Effect of Alkaline Mineral Complex Buffer Supplementation on Rumen Fermentation, Rumen Microbiota and Rumen Epithelial Transcriptome of Newborn Calves

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Wang et al. studied the effects of AMCB on serum immunity indexes, rumen fermentation, and microbiota of newborn calves. The serum analysis shows increased total protein and globulin levels in the treatment group. The RNA seq analysis shows that AMCB can improve the immunity and rumen pH of newborn calves.  The authors say that by regulating some gene expressions like FGB, MIF, ATF3, 436 MANF, and AOX1  in rumen epithelial tissue, rumen immune function was enhanced, and rumen health was maintained. The results are interesting. 

1. I recommend the authors to look at the article title. Doesn't look grammatically right.  

2. We should write as calves or calfs?

3. GLB expansion is missing.

4. Give the RNA seq DEG data in a supplementary file.

5. Did the authors check the gene's differential regulation from RNA seq with qPCR analysis?

6. The article needs to be checked with a native English-speaking person.

 

 

Comments on the Quality of English Language

The article needs to be checked with a native English-speaking person.

Author Response

Dear reviewer

Thank you very much for taking the time to review our manuscript and letting us know that you approve our experimental results. We have revised and supplemented our manuscript according to your suggestions, and the revised contents are detailed in the new manuscript. The following is our response to your suggestions.

1. I recommend the authors to look at the article title. Doesn't look grammatically right.  

6. The article needs to be checked with a native English-speaking person.

According to your suggestion, we have commissioned a professional polishing company to revise the grammar of our manuscript again. The previous manuscript polishing instructions have been uploaded in the supplementary file.

2. We should write as calves or calfs?

We have revised the title and relevant expressions in the manuscript, as detailed in the manuscript.

3. GLB expansion is missing.

The serum GLB content is equal to the TP content minus the ALB content. We have added instructions in the manuscript.

4. Give the RNA seq DEG data in a supplementary file.

We have submitted the raw data for rumen transcriptome analysis in the NCBI system and provided the NCBI serial number in the data description of the article. Due to the large data capacity of DEGs, uploading in the supplementary file is easy to interrupt. If you are interested in our research, please kindly download it in the information we provide, thank you for your understanding. The data presented in the study are deposited in the NCBI Sequence Read Archive (SRA) database repository, accession number PRJNA937435 (rumen tissue transcriptome), PRJNA972886(rumen fluid 16s rDNA).

5. Did the authors check the gene's differential regulation from RNA seq with qPCR analysis?

Thank you for your careful examination of our manuscript and let us know that our experimental analysis still has some shortcomings. Unfortunately, The number of rumen tissue samples was no longer sufficient to supplement relevant data. However, please rest assured that we ensure the authenticity of the experiment. We will also conduct research on lambs in the future. Your suggestions will be constructive to our data analysis, and we look forward to your continued attention to our follow-up research. Thank you very much for your suggestions.

 

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript is excellent, many variables evaluated, with emphasis on microbiota. I consider it an excellent experiment, however, but the statistical analysis was not adequate. The authors have repeated measurements, so the authors must seek a statistical analysis that allows evaluating the treatment x day interaction. After this analysis, interpretations may change, therefore, I will carry out a more detailed analysis of the manuscript in the next round of review.

Author Response

Dear reviewer

 

Thank you very much for taking the time to review our manuscript and letting us know that you have further modification suggestions for our data, and thank you for complimenting our experimental design. There is no doubt that your contribution will make this manuscript more perfect for others to refer to. Thank you again for your participation. According to your suggestion, we established a model for the analysis of blood indexes and rumen fermentation parameters. The following model indicates the interaction effects between treatment and group: Y = µ + T_i + G_j + TG_ij + E_ijk_l. Where Y is the dependent variable, µ is the overall mean, T_i is the time effect, G_j is the group effect, TG_ij is the interaction effect between T and G, and E_ijk_l is the random residual error. The analysis results are detailed in the manuscript.