-->PONE-D-25-25129-->-->Pest consumption by common bats in diverse landscapes-->-->PLOS ONE

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Reviewer #1: Journal: PLOS ONE

Manuscript ID: PONE-D-25-25129

Pest consumption by common bats in diverse landscapes

This manuscript explores how bats provide essential ecosystem services in the form of insect pest consumption. Using Rhinolophus bats as focal species, the authors used insect DNA found in fecal samples to estimate the impact bats were having on insect pest species in the region.

Main Comments:

• The introduction introduces the material well, but there are parts where the language is vague and there are not good connections between paragraphs. I recommend spending more time linking the concepts to why the focal species is a good model a bit more. Additionally, spend more time explaining on how insects are considered pest species in the region and connecting it to your focal species.

• Overall there is excellent information herein, but there is a lot of vague language used and there needs to be more time spent connecting concepts together.

• There are some changes needed for the tables and figures to make them cleaner and easier to understand. Always remember that the reader should be able to just read the captions and know what the figure is about without ever seeing it.

• Because the reader may be unfamiliar with the region, I recommend including a map with where the site are located/where samples were collected.

• The abstract states that the study is during the breeding season, but does not explain well more of the life history of the focal species. The study covers 7 months, which may encompass the breeding season and then some. More care should be given introducing the focal species and what they are a good model for this study (which I do think they are).

Specific suggestions and remarks

Introduction

37-38: Reduce sentence length “…feed on a wide variety of arthropods, including many pest species such as …”

40: Vague language; “This makes bats vital…”

42: Bats can offer; leave the potential open ended so that it gives the reader the sense that your work is adding to the collection of work

46: Plasticity of what? Be more specific and then give examples of those.

51-52: Avoid vague words such as “this”. Consider rephrasing “Because of bats ability to adjust their diets and foraging habitat, they are highly effective pest suppressors…”

60-61: Inappropriate to call other studies “run of the mill”. It gives the wrong impression about their value

60-67: I would consider reframing this paragraph to focus on how fecal DNA is a great tool to understand bat diets, and thus the pests they consume.

68-79: These two paragraphs do not connect to one another well. Maybe spend time on why it is important to use abundant and widely distributed species can help answer the question, and then go into your focal species as a model.

Methods

82: Be more specific as to where this study is occurring. It may be worth including a map of the region, and where samples were collected.

90-99: Inconsistent with time periods. Some state the week collection occurred, others just the month/day. Be consistent.

100-105: You introduce “OUT”s here, but do not explain what it is. It is explained later on, but currently the reader does not know what OTUs are. I suggest either expanding this section, or moving it further down to be aligned with the description of OTU.

145: If it is unpublished, how are the data accessible?

151: If focusing on these uncertainties, spend more time in the intro about the specifics.

156-170: It says there is a formula, but I do not see one written out.

Results

174: Were the samples pooled? Or were they tested individually? Is this proportion just the samples that tested for any pest species? What is the proportion of pest species within the positive samples?

192-193: Linked how? Be more specific.

193-195: Species of bat or of pest insect? Be more specific and how they are linked to the plant species.

230-232: How do we, the reader, know that is the usual time pest species emerge? What does “typical emergence pattern” mean? Is it species specific?

243-245: State earlier species general differences, such as their sizes.

251-252: Why do they have lower energy costs? If this is an assumption because of size, I suggest removal of the sentence. If it is supported in the literature, then cite the sources.

Discussion

257: That some species of horseshoe bats.

257-258: Get rid of “because of their remarkable adaptability”. Suggest it later in the discussion if appropriate.

260: Remove “in this study”

261-263: Combine sentences; “…resulting in few total occurrences, while other insect species are heavily consumed and form what is known as “core diet”..”

