 (2009): 632–642 (https://doi.org/10.1016/j.ympev.2009.05.036), Molecular Phylogenetics and Evolution 54 (2010): 726–745 (https://doi.org/10.1016/j.ympev.2009.10.033), BMC Biology 10 (2012): 12 (https://doi.org/10.1186/1741-7007-10-12), Methods in Ecology and Evolution 8 (2017): 1276–1283 (https://doi.org/10.1111/2041-210X.12729), PLoS One 16 (2021): e0240770 (https://doi.org/10.1371/journal.pone.0240770), Sato & Wolsan 2022 (Molecular systematics of the caniform Carnivora and its implications for conservation; pp. 41–59 in E. Do Linh San et al., eds., Small Carnivores: Evolution, Ecology, Behavior, and Conservation; Wiley-Blackwell, Hoboken; https://doi.org/10.1002/9781118943274.ch2), Chemical Senses 47 (2022): bjac033 (https://doi.org/10.1093/chemse/bjac033), Chemical Senses 50 (2025): bjaf045 (https://doi.org/10.1093/chemse/bjaf045), which are entirely omitted from this manuscript. These omitted articles are relevant to this manuscript and should be referred to where appropriate. It is of note that from among the omitted articles, the PLoS One 16 (2021), Chemical Senses 47 (2022), and Chemical Senses 50 (2025) articles are of particular importance given that the first (in PLoS One) publishes complete mt genomes of Galictis vittata, and the other two (in Chemical Senses) report novel and important molecular results from Lyncodon patagonicus.

Response: Absolutely all the citations mentioned by reviewer 4 have been incorporated into the text. Many of them are vital for generating new hypotheses about the origin of Lyncodontini in the Neotropics, especially Wolsan and Sato (2022) and Wolsan and Sato (2025).

3-Reviewer 4 said: Contrary to line 74, Mustelinae contains only 2 extant genera, Mustela and Neogale (Neovison is a junior synonym of Neogale), and Ictonychinae contains 6, rather than 5, extant genera (Poecilictis needs to be added; for rationale, see African Zoology 61 [2026]: 55–62, https://doi.org/10.1080/15627020.2026.2638562).

Response: Following reviewer 4's comment, this paragraph has been corrected.

4-Reviewer 4 said: Line 77: “Those same studies” here is misleading because what is written following this phrase (lines 77–100) is heavily biased toward the phylogenetic hypothesis of Koepfli et al. 2008, whereas the more widely and better supported hypothesis that places Ictonychinae sister to Lutrinae (see Fulton & Strobeck 2006, Wolsan & Sato 2010, Sato et al. 2012, all cited in this manuscript; and Nyakatura & Bininda-Emonds 2012—https://doi.org/10.1186/1741-7007-10-12 and Hassanin et al. 2021—https://doi.org/10.1371/journal.pone.0240770, which are omitted from this manuscript) is ignored. Instead this biased treatment, inform the reader about the two alternative hypotheses and provide the adequate references.

Response: The present version of the manuscript is no longer biased in favor of the work of Koepfli et al. 2008. The new results have been compared not only with those of Koepfli et al. (2008) but also with those of Wolsan and Sato (2010), Sato et al. (2012), Nyakatura & Bininda-Emonds (2012), and Hassanin et al. (2021), in particular. Therefore, the comparison of the results obtained has been broadened to include all works published to date on the topic studied.

5-Reviewer 4 stated: Contrary to line 85, libyca is of Poecilictis, rather than Ictonyx (see, e.g., Hayder et al. 2026—https://doi.org/10.1080/15627020.2026.2638562).

Response: Corrected following reviewer 4's comment.

6-Reviewer 4 said: Lines 90–91: Replace “The most outstanding” with “Two”.

Response: Corrected following reviewer 4's comment.

7-Reviewer 4 said: Correct “patagonian” to “Patagonian” (line 98), add the name of the New World clade (Lyncodontini), replace “ictonychins” (which refers to Ictonychinae, rather than Ictonychini) with Ictonychini, and end the sentence with the proper reference (Sato et al. 2012).

