Corn Straw Total Mix Dietary Supplementation of Bacillus Subtilis-Enhanced Growth Performance of Lambs by Favorably Modulating Rumen Bacterial Microbiome
Round 1
Reviewer 1 Report
Moderate English changes required
L5： "Li", not "L I"
L15: Delete "species"
L16: "the amount of " should be " the precentage of"
L17: "better cellulose degradation", this study did not measure the cellulose degradation, therefore, this is an inference, not a result.
L23 more reference are need here, such as:
Li, Z., H. Bai, L. Zheng, H. Jiang, H. Cui, Y. Cao, and J. Yao. 2018. Bioactive polysaccharides and oligosaccharides as possible feed additives to manipulate rumen fermentation in Rusitec fermenters. International Journal of Biological Macromolecules 109:1088–1094. doi:10.1016/j.ijbiomac.2017.11.098. Li, Z., X. Lei, X. Chen, Q. Yin, J. Shen, and J. Yao. 2021. Long-term and combined effects of N-[2-(nitrooxy)ethyl]-3-pyridinecarboxamide and fumaric acid on methane production, rumen fermentation, and lactation performance in dairy goats. Journal of Animal Science and Biotechnology 12:125. doi:10.1186/s40104-021-00645-4. L30-32: References are need here.
L54: NC was better for control than C
L114：“Single-end reads”?
Table 3: There was 3 times difference between groups for the Observed OTUs， What makes such a big difference？
Figure 2. delete the "venn2" on the figure
L179, and No need for italics
L187：Firmicutes
L188: group, respectively
Figure 4: the figure lenged need reword
Table 4 and 5: the Table lenged need reword
The information in Table 4 and 5 can be incorporated into Figure 3 and 5，respectively.
L224: compared with
L245: "indicating more bacteria species were observed from BS group lambs than from C group lambs", This is the result, not an inference.
Author Response
Point 1:Moderate English changes required
L5： "Li", not "L I"
Response 1：It has been modified, see the attachment for details
Point 2:L15: Delete "species"
Response 2：It has been modified, see the attachment for details
Point 3:L16: "the amount of " should be " the precentage of"
Response 3：It has been modified, see the attachment for details
Point 4:L17: "better cellulose degradation", this study did not measure the cellulose degradation, therefore, this is an inference, not a result.
Response 4：It has been modified, see the attachment for details
Point 5:L23 more reference are need here, such as:
Li, Z., H. Bai, L. Zheng, H. Jiang, H. Cui, Y. Cao, and J. Yao. 2018. Bioactive polysaccharides and oligosaccharides as possible feed additives to manipulate rumen fermentation in Rusitec fermenters. International Journal of Biological Macromolecules 109:1088–1094. doi:10.1016/j.ijbiomac.2017.11.098. Li, Z., X. Lei, X. Chen, Q. Yin, J. Shen, and J. Yao. 2021. Long-term and combined effects of N-[2-(nitrooxy)ethyl]-3-pyridinecarboxamide and fumaric acid on methane production, rumen fermentation, and lactation performance in dairy goats. Journal of Animal Science and Biotechnology 12:125. doi:10.1186/s40104-021-00645-4.
Response 5：It has been modified, see the attachment for details
Point 6:L30-32: References are need here.
Response 6：It has been modified, see the attachment for details
Point 7:L54: NC was better for control than C
Response 7：More inclined to use C expression control
Point 8:L114：“Single-end reads”?
Response 8：A new method, specific reference to the following literature
Kechin A, Boyarskikh U, Kel A, Filipenko M. cutPrimers: A New Tool for Accurate Cutting of Primers from Reads of Targeted Next Generation Sequencing. J Comput Biol. 2017 Nov;24(11):1138-1143. doi: 10.1089/cmb.2017.0096. Epub 2017 Jul 17. PMID: 28715235.
