-->PONE-D-26-00846-->-->Zoonotic endoparasites and Toxoplasma gondii   seropositivity in free-roaming cats (Felis catus  ) from an urban environment-->-->PLOS One

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Reviewer #1: Partly

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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Reviewer #1: Yes

Reviewer #2: Yes

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-->5. Review Comments to the Author

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Reviewer #1: Dear Authors,

Your manuscript entitled “Zoonotic endoparasites and Toxoplasma gondii seropositivity in free-roaming cats (Felis catus) from an urban environment” is a valuable contribution to the necessary surveillance of parasitic infections in animals living in urban settings. Studies of this type should be conducted regularly in order to accumulate important epidemiological data.

I believe that your manuscript is worthy of publication after addressing the following points:

1. Unfortunately, the reference to EPG measurement is not appropriate, as no quantitative method (e.g., McMaster) was applied. I understand that an estimation of the relative parasite burden was performed by counting parasite eggs across the entire preparation; however, this is not sufficient to be expressed using the strictly quantitative EPG metric. Therefore, I would recommend removing the EPG measure throughout the manuscript and, to utilise your approximate estimate of parasite burden, adopting an ad hoc metric, such as “number of eggs per preparation.”

2. I would suggest adding the study area to the title, namely, New York City. This is simply a suggestion, and I leave it to your judgment and preference.

3. Line 118. Please change “Occult Heartworm ELISA test was performed…” to “An ELISA test was performed…”. Note that “occult” refers to a heartworm infection in which microfilariae are not detected in the blood (not investigated in the present study).

4. Line 216. Please change “variant” to “species”.

5. Line 294. Please change “While our results suggest that patent adult heartworm infection is not currently hyper-endemic in NYC feral cats” to “While our results suggest that heartworm infection is not currently endemic in NYC feral cats.”

6. Please add to the Discussion the limitation of not performing the Knott’s test in the investigation of heartworm infection. Although heartworm infection in cats is typically amicrofilaraemic (i.e., occult), microfilaraemia does occur. Given that, a) the number of heartworms in cats is usually very low, b) that a single mature pair of adult heartworms can produce microfilariae, but c) would most probably result in a negative serological test (too little antigen to be detected), laboratory diagnosis in cats should include all possible tests, i.e. the Knott test, Ag detection and Ab detection.

Reviewer #2: The Manuscript Number PONE-D-26-00846, entitled “Zoonotic endoparasites and Toxoplasma gondii seropositivity in free-roaming cats (Felis catus) from an urban environment” show interesting and relevant results on zoonotic endoparasites in New York, particularly a high number of infected animals by Toxocara cati. The results, considering the results, reinforce the strategies employed to deal with parasites of free-roaming cats. However, authors are recommended to carefully revise and to include some information.

I hereby would like to present some recommendations for improvement of the current version of the manuscript.

Major comments

## Materials and Methods

Please, give some details regard to the following procedures:

- kits used for extracting genetic material from faeces.

- Describe the primers adopted in the trials (even in a supplementary file).

- Describe, briefly, the ELISA test protocol for detection of Occult.

## Discussion

Despite the main aim of the researchers was to diagnostic zoonotic endoparasites in free-roaming cats, I suggest the inclusion of a paragraph/sentence concerning human toxocariasis in The United States emphasizing the Human Development Index of |New York citizens as well as the high risk of infection by children playing in public places.

## Conclusion

I would be glad to verify the conclusion after the revised version of the manuscript, considering the recommendations.

Minor comments

In our opinion, the epg count is less relevant than the frequency of infected cats. A unique T. cati for example, may shed a high number of eggs/day (more than 100.000). Then, presence of eggs may not mirror the infection burden of parasites. Nevertheless, the number of infected cats is a surprisingly and undoubtedly an “extraordinary” result in a developed country.

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Reviewer #1: No

Reviewer #2: No

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Zoonotic endoparasites and Toxoplasma gondii   seropositivity in free-roaming cats (Felis catus  ) from New York City Boroughs

PONE-D-26-00846R1

Dear Dr. Kafle,

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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Kind regards,

Balbir B. Singh, Ph. D

Academic Editor

PLOS One

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