Effect of Dioscorea Opposite Waste Supplementation on Antioxidant Capacity, Immune Response and Rumen Microbiome in Weaned Lambs
Round 1
Reviewer 1 Report
This manuscript aimed to determine how adding Dioscorea opposite waste in weaned lambs diet affects the rumen microbiome, antioxidant capacity, and immune response. The research work has been executed well and the overall premise looks promising. Nevertheless, the manuscript requires extensive revisions, and several concerns need to be addressed as follows:
1) L37-39: The conclusion section should be rewritten, as in the current form, it is only a general statement. The conclusions should answer the aims of the study. Also, it needs connections with the recommended level to take home massage for both scientific and practical.
2) Are there statistics on the quantities produced from DOW globally or in China? please add to the introduction section.
3) Could DOW production meet the ruminant feed industry demand?
4) L98-99: The authors had to justify on what basis they selected the level of the tested Dioscorea opposite waste
5) L105: Please specify whether the lambs received the diets in groups or individuals within the same treatment.
6) L44: please show what anticoagulant is used and its concentration.
7) For blood analyses, it is highly recommended to mention the catalog number of the kits used.
8) L164: centrifugation speed and duration are not mentioned.
9) L229: Describe the statistical model used.
10) Why were the animal's body weight, growth performance and feed quantity not evaluated? Such measurements would have given the work more strength from an applied point of view.
11) Throughout the manuscript, the writing style should be formal from the third-person perspective. Do not use “we” (E.g. lines 81, 83, 384, 486, and 508… etc) or “our” (E.g. lines 37, 430, 506, … etc ).
12) The citation of the references should be revised according to the journal instructions. Please see some errors in lines 62, 99, 118, 128, and 132.
Author Response
Dear Reviewer,
We would like to express sincere appreciation to you and the anonymous reviewers who had given these constructive comments and suggestions. These comments and suggestions are all valuable and very helpful for revising and improving our manuscript entitled “Effect of Dioscorea Opposite Waste Supplementation on Anti-oxidant Capacity, Immune Response and Rumen Microbiome in Weaned Lambs”, as well as the important guiding significance for our researches.
We have revised the manuscript according to your kind advices and reviewers’ detailed suggestions. We sincerely hope this manuscript will be finally acceptable for publication on Fermentation. Thank you very much for all your help and looking forward to hearing from you soon.
Reviewer 1:
Point 1: L37-39: The conclusion section should be rewritten, as in the current form, it is only a general statement. The conclusions should answer the aims of the study. Also, it needs connections with the recommended level to take home massage for both scientific and practical.
Response 1: Thanks for your suggestions. We have checked and corrected the conclusions in the Abstract section. We add the main findings and aims of this trial, as well as the connections between the results and the recommended levels.
Point 2: Are there statistics on the quantities produced from DOW globally or in China? please add to the introduction section.
Response 2: Thanks for your valuable comments and suggestions. We regret that there are no references have been available for DOW yield in DOW globally or in China, because DOW is an emerging non-conventional feed. Nevertheless, The quantities produced of Dioscorea opposite in China were added in the Introduction section.
Point 3: Could DOW production meet the ruminant feed industry demand?
Response 3: Thanks for your valuable comments and suggestions. We are sorry that no references have been available on the application of DOW on ruminant. Therefore, further research is necessary to investigate the optimum amount of DOW to be added to ruminant diets and whether its yield can meet the needs of ruminant production.
Point 4: The authors had to justify on what basis they selected the level of the tested Dioscorea opposite waste?
Response 4: Thanks for your valuable comments, the level of DOW supplemented to the basal diets were added in the Introduction section. To the best of our knowledge, no reports on supplementing doses are available in weaned lambs or other ruminants. At the beginning of our study, the highest supplementing level of DOW was 30 %. But we found that the feed is difficult mixed and the lambs are picky eaters. Then we decreased the highest supplementing level to 20 % and the feed is well mixed and the sheep eating returned to normal. Therefore, we supplementing weaned lambs with DOW at 0, 10 %, 15 % and 20 % as experimental treatments in the study.
Point 5: Please specify whether the lambs received the diets in groups or individuals within the same treatment.
Response 5: Thanks for your comments. in the present study, the lambs received the diets in groups, and we have added the content to the Materials and Methods.
Point 6: please show what anticoagulant is used and its concentration.
Response 6: Thanks for your suggestion. The 5-mL vacuum blood collection tubes anticoagulant tubes containing 10 mg K2EDTA and we have added the result to the Materials and Methods.
Point 7: For blood analyses, it is highly recommended to mention the catalog number of the kits used.
Response 7: Thanks for your positive comments. We have added more details about the kits including the catalog number of the kits.
Point 8: centrifugation speed and duration are not mentioned.
Response 8: Thank you for your suggestion. In this experiment, the rumen fluid was only kept in centrifuge tubes, but not centrifuged and quickly transferred to -80℃ for further microbiome analysis.
Point 9: Describe the statistical model used.
Response 9: Thank you for your suggestion. The analytical model used in the present study was: Yijk=Mean+trt+Error, where, trt is a fixed effect. We have added the analytical model in the Statistical Analysis section.
Point 10: Why were the animal's body weight, growth performance and feed quantity not evaluated? Such measurements would have given the work more strength from an applied point of view.
Response 10: Thank you for your suggestion. Due to the aim of this experiment was conducted to investigate the effect of DOW on the physiological metabolism of weaned lambs, so we are sorry for not detected the growth performance and feed quantity in the present study. Therefore, further studies need to be designed to verify the effect of DOW on their growth performance and feed conversion efficiency.
