Antifungal Activity of Cell-Free Supernatants from Lactobacillus pentosus 86 against Alternaria gaisen

Round 1

Reviewer 1 Report

The manuscript entitled: "Antifungal activity of cell‐free supernatants from Lactobacillus pentosus 86 against Alternaria gaisen" is an interesting study related to the production of secondary metabolites by a lactic acid bacterium with antifungal activity against Alternaria gaisen.

The work is very valuable and well developed, but with little novelty and I consider that it should be published after some improvements.

I enclosed the manuscript with my revision and comments, but I think that it is very important that the authors described how and why did they select this specific strain.

Moreover, in the discussion section, I consider that information related to Alternaria fungal inhibition using LAB strains need to be included, and in particular to compare action mechanims.

I congratulate the authors for its interenting work.

 

Comments for author File: Comments.pdf

Author Response

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Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript “Antifungal activity of cell‐free supernatants from Lactobacillus 2 pentosus 86 against Alternaria gaisen” presented by Hua Ying Liu and colleagues (horticulturae-2528861) foccusses a major problem related to fresh fruits in the post harvest period, caused by fungi spoilage. The authors highlight the importance of biological control, rather than the use of chemical fungicides to avoid resistance problems. Therefore, they propose the use of cell- free supernatants from a food-friendly bacterium as a natural preservative.

 

ABSTRACT

 

The authors must explain what is LAB in the abstract, to help the readers to promptly identify what they are talking about.

 

1. INTRODUCTION

The introduction clearly presents an overview of the problem addressed by the work, and the aim of the research. This part cites the recent literature, showing that the work foccuses a ongoing problem.

 

2. MATERIALS AND METHODS

 

This section is well detailed. One of the positive features of this work is the evaluation of spore germination, cellular leakage, morphological alterations, and membrane integrity. These experiments are an important advancement to reach suitable conclusions in addition to the antifungal activity by 96-well plate assay.

 

3. RESULTS

Results are well presented, with tables and figures that were helpful to follow the results presentation. The authors presented results and discussion in two different sections, making the results presentation very clear.

Line 231 – In Figure 5, it is possible to observe differences in the diameter of the hypha in the different images, A, B, C and D. Therefore, it should be better to show images that could be compared to each other. The magnification should be clearer, since I only could see it after increasing the images.

 

In addition, the authors mention (lines 240-241) that “The cell surface was not smooth and attached substances appear (Figure 6 B) ”. The presence of “attached substances ” was already observed in the non-treated cells (lines 223 and 224) “There was little attached substance.”, and it is not possible to observe an increase of attached substances in the images of Figure 6. This part should be rephrased, and the characteristics and alterations mentioned should have beem pointed by arrows or in other way, so that the reader can follow the results descriptions.

 

4. Discussion

 

In lines 226-227, authors state that: “The cell inclusions were disappeared, while collapsed and shriveled hyphae was left”. The meaning of this conclusion is not clear. What do they mean by “cell inclusions” ? Please, inform that and point them in the images.

 

In lines 280-281, “The present study indicated that the CFS of L. pentosus 86 displayed excellent thermostability and enzymatic stabilities, and was stable in an acidic environment ”, I suggest mentioning a literature reference to support the “excellent” behavior cited by the authors.

 

5. CONCLUSIONS

This section is ok, with a quick overview of the results and concluding ideas.

 

Overall, it is an interesting work. I suggest some minor aspects below.

line 16. Correct to “preservative”

line 33-34: the ver is missing

lines 34, 35, 38...: Add italics for Alternaria

Line 67. Add italics for Pyrus bretschneideri

Line 70. Correct “Dextrose”

Line 78: Please, verify the correctness of 120 r/min

Line 151: Please, correct “48 h”

Lines 162.0163: Please, add a space before the unity (°C). The same in Table 1.

Table 1. I suggest using “Control group (CK)” in the heading for clarity, instead of CK only. Or, add the meaning of CK as a footnote in the Table, so it becames self explanaotory

Figure 8. What does FL1_H stand for? In the axis, add the unity. In addition, this figure does not have a good resolution and authors should try to improve it.

 

Author Response

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Author Response File: Author Response.docx

Reviewer 3 Report

This manuscript described Lactobacillus pentosus 86 cell-free supernatants (CFS) demonstrated excellent antifungal activity (over 85%) against Alternaria gaisen. The CFS was thermostable, showed enzymatic stability, and retained antifungal efficacy under various conditions. It damaged hyphae and spores, making it a potential natural preservative against A. gaisen-induced fruit diseases.

 

1.     Can authors provide a picture of the colony growth of Alternaria gaisen, which represent in Figures 1 and 3? 

2.     Always use italics for scientific names in your scientific papers, reports, or any formal writing related to biology, ecology, taxonomy, or other fields that involve naming and classifying organisms.

3.     Please consider rewriting the last paragraph. Keep this paragraph concise and focused on the essential aspects of the study design and objectives. The goal is to set the stage for the following section, where you will provide detailed information on the methods and materials used to conduct the research.

4.     In scientific writing, including in manuscripts, the symbol "p" representing the p-value should be italicized, along with other statistical symbols.

5.     Added a space before and after "°C" to separate it from the number properly (Line 162-164)

6.     When presenting figures in a scientific manuscript, it is essential to include relevant information, such as the magnitude of the microscope used, in the figure title or caption. This additional information helps readers understand the scale and size of the objects or features represented in the figure.

7.     When presenting images or illustrations of fungi or any other biological entities in a scientific manuscript, it is essential for the authors to label and indicate the various cellular components accurately. This labeling helps readers understand the structures and components of the fungi and aids in interpreting the study's findings. For example, if the image is a microscopic view of a fungal cell, the authors should label the different organelles present, such as the nucleus, Golgi complex, mitochondria, vacuole, endoplasmic reticulum, etc. The figure title or caption should also include a description of the labeled organelles for clarity.

8.     There are some typos and mistakes such as: 

Line 33: Please consider rewriting the sentence. 

Line 78: r/min to rpm

Line 85: spores/mL

Line 148: space between to and 96 h

Line 151: space between 48 and h

 

Author Response

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Author Response File: Author Response.docx