The Low Dose of Saccharomyces cerevisiae Is Beneficial for Rumen Fermentation (Both In Vivo and In Vitro) and the Growth Performance of Heat-Stressed Goats
Takehiro Nishida
Round 1
Reviewer 1 Report
Overall, well written, but I have a few questions.
L40; ruminal volatile fatty acid (TVFA) --- ruminal total volatile fatty acid (TVFA) ?
L57; cerevisiaesupplemented --- insert space
Table 1; Ca2HPO4 --- OK ?
L88-100; I don't understand the dates of the experiment. what is the relationship between HS and 4x4?
L104; dry matter (DM) --- DM
L120-140; I am not sure of the schedule for blood and rumen juice sampling. Isn't it common practice to collect them before the morning feed?
L146; Waltham, USA) --- Waltham, MA, USA)
L162; in vitro --- Italic
L165; Scientific, USA) --- Waltham, MA, USA)
L177; Soest (1970) [33]. ---Soest [33].
L193, 194; Hsp --- HSP ?
L203; Hsp --- HSP ?
L224; p --- Italic
Figure3, 4; Because of the large error, we can see that the SC2A measurements are varied. Is there a significant difference between the other SC treatments. I understand that there is a difference between HS2 and SC2A in TVFA and ADG, but there appears to be no difference in the others.
L260; (3, 4, 25) --- [3, 4, 25]
L296-299; It is curious that the effect is seen only at 0.6% and 0.06%; why is there no effect at 0.12% between 0.6% and 0.06%?
L253-312; The dosing of Saccharomyces cerevisiae was not consistent with the results, so I have no idea what to choose and how much to dose.
Author Response
Dear reviewer,
Thanks for your thoughtful comments and suggestions. As shown in the manuscript, we have made careful modifications to the manuscript. We have made thorough revisions to improve the English as needed. Below you will find our general reply and point-by-point responses to the reviewers’ comments/ questions.
Point 1: L40; ruminal volatile fatty acid (TVFA) --- ruminal total volatile fatty acid (TVFA) ?
Response 1: “ruminal volatile fatty acid (TVFA)” has been changed into “ruminal volatile fatty acids (VFAs)” at line 40.
Point 2: L57; cerevisiaesupplemented --- insert space
Response 2: The sentence has been rewrittenat lines 55-57.
Point 3: Table 1; Ca2HPO4 --- OK ?
Response 3: Ca2HPO4 is the source of Ca2+. It is safe and effective.
Point 4: L88-100; I don't understand the dates of the experiment. what is the relationship between HS and 4x4?
Response 4: HS is the first phase in this study to obtain heat-stressed goats. The probiotic feeding experiment started when the experimental heat-stressed goats were well prepared. That is, HS is the basis for completing the 4 × 4 Latin square experiment.
Point 5: L104; dry matter (DM) --- DM
Response 5: “dry matter (DM)” has been changed into “DM” at line 104.
Point 6: L120-140; I am not sure of the schedule for blood and rumen juice sampling. Isn't it common practice to collect them before the morning feed?
Response 6: Blood samples were collected in the morning after overnight fasting (24 fasting). Rumen fluids were collected in the morning after 4 h of feeding. It is common practice to collect these samples in goat studies, and in our several previous studies adopted this method to collect blood and rumen fluid samples.
Point 7: L146; Waltham, USA) --- Waltham, MA, USA)
Response 7: “Waltham, USA)” has been changed into “Waltham, MA, USA)” at line 146.
Point 8: L162; in vitro --- Italic
Response 8: “in vitro” has been changed into “ in vitro” at line 163.
Point 9: L165; Scientific, USA) --- Waltham, MA, USA)
Response 9: “Scientific, USA)” has been changed into ‘Waltham, MA, USA)” at line 167.
Point 10: L177; Soest (1970) [33]. ---Soest [33].
Response 10: “Soest (1970) [33]” has been changed into “Soest [33]”at line 179.
Point 11: L193, 194; Hsp --- HSP ?
Response 11: “Hsp” has been changed into “HSP” at line 195.
Point 12: L203; Hsp --- HSP ?
Response 12: “Hsp” has been changed into “HSP” at line 205.
Point 13: L224; p --- Italic
Response 13: “p” has been changed into “p” at line 226.
