Morphological Integration on the Calcaneum of Domestic Sheep (Ovis aries Linnaeus, 1758)—A Geometric Morphometric Study

Round 1

Reviewer 1 Report

Dear authors

I have few suggestions:

Comments:

1.Page 5-6

Line 242 251

Results present as a table.

 2.Tabela 1

Delete Table 1.

Add breeds and Number and the specimens of sheep in text Merina branca (n=2), Merina preta (n=4), Merina (n=7), Soay (n=1), and Unknown (n=18), (insert in Page 3, Line 131)

3.I also think it would be interesting to do and add a CVA analysis.

Add Canonical variate analysis (CVA) output of the sheep calcaneum comparison between breeds and shape of the sheep calcaneum for each axis according the wireframe. This analyses would show the separation of the breeds according to the shape of the calcaneum (different color or symbol of breeds). Add the appropriate text in all parts of the manuscript (M and M, Results and Discussion).

 4.Figure 1

Do you have an image with better resolution of the sheep calcaneum?

Author Response

Dear Reviewer.

Thanks for your suggestions.

Original table 1 for breeds samples have been deleted and a new table 1 (Procrustes ANOVA) has been included.

To add a CVA analysis seems us of no interest as our goal was not to compare breeds. In adition, this analysis was done in our previous study in Animals

All three figures have been improved in resolution and Figure 1 have been modified for a new photograph of a new specimen

Author Response File: Author Response.docx

Reviewer 2 Report

The article by P.M. Parés-Casanova et al. 'Morphological integration on the calcaneum of domestic sheep ...' is devoted to a problem of morphological integration and modularity of morphological structures on the example of the calcaneum in sheep using methods of geometric morphometrics (GM). The formulation of the research task is quite legitimate, and the study of the integration and modularity of the calcaneum of sheep using the arsenal of GM methods was carried out by the authors for the first time. The authors correctly performed all the elements of the study, strictly following the recommendations of leading experts in the field of GM. Conclusions about the relative independence of the formation and variation of calcaneal bone modules are statistically justified.

However, while reading the article, there were some small comments. Firstly, at the beginning of the introduction, the authors of the article gave definitions of the concepts of morphological integration and modularity without references to the authorship of other authors (in particular, C.P. Klingenberg is not mentioned). Therefore, when reading, an erroneous impression may arise, as if the authors of the article themselves were the first to come to these ideas and methods of allocating modules using GM and PLS. In the future, with explanations of a methodological nature, a bibliographic reference to the founder of the method appears in the text, but only as a private mention. In order for the readers of the article not to get the wrong impression about the priority of the authors of the article in the development of ideas about morphological integration and modularity, they should mention from the beginning the key article by C.P. Klingenberg (2014) and pay a little more attention to this aspect in the introduction. Secondly, the text of the article does not sufficiently disclose its applied aspect.  The authors could additionally write about this in the introduction or in the conclusion. Applying the methods of geometric morphometrics, the authors for the first time revealed the phenomenon of relative independence of the development of two modules of the calcaneus. It is desirable to explain in more detail what is the main applied aspect of the work? If the morphogenesis of the sheep calcaneum was previously poorly studied, and the authors' research has brought something new to its understanding, then it needs to be said in more detail in the text.

The text of the work is well verified and there are few typos in it, which, however, should be corrected. In lines 247-248, it’s  need to align the text, on line 280, remove the unnecessary repeated word "for", in line 390, probably need to specify the pages to the authors' article or DOI.

In general, the work may be interesting and useful to specialists in the field of geometric morphometrics as a concrete example of assessing morphological integration and modularity in sheep. Therefore, after making appropriate edits to the text, it can be recommended for publication in a special issue of the journal.

Author Response

Dear Reviewer.

Thanks for your comments.

Klingenberg's reference and others have been added and the rest of minor suggestions have been also solved.

Due to the scaricty of similar studies, discussion remains very limited, although future works are outlined.

Author Response File: Author Response.docx

Reviewer 3 Report

Comments

1.      Title: Title is very catchy and concisely states the main subject of the study cases.

2.      Abstract: Abstract need to clarify the significance of the study's findings.

3. Introduction:

a)      The first sentence of the introduction could be made clearer to better convey the definition of morphological integration.

b)      The introduction could be broken down into smaller paragraphs for better readability.

c)      The terms "morphological modules" and "functional whole" could be defined in more detail.

d)      The introduction could provide a brief overview of why the Partial Least Squares (PLS) and Escoufier RV coefficient are commonly used to study morphological integration, rather than diving directly into the technical details of these methods.

4.  Material and Methods: Sufficient and well explained

5. Results and discussion: both sections were well structured but the authors could benefit from including some more references to support their arguments.

