Effects of Additives on the Fermentation Quality, In Vitro Digestibility, and Aerobic Stability of Amaranth (Amaranthus hypochondriacus) and Wheat Bran Mixed Silage
Round 1
Reviewer 1 Report
The article submitted for review relates to the field of improving the quality of agricultural animal feeds, the prospects of increasing their nutritional value and safety through the use of lactic acid bacteria and cellulase enzyme.
I have significant comments.
In connection with the new classification of lactobacilli, Lactobacillus plantarum LP1 should be called Lactiplantibacillus plantarum LP1.
The authors already have a similar paper, but with a different object of study (Yi, Q.; Wang, P.; Tang, H.; Yu, M.; Zhao, T.; Sheng, Z.; Luo, H. Fermentation Quality, In Vitro Digestibility, and Aerobic Stability of Ensiling Spent Mushroom Substrate with Microbial Additives. Animals 2023, 13, 920. https://doi.org/ 10.3390/ani13050920). In this regard, the article would like to see a feature of this study, especially since the metabolism of strain Lactobacillus plantarum LP1 in amaranth silage is different from Mushroom silage.
Due to the fact that the paper is submitted to the journal Fermentation the authors need to focus more on the fermentation characteristics due to the presence of different additive. It would be very interesting to compare the differences in the production of lactic acid (LA), acetic acid (AA), propionic acid (PA) and butyric acid (BA) depending on the plant material, what this is related to. The authors have already used this composition of lactobacillus and cellulase in other works. The cycle of work of the authors allow them to analyze the changes
The reference by Zhang et al. [61] is not used correctly. This paper did not specifically use cellulase enzyme, citing " (3) 1 g fibrolytic enzyme per 1 kg FM (EN; 10,000 U/g activity, XS biotechnology Co., Ltd., Beijing, China);". In the peer-reviewed paper, the authors write “The best aerobic stability (153 h) of AWB mixed silage was obtained with the addition of lactic acid bacteria and cellulase mixture at 60% MC, indicating that the addition of lactic acid bacteria and cellulase mixture was more favorable to prolong the aerobic stability of silage, which was consistent with the results of Zhang et al. [61]. Zhang et al. [61] showed that the lactic acid bacteria and cellulase mixture treatment group had higher LA and AA contents, lower pH and yeast counts, reduced microbial diversity and improved aerobic stability of the silage compared to the lactic acid bacteria treatment group and cellulase treatment group”. There is no mention of cellulase activity of this enzyme preparation in the work of Zhang et al. [61].
Line 396, 537 - of homozygous fermentative lactic acid bacteria - it's not correct! Should use the term homofermentative!
Line 418 - with homotypic fermentative lactic acid bacteria - it's not correct! Should use the term homofermentative!
Line 418 - heterotypic fermentative lactic acid bacteria - it's not correct! Should use the term heterofermentative!
Lines 389, 390, 392, 431, 590 - Latin names should be italicized.
415-416 with Lactobacillus heterotrophic – need ”with heterotrophic lactobacilli
Comments for author File: 
Comments.pdf
Author Response
Dear reviewer, thank you for your encouraging and warm comments and suggestions, all of your suggestions are very important, and they all have important guiding significance for our future research work. Based on this we have revised and (we think) strengthened our paper.
Point 1: In connection with the new classification of lactobacilli, Lactobacillus plantarum LP1 should be called Lactiplantibacillus plantarum LP1.
Response 1: We fully appreciate your suggestion. We revised that section in the paper. (PDF:128)
Point 2: The reference by Zhang et al. [61] is not used correctly. This paper did not specifically use cellulase enzyme, citing " (3) 1 g fibrolytic enzyme per 1 kg FM (EN; 10,000 U/g activity, XS biotechnology Co., Ltd., Beijing, China);". In the peer-reviewed paper, the authors write “The best aerobic stability (153 h) of AWB mixed silage was obtained with the addition of lactic acid bacteria and cellulase mixture at 60% MC, indicating that the addition of lactic acid bacteria and cellulase mixture was more favorable to prolong the aerobic stability of silage, which was consistent with the results of Zhang et al. [61]. Zhang et al. [61] showed that the lactic acid bacteria and cellulase mixture treatment group had higher LA and AA contents, lower pH and yeast counts, reduced microbial diversity and improved aerobic stability of the silage compared to the lactic acid bacteria treatment group and cellulase treatment group”. There is no mention of cellulase activity of this enzyme preparation in the work of Zhang et al. [61].