Response: The corresponding correction was made. The term Lyncodontini has also been used throughout the text following the terminology of Sato et al. (2012).

8- Reviewer 4 said:

Line 187: Note that Sato et al. 2012 (cited in this manuscript) and Chem. Senses 47 (2022): bjac033 (https://doi.org/10.1093/chemse/bjac033) and Chem. Senses 50 (2025): bjaf045 (https://doi.org/10.1093/chemse/bjaf045), both not cited, provide exact localities for Lyncodon patagonicus.

Reply: Both studies have been incorporated into the new text as they are very important for generating new hypotheses regarding the origin of Lyncodontini. The geographic areas from which Lyncodon patagonicus samples were obtained in those studies have also been cited.

9-Reviewer 4 said: Line 188: Correct “neotropical” to “Neotropical”.

Reply: Corrected.

10-Reviewer 4 said: Contrary to line 225, there are only two extant grison species (Galictis vittata and Galictis cuja). Lyncodon patagonicus is not a grison.

Response: This has already been corrected throughout the text.

11-Reviewer 4: The paragraph from line 225 to 247 needs a serious revision to incorporate the relevant articles listed in Comment 2 above. Note that this paragraph lacks reference not only to the articles omitted from this manuscript (Comment 2), but also to two important molecular studies that are cited elsewhere in this manuscript, Fulton & Strobeck 2006 and Wolsan & Sato 2010. To compare these studies’ datasets to those discussed in this paragraph, the former study is based on 79 carnivoran species, with 20 genera and 36 species of mustelids (including Galictis vittata), and 5 nuclear markers totaling 2974 bp, while the latter study is based on a very large dataset of 52 musteloid species, with 22 genera and 43 species of mustelids (including Galictis vittata), and 25 nuclear and 29 mitochondrial markers totaling 27,965 bp.

Response: Both quotes have been introduced in the text and commented on in the place in the manuscript suggested by evaluator 4.

12-Reviewer 4 said: Lines 266–271: Revise this to make it clear to the reader that the problem with obtaining DNA from the three Galictis and Lyncodon species, which resulted in a very small DNA sample in this study, refers specifically to this study’s sampling, and not to the overall available sampling. Note that other authors managed to sequence multiple nuclear genes and much longer mitochondrial sequences from these species (see articles listed in Comment 2).

Response: It has already been mentioned, to make it clear in the text, that the difficulty in obtaining a good quantity and quality of DNA is specific to this study.

13-Reviewer 4 said: Lines 273 and 289: The relevant references need to be added (see Comments 2 and 11).

Response: All the citations mentioned by this reviewer have been included in the different parts of the text that he suggested.

14-Reviewer 4 said: Line 301: While this manuscript uses some GenBank sequences (generated by Bornholdt et al. 2013), one can wonder why other Galictis and Lyncodon sequences published by other authors and available in GenBank are not used, despite the fact that the DNA sampling used in this study is very small.

Response: Only GenBank sequences (especially from Galictis cuja) with well-confirmed geographic origins were used in this work. For example, the three Galictis vittata sequences from Bornholdt et al. 2013 were not used because the authors stated they were Peruvian but did not specify exactly where or when they were obtained. When compared to our Peruvian samples, which had precise geographic origins, the Bornholdt et al. 2013 sequences showed anomalous behavior. Furthermore, there is only one sequence of the mitochondrial gene Cyt-b for Lyncodon patagonicus in GenBank, and this is a gene that we did not use in this study.

15-Reviewer 4 said: Lines 310–313 (Table 1): Voucher information (reference numbers, collections) for the sequences obtained in this study needs to be added. Providing GenBank accession numbers for these sequences would also be useful.

Reply: All the samples used are from the wild, obtained directly by the authors, except for two Colombian samples of Galictis vittata, which come from a museum. The name of the museum and the corresponding code for these samples have been included. Regarding GenBank, the sequences will be uploaded as soon as the article is published.

16-Reviewer 4 said: Lines 849–850: Correct “ichthyonychin” to “ichtonychine”.

Reply: It was corrected.

17-Reviewer 4 said: Line 859: Correct “Mustelid” to “mustelid”.

Reply: It was corrected.

