PONE-D-25-16397Effects of woody plant encroachment on abundance of multiple tick species in the U.S. Great PlainsPLOS ONE

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Additional Editor Comments :

This manuscript has already been reviewed by two highly qualified experts, who have provided valuable feedback. Both reviewers recognize the potential of this study and find it appealing to the journal’s readership. At the same time, they have pointed out several issues, particularly regarding the need for more detailed descriptions in the Methods section and suggested revisions to the statistical analyses. Changes to the statistical methods, if implemented, may influence the study's results and subsequent discussion. I strongly recommend that the authors carefully address all of the reviewers’ comments and suggestions. The revised manuscript will be sent back to these expert reviewers for further evaluation. Whether the article will ultimately be accepted depends on the quality and completeness of these revisions.

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Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

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

Reviewer #1: No

Reviewer #2: Yes

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3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: This manuscript presents an interesting study on tick abundance across different stages of eastern redcedar (ERC) encroachment. I offer several suggestions to improve clarity and analytical rigor in the methods, statistical analysis, and results presentation. Key concerns include justifying the use of abundance over density, clarifying how seasonal variation was handled, and adopting more appropriate statistical tools such as the appropriateness of using AIC and model comparisons when there are no competing models except the null, using emmeans for post-hoc comparisons and interpretation of model outputs. I recommend using more descriptive labels for categorical variables (e.g., "early ERC" instead of "stage 2") and translating beta coefficients into meaningful effect sizes (e.g., percent changes) for better reader comprehension. Minor comments include consistency in formatting, spelling, terminology, and presentation of duplicated information.

Methods

Statistical Analysis.

Is there a reason for using abundance for the tick drags rather than density of ticks?

How are you accounting for the different times of the year that ticks are collected in your analysis?

Regarding AIC, do you low sample sizes, it might be more appropriate to report AICc, which corrects for bias associated with small sample sizes and is more accurate.

Line 247, how are you defining greater support. Is this delta AIC > 2?

Line 249-254: At first read, I wasn’t sure what you mean by stage. I have to reread. Could you use the actual category (grassland, early ERC, moderate ERC and mature ERC. This would make the job of the reader easier. Also you are considering the grassland as the the reference stage or the control. I’m not sure what are you running separate models with each of the other stages of ERC encroachment as the reference for the intercept. To perform post comparisons, it would be more appropriate to use emmeans package as this would adjust the p-values for multiple comparisions?

Results

Table 2: I’m sure what you are using AIC when there are only two models to compare, one of which is the null. Usually you would have multiple predictors that you compare. In doing this, it would make more sense to just report the p-values.

Figures: for the X axis, instage of putting Stage 1,2,3 can you use “Open grassland”, “early ERC”, etc.

When describing the results, can you put it in context, instead of just stating the beta values, transform this into a percentage change. For instance in lines 318-319, based on the beta values, and if you are using a log link, the beta value can be convert indicates that "A. americanum was about 5 times as abundant in stage 2 as in stage 1," or ~400% more abundant, relative to stage 1."

I’m not sure how you got the letter values for the figures. What is a coefficient analysis? See above comments about using emmeans.

Throughout the manuscript, you have a tendency to have the same information (AIC values, beta values) in the text that is in a table. Choose one,

Discussion.

The discussion is well written given the current results, I won’t provide speciific comments as it is unknown if it will change when the method/results change.

Minor Comments

Line 168L misspelling “centeral”

Lines 179: be consistent with the date format, earlier, you put the month day and on this line you put day month.

Be consistent with numbers, when using numbers less than 10, spell out the entire word.

Line 258-263: what there an actually analysis for this? Also, the tect could be a bit more concise “Measurements of ERC trees confirmed that our sampling captured distinct stages of encroachment: both tree height and horizontal percent cover increased progressively across stages. Average tree heights for stages 1–4 were 0 m, 3.6 m, 5.3 m, and 7.2 m, respectively, while average horizontal cover was 0%, 30.2%, 55.2%, and 89.3%.”

Line 276-267: can you combine the first two sentences. Maybe “ June 2024, with increased abundance in 2024 than 2023.”

Line 280: from instead of “form”

Reviewer #2: The manuscript “Effects of woody plant encroachment on abundance of multiple tick species in the U.S. Great Plains” reports the results of comparisons of field collections of ticks from native grassland habitats and habitats classified by three stages of wood plant encroachment. This is an important topic that will be of interest to a broad audience given the course of global change especially in regard to land use and climate. The study presented here is straightforward: ticks were collected in the four habitat classes using two established methods and abundances within species were compared, with two life stages of the most abundant species, Amblyomma americanum, examined separately. Overall, the manuscript is well written and to my judgement, scientifically sound. Most of my comments and suggestions are minor. One thing that is essential is to improve the detail in the Methods regarding the collection methods. This section should include details on the traps that were used, specifics about flagging, and citations of any field techniques or protocols that were used. The conclusions seem to focus on the results related to the two tick species that had higher abundances in habitats with woody plant encroachment, while the results for Amblyomma maculatum seem to be downplayed a bit by the focus on the other two. It is important to describe these results here, and in the Abstract and first paragraph of the Discussion, and to not generalize given the caveat that it was not all tick species that increased in abundance with wood plant encroachment. Overall though, this study is a valuable contribution, well-written, scientifically sound, and interesting.

