PONE-D-24-47355Use of a mouse model for the isolation of Borrelia puertoricensis from ticks collected in Merida, Yucatan, MexicoPLOS ONE

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Additional Editor Comments:

Title: I would suggest removing "collected in Merida, Yucatan, Mexico " as the mouse model could be used to isolate ticks from other areas

Line 61: remove “This includes wild animal nests or dens”

Lines 135 and 192: remove “Sequences were deposited in GenBank.”, being reported below. The same goes for the “Annotation pipeline….(accession numbers CP149102-CP149123)”

Lines 248-249: Rewrite this sentence “These results show that A. puertoricensis is present in Mexico likely parasitizing opossums in urban neighborhoods”,  since as it stands it seems like a comment and not a result

Lines 329-330:  Change to “such as the vector ecology tick misidentification, lack of diagnostic tests and due to the difficulty culturing the bacteria”

Reviewers' comments:

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Comments to the Author

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Reviewer #1: Yes

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

Reviewer #1: N/A

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Reviewer #1: Yes

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Reviewer #1: Yes

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5. Review Comments to the Author

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Reviewer #1: The present study investigates if Ornithodoros puertoricensis ticks might act as vectors for Borrelia puertoricensis, not being previously reported in Ciudad Caucel.

Line 122: Is there an explanation to analyse such a low number of ticks (n=3)? This number might be low to emphasize if a result is reliable and to further perform statistics in the future. Please explain and clarify.

Line 151: Please briefly describe how the exsanguination with a heparinized syringe to quantify and isolate the bacteria was performed

Line 155: Please briefly describe how the samples were analyzed to evaluate the antibody response against Borrelia

Line 156: Please specify what is a “chemically induced immunosuppression mouse model”, and how these animals were “prepared” for the experiments

Line 171: Please briefly justify why the choice of the temperature at 35ºC to cultivate the bacteria, once the ticks temperature may vary accordingly to the environment (being even higher/lower than 35ºC). Please specify if other temperatures were used to test the best culture conditions

Line 173: Please specify which was the expected concentration of bacteria considered “enough” for the subculturing and DNA isolation. Only saying that “when the bacteria were detected” may not be enough to the reader to repeat the procedures.

Line 274: I wonder whether if the extraction with the phenol-clorophorm might represent a good choice to perform genome sequencing. Where the information related to the quality of the analysis are showed? The authors are invited to clarify these points somewhere on the paper.

Line 331: Please specify which are the “Recent improvements in culture medium formulations” that have been made lately to clarify for the reader.

Line 332: Which may be the main reasons for the absence of which the accessible animals for the spirochete isolation? The authors are invited to describe these informations on the text.

Line 374: Please mention a reference that mention this information “There is increasing evidence that B. puertoricensis is more prevalent in Latin America”.

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Reviewer #1: No

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Use of a mouse model for the isolation of Borrelia puertoricensis from soft ticks

PONE-D-24-47355R1

Dear Dr. Ibarra,

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Maria Stefania Latrofa

Academic Editor

PLOS ONE