18-Reviewer 4 said: Lines 859–863 (Table 8): Add divergence time estimates from Sato et al. 2012.

Response: As the reviewer suggested, a column was added to Table 8 indicating the divergence times estimated by Sato et al. (2012).

19 - Reviewer 4 said: Lines 865–930: Note that comparing solely divergence time posterior means without considering 95% posterior intervals can be misleading and should be avoided in a serious comparison. Therefore, revise this text to avoid the subjective and unjustified overestimations of similarity with Koepfli et al.’s 2008 temporal estimates and dissimilarity with those of Sato et al. (2012).

Reply: The comparison has already been made, not only of the means but also considering the 95% intervals. Some values ​​showed no significant differences between the results obtained and those of Koepfli et al. (2008) and Sato et al. (2012). However, for some estimates, both the mean and the upper limit of the 95% interval were significantly higher in this study compared to the values ​​estimated by Sato et al. (2012).

20-Reviewer 4 said: Lines 957–963: The sampling of this study may be the largest with respect to the number of extant Lyncodontini individuals (Lyncodontini is the name of the Galictis–Lyncodon clade; use it) but is very small (as compared to previous studies) with respect to DNA. Make this clear and revise the last sentence (lines 959–963) to avoid the striking overestimation of this study’s importance for the “taxonomic, systematic and evolutionary knowledge of the extant Neotropical Ictonychinae species”. Given the current current knowledge in these areas (exemplified by the results of studies that are omitted in this manuscript; see Comment 2), the very small DNA sampling, and the fact that this sampling is only of mtDNA, this study predicts rather a limited contribution to the knowledge of supraspecific relations in the extant Lyncodontini). However, given a relatively large sampling of Galictis vittata individuals and the focus on mtDNA, this study's results are important at the infraspecific level. Instead of putting importance where it does not belong, focus on the real populational importance of this study (this actually refers to the whole manuscript).

Response: This evaluator's comment has been taken into consideration. For this reason, at the beginning of the Discussion the following has been written: "This is the study that, to date, has analyzed the highest number of samples for some mitochondrial genes in some Lyncodontini as G. vittata and L. patagonicus. Although the sample sizes are small, they are the largest used for both species to date. In this sense, this work aims to make known to the scientific community the large gaps that exist in the taxonomic, systematic and evolutionary knowledge of the extant Neotropical Lyncodontini species and aims to stimulate other research groups to obtain many more samples of these species for molecular genetic studies. A limitation of this study has to do with the very small DNA sampling, and the fact that this sampling is only of mtDNA, which may make this study predicts rather a limited contribution to the knowledge of supraspecific relations in the extant Lyncodontini.

21-Reviewer 4 said: Contrary to lines 1118–1120, this study is not “the largest genetic analysis” of Galictis vittata to date because previous studies examined much more DNA data of this species (Hassanin et al. 2021—https://doi.org/10.1371/journal.pone.0240770, which examined the entire mitochondrial genome of this species, only regarding mtDNA).

Response: The reviewer's comment is only partially accurate. It is true that Hassanin et al. (2021) generated a complete mitogenome from a single G. vittata specimen, but only from a single specimen with no clear geographic origin. Here, we present results from 13 G. vittata specimens distributed across Guatemala, Colombia, French Guiana, Ecuador, Peru, and Bolivia. Therefore, as we stated in the text: “recall that this is the study with the most G. vittata samples and the largest geographical area analyzed to date using molecular markers for this species"

22-Reviewer 4 said: Lines 1185–1188: This is speculation. This and the following taxonomic conclusions (lines 1191–1198) are not justified and should be discarded.

Reply: It is indeed speculation, but the new results are consistent with it. Therefore, we propose a new hypothesis that could change the taxonomic nomenclature of the Lyncodontini. As we stated in the text: "Although this aspect discussed is speculative, only future research with a larger number of samples from the three species and with a wider variety of genetic markers will be able to more precisely elucidate the taxonomy of the Lyncodontini." For this reason, we have retained that paragraph in the text.