263: “These” is vague; “The insects comprising the core species…”

265-266: Explain more the adaptability you document ahead of the ending sentence such as “Because we documented bats doing x,y,z, we argue that they have an adaptable diet and thus…”

270-271: This is not really what your study looked at, but your study does suggest it. Propose the ideas as potential hypotheses, not as fact.

278-281: This sentence seems out of place compared to the topic sentence.

299-305: I think this paragraph, while interesting, is not necessary and could be removed or incorporated elsewhere.

Tables and Figure Captions

Table 1:

Table 2: Add more to the table caption including writing out the full site names and species names.

Table 3: Use decimal points rather than commas.

Table 4: A bit confusing; the total diet is less than individual sites. Explain more in the caption so that the reported values make more sense.

Figure 1: Explain how sampling week is in relation to the week of the year somewhere.

Figure 2: Spell out full bat species name. The metric is needed (%?) for the wPOO. What are the units?

Figure 3: Units needed on (a); Kg of what? Put more on axis label

Figure 4: Is this for both species or just one? State it in the figure caption.

Figure 5: Spell out full bat species name. Add units on axis.

Reviewer #2: Peer Review: “Pest consumption by common bats in diverse landscapes” by Aldasoro

The manuscript, “Pest consumption by common bats in diverse landscapes”, details an important narrative of the ecosystem service that bats provide as pest consumers. Quantifying the abundance and diversity of pests that bats consume in varying agricultural settings throughout a season plays an important role in promoting bat conservation and overall ecological health. This manuscript supports this foundational knowledge by studying two common & abundant species in Spain and describing their pest consumption throughout the season. While the manuscript makes several claims without citations or data to support them, with further citations and specifications, this manuscript will benefit the body of literature well.

I think my biggest issue with this manuscript is that it makes the claim that the seasonality in consumption of different pests follows the pulses in pest abundance associated with hatches. However, there are no citations or data that support the seasonality of pulses in pest abundance. Without supporting data to show that the peak in species-specific pest abundance mirrors the peak in species-specific consumption, I don’t think you can make this claim. I believe that this is a highly-probable theory, but the diet data that you are analyzing doesn’t support this claim until you add additional data about seasonality of the pest abundance.

Additionally, you also make the claim that lower pest abundance comes from sites that have more diverse and natural surroundings, however you don’t provide any data to back this claim up. This seems like an easily quantifiable fix with land use imagery to quantity the amount of natural area to anthropisation around each site, but without the data to support this- its an unsubstantiated claim in the paper. Detailed in the comments below are more instances where more specifics are needed or claims are quite broad.

Specific Edits:

Line 27: “they can go one step beyond human capabilities” – Please be specific about what you mean here.

Line 29: “several tons of insects” – Give a range of values that you have found here.

Line 50: “frequently consume insects from outside their hunting grounds” – I would say that many bats do not have specific hunting grounds, but forage opportunistically as you mention. I think just rewording this phrase would be best.

Line 56: “The larger the population, the greater the collective consumption of pests” – This seems like an assumption to me and I could think of many ecological scenarios where abundance of bats does not correlate with pests consumed. I would rephrase this to accommodate some of this uncertainty.

Line 60: “Run-of-the-mill” – This seems a little derogative to the studies mentioned- I would choose a different descriptor.

Line 65: What method have these studies used to estimate pest consumption? You refer back to this later, so I think it would be helpful to be specific here.

Line 77-79: I think a clearer description of research question, study objective, and study importance would be beneficial here. The last sentence just feels tacked on without really incorporating it into the study importance. Just rephrasing this would create a much stronger introduction

Line 83: Double check your species names here- I think you can start using abbreviations for both species. This happens several times throughout the paper including with the pest species, but I am not going to list them all.

Table 1: Population sizes in colonies vary quite a bit over the course of the year as juveniles are born. Are these populations sizes pre- or post- birth pulse in abundance?

Line 97: “Up to 24 samples of 4-6 pellets” - How did you decide on how many samples per session? How did you did you decide on which pellets to pick up? Randomly? Off the top? Largest? Most whole pellets?

