Review Reports - Physiological and Ecological Correlates of the Cellular and Humoral Innate Immune Responses in an Insular Desert Bat: The Fish-Eating Myotis (Myotis vivesi)

Round 1

Reviewer 1 Report

The manuscript was well written. However, it is better to cite updated references instead of the old ones. For example, lines 75-76: "Bats are the second.... (21)". In fact, bat diversity has been changed rapidly over the past decade. Therefore, it would be better to cite updated references (e.g. Wilson et al. 2019. Handbook of the Mammals of the World. Lynx Edicions, Barcelona. ISBN: 978-84-16728-19-0: 1008 pp.)

Author Response

We agree with the suggestion of the reviewer and cite an update source of bat diversity:

Simmons, N.B., and Cirranello A.L. 2021. Bat species of the world: a taxonomic and geographic database. Accessed on 08/01/2022.

Reviewer 2 Report

Overall, I would like to congratulate the authors of the manuscript entitled “Physiological and ecological correlates of the cellular and humoral innate immune responses in an insular desert bat: the Fish-eating Myotis (Myotis vivesi)” for the clarity of the text, the basic idea behind the study, which is novel and interesting. I think this ms is ready for publication, congratulations!

Author Response

We thank the reviewer for the comments. We made several corrections along the text, including correcting Fig. 5.

Author Response File: Author Response.docx

Reviewer 3 Report

Abstract

The abstract needs attention.

You start with a background move to a research question, go back to a background, move again to a research question

The general statement is not directly related to your study except of the aridity “Aridity and geographical isolation are two factors expected to shape immune expression in animals”

No need: In general, the swelling response of female and male bats was lower in spring

Is it new information?

Both external morphological traits and hormone levels indicated that females were pregnant in spring and lactating in summer, and that males were reproductively active in autumn, when mating probably occurs.

L37, 38 Change from in year 1 of the study to first year of the study

Change male bats to males

L39 it is not significant you can not state it

On line 37 you talk about BKA and then you move to swelling and then back to BKA

Change from At least in males,

These results may suggest that….

Delete lines 49-51.

What is your take home message?

Introduction

L. 94: Add: In this study, we….

L107: Delete it into relevant to the introduction.

L113: This is the place to talk about the aridity aspect of the study. Does it have any relevance to your research questions as the opening statement in your abstract declares?

Also look at:

Pilosof, S., Korine, C. Moore, M. and Krasnov, B. 2014. Sewage-water contamination alters immune response in a desert bat. Mammalian Biology-Zeitschrift für Säugetierkunde, 79:183-188.

Korine, C. Pilosof, S. Gross, A. Morales-Malacara J.B. and Krasnov, B. 2017. The effect of water contamination and host-related factors on ectoparasite load in an insectivorous bat. Parasitology Research, 116:2517-2526.

Methods

We measured each individual´s body mass and forearm length to estimate the scaled mass 132 index (43) as an estimator of the body condition index (BCI);

Provide model of balance and caliper. Move ref 43 after BCI.

L154. You already discussed PHA in the introduction.

How many bats, and how many female, males?

How long this experiment lasted? Any signs of pain? Where the bats release in the place of the capture?

L171: The same questions as for the PHA experiment. – sample size, male, female.

Where was the experiment carried out?

Data analysis

L212: We compared indices”, indices of what?

Where is the statistical analysis for the steroid hormones?

Results

Line 252: the sentence is not clear in spring with significant differences with females in autumn

Line 262: “had higher value” values of what?

L283: Add the place and country of capture for all the figures and tables. Year if needed (Table 2 for example).

L319: The sentence is too long, please split.

L357: Delete significantly

Discussion

The discussion is too long, not focusing enough and consequently the take home messages are not always clear.

For example, lines 376-381 are all repetition of the results. Along the discussion there are parts where there is a repetition of the results.

Aridity is not discussed.

What is the take home message?

The end of the discussion includes topics that are not connected such as Mercery pollution.

Author Response

REVIEWER

Abstract

The abstract needs attention.

You start with a background move to a research question, go back to a background, move again to a research question

The general statement is not directly related to your study except of the aridity “Aridity and geographical isolation are two factors expected to shape immune expression in animals”

AUTHORS

We modified the 1^st sentence to : “Immune response is affected by aridity, but it has been rarely examined in desert-dwelling bats.” and eliminated this sentence “The Fish-eating Myotis feeds regularly on marine fish and crustacea and it is endemic to desert islands in the Gulf of California, Mexico.”

REVIEWER

No need: In general, the swelling response of female and male bats was lower in spring.

AUTHORS

We are not sure what the reviewer suggested. We erased “In general.”

REVIEWER

Is it new information?

AUTHORS

We erased this statement because it repeats information previously presented.

REVIEWER

L37, 38 Change from in year 1 of the study to first year of the study

AUTHORS

Done.

REVIEWER

Change male bats to males.

AUTHORS

Done.

REVIEWER

L39 it is not significant you can not state it

AUTHORS

We agree.

REVIEWER

On line 37 you talk about BKA and then you move to swelling and then back to BKA.

AUTHORS

We keep it the way it is because we organized the results in 4 sections for both swelling and PBA: 1) seasonality, 2) body condition, 3) parasite load, and 4) sex hormones.

REVIEWER

Change from At least in males,

AUTHORS

We eliminated this sentence: At least in males, Fish-eating Myotis with higher body mass and body condition might be in a better nutritional condition to respond to the PHA challenge.