Point 14: Figure3, 4; Because of the large error, we can see that the SC2A measurements are varied. Is there a significant difference between the other SC treatments. I understand that there is a difference between HS2 and SC2A in TVFA and ADG, but there appears to be no difference in the others.
Response 14: By double checking the original data, the results, which are finally present in these two figures were correct. There is no significant difference between the other SC treatments.
Point 15: L260; (3, 4, 25) --- [3, 4, 25]
Response 15: “(3, 4, 25)” has been changed into “[3, 4, 25]” at line 261.
Point 16: L296-299; It is curious that the effect is seen only at 0.6% and 0.06%; why is there no effect at 0.12% between 0.6% and 0.06%?
Response 16: We’re as curious about this as you are. We speculate that it results from the interaction between Saccharomyces cerevisiae and rumen microbiota. In our further study, we will analyze the effects of Saccharomyces cerevisiae supplemental dose on the composition and function of rumen microbiota.
Point 17: L253-312; The dosing of Saccharomyces cerevisiae was not consistent with the results, so I have no idea what to choose and how much to dose.
Response 17: This study is a continuation of our previous study because we believe excessive addition of probiotics may have adverse effects on rumen microbiota. Therefore, we drastically reduced the supplemental level of Saccharomyces cerevisiae. This study verified that this low dose of Saccharomyces cerevisiae achieved similar results to the previous study. In this study, combined with the parameters of rumen fermentation and growth performance, the appropriate supplement level of this probiotic is 0.60‰ of the dry matter concentration in the basal diet. We have summarized the above in the “ Conclusion” section,
Author Response File: 
Author Response.pdf
Reviewer 2 Report
ABSTRACT
The abstract should be a total of about 200 words maximum. Please reduce it
INTRODUCTION
Line 34 The development: write the in lowercase
Line 35 The longitude write the in lowercase
Line 48 nutritional studies: add references
Line 55 yeast supplementation: which type of yeast
Line 56 in the rumen: of which species?
Line 57 Saccharomyces cerevisiaesupplemented: add a space
Line 58 in the rumen of sheep: and in cattle?
Line 66: delete of our previous study
MATERIALS AND METHODS
Line 75 last year: write the year
Line 77 twelve: according to which statistical calculation was this number chosen?
Line 77 female: explain the choice of only female. what state were they in? had they ever given birth?
Line 78 Macheng Black × Boer: explain the choice of this breed
Point 2.2 and 2.3: the timing of the experimentation is not clear: in point 2.2 it says that there is a control period of 15 days and a stress model of 15 days. In the point 2.3 the experimental cycle lasted for 21 days
Line 102-110: explain the need to carry out the experiment in vitro
Line 140: add a scheme/figure showing the time line of all experiments
Point 2.5: add catalogue numbers for all kit and reagents used
RESULTS
Add in the text the reference to Figure 1 A
REFERENCES
Rewrite references following journal’s insructions:
Journal Articles:
1. Author 1, A.B.; Author 2, C.D. Title of the article. Abbreviated Journal Name Year, Volume, page range.
Books and Book Chapters:
2. Author 1, A.; Author 2, B. Book Title, 3rd ed.; Publisher: Publisher Location, Country, Year; pp. 154–196.
3. Author 1, A.; Author 2, B. Title of the chapter. In Book Title, 2nd ed.; Editor 1, A., Editor 2, B., Eds.; Publisher: Publisher Location, Country, Year; Volume 3, pp. 154–196.
Author Response
Dear reviewer,
Thanks for your thoughtful comments and suggestions. As shown in the manuscript, we have made careful modifications to the manuscript. Below you will find our general reply and point-by-point responses to the reviewers’ comments/ questions.
Point 1: ABSTRACT
The abstract should be a total of about 200 words maximum. Please reduce it
Response 1: Thank you for your suggestion! The “Abstract” section has been reduced to 197 words.
Point 2: INTRODUCTION Line 34 The development: write the in lowercase
Response 2: “The development” has been changed into “the development” at line 32.
Point 3: Line 35 The longitude write the in lowercase
Response 3: “The” has been changed into “the” at line 33.
Point 4: Line 48 nutritional studies: add references
Response 4: The reference [8] has been added at line 47.