Author Response

Dear Reviewer.

Thanks for your kind suggestions.

Some new references have been added. Introductory part has been "lightened"breaking it down into smaller paragraphs.

It is our opinion that more information on technical and statistical terms could be confusing for the general reader. In any case, we are obviously open to modify the corresponding paragraphs according to your comment.

Author Response File: Author Response.docx

Reviewer 4 Report

Dear authors,

Unfortunately, I just realised that majority of submitted work was published from the same group in MDPI family of journals (Animals). If you check previously published MS (https://pubmed.ncbi.nlm.nih.gov/36359069/, PMID: 36359069) you can see on the first view that was the same figure (specimen 1904). Moreover, If you check the page 4 of submitted study (line 148-183), actually that is the same text. Could you please check the line 175-177  “First, we repeated the photographs of the same specimen (the first author) ten times in the course of ten days, mounting and dismounting both camera and tripod each time.” and compare with the previously published MS (PMID: 36359069) “First, we repeated (the first author) the photographs of the same specimen ten times during ten days, mounting and dismounting both camera and tripod each time.” Moreover, MS was published in the same family (MDPI) few months ago.

Finally, anatomical nomenclature wasn’t adjusted based on Nomina Anatomica Veterinaria 2017 (NAV). Based on NAV anterior and posterior can be used on the eye only. On the appendicular skeleton (fore- and hindlimb), depending on the region following terms can be used: proximal, distal, cranial, caudal, medial, lateral, palmar, plantar, dorsal, axial, abaxial.

Kind Regards

Ivan

Author Response

Dear Reviewer.

Thanks for you comments. 

As pictures collection and photographic procedure were the same as the refered our published article, methodology it is exactly the same (as it is stated), in fact, and it has been expressed in a similar way (although changes have been done to avoid an absolute similarity).

For anatomical references, strict terms from NAV appear in italics. English directional/positional terms have been changed according to your comment.

Author Response File: Author Response.docx

Reviewer 5 Report

Abstract

1. Please  explain methods in this  section.

Introduction

1. please include  information  about  other  animal in this  part.

Material  and methods

1. Please include information  about animal such  as   age  and .....

Discussion

1.Please explain and compare the clinical applications of this method

2. please explain  about limitation of this  study.

Author Response

Thanks a lot for your kind suggestions.

Some paragraphs have been rewritten in order to fulfill your comments.

Author Response File: Author Response.pdf

Round 2

Reviewer 4 Report

Dear author,

line 157.......geometric morphometric......Geometric is not necessary word. If we know what morphometry is!

If you have 47 animals x 10 figgures = 470 figures. If is that true, why you showed the same figure as in previously published MS (PMID: 36359069) (Animal: 1904).

In the supplementary data, I would like to see all raw figures (470)!!!!!

Author Response

Than you for your suggestions

We have added a new photograph from a new specimen in Figure 1

We have removed the term "Geometric" in line 157

We did a total of 47 different photographs from the 47 calcanea, to carry out our study. However, previously to start with this analysis, we evaluated intra-observer error. This was done repeating photographs of two specimens, from the 47, ten times during ten different days (20 photos in total). During this period, when each repeated photograph was taken, the tripod and the camera were mounted and dismounted each time.  In addition to this, we also randomly selected ten photographs of ten different specimens from the 47 calcanea (10 photos in total). Then, on the thirty photographs (20 repeated photos and 10 photographs randomly selected from the 47 calcanea images) the 20 landmarks were located twice in a ten-day period.  After digitalization we used a Procrustes ANOVA test both among and within repeated calcanea photographs in order, to statistically evaluate the measurements consistency (Lloveras et al., 2022). This has been explained in lines 204 to 227.

We would not like to publish our 47 images of the calcanea. However, if you like we can sent to you these 47 images.

Author Response File: Author Response.docx

Round 3

Reviewer 4 Report

Dear Author,

the same figure cant be published in two different MS, or if you do that, that MUST be quoted properly!!!!!!! First lesson on the first day of PhD program!

Author Response

Dear Reviewer,

You have not paid attention to the reviewed manuscript submitted nor to the Figure 1. If you had, you would have realized that the figure 1 included is not the same that the previous. The first Figure 1 correspond to the image of the calcaneum of the individual 1904 and the new Figure 1 correspond to the image of the calcaneum of individual 1906. As it is explained in the Materials and Methods of the paper, the calcanea comes from the osteological collection of Laboratório de Arqueociências in Lisbon and because of this, the different individuals (and each of its elements) are numbered. The style of the numbers, of course, are similar because they belong to the same collection. If you pay attention in both figures you will realise that the specimens are different (although they are sheeps, they have differences in form) and have different numbers.