Mixed Strains of Bacillus velezensis PBS-17, Lactiplantibacillus plantarum J-135, and Saccharomyces cerevisiae DAE-4 Increase the Storage Capacity of Fermented Feed and Silage
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsThe manuscript fermentation-3340636 is devoted to the production of silage using Bacillus velezensis PBS-17, Lactiplantibacillus plantarum J-135 and Saccharomyces cerevisiae DAE-4 cultures isolated from various natural sources in the Republic of Korea. The work is relevant. The work corresponds to Section Industrial Fermentation, Journal Fermentation. The merit of the work is the scale of the research, the mass of the samples was 500 kg, the duration of the tests was 3 weeks.
There are a number of methodological issues.
1) It should be added in the abstract that the ratio of Bacillus velezensis PBS-17, Lactiplantibacillus plantarum J-135 and Saccharomyces cerevisiae DAE-4 was (1:1:1).
2) The ratio (1:1:1) cannot be considered justified, since there is no data on ensiling the same substrate with individual strains, or two of the presented strains, or three strains, but in a different ratio, for example (1:3:1), since Lactiplantibacillus plays a leading role in ensiling.
3) It is not clear from the abstract, it is not clear from the materials and methods, which commercial microorganisms were used for control group 1 and control group 2. In the chapter "materials and methods" the genera and species, strains should be listed, the manufacturer of commercial microorganisms should be indicated. The application dosage should be indicated.
4) The authors' assumption in the introduction that a mixed culture of Bacillus velezensis PBS-17, Lactiplantibacillus plantarum J-135 and Saccharomyces cerevisiae DAE-4 will surpass individual strains is not substantiated. 5) The introduction should indicate what is new about the research.
6) In paragraph 2.6, add the volume of the silo machine, its name and manufacturer.
7) In paragraph 2.7, add the volume of the fermenter, its name and manufacturer.
8) In the discussion of the results, compare your results with those described in the literature.
Author Response
Comment 1: It should be added in the abstract that the ratio of Bacillus velezensis PBS-17, Lactiplantibacillus plantarum J-135 and Saccharomyces cerevisiae DAE-4 was (1:1:1).
Response 1: The mixing ratio of the microbial mixture was added to Abstract.
Comment 2: The ratio (1:1:1) cannot be considered justified, since there is no data on ensiling the same substrate with individual strains, or two of the presented strains, or three strains, but in a different ratio, for example (1:3:1), since Lactiplantibacillus plays a leading role in ensiling.
Response 2: Among various microbial ratios, the efficiency of fermented feed was excellent at the 1:1:1 ratio. For example, at other rates, the number of certain microorganisms decreased sharply or the antibacterial activity decreased sharply during the growth period.
Comment 3: It is not clear from the abstract, it is not clear from the materials and methods, which commercial microorganisms were used for control group 1 and control group 2. In the chapter "materials and methods" the genera and species, strains should be listed, the manufacturer of commercial microorganisms should be indicated. The application dosage should be indicated.
Response 3: Each microbial agent provided by the Agricultural Technology Center was used, control 1: single-strain probiotic (Bacillus substilus), control 2: mix-strain probiotic (Bacillus, yeast, LAB)
Comment 4: The authors' assumption in the introduction that a mixed culture of Bacillus velezensis PBS-17, Lactiplantibacillus plantarum J-135 and Saccharomyces cerevisiae DAE-4 will surpass individual strains is not substantiated.
Response 4: Research suggests that bacillus bacteria promote the growth of LAB, and yeast has a significant impact in forming aerobic conditions for LAB growth.
Comment 5: The introduction should indicate what is new about the research.
Response 5: The purpose is to isolate microorganisms with excellent antibacterial activity and develop complex microbial additives with high antibacterial activity. Added to the introduction.
Comment 6: In paragraph 2.6, add the volume of the silo machine, its name and manufacturer.
Response 6: Added silage machine information to paragraph 2.6.
Comment 7: In paragraph 2.7, add the volume of the fermenter, its name and manufacturer.
Response 7: Microbial fermentation was conducted on a laboratory scale.
Comment 8: In the discussion of the results, compare your results with those described in the literature.
