Dietary Supplementation of a Mixture of Nucleotides, β-Glucan and Vitamins C and E Improved the Growth and Health Performance of Olive Flounder, Paralichthys olivaceus
Round 1
Reviewer 1 Report
L117-121 It is understandable that this study conducted to evaluate commercial functional immunostimulant mixture, but in the experimental design, a control group using nucleotides, β-glucan, vitamin C, or vitamin E alone should be designed to fully demonstrate the necessity of using four compounds in combination.
L119-120 What is the basis for the ratio of nucleotide, β-glucan, vitamin C and vitamin E in functional additives?
L143-158 Please readjust the line spacing.
L180 “Fish were counted and weighed and lengths were measured individually...” Please rewrite this sentence.
L353 Parentheses are missing.
Discussion
Most of the author's discussion is devoted to explaining the effects of nucleotides and β-glucan on the growth, the innate immune parameters, phagocytic activity, antioxidant capacity, upregulating anti-inflammation and immune-related gene expressions. but there is a lack of discussion of the necessary for vitamin C and E combinations in the FIM.
No.
Author Response
Dear Editor In-Chief,
We would like to thank you and reviewers for the thoughtful suggestions and insightful comments on our manuscript (fishes-2398942). We have carefully revised the manuscript according to the comments/suggestions and provided our point-by-point response below. The revised contents in the manuscript are indicated with track changes.
We eagerly anticipate your response and hope you will find our manuscript suitable for publication in Fishes Journal.
Sincerely yours,
Kyeong-Jun Lee, Ph.D.
Response Letter - fishes-2398942
Dietary supplementation of a mixture of nucleotides, β-glucan and vitamins C and E improved the growth and health performance of olive flounder, Paralichthys olivaceus - fishes-2398942.
Responses to the comments of reviewer #1.
L117-121: It is understandable that this study conducted to evaluate commercial functional immunostimulant mixture, but in the experimental design, a control group using nucleotides, β-glucan, vitamin C, or vitamin E alone should be designed to fully demonstrate the necessity of using four compounds in combination.
Answer: We agree with reviewers’ suggestion. Though we did not include individual diets to evaluate individual nutrient components in this study, we referred and previously conducted experiments in our laboratory and other laboratories to identify the immunostimulant effects of those individual nutrients and optimum levels of them for olive flounder (references are mentioned below). Therefore, we designed a study to evaluate the effect of the combination of those four nutrients as an immunostimulant and to identify the optimum level of the mixture. To the best of our knowledge, there is a lack of evidence on studies that have been conducted to evaluate mixtures of immunostimulants for olive flounder. Accordingly, we included this information in the introduction section (Line no: 110-115)
References: -
Nucleotides; Song, J. W., Lim, S. J., & Lee, K. J. (2012). Effects of dietary supplementation of inosine monophosphate on growth performance, innate immunity and disease resistance of olive flounder (Paralichthys olivaceus). Fish & shellfish immunology, 33(4), 1050-1054. https://doi.org/10.1016/j.fsi.2012.07.011
β-glucan; Hasan, M. T., Jang, W. J., Lee, S., Kim, K. W., Lee, B. J., Han, H. S., ... & Kong, I. S. (2018). Effect of β‐glucooligosaccharides as a new prebiotic for dietary supplementation in olive flounder (Paralichthys olivaceus) aquaculture. Aquaculture research, 49(3), 1310-1319.
Vitamin C; Wang, X., Kim, K., & Bai, S. C. (2002). Effects of different dietary levels of L‐ascorbyl‐2‐polyphosphate on growth and tissue vitamin C concentrations in juvenile olive flounder, Paralichthys olivaceus (Temminck et Schlegel). Aquaculture Research, 33(4), 261-267.
Vitamin E; Kim, M. S., Hur, D. H., Lee, S. M., Jeoung, E., Jeong, H. D., & Hong, S. (2017). Effects of vitamin E with different levels or sources of dietary lipid on the growth and expression of inflammatory, oxidative stress, and apoptotic genes in the head kidney of olive flounder, Paralichthys olivaceus. Journal of the World Aquaculture Society, 48(3), 518-530.