Point 11: Throughout the manuscript, the writing style should be formal from the third-person perspective. Do not use “we” (E.g. lines 81, 83, 384, 486, and 508… etc) or “our” (E.g. lines 37, 430, 506, … etc).
Response 11: Thank you for your suggestion. We checked and corrected the expressions of the manuscript and changed some the non-standard first -person to the third-person.
Point 12: The citation of the references should be revised according to the journal instructions. Please see some errors in lines 62, 99, 118, 128, and 132.
Response 12: Thank you for your suggestion. We have checked the references in the manuscript and corrected the errors therein.
Reviewer 2 Report
Effect of Dioscorea Opposite Waste Supplementation on Anti- oxidant Capacity, Immune Response and Rumen Microbiome in Weaned Lambs by Ruochen Yang et al.
The paper aimed to identify and explore the potential of alternative non-conventional feed sources: this is a very important area of research involving economical, environmental and social aspects.
The authors performed a huge work, particularly concerning the rumen microbiome, of which they very well described the adopted methods and discussed the results. However, some modifications and or claryfication need to improve the paper, as follows:
Line 60: please, provide the meaning of SD
Line 81: please replace “we” with “We”
Line 93: 1.2 – 13.5 °C: it is correct? Please clarify
Table 1 – I am not sure that replacing corn grain with DOW (11.8% CP /DM as reported in table 2) and decreasing soybean meal in the DOW’s diets arises all isoprotein diets. On the other hand the authors, on line 414-151, to discuss their results concenrning UA decrease have written as follows: “Therefore, the changes in UA concentration after lambs were fed DOW-supplemented diet are probably associated with the lower protein content in DOW”. Please, clarify
Table 2: Crude protein, ADF, NDF etc are not ingredients, please correct
Line 160: please clarify why the rumn fluid has been collected only for CCON and DOW 3 diets
Line 435: please, after “fructooligosaccharides in DOW” add the following sentence; “as demonstrated also in other ruminants species with other plants (Lombardi et al., 2017, Buffalo Bulletin, Vol.36 No.3, 497-503).
Author Response
Dear Reviewer,
We would like to express sincere appreciation to you and the anonymous reviewers who had given these constructive comments and suggestions. These comments and suggestions are all valuable and very helpful for revising and improving our manuscript entitled “Effect of Dioscorea Opposite Waste Supplementation on Anti-oxidant Capacity, Immune Response and Rumen Microbiome in Weaned Lambs”, as well as the important guiding significance for our researches.
We have revised the manuscript according to your kind advices and reviewers’ detailed suggestions. We sincerely hope this manuscript will be finally acceptable for publication on Fermentation. Thank you very much for all your help and looking forward to hearing from you soon.
Reviewer 2:
Point 1: please, provide the meaning of SD
Response 1: Thanks for your valuable question. SD rats are a strain of rats (rat; rattus norvegicus), bred from Wistar rats at Sprague Dawley Farm in the United States, and are widely used in pharmacological, toxicological, pharmacodynamic and GLP experiments. So, SD is an abbreviation of “Sprague Dawley”.
Point 2: Line 81: please replace “we” with “We”
Response 2: Thank you for your suggestion. We have revised and corrected it.
Point 3: 1.2 – 13.5 °C: it is correct? Please clarify
Response 3: Thanks for your valuable question. This experiment was conducted from February to May, with nighttime temperatures from -1 to 1℃ and daytime temperatures from 9 to 13 ℃.
Point 4: Table 1 – I am not sure that replacing corn grain with DOW (11.8% CP /DM as reported in table 2) and decreasing soybean meal in the DOW’s diets arises all isoprotein diets. On the other hand the authors, on line 414-151, to discuss their results concerning UA decrease have written as follows: “Therefore, the changes in UA concentration after lambs were fed DOW-supplemented diet are probably associated with the lower protein content in DOW”. Please, clarify
Response 4: Thanks for your valuable question. The results in table 2 showed that the concentration of CP in DOW is 11.8%, which is greater than that of corn grain (8.5%) and lower than that of soybean meal (39.5 %). When formulating rations, we follow the principle of isoenergy and isonitrogen, so DOW replaces more corn and less soybean meal, thus maintaining a dietary protein content at 16.7. The linearly decreased in plasma UA levels after lambs fed DOW diet. We have corrected the interpretation in the manuscript to suggest that the decrease in plasma UA may be related to the saponins in DOW based on the previous results.
Point 5: Crude protein, ADF, NDF etc are not ingredients, please correct
Response 5: Thank you for your suggestion. We have revised and corrected it.
Point 6: please clarify why the rumen fluid has been collected only for CON and DOW 3 diets
Response 6: Thanks for your valuable question. In the present study, we first analyzed the blood indicators of each group, including antioxidant and immune indicators, and we found that the lambs in the DOW3 group had the most significant improvement in immune and antioxidant capacity. Meanwhile, Previous studies have shown that Dioscorea has the function to improve intestinal health and regulate the intestinal microbiota. So, we collected the rumen fluid from CON and DOW 3 group for microbiome sequencing.
Point 7: please, after “fructooligosaccharides in DOW” add the following sentence; “as demonstrated also in other ruminants species with other plants (Lombardi et al., 2017, Buffalo Bulletin, Vol.36 No.3, 497-503).
Thank you for your suggestion. We have added the sentence and related literature into the manuscript.
Round 2
Reviewer 1 Report
No further comments are to be addressed