Response 8: In the search for microorganisms, we focused on comparing the activity of microorganisms isolated from the same area, and predicted their activity by referring to other literature. Numerical comparison is judged to be difficult
Author Response File: 
Author Response.pdf
Reviewer 2 Report
Comments and Suggestions for Authors1- The introduction section lacks a comprehensive literature review, which is essential to provide context and demonstrate the relevance of the study. I strongly recommend the authors include at least 10 references from recent research studies that address similar objectives or methodologies. This will strengthen the scientific foundation of the manuscript, highlight the novelty of the work, and situate the findings within the broader field.
2- The introduction does not clearly present the novelty and specific contribution of this research. It is strongly recommended that the authors explicitly explain the unique aspects of this study and how it advances the field. Additionally, the steps of the research should be outlined and compared with existing literature to highlight how this work addresses gaps or improves upon prior studies. This will help clarify the significance and originality of the research within the context of the current body of knowledge.
3- In Section 2.1, please provide a reference for the statement: "One gram of soil was mixed with 100 mL of 0.85% NaCl solution and then spread onto Luria-Bertani (LB) agar using the serial dilution method." Additionally, clarify how this specific combination was used in the experimental setup and its relevance to the objectives of the study. Providing more details will enhance the reproducibility and understanding of the methodology.
4- In the Materials and Methods section, it is essential to provide references for all parts of the methodology that are adapted from existing studies. I strongly recommend referencing all parts and clearly specifying which parts were developed by the authors. The methods developed by the authors must be thoroughly explained in detail, considering the background and relevance of the research, to ensure they are scientifically acceptable and reproducible. In addition, creating a diagram to visually explain the methodology is recommended, as it will enhance the clarity and understanding of the experimental approach.
5- In the conclusion section, it is recommended to clearly explain the results and their significance in contributing to the field. Include specific numerical results to provide concrete evidence of the study's findings and their impact. This will help emphasize the relevance and practical applications of the research outcomes.
Author Response
Comment 1: The introduction section lacks a comprehensive literature review, which is essential to provide context and demonstrate the relevance of the study. I strongly recommend the authors include at least 10 references from recent research studies that address similar objectives or methodologies. This will strengthen the scientific foundation of the manuscript, highlight the novelty of the work, and situate the findings within the broader field.
Response 1: References have been added to the introduction and method section.
Comment 2: The introduction does not clearly present the novelty and specific contribution of this research. It is strongly recommended that the authors explicitly explain the unique aspects of this study and how it advances the field. Additionally, the steps of the research should be outlined and compared with existing literature to highlight how this work addresses gaps or improves upon prior studies. This will help clarify the significance and originality of the research within the context of the current body of knowledge.
Response 2: Explanation of fermented feed and silage and emphasis on the importance of microbial agents during the fermentation process.
Comment 3: In Section 2.1, please provide a reference for the statement: "One gram of soil was mixed with 100 mL of 0.85% NaCl solution and then spread onto Luria-Bertani (LB) agar using the serial dilution method." Additionally, clarify how this specific combination was used in the experimental setup and its relevance to the objectives of the study. Providing more details will enhance the reproducibility and understanding of the methodology.
Response 3: References for experimental methods are provided.
Comment 4: In the Materials and Methods section, it is essential to provide references for all parts of the methodology that are adapted from existing studies. I strongly recommend referencing all parts and clearly specifying which parts were developed by the authors. The methods developed by the authors must be thoroughly explained in detail, considering the background and relevance of the research, to ensure they are scientifically acceptable and reproducible. In addition, creating a diagram to visually explain the methodology is recommended, as it will enhance the clarity and understanding of the experimental approach.
Response 4: Conduct research, design experiments, and submit with reference to methods.
Comment 5: In the conclusion section, it is recommended to clearly explain the results and their significance in contributing to the field. Include specific numerical results to provide concrete evidence of the study's findings and their impact. This will help emphasize the relevance and practical applications of the research outcomes.
Response 5: Emphasis was placed on the development of complex microbial preparations to prevent bacterial contamination and spoilage, which are problems in the production of silage and fermented feed.
Author Response File: Author Response.pdf