L119-120: What is the basis for the ratio of nucleotide, β-glucan, vitamin C and vitamin E in functional additives?
Answer: The ratio of nucleotide, β-glucan, vitamin C and vitamin E were decided according to previous studies that were conducted to identify the effects and optimum levels of those nutrients for olive flounder. ROVIMAX® HB Ultra contains nucleotides and β-glucan at adequate levels. However, vitamins C and E were added through the vitamin mixture to fulfill the lacking amounts of the nutrients.
L143-158: Please readjust the line spacing.
Answer: Thank you for your valuable suggestion. Line spacing was adjusted according to the journal requirements (Line no 157-158).
L180: “Fish were counted and weighed and lengths were measured individually...” Please rewrite this sentence.
Answer: According to reviewers’ suggestion, the sentence was rewritten to improve the clarity (Line no 186-187).
L353: Parentheses are missing.
Answer: Thank you for correcting the mistake. Parenthesis was added and mistake was corrected (Line no, 360).
Discussion
- Most of the author's discussion is devoted to explaining the effects of nucleotides and β-glucan on the growth, the innate immune parameters, phagocytic activity, antioxidant capacity, upregulating anti-inflammation and immune-related gene expressions. but there is a lack of discussion of the necessary for vitamin C and E combinations in the FIM.
Answer: Thank you for reviewer’s valuable suggestion. Discussion was modified accordingly (Discussion, Line no; 25-28, 37-40, 62-63, 94-96 and 142-147)
Author Response File: 
Author Response.docx
Reviewer 2 Report
The manuscript presents a scientifically sound and well-written study that is both interesting and relevant to the readership of the journal. The research methods are well-described and the results are clear and well-supported by the data presented. The authors have also provided a comprehensive discussion of their findings and their implications, highlighting the significance of the using a functional immunostimulant mixture (FIM; nucleotides, β-glucan and 11 vitamins C and E) to improve growth and health performance of olive flounder.Overall, I believe that the manuscript would make a valuable contribution to the journal and would be of interest to the journal's readership. I have no major concerns or criticisms and I recommend this manuscript for publication at Fishes in its current form.
Author Response
Dear Editor In-Chief,
We would like to thank you and reviewers for the thoughtful suggestions and insightful comments on our manuscript (fishes-2398942). We have carefully revised the manuscript according to the comments/suggestions and provided our point-by-point response below. The revised contents in the manuscript are indicated with track changes.
We eagerly anticipate your response and hope you will find our manuscript suitable for publication in Fishes Journal.
Sincerely yours,
Kyeong-Jun Lee, Ph.D.
Response Letter - fishes-2398942
Dietary supplementation of a mixture of nucleotides, β-glucan and vitamins C and E improved the growth and health performance of olive flounder, Paralichthys olivaceus - fishes-2398942.
Reviewer #2
Comments and Suggestions for Authors:
The manuscript presents a scientifically sound and well-written study that is both interesting and relevant to the readership of the journal. The research methods are well-described and the results are clear and well-supported by the data presented. The authors have also provided a comprehensive discussion of their findings and their implications, highlighting the significance of the using a functional immunostimulant mixture (FIM; nucleotides, β-glucan and 11 vitamins C and E) to improve growth and health performance of olive flounder. Overall, I believe that the manuscript would make a valuable contribution to the journal and would be of interest to the journal's readership. I have no major concerns or criticisms and I recommend this manuscript for publication at Fishes in its current form.
Thank you very much for your valuable comment on our manuscript. We are happy to give our best for the publication.
Author Response File: Author Response.docx
Reviewer 3 Report
I have read the manuscript completely. As far as I am concerned, the manuscript is well written and well structured. The results were presented clearly. I also appreciate the authors for taking time to write the manuscript neatly and represented the results systematically. But I would suggest the authors to consider some points discussed below and revise the manuscript accordingly.