Line 101: How many samples were considered contaminated from the 413 and 377 samples?

Line 102: Here you say you are analyzing them based on sampling month, but some of your figures are divided up by sampling week? Is that just for R. ferrumquinum? I’m a little confused about this part.

Line 142: How did you decide what was potential prey and what wasn’t?

Line 154: I would love to see a map of site locations and this general region in Spain- I think this would help delinate some of the later population size estimates.

Line 156: You outline your variables here, but I never see an actual formula about how you calculate these together?

Line 160: It would be good to know the where the 30% comes from in these studies as compared to the 70% in c1. Why such a big difference between the two? Are the studies for c2 also during pregnancy and lactation? Or males? If you just chose the min and max values from the literature, I would just state it as such.

Line 163: “Basque Country of Aihzartza (2001)” – Reword: it sounds like there is a country in Basque named Aihzarta?

Table 3: I understand that you are using wPOO here, but it feels a little nebulous to me. If the max that a wPOO can be is 100% (where they only eat one thing), then 8.5% of insects that R. hipposideros eats is a pest seems rather low?

Figure 2a: I would reduce the y-axis range down to 1500 wPOO. Since there is only one main high point in that range, it would be more easier to read the smaller values.

Line 232: I think this is a key point in your paper, but it would be a lot stronger to have data to back this up. Does the Integrated Pest Management Guides of the Spanish Government have temporal data to cite for an abundance comparison with the peak consumption? Otherwise, you are stating this with no citation or evidence to back the claim up.

Line 245 & 247: You now use the common name here- I would stick to either Latin name or common name.

Line 248: I think the map part may help a bit here, but how far of a spatial scale would the 79.28kg eaten be covering? An estimate of the home range of the colonies would be beneficial here. Would you expect this estimate to vary between colonies? Are the ranges in colonies overlapping?

Line 254: This is the first time that you mention anthropisation in the paper. How did you calculate this? Which sites are higher or lower in terms of anthropisaion?

Line 257: Based on what you just presented, it seems like they eat more than “several” agroforestry pests?

Line 264 &267-268: Again, I think your work would be a lot stronger if you had data to compare the peak outbreaks of pests with the peaks in consumption and showed that bats are moving from one pest to the next and are truly following the varying peaks in abundance. Right now, you just show us that consumptions changes throughout the season without proving to us that it follows peak abundance. I get that you may not have data to prove localized abundance, but even literature supporting the timing of these peaks would be strengthen your work.

Line 269: “typical general” – I think you only need one of these words here.

Line 274: Again, a map to see these rivers would be great. I also want to emphasize again that you haven’t provided us any context or data to differentiate between sites in the amount of “diverse natural habitat” nearby. Maybe in Table 1- you could put a percentage of natural habitat vs anthropogenic habitat? Its hard to know if you consider a pine plantation as natural or anthropogenic otherwise or what the difference is between Mediterranean bush vs shrubland?

Line 279-281: “bats’ impact can exceed their foraging areas”- How do we know they aren’t foraging down there? Are the insects moving to the bats or vice versa? Or do the rice pests utilize other plants as well?

Line 297: “horseshoe bats are among the most important bat species for pest suppression” – This seems like a bit of a stretch to me for the subject of this paper. How do we know that other species do not eat higher rates of pests? Are they the biggest bat in the region in which case, you could rephrase this comment? Maybe caveat it with “Given that horseshoe bats are one of the most common and can be one of larger-bodied bats in the region, …”

Line 314: “one step beyond human capabilities” – Again, I feel like this phrasing is a little weird. I would reword since I can see what you are saying here.

Line 326: I also think a line in here about education on the importance of bats and their ecosystem services within the public would also be important for maintaining bat populations and positive bat sentiments.

