Fermentation Quality and Bacterial Diversity of Broussonetia papyrifera Leaves Ensiled with Lactobacillus plantarum and Stored at Different Temperatures

Round 1

Reviewer 1 Report

The author adequately presents the research conducted on the fermentation segment of Broussonetia papyrifera Leaves silage, mainly by screening the bacterial diversity for the samples with or without Lactobacillus added, at different stages in time and temperatures.

Ensiling is currently the most effective technique for feedstuff preservation as high-quality forage, being able to conserve 93% of crops gross energy when inoculated with lactic acid bacteria (LAB). Lactobacillus was introduced into the US silage market in 2002. Since then, many studies, associated with many citations, have documented its effects of increasing acetic acid and reducing the numbers of yeasts and molds, on different plant species, thus improving aerobic and duration stability, so the topic is not original by itself.  

However, the main trends in silage making today are a return to the basic principles of silage fermentation from two decades ago, namely the proper supplementation of the silage to ensure maximum economic utilization. Traditionally, LAB starter cultures were applied to lower the pH by producing lactic acid and to inhibit the growth of undesirable epiphytic microorganisms by competing for nutrients. Current trends show a focus on lactic acid bacteria inoculants, as an effective tool for creating microbial quality of silages and selecting new species with extraordinary features, topic addressed by the current paper. While the contemporaneous papers (many, but with decent views and few cites in Open Access libraries) focus on identifying new LAB cultures mixed or without other bacteria, their interactions and metabolome dynamics, the author focuses on effective measures to inhibit undesirable microorganisms when ensiling B. papyrifera, due to the fact that more than 5 million hectares are currently planted within an anti-poverty project. Due to the scale of the project, it is likely that the results will be used in the implementation area in several domains.

The paper has a good reading flow, presents detailed statistical analysis, the results are consistent and the conclusions and correlated with the arguments presented and the main theme. 

Small corrections are needed to improve the paper:

• Line 75 – Could the author please remove “The” since the definite article is used when the nouns are specific and here is not the case and add “s” at the end for herbicides and fertilizers
• Line 123 – please delete la last part of the sentence “for quantification” since It is already mentioned in the previous line
• Line 129 - Could the author please move the link to the Reference section
• Line 131 - Could the author please move the link to the Reference section
• Line 165-166- Could the author please move the verb in the correct order to create more sense for the reader. The lowest DM loss in the LP treatment at 30℃ (0.62 g/kg FM) was recorded – to “The lowest DM loss in the LP treatment was recorded at 30℃ (0.62 g/kg FM)“
• Line 327-328 - Could the author please replace the construction “Studies have shown that the addition of Lactobacillus to silage” with “while other studies that” or similar to avoid repetition
• Line 350 - Could the author please replace the “,” with “.” to create sense to the text.
• Line 383- Could the author please remove the “all” since all the subjects were already mentioned in the sentence
• Line 387-388 - Could the author please remove the sentence “In conclusion, the addition of LP and stoing at 30℃ could effectively improve the fermentation quality of B. papyrifera leaves silage.” since the “Conclusion” section already has a very similar sentence in the beginning of the paragraph

Comments for author File: Comments.pdf

Author Response

Dear reviewer,

Thank you very much for your approval of our manuscript. I have revised the manuscript in the light of your comments and have marked it in red.

The details of the changes are shown as follows,

Line 4, replaced "," with "and"

Line 80, replaced " The herbicide and fertilizer " with " Herbicides and fertilizers "

Line 131, deleted "for quantification"

Line 137 and 139, we are not certain in what format the link will be moved to the reference section

Line 178-179, replaced "at 30℃ (0.62 g/kg FM) was recorded" with " was recorded at 30℃ (0.62 g/kg FM)."

Line 191-193, rephrase the original sentence as “With the ensiling days prolonged, the count of LAB significantly decreased at 30℃ (P < 0.01) and the count of LAB also decreased significantly (P < 0.01) in LP treatment at 15℃.”

Line 194, added “at”

Line 306, rephrase the original sentence as “The pH was 4.49 in the LP treatment at 30℃.”

Line 350-351, replace the construction “Studies have shown that” with “while other studies that”

Line 375, replace the “,” with “.”

Line 410, remove the “all”

Line 414, remove the sentence “In conclusion, the addition of LP and stoing at 30℃ could effectively improve the fermentation quality of B. papyrifera leaves silage.”

Kind regards,

Yours sincerely,

MingYang Zheng

Author Response File: Author Response.docx

Reviewer 2 Report

This paper gives the interesting results of influence of Lactobacillus plantarum addition during ensiling Broussonetia papyrifera leaves on quality of obtained silages. More importantly, the results showed the influence of two different storage temperatures on fermentation quality of Broussonetia papyrifera leaves silage, which has not been previously reported.

The layout of the paper is sufficiently good.  The English is also sufficiently good. The methodology is clear and detailed but it would be useful that authors explained if the same methods were used for analyzing quality of raw Broussonetia papyrifera leaves (presented in Table 1) as for the samples of silage. The results are presented in manner so that potential readers can clearly follow the given discussion. However I could not find any figures that are mentioned in manuscript, not even as a supplementary data. Beside tables there was no any other document supplemented to a manuscript. I could only find titles of four figures. For better understanding of results and it would be mandatory that figures be included.

The authors showed clear influence of temperature on quality of fermentation and silage process of Broussonetia papyrifera leaves which is interesting topic that can be further investigate. I would like to recommend this article for publishing after minor revision.

Author Response

Dear reviewer,

Thank you very much for your approval of our manuscript. It is possible that the figures were missed due to unfamiliarity with the submission system. We are now attempting to resubmit the figures.

Kind regards,

Yours sincerely,

MingYang Zheng

Author Response File: Author Response.docx