REVIEWER

These results may suggest that….

AUTHORS

See end of abstract.

REVIEWER

Delete lines 49-51.

AUTHORS

Agree. Done.

REVIEWER

What is your take home message?

AUTHORS

We added some our conclusion at the end of the abstract (lines 41-45).

REVIEWER

Introduction

94: Add: In this study, we….

AUTHORS

Done.

REVIEWER

L107: Delete it into relevant to the introduction.

AUTHORS

We think that the information is relevant to the introduction.

REVIEWER

L113: This is the place to talk about the aridity aspect of the study. Does it have any relevance to your research questions as the opening statement in your abstract declares?

Also look at:

Pilosof, S., Korine, C. Moore, M. and Krasnov, B. 2014. Sewage-water contamination alters immune response in a desert bat. Mammalian Biology-Zeitschrift für Säugetierkunde, 79:183-188.

AUTHORS

We rearrange this section and include the suggested references (lines 136-143).

REVIEWER Methods

We measured each individual´s body mass and forearm length to estimate the scaled mass 132 index (43) as an estimator of the body condition index (BCI);

Provide model of balance and caliper. Move ref 43 after BCI.

AUTHORS

Both suggestions were included. Ref. 43 is ref.48 now.

REVIEWER

L154. You already discussed PHA in the introduction.
AUTHORS

We agree and deleted this part.

REVIEWER

How many bats, and how many female, males?

AUTHORS

We added that information.

REVIEWER

How long this experiment lasted? Any signs of pain? Where the bats release in the place of the capture?

AUTHORS

The length of the experiment is already pointed (12 hours after the injection). Since some of the information requested applies to all sampling procedures, we added the following in lines 208-212: “All the procedures were minimally invasive, and the bats were released in the place of capture within an hour after the last measurement or sample collection after being rehydrated with fresh water and fed with a small piece of fresh fish.”

REVIEWER

L171: The same questions as for the PHA experiment. – sample size, male, female.

AUTHORS

Samples size for each sex was already in Results. We moved that to Methods.

REVIEWER

Where was the experiment carried out?

AUTHORS

We added that information (lines 242-243).

REVIEWER Data analysis

L212: We compared indices”, indices of what?

AUTHORS

Swelling indices (added).

REVIEWER

Where is the statistical analysis for the steroid hormones?

AUTHORS

That was already presented in lines 303-310.

REVIEWER

Results

Line 252: the sentence is not clear in spring with significant differences with females in autumn.

AUTHORS

We corrected to: “..females had lower BCI in spring than in autumn (p = 0.04).”

REVIEWER Line 262: “had higher value” values of what?

AUTHORS

Of progesterone content. Fixed.

REVIEWER

L283: Add the place and country of capture for all the figures and tables. Year if needed (Table 2 for example).

AUTHORS

Place and country added in all tables and figures. We do not present year except where needed (Fig. 4A)

REVIEWER

L319: The sentence is too long, please split.

AUTHORS

We modified to (lines 424-431):

“In females (n = 57), progesterone content was not correlated with any inflammation index (S_maxper: r = 0.12, p =0.37; S_maxindiv: r = 0.20, p = 0.14); Figs. 3A-B). In contrast, estradiol content was significantly correlated with S_maxindiv (r = 0.32, p = 0.02; Fig. 3C) and it had a nearly significant correlation with S_maxper (r = 0.25, p = 0.07; Fig. D).”

REVIEWER

L357: Delete significantly.

AUTHORS

Done.

REVIEWER

Discussion

The discussion is too long, not focusing enough and consequently the take home messages are not always clear.

AUTHORS

We modified parts of the discussion.

REVIEWER

For example, lines 376-381 are all repetition of the results. Along the discussion there are parts where there is a repetition of the results.

AUTHORS

We reduced the reference to results to the minimum needed to place in context the discussion.

REVIEWER

Aridity is not discussed.

AUTHORS

We added some thoughts about this subject at the end of the discussion (lines 653-658).

REVIEWER

What is the take home message?

The end of the discussion includes topics that are not connected such as Mercery pollution.

AUTHORS

We modified the discussion (lines 648-653). We eliminated the microbiota part of the story.

Round 2

Reviewer 3 Report

The authors adequately replied to all my comments and questions.

I have few minor corrections that should be made before accepting the MS for publication.

L92 change from with other to on

Lines 103-106 change to past

Discussion

You repeat in detail the results on line 400 and on lines 413-417 and there is no need for that,

Line change from the body mass to body mass

Author Response

We addressed the comments as follows:

1-L92 change from with other to on.

We did so (line 141).

2-Lines 103-106 change to past.

We changed to past (lines 168-172).

3-You repeat in detail the results on line 400 and on lines 413-417 and there is no need for that.

We partly agree because we consider that the discussion needs some context. We shortened the sentences:

Lines 519-521: "Higher testosterone levels were measured in autumn, but the two males on which we measured testosterone in spring had similarly high values (19.6 and 14.6 ng/ml)."

Lines 547-549: "BKA in year 1 was lower in spring and summer than in winter, and that in spring and summer in year 2. Females collected in spring in year 1 were probably in early pregnancy, whereas females in year 2 were in a more advanced pregnancy stage."

4-Line change from the body mass to body mass.

We changed that (line 573).