Point 9:Table 3: There was 3 times difference between groups for the Observed OTUs， What makes such a big difference？
Response 9：The addition of Bacillus subtilis increased the species diversity of rumen contents and the Observed species increased
Point 10:Figure 2. delete the "venn2" on the figure
Response 10：It has been modified, see the attachment for details
Point 11:L179, and No need for italics
Response 11：It has been modified, see the attachment for details
Point 12:L187：Firmicutes
Response 12：It has been modified, see the attachment for details
Point 13:L188: group, respectively
Response 13：It has been modified, see the attachment for details
Point 14:Figure 4: the figure lenged need reword
Response 14：It has been modified, see the attachment for details
Point 15:Table 4 and 5: the Table lenged need reword
Response 15：It has been modified, see the attachment for details
Point 16:The information in Table 4 and 5 can be incorporated into Figure 3 and 5，respectively.
Response 16：Table 4 and 5 are supplementary instructions to Figure 3 and 5, and it is necessary to retain them
Point 17:L224: compared with
Response 17：It has been modified, see the attachment for details
Point 18:L245: "indicating more bacteria species were observed from BS group lambs than from C group lambs", This is the result, not an inference.
Response 18：It has been modified, see the attachment for details
Author Response File: 
Author Response.pdf
Reviewer 2 Report
The manuscript titled “Corn straw total mix dietary supplementation of Bacillus subtillus enhanced growth performance of lambs by favorably modulating rumen bacterial microbiome” showed a novel finding that B.subtilis to feed boosted the amount of Bacteroidetes and other microbial flora in the rumen of meat sheep significantly and resulting in better cellulose degradation, for higher energy efficiency. The experiment design is simple, while the manuscript was well-prepared, the English writing was good, there is still some parts need to be improved as following:
Introduction: There is still some part to explain that why you use the B.subtilis on modulating rumen bacterial microbiome in lambs.
Materials and methods: why you use the 300 g/ton B.subtilis. How you added this dosage of B.subtilis. Table 1 shall also showed the analyzed value of these diet. Please upload the microbial community raw data in NCBI.
Results and Discussion: Please do the relationship between microbiota and other related parameters (VFA, NH3-N and so on).
Conclusion: Please confirm that “The dietary concentration of 3×108 CFU/kg”, since in the M & M, the author did not give the level of B.subtilis.
There is also some other mistakes:
Line 98: Beijing, China;
Line 108 and 110，Thermo Scientific，please add which city, which country?
Author Response
Point 1: Introduction: There is still some part to explain that why you use the B.subtilis on modulating rumen bacterial microbiome in lambs.
Response 1: It has been modified, see the attachment for details.
Point 1: Materials and methods: why you use the 300 g/ton B.subtilis. How you added this dosage of B.subtilis. Table 1 shall also showed the analyzed value of these diet. Please upload the microbial community raw data in NCBI.
Response 1: The amount of subtilis spores was referred to the following literature.
Zheng D Z. Application of Calsporin Bacillus subtilis in livestock feed [C]//.2007 Shandong Feed Science and Technology Exchange Conference Proceedings.,2007:103-107.
B.subtilis C-3102 is used in the commercial product Calsporin®ï¼Œthe microbial community raw data cannot be uploaded.
Point 2: Results and Discussion: Please do the relationship between microbiota and other related parameters (VFA, NH3-N and so on).
Response 2: It has been modified, see the attachment for details.
Point 3: Conclusion: Please confirm that “The dietary concentration of 3×108 CFU/kg”, since in the M & M, the author did not give the level of B.subtilis.
Response 3: It has been modified, see the attachment for details.
Point 4: There is also some other mistakes:
Line 98: Beijing, China;
Line 108 and 110，Thermo Scientific，please add which city, which country?
Response 4: It has been modified, see the attachment for details.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
L16-19, This sentence need be reworded.
L25 :ref 1-6 all focused on poultry, not ruminant. The cited references need relevant to the research.
L118: "Single-end reads" or "paired-end reads"
Author Response
Point 1:L16-19, This sentence need be reworded.
Response 1：It has been modified, see the attachment for details
Point 2:L25 :ref 1-6 all focused on poultry, not ruminant. The cited references need relevant to the research.
Response 2：It has been modified, see the attachment for details
Point 3:L118: "Single-end reads" or "paired-end reads"
Response 3：It is Single-end reads
