The Fermentation Quality, Antioxidant Activity, and Bacterial Community of Mulberry Leaf Silage with Pediococcus, Bacillus, and Wheat Bran

Round 1

Reviewer 1 Report

A brief Summary

In this manuscript authors report about the investigation of effects of different strains and wheat bran on the fermentation quality, antioxidant activity, and bacterial community of mulberry leaf silage. Mulberry trees exhibit adaptability to diverse environments. Another advantage of this woody plant is that mulberry leaves have very comprehensive and balanced nutrient content. Other natural bioactive compounds exhibit also antibacterial, antioxidant, and anti-inflammatory effects. Mulberry leaves are employed as a high-quality protein feed material in animal production.

Despite many benefits, a problem exists with high moisture and protein content in mulberry leaves, which causes difficulties with long-term storage. Authors have proved that addition of Lactobacillus and fungi can improve silage quality.

General concept comments

The main question addressed by this specific research is relevant and interesting. Fast growth in animal production in China, as well as in many other places of the world, causes a scarcity of feed resources. Thus, a need for new unconventional feed resources is more than obvious.  

Specific comments

The title of the manuscript properly reflects the subject of the paper.

Abstract provides an accessible summary of the paper.

Keywords accurately reflect the content of this research.

The manuscript is clear, relevant for the field and is presented in a well-structured manner. Most of the cited references (36 aut of 46) are recent publications (within the last 10 years) and relevant to the field. There are no self-citations included. The manuscript is scientifically sound, and the experimental design was appropriate to test the hypothesis, which was completely confirmed. After proper methodology (addition of Lactobacilus and Bacillus) was used, the results show the improved fermentation quality of mulberry leaf silage in comparison with other treatments. In my opinion the manuscript’s results are reproducible. There are enough details given in the Materials and methods section.

Authors are kindly asked to add a short explanation about the content of water-soluble carbohydrates (WSC) which reflects the fermentation quality. This factor is firstly mentioned in Section 2.3 (line 112). Then it was used and explained in Discussion section (line 259). I think it would be better to transfer this explanation to Section 2.3.

All four tables are clear. All four figures properly show the data. Anyway, I would recommend bigger letters, as at present situation it is difficult to read legends and titles of x and y-axis. Nevertheless, data are interpreted appropriately and consistently throughout the manuscript. Discussion section is precise and extensive. The chemical composition and microbial population, active components and antioxidant activity, bacterial diversity and abudance of fresh mulberry leaves and mulberry leaf silage were compared.

Statistical analysis was successfully used. Obtained data were analyzed with SPSS 26.0 software, employing One-way ANOVA and Duncan's method to determine statistical significance at the p < 0.05 level.

Conclusion section is short, but precise. It is consistent with the evidence and arguments presented.

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

These are my remarks and suggestions:

·       The title should be rephrased for clarity and to better explain the purpose of the study. Also, was the study conducted with Lactobacillus spp ??? and Bacillus spp ? In material and methods, in the strains used, the Lactobacillus spp aren’t presents

·       Lines 14-15. Please remove the (Lactobacillus and Bacillus) from the abstract

·       Line 81. Bacillus licheniformus- italic

·       Lines 14-15,64, 72-73 and all over the text. Add after the genus spp. (eg. Bacillus spp.)

·       Line 72-75. Please rephrase

·       Line 79. I suggest rephrasing in Bacterial strains and culture conditions. Also,  provide information about the strains used (were they isolated or purchased?)

·       I suggest a separation of subchapter 2.3 as follows: determination of chemical composition, Fermentation parameters detection and analysis, Microbiological evaluation

·       For the microbiological analysis provide more details or add a reference

·       Line 131 Eosin-Methylene Blue agar medium instead of Eosin-Methy Blue agar medium

·       Lines 174-177. Please rephrase (the information is repeated from the table 1)

·       Tables 2 and 3 Please provide SEM or SD for each treatment. How were the results expressed? As a mean of triplicate measurements ± standard deviation??? Please provide this information in the 2.6 subchapter

Author Response

Please see the attachment.

Author Response File: Author Response.pdf