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-->PONE-D-25-25129R1-->-->Pest consumption by common bats in diverse landscapes-->-->PLOS One

Dear Dr. Aldasoro,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.-->--> -->-->Overall, this manuscript is vastly improved from the original version, and the authors are to be commended for engaging with the reviewers to improve their work. There remain a few outstanding details that Reviewers 1 & 2 identify, and, overall, a careful proofread to address errors in spelling/grammar/syntax/punctuation. Once these issues are resolved, the manuscript will be acceptable for publication in PLoS ONE.-->-->

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Graciously, the original two reviewers agreed to review this resubmission. The reviewers agree and I concur that the manuscript has vastly improved since the original version, however their recommendations were split. Reviewer 1 identifies a few remaining issues that need to be address, but argues for minor revisions. Reviewer 2 identifies several areas where additional nuance and context is required, but argues to accept. Both reviewers note that the manuscript, overall, needs a careful revision to address errors in grammar/spelling/syntax/punctuation. Once these minor revisions are complete, this manuscript should be ready for publication in PLoS one.

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Reviewer #1: All comments have been addressed

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Reviewer #1: Journal: PLOS ONE

Manuscript ID: PONE-D-25-25129

Second Review

Pest consumption by common bats in diverse landscapes

This manuscript explores how bats provide essential ecosystem services in the form of insect pest consumption. Using Rhinolophus bats as focal species, the authors used insect DNA found in fecal samples to estimate the impact bats were having on insect pest species in the region.

Main Comments:

• The authors did a diligent job on addressing the comments and critiques from the first review. The manuscript has greatly improved, and I only have a few small recommendations. The manuscript is greatly improved and provides much better context for the work and findings.

Specific suggestions and remarks

Introduction

75-76: Consider separating into two paragraphs with the species info in the next

Methods

88-89: Be more specific as to where this study is occurring; state the country.

91: Expand the figure description. What is the inset map? What is it focusing in on?

122-128: Move the ethics statement up further

Reviewer #2: Thank you for your revision & thoughtful responses to reviewer comments. I believe that your manuscript is stronger because of these revisions. There are still some grammatical and spelling errors throughout the manuscript. Mainly this is a topic with a lot of nuances and I have listed out a couple spots below where I feel the nuance may still need to be addressed.

Line 171: I would organize these descriptions based on the order they appear in your equation. Also your equation uses ‘pop’ and your description uses ‘pop size’ – these should be identical. I would also point out that you are using a population estimate from 2001 which is 25 years ago. Do you expect these population estimates to have changed at all? You also report that ‘other studies’ report similar population sizes between the two, but don’t cite these studies.

Line 287: I still feel a little conflicted about something that occurs in 5% of the total samples across all colonies being considered as “heavily consumed”. I understand that you have a citation to support this, but to me it still seems like it should be higher to be considered the “core diet”. Almost all of the individual pest species have a wPOO of less than 1% abundance across all sites….

Line 293: Again, “manage pest populations effectively” is a bit strong.

Line 324: You didn’t prove that there R.hi had a significant impact on pest species- just that they ate a lot of them. Their consumption may have been a drop in the bucket of O. sacchari abundance levels- we just don’t know.

Figure 1. I think some of these site names are mixed up. Shouldn’t the labels read as Matxinbenta (MA) and Balmeseda (BA) instead of the species names? Or if that is correct, I don’t understand why and as such this should be added to the figure caption.

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Reviewer #1: No

Reviewer #2: No

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<p>Pest consumption by common bats in diverse landscapes

PONE-D-25-25129R2

Dear Dr. Aldasoro,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

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Kind regards,

Mark A. Davis, Ph.D.

Academic Editor

PLOS One

Additional Editor Comments (optional):

After reviewing the Responses to Reviewrs and the revised manuscript, it's clear that the Authors have been thorough and diligent in their revisions. This manuscript now meets PLoS ONE's critera for publication, and I look forward to seeing it published.

Reviewers' comments: