PONE-D-20-16747

The masked seducers: Lek courtship behavior in the Wrinkle-faced bat Centurio senex (Phyllostomidae)

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Reviewers' comments:

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Reviewer #1: Yes

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

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Reviewer #1: The authors described for the very first time Lek mating behavior in the bat Centurio senex, this is a valuable and exciting finding. Furthermore, the manuscript is expertly prepared. It is extremely rare to read an article that so thoroughly anticipates and answers questions/concerns. Both the topic, the evaluation of the data, and the quality of the writing, made this a true pleasure to review.

My only very minor comments are as follows:

- Even if its just observational, as the bats weren't individually marked, do the authors have an idea if the same roosts were used by the same individuals on consecutive days (that is: had some of them maybe some natural markings that made them identifiable)?

- Although all 53 perches were marked, I am assuming not all were fitted with cameras & microphones, thus I would encourage the authors to acknowledge this and discuss the possibility that other copulation events might have been missed. The authors present compelling evidence that this is in fact mating behaviors but with only one recorded event I believe it necessary to address this either lack of success by the males or impossibility to survey all perches simultaneously allowing for some missed events.

- A table or set of graphs that shows the statistical comparison of the features of perched and in flight echolocation would be very useful. Also, the metabolic cost of perched echolocation and potential uses in communication has been discussed for other species (for example: Front Physiol. 2013; doi: 10.3389/fphys.2013.00066. Metabolic costs of bat echolocation in a non-foraging context support a role in communication. D. Dechmann, M. Wikelski, H. van Noordwijk, C. Voigt, S. L. Voigt-Heucke.)

- The authors discuss the differences in the composition of the songs in regards to the number of elements, but, were there differences in the spectro-temporal signatures of the syllables among the bats that may allow for identification of individuals? (Bohn KM, Schmidt-French B, Schwartz C, Smotherman M, Pollak GD (2009) Versatility and Stereotypy of Free-Tailed Bat Songs. PLoS ONE 4(8): e6746. https://doi.org/10.1371/journal.pone.0006746)

- For the behavioral observations it would be useful to have percentages of time, for example the authors describe "94.1 ± 13.89 min of total observation time" which one needs to go back further up to check how long was the total observation time. Adding % would aid the reader to quickly understand if this is a significant amount of time.

- Are all visitors female? The authors mention this question in the discussion do not explain why this is not something that can be know with the collected data. What is the limitation to answering this question with the collected data? viewing angle for the visiting bat? few perch sites being recorded and this identification being hard to do with the naked eye? Any or all of this are valid, it just would be good to explain why this is not known.

- The authors mention that sound production with the wings in bats is rare and this is true, yet they might want to cite a paper that shows a species of fruit bats that use wing beat sounds for sonar (Nonecholocating Fruit Bats Produce Biosonar Clicks with Their Wings. Arjan Boonman, Sara Bumrungsri, Yossi Yovel. https://doi.org/10.1016/j.cub.2014.10.077)

I would also recommend citing the two following review articles:

- This one reviews auditory processing of communication in bats and points out which species until the data it was published have been described to show songs. This is relevant the introduction. (Salles, A., Bohn, K. M., & Moss, C. F. (2019). Auditory communication processing in bats: What we know and where to go. Behavioral Neuroscience, 133(3), 305–319. https://doi.org/10.1037/bne0000308)

- This other one more broadly reviews social communication in bats and includes detailed descriptions of acoustic, visual, tactile and olfactory information different species obtain through these sensory modalities. (Chaverri, G., L. Ancillotto, and D. Russo. 2018. Social communication in bats. Biological Reviews, 93: 1938-1954).

Reviewer #2: Review for: Rodriguez-Herrera et al.: The masked seducers: Lek courtship behavior in the Wrinkle-faced bat Centurio senex (Phyllostomidae)

The authors report on an observational study on the bat species Centurio senex in Costa Rica. The bat is famous for its rather strange looking facial mask. Beyond, not much is known about the species. This is also true for lek courtship behavior in bats.

Overall, I very much enjoyed reading the manuscript. The fact that the study is more or less purely descriptive and does not contain many hypothesis, predictions and experiments is in this case a plus. It allows the authors to tell the readers about their observations free from any bias. Moreover, especially in the discussion the authors address many questions and give ideas for future studies, which can then be conducted based on hypothesis and experiments. The manuscript thus can be seen as an important starting point for further studies on C. senex, lek behaviour in bats and more generally courtship behaviour in animals.

I only have some minor points, which I will address in the following:

Introduction

“Additionally, variables such as time and place where bats will show courtship behavior are for many species extremely difficult or impossible to predict” … and to access, I would say.

Hypothesis: The only hypothesis in the paper refers to the facial mask. However, I am missing some predictions that explain this hypothesis. What might be the function of the mask? In example: Might certain features of the mask (e.g. size) act as a sign for good genes? Might it be a protection against competing males (as the authors suggest in the discussion)?

Materials and Methods

“On October 3rd, perches were individually marked.” Why only on this date? Or do the authors mean: From October 3rd on, perches were individually marked?

Ultrasound recording

Regarding the equipment for the USG 116Hm: Which microphone did the authors use? The authors write that the microphone was placed “as close as possible”? How close was this on average?

Did the authors trigger manually, automatically or did they choose continuous recordings? If the recording was done automatically, which settings did they choose?

“Additionally, we recorded also echolocation calls from bats flying in the study area.” Which bats were recorded? Does this refer to C. senex or any bat species flying in the study area?

Results

Were there any obvious reasons, why the number of perching males started to decrease in October? Could this have been the end of mating season, rainy/dry season or due to monitoring? If so, it should be discussed in the Discussion section.

Average height of the perches: In the M&M section it is missing how these measures were done.

Echolocation behavior: For the bandwidth the SD is missing in the text.

Discussion

Vision: Although this cannot be proven, I would guess that the males’ facial mask itself might be an indication that females select males based on visual information.

Olfaction: This is just an idea for future tests that came to my mind. Might the facial mask have an olfactory function similar to the pouches of Saccopteryx? Secreted fluids would than be protected as long as the mask is lowered. When the mask is raised, pheromones could be spreaded through the air.

Best wishes

Michael Schöner

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Reviewer #1: No

Reviewer #2: Yes: Michael Schöner

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The masked seducers: Lek courtship behavior in the Wrinkle-faced bat Centurio senex (Phyllostomidae)

PONE-D-20-16747R1

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Kind regards,

Vivek Nityananda

Academic Editor

PLOS ONE

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