L40: Mention results for Amblyomma maculatum

L56: Capitalize initial letters of words in names only when proper noun

L68: “effects” should be “affects”

L79: “Eastern redcedar” should be “eastern redcedar”; be consistent with the use of common names and capitalization of initial letters. My suggestion would be that the only words in common names to have an initial capitalized letter should be proper nouns e.g., Bourbon virus, West Nile virus, Bachman’s sparrow, otherwise, they should be, for example, white-tailed deer, white-footed mouse, etc. The alternative would be to capitalize each initial letter e.g., White-tailed Deer. But, it should never be White-tailed deer. Whichever you prefer, be consistent throughout the manuscript. Double check that all other common names are correctly written.

L88: Suggest inserting “those of” between “including” and “birds”

L120: The Methods are well presented, but could use additional detail in some areas. In particular, details about the trapping and flagging need to be included.

L133: If abbreviations are to be used, be consistent with their use. Here, be consistent with use of ERC v. eastern redcedar, and check throughout the manuscript. Also check WPE. Once defined, subsequent mentions of the phrase should be abbreviated.

L136-145: Consider presenting this information as a table

L179: Awkward wording, suggest rewording: “periods when tick activity averages lower than during early summer”

L176-179: Be consistent with date formatting: April 1st – July 15th vs. 16 July – 31 Oct)

L193: Is Ixodes abbreviated here as Ix. for a reason?

L194: Elaborate on the CO2 traps and flagging methods with citations. Were traps commercially purchased or built? Give specifics about flagging. How long were traps left in place and was this consistent across site visits? Were transects linear?

L200: Cite keys that were used.

L209-213: Can this be reworded for clarity? I think I understand what is being communicated, but it took a moment to interpret, especially L210

L221: Could statistical comparisons not be performed on mean tree height and horizontal percent cover between encroachment stage categories?

L228: Were larvae collected?

L240: Spell out Akaike Information Criterion here, put abbreviation in parentheses

L268-270: For consistency, abbreviate names of genera after the first mention of a species name

L280: Change “form” to “from”

L284: Table 1 legend – for clarity, insert “(stage 1)” after “open grasslands”

L308: Would it be meaningful to run this analysis for total Amblyomma americanum? (i.e., Amblyomma americanum adults + nymphs, with trapping method counts combined, as was done for Amblyomma maculatum and Dermacentor variabilis.) Consider including that if it would be meaningful.

L362: Insert “(stage 1)” after “grasslands”

Discussion: The Discussion section is well written, and I appreciate the inclusion of the study’s limitations, especially the potential that trap placement could have biased results. The only weak part of the Discussion is the first paragraph. The language here could be adjusted to make references to trapping methods and stages more clear. It could also be helpful to split this into two paragraphs, with the first focusing summarizing the most important results, (briefly) reiterating the question and the work that was done, and the second explaining the details described in L372-378.

L368: Consider including in this first sentence a statement referencing that Amblyomma maculatum abundances were lower in the Stage 2-4 samples. This result is similarly important as those for the other two species.

L373: Suggest inserting “(overall)” after D. variabilis

L375, 376: For clarity, suggest replacing “collected by trapping” to “collected in CO2-baited traps”

L398: Juniper, or redcedar?

L488: I would include in this initial sentence mention of your results for Amblyomma maculatum as well. It is certainly important that abundance was higher for Amblyomma americanum and Dermacentor variabilis in grasslands with woody plant enchroachment, but it is equally meaningful that other tick species, Amblyomma maculatum, was more abundant in native grasslands. Given that, I would also reword this first sentence a bit – rather than stating that this encroachment facilitates increased abundance of ticks, broadly, I think it would be more accurate to describe this is “some species of ticks”. It is a bit misleading without that caveat.

L493: Would it be worthwhile to rerun your analysis with all three tick species combined? If that were done, you could generalize about ticks a bit more broadly.

L500: Should ERC be WPE?

L509: Was anything known about the fire regime at the study sites? Consider including a note regarding this in the Methods or discussing as a limitation in the Discussion.

L511: “of some species” should be inserted after “increased tick populations”

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Reviewer #1: Yes:  Samniqueka Halsey

Reviewer #2: No

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Effects of woody plant encroachment on abundance of multiple tick species in the U.S. Great Plains

PONE-D-25-16397R1

Dear Dr. Propst,

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,

Xiao Guo, Ph.D.

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

The authors have adeptly and thoroughly addressed the reviewers' concerns, and the manuscript's overall quality has been substantially enhanced. The paper is well-structured, its arguments are lucid and coherent, and I therefore recommend that it be accepted for publication.

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #1: All comments have been addressed

Reviewer #2: All comments have been addressed

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2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Yes

Reviewer #2: Yes

**********

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

Reviewer #1: Yes

Reviewer #2: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

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5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: Yes

Reviewer #2: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: Thank you for addressing the review comments. I have no further concerns at this time and believe the manuscript is suitable for publication.

Reviewer #2: (No Response)

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7. PLOS authors have the option to publish the peer review history of their article (what does this mean? ). If published, this will include your full peer review and any attached files.

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Reviewer #1: Yes:  Samniqueka Halsey

Reviewer #2: No

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