23-Reviewer 4 stated: Lines 1210–1211: Use either “two studies” or “three most comprehensive studies” and also discuss Wolsan & Sato 2010 because Koepfli et al. 2008 is less comprehensive than Wolsan & Sato 2010. However, it would be more useful and ethical if all relevant publications (some of those listed in Comment 2) were referenced in support of the discussed alternative hypotheses.

Response: In the current version of the manuscript, all the phylogenies obtained by all the authors mentioned by the reviewer have been discussed, not just that of Koepfli et al. (2008).

24-Reviewer 4 said: Line 1221: The late Oligocene to early Miocene European Plesictis is the oldest known mustelid (technically it is the oldest known stem mustelid), its oldest known species is Plesictis plesictis, and the oldest known occurrence of this species dates to 24.7–23.3 million years ago (Ref. 144 plus Acta Palaeontologica Polonica 44 [1999]: 223-230 [https://doi.org/10.1016/j.ympev.2009.08.019]). North American fossils reported as “Plesictis” by Ref. 145 do not represent Plesictis.

Response: All changes related to paleontological data have been corrected following the comments of this reviewer, who is a specialist in Mustelidae paleontology. All these citations have been included in the text.

25-Reviewer 4 said: Contrary to lines 1222–1223: The “32.4 to 30.9 mya in Asia” date is for the origin of Musteloidea, not Mustelidae, and Kollias & Fernandez-Moran (not solely Kollias) cited this date after Sato et al. 2012.

Response: It was corrected.

26-Reviewer 4 said: Contrary to line 1225, Promartes and Plesiogale are very distantly related musteloids, and neither is closely related to Guloninae.

Answer: It was deleted from the text.

27-Reviewer 4 said: Lines 1252–1263: Do not overestimate the differences that actually may not exist (see Comment 19), compare hypotheses rather than publications, and provide all relevant references in support of the discussed hypotheses (see Comment 4).

Answer: It was done following the suggestions of reviewer 4.

28-Reviewer 4 said: Lines 1291–1292: Results of Sato et al. 2012 are here omitted despite the fact that this study performed a more comprehensive and better supported biogeographic analysis than that of Koepfli et al. 2008. This and many other examples throughout this manuscript show that, for any reason, Koepfli et al. The 2008 study is favored over other relevant and often better-supported studies, which benefits neither this manuscript nor the field of science. Please revise this manuscript accordingly to address this issue.

Response: Throughout the text, the results obtained in this research have been compared not only with those of Koepfli et al. (2008, 2017), but also, and especially, with those of Wolsan and Sato (2010), Sato et al. (2012), Sato and Wolsan (2022), and Hassanin et al. (2021).

29-Reviewer 4 said: Line 1294: Add reference to the most recent relevant study of Baranogale (Annales Societatis Geologorum Poloniae 94 [2024]: 255–272, https://doi.org/10.14241/asgp.2024.09).

Response: The citation suggested by reviewer 4, which is important in the context of the paleontological knowledge of the mustelid group studied, was introduced and discussed.

30-Reviewer 4: Lines 1304 ff.: Revise this using information from Journal of Systematic Palaeontology 22 (2024): 2348032 (https://doi.org/10.1080/14772019.2024.2348032) and references within.

Response: The information suggested by this reviewer from the Journal of Systematic Palaeontology 22 (2024) was reviewed and incorporated.

31-Reviewer 4 stated: Line 1374 ff.: Discuss the hypothesis of semiaquatic origin for Lyncodontini (see Chemical Senses 50 [2025]: bjaf045, https://doi.org/10.1093/chemse/bjaf045).

Reply: This is perhaps the most significant and vital contribution made by Reviewer 4. The hypothesis that Lyncodontini had a predecessor with a typically aquatic diet and habitat is extremely important. This opens up new possibilities for the hypotheses we present in this work regarding the origin of Neotropical Lyncodontini and how they colonized South America. We are extremely grateful to the reviewer for this particular contribution and for all the others in general.

The authors are deeply grateful to this reviewer and the three previous ones for their interesting contributions that helped to improve the quality of this manuscript.

Sincerely Yours,

Prof. Manuel Ruiz-García. PhD

Round 2

Reviewer 4 Report

Comments and Suggestions for Authors

I have no further comments.