Point 5: Line 55 yeast supplementation: which type of yeast
Response 5: The expression of this sentence is not accurate enough; we have rewritten the sentence. Saccharomyces cerevisiae is included in the yeast group, so it does not have to be placed alongside yeast (at lines 55-56).
Point 6: Line 56 in the rumen: of which species?
Response 6: The rumen of goats, cattle, and bulls and this species have been added at line 55.
Point 7: Line 57 Saccharomyces cerevisiaesupplemented: add a space
Response 7: The sentence has been rewritten at lines 55-57.
Point 8: Line 58 in the rumen of sheep: and in cattle?
Response 8: Both in the rumen of sheep and cattle.
Point 9: Line 66: delete of our previous study
Response 9: “our previous study” has been deleted.
Point 10: MATERIALS AND METHODS Line 75 last year: write the year
Response 10: The year has been added at line 74.
Point 11: Line 77 twelve: according to which statistical calculation was this number chosen?
Response 11: According to our previous studies:
Cai, L.Y.; Yu, J.K.; Hartanto, R.; Qi, D.S Dietary supplementation with Saccharomyces cerevisiae, Clostridium butyricum and their combination ameliorate rumen fermentation and growth performance of heat-stressed goats. Animals 2021, 11, 2116.
Cai, L.Y.; Hartanto, R.; Zhang, J.; Qi, D.S. Clostridium Butyricum improves rumen fermentation and growth performance of heat-stressed goats in vitro and in vivo. Animals. 2021, 11: 3261.
Cai, L.Y.; Hartanto, R.; Xu, Q.B.; Zhang, J.; Qi, D.S. Saccharomyces cerevisiae and Clostridium butyricum could improve B-vitamin production in the rumen and growth performance of heat-stressed goats. Metabolites. 2022, 12, 766.
Point 12: Line 77 female: explain the choice of only female. what state were they in? had they ever given birth?
Response 12: Because in this study, only female goats met the screening criteria for our experimental animals (same birthday). They are kept in Xiangyang city of China. They never gave birth.
Point 13: Line 78 Macheng Black × Boer: explain the choice of this breed
Response 13: This goat breed is widely raised for meat in the Jianghuai region of China.
Point 14: Point 2.2 and 2.3: the timing of the experimentation is not clear: in point 2.2 it says that there is a control period of 15 days and a stress model of 15 days. In the point 2.3 the experimental cycle lasted for 21 days
Response 14: These two points were to describe the different experiments, respectively in this study. In point 2.2, the purpose is to obtain the heat-stressed goats, while in point 2.3, the heat-stressed goats were used as experimental animals for probiotics feeding experiment.
Point 15: Line 102-110: explain the need to carry out the experiment in vitro
Response 15: In vitro fermentation test is necessary to determine the effects of probiotics on rumen fermentation to determine the supplemental level of probiotics in the subsequently feeding experiment.
Point 16: Line 140: add a scheme/figure showing the time line of all experiments
Response 16: Thanks for your suggestion, but we consider that the experiment process has been fully introduced in the “Materials and methods” section, so there is no need to add an extra scheme or figure to explain it.
Point 17: Point 2.5: add catalogue numbers for all kit and reagents used
Response 17: Catalogue numbers have been added for all kits (at lines 141, 144, 145, 149, and 154).
Point 18: RESULTS Add in the text the reference to Figure 1 A
Response 18: Thanks for your reminder! The result related to Figure 1A has been added at line 192.
Point 19: REFERENCES Rewrite references following journal’s insructions:
Journal Articles:
- Author 1, A.B.; Author 2, C.D. Title of the article. Abbreviated Journal Name Year, Volume, page range.
- Books and Book Chapters:
- Author 1, A.; Author 2, B. Book Title, 3rd ed.; Publisher: Publisher Location, Country, Year; pp. 154–196.
- Author 1, A.; Author 2, B. Title of the chapter. In Book Title, 2nd ed.; Editor 1, A., Editor 2, B., Eds.; Publisher: Publisher Location, Country, Year; Volume 3, pp. 154–196.
Response 19: Thanks for your suggestion! The “References” section has been rewritten accroding to the journal’s instructions.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
I have no further comments.