Line 51-52: metabolically costly? Revise the words
Line 68: remove “Vitamin C” word
Line 224: remove “and” after reference 53
Line 351: Instead of giving (nmol min-1ml-1) it is better to give nmol/min/ml to follow the uniformity in units representation.
Line 353: check the spelling for expression.
Line 356-358: The IL- and TNF were downregulated but not with increasing dietary levels. I accept there is no significance but not decreasing with increasing levels of diet. Please check the line with corresponding figure.
My suggestion is to give the gene expression values in graphs separately rather than giving in Tables. It is feasible for the readers to identify the gene expression differences between the groups. It is not mandatory but my suggestion.
Please explain in introduction why you have chosen already commercially available rovimax i.e available in markets to improve immunity in aquaculture species ?
Author Response
Dear Editor In-Chief,
We would like to thank you and reviewers for the thoughtful suggestions and insightful comments on our manuscript (fishes-2398942). We have carefully revised the manuscript according to the comments/suggestions and provided our point-by-point response below. The revised contents in the manuscript are indicated with track changes.
We eagerly anticipate your response and hope you will find our manuscript suitable for publication in Fishes Journal.
Sincerely yours,
Kyeong-Jun Lee, Ph.D.
Response Letter - fishes-2398942
Dietary supplementation of a mixture of nucleotides, β-glucan and vitamins C and E improved the growth and health performance of olive flounder, Paralichthys olivaceus - fishes-2398942.
Responses to the comments of reviewer #3
I have read the manuscript completely. As far as I am concerned, the manuscript is well written and well structured. The results were presented clearly. I also appreciate the authors for taking time to write the manuscript neatly and represented the results systematically. But I would suggest the authors to consider some points discussed below and revise the manuscript accordingly.
Line 51-52: metabolically costly? Revise the words
Answer: The term “metabolically costly" was revised according to authors suggestion (Line no 52-53).
Line 68: remove “Vitamin C” word
Answer: Thank you to correct the mistake. The word “Vitamin C” was removed (Line no 69).
Line 224: remove “and” after reference 53
Answer: Thank you for correcting the mistake. The word “and” was removed and sentence was modified accordingly (Line no 225 – 227).
Line 351: Instead of giving (nmol min-1ml-1) it is better to give nmol/min/ml to follow the uniformity in units representation.
Answer: Thank you for the valuable suggestion. Unit was corrected accordingly (Line no 358).
Line 353: check the spelling for expression.
Answer: Thank you to correct the mistake. Spelling was corrected (Line no 360).
Line 356-358: The IL- and TNF were downregulated but not with increasing dietary levels. I accept there is no significance but not decreasing with increasing levels of diet. Please check the line with corresponding figure.
Answer: We agree with the reviewer’s valuable suggestion. According to suggestion, the relevant sentence was changed (Line no 364-365).
Q1; My suggestion is to give the gene expression values in graphs separately rather than giving in Tables. It is feasible for the readers to identify the gene expression differences between the groups. It is not mandatory but my suggestion.
Answer: Most of publications have given gene expression results as graphs as reviewers said. But in this study, we expected to evaluate the effect of immunostimulant under different criteria such as growth, non-specific immune responses and inflammation. Therefore, we hope to give gene expression results in relevant tables.
Q2; Please explain in introduction why you have chosen already commercially available rovimax i.e available in markets to improve immunity in aquaculture species?
Answer: We included below paragraph to introduction section as a justification (Line no: 110-115). “Although previous studies identified the effects of individual immunostimulants and optimum levels, limited studies evaluated the effect of combinations of immunostimulants. ROVIMAX® HB Ultra is a commercially available immunostimulant mixture containing nucleotides, Saccharomyces cerevisiae derived β-glucan, vitamin C and vitamin E at optimal levels for olive flounder”.
Author Response File: Author Response.docx
Round 2
Reviewer 1 Report
No.
No.
