PONE-D-20-18942

Diversity of mosquitoes (Diptera: Culicidae) collected in different types of larvitraps in an Amazon rural settlement

PLOS ONE

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Belchior nº 00812016 - PAIC - AM – 2016. We would also like to thank Fiocruz / VPEIC

for the scholarship used to carry out the study and the post-graduate course in Condições

de Vida e Situações de Saúde na Amazônia of the Instituto Lebônidas e Maria Deane -

Fiocruz Amazônia, for the training and qualification of Master in Public Health."

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"This work is funded by the Fundanção de Amparo a Pesquisa do Estado do Amazonas – FAPEAM through the Programa de Excelência em Pesquisa Básica e Aplicada em Sáude (public call nº 004/2014). The article is a by-product of the umbrella project "Arboviroses emerging in the Amazon: Alphavirus incidence risk factors with an emphasis on Mayaro, on the agricultural frontier in Central Amazon ", under the coordination of FACP. Information for this grant can be found at http://www.fapeam.am.gov.br/wp-content/uploads/2014/07/DecCD-159_2014.pdf. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."

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Reviewers' comments:

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Comments to the Author

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Reviewer #1: Partly

Reviewer #2: Partly

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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: No

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Reviewer #1: Yes

Reviewer #2: No

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5. Review Comments to the Author

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Reviewer #1: The MS “Diversity of mosquitoes (Diptera: Culicidae) collected in different types of larvitraps in an Amazon rural settlement” is relevant and agrees with the purposes of the Journal. However, I recommend 'Major Revision' modifications, as follows:

Comments

Line 33. Change "Trichoprosopom digitatum" to "Trichoprosopon digitatum". Melanoconion in “Cx. (Melanoconion) sp.”, written in italics, please.

Lines 92 and 93. Do you mean "by" here? Change "Koppen" to "Köppen” climate classification".

Lines 101-102. Include in the text a reference about these deforestation data in the sampling area.

Lines 103-106. Authors need to include the geographic coordinates of the collection points and characterize the different collection points as well as information on the vegetation cover of the sampling area.

Line 109. Sampling was carried out for a short period. I do not consider that it allowed a conclusion on distribution patterns and knowledge of species diversity.

Line 110. What is the size of the container used? Were they plastic containers?

Lines 110-11. Were the traps (bamboo internodes, tires, and plastic containers) reused? If the answer is positive, please make it clear how the treatment was performed for relocation at the sampling points?

Line 120. Specify the type of plastic transport bag for the immature.

Line 123. How were the larvae fed?

Line 126. Incorrect reference citations, actually, there are two volumes:

Lane, J. 1953a. Neotropical Culicidae - 1st Volume. University of São Paulo, São Paulo. (and)

Lane J. 1953b. Neotropical Culicidae - 2nd Volume. University of São Paulo, São Paulo.

Line 128. In which entomological collection were the specimens collected deposited?

Lines 126-128. Authors must include the following bibliographic reference: J F Reinert. J Am Mosq Control Assoc. 2000 Sep;16(3):175-88. New Classification for the Composite Genus Aedes (Diptera: Culicidae: Aedini), Elevation of Subgenus Ochlerotatus to Generic Rank, Reclassification of the Other Subgenera, and Notes on Certain Subgenera and Species

Line 147. Melanoconion in “Cx. (Melanoconion) sp.” written in italics, please.

Lines 154-156. Why did you use "and" in the text to separate names of the authors of species and here, in the table you used "&"? You must see the instructions of the Journal, and uniform it.

Table 1-4. Inform the sampling period.

Table 1. Sa. chloropterus Von Humboldt. It is double wrong: firstly, it should be written "von" with the initial letter in lower case; secondly, it is a German a preposition between the name and the surname (as "de" in Portuguese in names such as "João de Almeida"); therefore, the citation of the surname MUST NOT BEGIN with the preposition, just with the surname: Humboldt.

Table 1. Toxorhynchites haemorrhoidalis haemorrhoidalis (Fabricius). In order to make it more concise, in the table, since it is the nominotypical subspecies, you could abbreviate this specific name, which is the same as the subespecific.

Line 185. Tx. haemorrhoidalis haemorrhoidalis. In order to make it more concise, since it is the nominotypical subspecies, you could abbreviate this specific name, which is the same as the subespecific.

Line 232. Cx. (Melanoconion) spp.: Melanoconion in italics; it is a subgeneric name!!!!!

