Round 1

Reviewer 1 Report

In this study, Chen et al. investigated “A new C-type of Ruditapes philippinarum and its potential role in feeding selection”. Listed below are some comments and suggestions:

- Title, “Ruditapes philippinarum” should be italicized

- Line 17, “QPCR”, please correct, there is no QPCR with gene expression. And Rpcl gene should be italicized

- Abstract section, what is the final conclusion of this study? Please clarify.

- Citations in the text vary from those in the Reference section. The order is also distinct. The authors should adhere to the journal's instructions. Please update all citations, since they are all outdated.

- Introduction section, The author neglected to identify the primary purpose of this research. The information is not interconnected in particular the first paragraph

- Materials and Methods, “The adult R. philippinarum with about 5–7 g weight”, please check English grammar.  

- Feeding selection is the main point of this study. However, the authors state that “The feeding experiment was arranged by buying R. philippinarum from the market and soaking them in seawater”. What is the significant meaning of this study?

- The gene name is “β-actin”, not “β-action” in Table 1.

- The methods described in this study were not cleared (sections 2.1, 2.2, 2.3) and logical. Information is mixed. For RNA isolation, how many grams of sample was collected? Which protocol was used to isolate total RNA? How much total RNA was used for cDNA synthesis? Were the primers designed by the authors? It should be clarified also. How about technical replicates and biological replicates for qPCR?

- Lines 127-128, please check English grammar.

- Line 174, please correct “Statisticalmethods”

- Full-length cDNA of Ruditapes philippinarum C-type lectin is 929 bp. However, it was 930 bp in this study. Do the author use the same source?

- Experimental feeding should be included.

- Results, there is no scale bar in figure 3. What is/are outgroup(s) used for generating this phylogenetic tree? The figure legend should be rewritten.

- In Figure 5, the relative expression of gill tissue is approximately 1500. It is super amazing. What is the normal range of the Ct value of β-actin and investigated genes? The significant letter order is wrong.  

- Three biological replicates (n = 3) were used for qPCR, limited number of samples. The results may not be reliable.

Based on the findings in this study, the authors failed to explain the probable involvement of C-type lectin of Ruditapes philippinarum in feeding selection. How does it influence and function? The authors need to emphasize how the reference provides support to your research.

 

- A decent review of English is required.

Author Response

Dear reviewers,

Hello, thank you for your valuable comments, the revision has been completed.

Yours,

Chensentao

Author Response File: Author Response.pdf

Reviewer 2 Report

The manuscript with ID (fishes-2092059) by Chen and coauthors has reported a novel C-type lectin derived from a bivalve mollusk known as Ruditapes philippinarum and evaluated its potential roles in feeding selection. The study is interesting; however, there are several major revisions should be revised by the authors before the manuscript considered for publication in Fishes. The authors have to prepare a point-by-point response the points raised by the anonymous reviewer.

Q1: I suggest changing the title to be

“A novel C-type lectin and Its Potential Role in Feeding selection in Ruditapes philippinarum”

Q2: I made several track changes and highlights in the PDF supplied with my reviews. Authors have to address them and revise all points very carefully.

Q3: Authors should use MDPI reference style throughout the whole manuscript. I suggest using EndNote or any reference management program.

Q4: Line 63: Add the citation here in this position (Pales Espinosa & Allam, 2013).

Q5: Line 67: which oyster type?

Q6: Lines 69-70: revise 2010a or 2010b

Q7: Line 74: In this position, authors have to add a brief information on nutrigenomics in crustaceans. I suggest using this reference “Nutrigenomics in crustaceans: Current status and future prospects”

Q8: Line 95: add details on kits used.

Q9: Table 1: Please add the following: Tm, T annealing, NCBI GenBank accession numbers, Primer efficiency %, and Product size

Q10: Line 130: You should cite this reference in this position

Livak, K. J., & Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 25(4), 402-408.‏

Q11: Reference section: Latin names should be written italic

Comments for author File: Comments.pdf

Author Response

Dear Reviewer,

Hello, thank you for your valuable comments, I have completed the revision. I am very honored to have you as my first article reviewer, and you are definitely the best reviewer. I wish you a happy life, stay away from the new crown, and a successful career.

Your fans，

ChenSentao

Author Response File: Author Response.pdf

Reviewer 3 Report

This research highlights the importance of lectins in feeding performance of ruditapes philippinarum. It may benefit aquatic species culture and differentiate the effect of baited and toxic macroalgae useful for application in mariculture as well . However, No explanation of figure 6 on the result section, remove if necessary. Proper axis title in needed on Fig. 8 & 9.  Minor changes is recommended.

Author Response

Dear Reviewer,

Hello, thank you for your valuable comments, I have completed the revision. I wish you a happy life.

Yours,

ChenSentao

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

Dear Authors,

I have not seen any responses from the authors. The attached file just includes the revised manuscript version. I have no idea about it.

Cheers,

Author Response

I am very sorry for my mistake operation and hope to get the understanding of the reviewer. Here is the latest response I uploaded. Thank you!

Yours,

Sentao Chen

Author Response File: Author Response.pdf

Reviewer 2 Report

There are several minor points and comments that should be revised by the authors.

Line 2: Ruditapes philippinarum – write italic

Line 7: “Lectins belong to a carbohydrate recognition receptor” – Remove this sentence.

Line 8: is a hot topic in feeding choice studies of bivalve mollusks.

Line 16: The qPCR

Line 17: observed in all examined tissues

Line 18: The in vitro

Line 22: Rpcl expression in the lip and gill of clam fed with

Line 24: selection process in R. philippinarum

Line 28: a deterministic role in feeding of R. philippinarum

Line 35: add appropriate reference

Line 38: an important ecological position among other bivalves.

Line 41-42: Lectins are widely distributed in bacteria, viruses, fungi, protozoa, as well as plants 41 and animals. Authors should add appropriate reference. I suggest the below references.

Lis, H., & Sharon, N. (1998). Lectins. In P. J. Delves (Ed.), Encyclopedia of Immunology (Second Edition) (pp. 1535-1541). Oxford: Elsevier.

Sharon, N., & Lis, H. (2013). Lectins. In W. J. Lennarz & M. D. Lane (Eds.), Encyclopedia of Biological Chemistry (Second Edition) (pp. 701-705). Waltham: Academic Press.

Line 62: Oyster - not italic

Line 63-64: Espinosa et al. (2010) – Use MDPI reference style

Line 81: remove “in the field”

Line 85: in vitro - write italic

Line 93-94: remove “as parallel samples”

Line 407-408: Oreochromis niloticus - write italic

Line 418: Sinonovacula constricta - write italic

Line 482: Heterocapsa circularisquama - write italic

Author Response

Dear Reviewer,

Thank you very much for your second revision. I took your comments very seriously and then revised them in the text, but in the middle where I needed to insert the literature I did not use the literature provided by the teacher, but chose to insert my fourth reference, which is mentioned in my fourth literature, I hope the teacher can understand. I hope that you will understand. The revised part is in red, and I hope to receive the manuscript soon. Thank you for the reviewer's hard work!

Your fans，

Sentao Chen

Author Response File: Author Response.pdf

Round 3

Reviewer 1 Report

Accept the present form

Author Response

Dear reviewers,

I would like to thank the reviewer for repeatedly optimizing my article. This is the final version of the submission. I hope it will be accepted soon.

Yours,

Chensentao

Author Response File: Author Response.docx