Response 2: We fully appreciate your suggestion. We revised that section in the paper. (PDF:607-611)
Point 3: Line 396, 537 - of homozygous fermentative lactic acid bacteria - it's not correct! Should use the term homofermentative!
Response 3: We fully appreciate your suggestion. We revised that section in the paper. (PDF:398, 543)
Point 4: Line 418 - with homotypic fermentative lactic acid bacteria - it's not correct! Should use the term homofermentative!
Response 4: We fully appreciate your suggestion. We revised that section in the paper. (PDF:421-422)
Point 5: Line 418 - heterotypic fermentative lactic acid bacteria - it's not correct! Should use the term heterofermentative!
Response 5: We fully appreciate your suggestion. We revised that section in the paper. (PDF: 421-422)
Point 6: Lines 389, 390, 392, 431, 590 - Latin names should be italicized.
Response 6: We fully appreciate your suggestion. We revised that section in the paper. (PDF:391, 392, 394, 434, 595)
Point 7: 415-416 with Lactobacillus heterotrophic – need ”with heterotrophic lactobacilli
Response 7: We fully appreciate your suggestion. We revised that section in the paper. (PDF:417-418)
Thank you again for your suggestions and hope to learn more from you.
Author Response File: Author Response.pdf
Reviewer 2 Report
The authors mixed residues of amaranth and wheat bran in three different concentrations. After 60 days, they tested how the addition of Lactic Acid Bacteria, cellulase and LAB plus cellulase influence the fermentation parameters, aerobic stability and silage digestibility. Adequate analysis was performed and the data compared between each treatment and controls. The results were comparable to experiments with other raw materials developed by diverse authors. The data support that low-moisture-content treatment and the addition of LAB and cellulase improved the parameters tested representing a good silage product that need to be tested in vivo.
Title – Correct the H to h in the name of the species
Abstract – Is the abstract according to the fermentation rules? I think it is too long. Review it and try to short it down. For example:
in lines 16-17, instead of “with amaranth as the raw silage material, and the moisture content of the raw silage material was 16 adjusted to 60%, 65%, and 70% using wheat bran.”
you can write … with amaranth as the raw silage material, and the moisture content was 16 adjusted to 60%, 65%, and 70% using wheat bran.
in lines 21-23, instead of “The results showed that under the same L group, the pH in the 60% moisture content (MC) group was lower than that in the 70% MC group, and the lactic acid (LA) content in the 60% MC group was higher than that in the 70% MC group.”
you can write … The results showed that under the same L group, the pH and the lactic acid (LA) in the 60% moisture content (MC) group were lower and higher, respectively, than in the 70% MC group.
Other comments
1. Line 46 – Correct the value presented (106 instead 106).
2. Lines 56, 73, 98, 100, 371, 389, 390, 392, 415, 431, 453, 454, 475, 489-491, 590 - Apply the italic format to the name of the genus and species.
3. Line 73, and several other lines – It is not correct to write clostridium as a genus name. If you want to write the genus name it must be written with a capital C and in the italic format. However, if you want to refer the bacteria as a group you must write clostridia.
4. Line 102 – “rarely been reported.” – Add the references you found.
5. Line 146, 160, 205, 208, 312 – Change hours to h
6. Line 147-148 – Change “qualitative filter paper to obtain the extracts, and the pH of the extracts was determined by” to … qualitative filter paper. The pH of the final extracts was determined by …
7. Line 182 – Change Kg to kg
8. Lines 212-214 – The formula as it is it is not very clear. The minus it is difficult to see. Maybe the formula should be written in a more graphic way.
