Ice-Nucleating Gut Microbes in Insects: A Scoping Review

Round 1

Reviewer 1 Report

This kind of studies are relevant for scientific community.

This manuscript needs extensive reformulation. The conclusions are confused and no data suports them. There is no evidence for the assumptions taken.

English and technical aspects need to be reviewed.

This kind of studies are relevant for scientific community.

This manuscript needs extensive reformulation. The conclusions are confused and no data suports them. There is no evidence for the assumptions taken.

English and technical aspects need to be reviewed.

Author Response

This kind of studies are relevant for scientific community.

-Thank you!

This manuscript needs extensive reformulation. The conclusions are confused and no data suports them. There is no evidence for the assumptions taken.

-That's rather vague, and does not match the other three reviews. I did add more information for the conclusions, which should make things clearer, but I think my evidence is well laid out.

English and technical aspects need to be reviewed.

-I am a native English speaker. With respect, given the grammar errors in your own review comments, I do not think you are qualified to assess the quality of the English in my paper.

Reviewer 2 Report

Review with a limited scientific value, but useful for the topic and worthy of being published

 Minor editing of English language required

Author Response

Review with a limited scientific value, but useful for the topic and worthy of being published

-Thank you

Minor editing of English language required

-I checked for typos and corrected them.

Reviewer 3 Report

This manuscript summarized the studies of ice nucleation by gut microbes in insects. Although a review of insect freeze tolerance was recently published in 2023 by Morgan-Richards et al., this manuscript focuses on ice nucleation microbes, a topic that remains relevant and of interest to the scientific community. The authors note that only a few research groups have studied these microbes and suggest their potential use as biopesticides. The paper is well-structured and well-written. I recommend it for publication.

Author Response

Thank you for the 5-star review!

Reviewer 4 Report

The reviewed manuscript is devoted to a completely new problem – the accumulation of data on the effect of ice nucleating microorganisms in the digestive tract of insects on their survival at extra low temperatures (on the freeze-tolerant of insects). Despite the fact that the authors have put a lot of effort into finding information related to this issue and systematizing it using advanced approaches, I believe that this article needs significant revision before it can be published.

1.    It would be more convenient to perceive the information presented in the text if you provide a diagram illustrating the mechanism of preventive action of ice nucleating microorganisms on insect’s freeze-tolerant.

2.    You have a significant number of sources from 20 years ago in your list of references (more than 60% of sources are older than 2000). Maybe this study has lost its relevance today or you should perform a detailed search among more recent publications using other keywords

3.    Tables 1 and 2 show the names and species of insects and the species/strains of microorganisms, but since you have a review and analytical article, it’s would be better to add more information in the tables, for example: properties of microorganisms, their effect on the physiology of insects, etc.

4.    You need to add conclusions! In addition to the general conclusion on your material, give suggestions on the use of these properties of microorganisms in various industries and prospects for future research.

Author Response

1. It would be more convenient to perceive the information presented in the text if you provide a diagram illustrating the mechanism of preventive action of ice nucleating microorganisms on insect’s freeze-tolerant.

Okay. We have added such a diagram.

2. You have a significant number of sources from 20 years ago in your list of references (more than 60% of sources are older than 2000). Maybe this study has lost its relevance today or you should perform a detailed search among more recent publications using other keywords

We did not reject any papers relating to ice nucleation, as there are relatively few overall. I would not say the field has lost its relevance, but maybe its sexiness. I have added some discussion of this to the discussion section. I do not think the search string can be improved or that another search is warranted, but I will note that another paper on this topic was made available online ("in press") while this manuscript was under peer review [DOI: 10.1093/ee/nvae027]. I have added this brand new paper to the discussion.

3. Tables 1 and 2 show the names and species of insects and the species/strains of microorganisms, but since you have a review and analytical article, it’s would be better to add more information in the tables, for example: properties of microorganisms, their effect on the physiology of insects, etc.

-A good suggestion. I have added a column to Table 1 stating if the insect is freeze tolerant or freeze susceptible, and a column to Table 2 about the effect of the microbe on the physiology of the insect in the experiment. There is not much to say about the properties of the microbes, other than that they are all ice nucleating active.

4. You need to add conclusions! In addition to the general conclusion on your material, give suggestions on the use of these properties of microorganisms in various industries and prospects for future research.

-Very well, I have added some more information.

Round 2

Reviewer 4 Report

The authors of the reviewed article have performed significant work to improve the quality of the manuscript. In general, I suppose that the article is suitable for publication in Microbiology Research and meets all the established requirements

But I would like to draw the attention of the authors to some points:

For Figure 1. You have an overly detailed and extended note. I suppose it can be moved to the text of the article, and leave only a brief explanation.

In table 1, you provide links to references in column 3, although you have the last column selected for all citations.

Author Response

Thank you for the excellent suggestions!

-I have deleted redundant text from the caption of figure 1 that was explained in the main text. It is now half as long as it was before.

-I edited the caption to clarify this. The last column is the citation of the INA paper described in that row or rows, while the citation in column 3 means the information on whether it is freeze-susceptible or freeze-tolerant came from a different paper than the INA experiment paper cited in the last column. I hope this edit is sufficient. I prefer to keep things separate because some organisms are associated with multiple papers.