Line 235. Did the authors obtain male specimens to identify at a specific level?

Line 255. Toxorhynchites haemorrhoidalis haemorrhoidalis, why not abbreviate the name of this genus if you are doing it for all other genus?

Discussion: The authors poorly discussed the medical importance of the mosquito species found in this study.

Lines 306-310. The conclusion was not clear, the sampling was carried out in a short period of collection. Therefore, how is it possible to associate a change in the oviposition behavior of some vector species of etiologic agents?

Line 391. Aedes (Stegomyia) aegypti: Stegomyia in italics; it is a subgeneric name!!!!!

Line 411. Change "Neotropical Culioidae" to "Neotropical Culicidae".

Reviewer #2: In general, the manuscript is sound and has new biological information. There are some weak parts that demand more work and time from authors. I will mention a list of points that they need to consider before the MS is accepted for publication.

1- Introduction - "Rural settlements are established in places that are favorable for human-vector interaction due to their proximity to extensive forest areas and the lack of adequate infrastructure for the residents who live there." I partially disagree with authors about their argument. The places are favorable or become favorable because of anthropogenic changes in the forest environments?

2- What do authors mean by "the habitats of residents? Why habitat? Can it be socioecological conditions?

3- Authors need to adjust the use of some words. For instance, it is not common to say "Mosquitoes are characterized by their flexible use of a wide variety of breeding sites," The word flexible is not adequate, there are better word to use. i.e. generalist. Breeding sites can be replaced by larval habitats, adult habitat.

4- Please consider a better way to mention "from larges found at the soil level", ground pools, standing water, others? Authors must check the terminology used in mosquito ecology.

5- Authors must consider to use the correct ecological term for "small pools of water stored in leaves in the canopy of trees." Can it be phytotelma?

6- Immature forms of Anopheles species have been recorded in artificial containers [16,17], including An. darlingi Root, which was found in an artificial lagoon in the urban region of Manaus in the state of Amazonas. Please rewrite this part, because an artificial lagoon is not an artificial container.

7- In the last paragraph of the introduction, authors say "Due to the importance of sanitary conditions and the ecological evolution that has led to the use of water containers for breeding by female mosquitoes, this work aimed to identify mosquito species that colonize different types of artificial breeding places, located in environments with different levels of anthropization within the state of Amazonas."

This entire paragraph needs to be rewritten. What do authors mean when they say sanitary conditions and ecological evolution? What is a water container? Why breeding by females? Can female breed on water containers? Can you replace artificial breeding places by larval habitats? Because your investigation is limited to a single settlement in Presidente Figueiredo, I consider it is incorrect to say, "within the state of Amazonas". Your sample is small, and it is does not represent the immense diversity found in the Amazon. Please considering the focus of your study, rewrite this part.

Materials and Methods

1- Can you please provide better details of the climate in the region studied?

2- Please clarify your procedure to choose locations to install your traps. Did you used satellite images to select locations within the settlements area? How did you select locations within the settlement? You said you installed 9 traps per sampling point, please define what you mean by sampling point. Please, add this information, they are essential for an ecological study.

3- Considering you used tap water to fill the traps, do you think your sampling was biased based on the female choice that depends on several molecules present in the habitats, including pheromones. How the water characteristics may have affect your sampling diversity and abundance and occurrence of mosquitoes?

4- What do you mean by material collection? Please clarify.

5- How did you define the 15 days interval between installation and collection days?

6- I assume you installed 162 traps each field collection. Did you install all traps in a single day?

7- How did you avoided cross contamination among traps?

8- A believe the larvae were in early stages and most have died during the trip and in the lab. Have you counted and identified all of them? This needs to be clarified because of your statistical analyses and diversity indexes you used.

9- Figure 3 legend needs to be together with the graphic.

Discussion

1- It is of paramount importance to explain why authors consider that the larvitraps were good to sample the most common mosquito species in the Rio Pardo agrovillage. Can you please provide reference (in addition to the thesis you cited) of other published studies on the same agrovillage? How can you support your argument that traps recovered the most common species? Please add reference. How can you be sure your trap collections in artificial habitats were able to return most species present in the region?

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Reviewer #1: No

Reviewer #2: No

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Diversity of mosquitoes (Diptera: Culicidae) collected in different types of larvitraps in an Amazon rural settlement

PONE-D-20-18942R1

Dear Dr. Almeida,

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.

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Rafael Maciel-de-Freitas

Academic Editor

PLOS ONE