9. Lines 224-233 – When the authors refer to x.x times higher you should use values with two decimal figures. As an example, 86.58 x 1.1 = 95.24 instead of 93.86 if 1.08 is used.
10. Tables – The values in the tables should be aligned to make the reading easier.
11. Lines 389-390 – According to the taxonomic rules, one same text should not have one same abbreviation to different genera.
12. Lines 416-419, 537 – Homotypic, Heterotypic, Heterotrophic, Homozygous are used incorrectly. Homotypic and Heterotypic refers to nomenclatural issues. Heterotrophic refers to nutrition. Homozygous is related with sexual reproduction in fungi. I think the authors want to use Homofermentative or Heterofermentative. Please check each case and correct it according.
13. Lines 489-492 – To remove the repeated idea, change “Cellulase promoted the degradation of lignocellulose in Pennisetum sinese silage, reduced the crystallinity of lignocellulose, and improved the conversion of lignocellulose in Pennisetum sinese silage, indicating that cellulase was a very useful silage pretreatment method [44].” To … In Pennisetum sinese silage, Cellulase reduced the crystallinity of lignocellulose and improved it conversion, indicating that it was a very useful silage pretreatment method [44].
14. Lines 494-495 – Change “findings of Yi et al. [31]. Yi et al. [31] found” to “findings of Yi et al. [31]. These authors also found …
15. Line 509 – Change microflora to microbiota
16. Lines 577-579 – The reference in the end of the sentence it is not correct. Apply it after … to 128 h [55], …
17. Lines 602-603 – Change “was consistent with the results of Zhang et al. [61]. Zhang et al. [61] showed” to … was consistent with previous results [61]. Zhang et al. [61] also showed …
18. Line 612 – Apply the italic format to in vivo
19. References 1, 2, 4, 7, 9, 11, 21, 22, 26, 32, 34, 36, 38, 41, 44, 48, 55, 56, 59 – In these references check and correct the names of the genera and species. Sometimes they are not in the italic format, others the name of the genus it is not written with capital letter and others both must be corrected.
.
Author Response
Dear reviewer, thank you for your encouraging and warm comments and suggestions, all of your suggestions are very important, and they all have important guiding significance for our future research work. Based on this we have revised and (we think) strengthened our paper.
Point 1: Title – Correct the H to h in the name of the species
Response 1: We fully appreciate your suggestion. We revised that section in the paper. (PDF:4)
Point 2: Abstract – Is the abstract according to the fermentation rules? I think it is too long. Review it and try to short it down.
Response 2: We fully appreciate your suggestion. We revised that section in the paper. (PDF:12-46)
Point 3: Line 46 – Correct the value presented (106 instead 106).
Response 3: We fully appreciate your suggestion. We revised that section in the paper. (PDF:53)
Point 4: Lines 56, 73, 98, 100, 371, 389, 390, 392, 415, 431, 453, 454, 475, 489-491, 590 - Apply the italic format to the name of the genus and species.
Response 4: We fully appreciate your suggestion. We revised that section in the paper. (PDF:63, 80, 105-107, 373, 391, 392, 394, 417-418, 434, 456, 478, 492, 595)
Point 5: Line 73, and several other lines – It is not correct to write clostridium as a genus name. If you want to write the genus name it must be written with a capital C and in the italic format. However, if you want to refer the bacteria as a group you must write clostridia.
Response 5: We fully appreciate your suggestion. We revised that section in the paper. (PDF:80, 376, 458)
Point 6: Line 102 – “rarely been reported.” – Add the references you found.
Response 6: We fully appreciate your suggestion. The meaning of this sentence is not clearly expressed. Corrected to: “Studies on the addition of lactic acid bacteria and cellulase to amaranth and wheat bran (AWB) mixed silage have not been reported”. (PDF:109)
Point 7: Line 146, 160, 205, 208, 312 – Change hours to h
Response 7: We fully appreciate your suggestion. We revised that section in the paper. (PDF:153, 167, 207, 210, 314)
Point 8: Line 147-148 – Change “qualitative filter paper to obtain the extracts, and the pH of the extracts was determined by” to … qualitative filter paper. The pH of the final extracts was determined by …
Response 8: We fully appreciate your suggestion. We revised that section in the paper. (PDF:154-155)
Point 9: Line 182 – Change Kg to kg
Response 9: We fully appreciate your suggestion. We revised that section in the paper. (PDF:184)
Point 10: Lines 212-214 – The formula as it is it is not very clear. The minus it is difficult to see. Maybe the formula should be written in a more graphic way.
Response 10: We fully appreciate your suggestion. We revised that section in the paper. (PDF:215)
Point 11: Lines 224-233 – When the authors refer to x.x times higher you should use values with two decimal figures. As an example, 86.58 x 1.1 = 95.24 instead of 93.86 if 1.08 is used.
Response 11: We fully appreciate your suggestion. We revised that section in the paper. (PDF:228-235)
Point 12: Tables – The values in the tables should be aligned to make the reading easier.
Response 12: We fully appreciate your suggestion. We revised that section in the paper. (PDF:236, 255, 273, 294, 302)
Point 13: Lines 389-390 – According to the taxonomic rules, one same text should not have one same abbreviation to different genera.
Response 13: We fully appreciate your suggestion. We revised that section in the paper. (PDF:391-392)
Point 14: Lines 416-419, 537 – Homotypic, Heterotypic, Heterotrophic, Homozygous are used incorrectly. Homotypic and Heterotypic refers to nomenclatural issues. Heterotrophic refers to nutrition. Homozygous is related with sexual reproduction in fungi. I think the authors want to use Homofermentative or Heterofermentative. Please check each case and correct it according.
Response 14: We fully appreciate your suggestion. We revised that section in the paper. (PDF:398, 421-422, 543)
Point 15: Lines 489-492 – To remove the repeated idea, change “Cellulase promoted the degradation of lignocellulose in Pennisetum sinese silage, reduced the crystallinity of lignocellulose, and improved the conversion of lignocellulose in Pennisetum sinese silage, indicating that cellulase was a very useful silage pretreatment method [44].” To … In Pennisetum sinese silage, Cellulase reduced the crystallinity of lignocellulose and improved it conversion, indicating that it was a very useful silage pretreatment method [44].
Response 15: We fully appreciate your suggestion. We revised that section in the paper. (PDF:492-494)
Point 16: Lines 494-495 – Change “findings of Yi et al. [31]. Yi et al. [31] found” to “findings of Yi et al. [31]. These authors also found …
Response 16: We fully appreciate your suggestion. We revised that section in the paper. (PDF:499)
Point 17: Line 509 – Change microflora to microbiota
Response 17: We fully appreciate your suggestion. We revised that section in the paper. (PDF:514)
Point 18: Lines 577-579 – The reference in the end of the sentence it is not correct. Apply it after … to 128 h [55], …
Response 18: We fully appreciate your suggestion. We revised that section in the paper. (PDF:583-584)
Point 19: Lines 602-603 – Change “was consistent with the results of Zhang et al. [61]. Zhang et al. [61] showed” to … was consistent with previous results [61]. Zhang et al. [61] also showed
Response 19: We fully appreciate your suggestion. When we checked the paper, we found the citation to be inaccurate and removed the section. (PDF:607-611)
Point 20: Line 612 – Apply the italic format to in vivo
Response 20: We fully appreciate your suggestion. We revised that section in the paper. (PDF:617)
Point 21: References 1, 2, 4, 7, 9, 11, 21, 22, 26, 32, 34, 36, 38, 41, 44, 48, 55, 56, 59 – In these references check and correct the names of the genera and species. Sometimes they are not in the italic format, others the name of the genus it is not written with capital letter and others both must be corrected.
Response 21: We fully appreciate your suggestion. We revised that section in the paper. (References 1, 2, 4, 7, 9, 11, 21, 22, 26, 32, 34, 36, 38, 41, 44, 48, 55, 56, 59)
Thank you again for your suggestions and hope to learn more from you.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
The authors have taken into account almost all comments.
