Go West: Hirudins and Decorsin/Ornatin-like Platelet Aggregation Inhibitors in Two Representatives of American Hematophagous Leeches
, Bernhard H. Rauch 4 and Christian Müller 1,*Round 1
Reviewer 1 Report
REVIEW of the article: “Go West: Hirudins and decorsin/ornatin-like platelet aggregation inhibitors in two representatives of Middle and South American hematophagous leeches” submitted to the Parasitologia.
In this article, the authors provide identification, molecular and functional characterization of biologically active substances from leeches salivary glands. Basing on the re-investigation of the freely accessible raw transcriptome datasets of L. mexicana (SRX5688711) and H. vizottoi (SRX960561), new findings were done.
I would say well done!
The introduction provides sufficient background and includes all relevant references.
The research is well designed and illustrated.
The used methodology was adequately described.
The conclusions are supported by the results.
The manuscript is of potential interest to a wide range of readers, including scientists of various specialties (biology, physiology, biochemistry, biomedicine, etc.).
The manuscript is written, as far as I can judge, in good literary English.
Author Response
Comments and Suggestions for Authors
REVIEW of the article: “Go West: Hirudins and decorsin/ornatin-like platelet aggregation inhibitors in two representatives of Middle and South American hematophagous leeches” submitted to the Parasitologia.
In this article, the authors provide identification, molecular and functional characterization of biologically active substances from leeches salivary glands. Basing on the re-investigation of the freely accessible raw transcriptome datasets of L. mexicana (SRX5688711) and H. vizottoi (SRX960561), new findings were done.
I would say well done!
The introduction provides sufficient background and includes all relevant references.
The research is well designed and illustrated.
The used methodology was adequately described.
The conclusions are supported by the results.
The manuscript is of potential interest to a wide range of readers, including scientists of various specialties (biology, physiology, biochemistry, biomedicine, etc.).
The manuscript is written, as far as I can judge, in good literary English.
Answer:
Thank you very much for the kind review!
Reviewer 2 Report
This manuscript presents a reanalysis of publicly available datasets to identify and characterize the molecular and functional profiles of anticoagulant transcriptomes of two species of American leeches, Limnobdella mexicana andHaementeria vizottoi. The paper is well-organized, concise, and well-written. Overall, my comments and suggestions are minor and I expect this will not need extensive revisions to be ready for publication.
One reoccurring point that needs to be reconsidered is that Limnobdella mexicana is considered a North American species since it is distributed broadly across Mexico (although the species needs to be revised this will ultimately be refined). I don’t think this will change the ideas presented in the paper, but Limnobdella species are not only a Middle American or South American species. In the Introduction (pg 2, line 91), it would be helpful to provide a citation for the feeding behavior and host preferences of Limnobdella species. I am not aware of records of Limnobdella species parasitizing birds. They are most commonly found on mammals (e.g. cattle) and amphibians, usually frogs.
In the abstract, it says “The leech species represent the two basal orders of leeches,”. The orders Rhynchobdellida and Arhynchobdellida are not basal in any way. If Acanthobdellida is recognized as the third order of leeches (this is a relatively minor point of taxonomic debate), it would be basal to Rhychobdellida and Arhynchobdellida. If Acanthobdellida is not considered a separate order within Hirudinida, then there are only two orders and one is not basal to the other. It is best in this case to remove the word “basal” from the abstract.
A very important point comes up at the end of section 2.3 Functional characterization I: thrombin inhibition. In the final paragraph, the names of the factors do not match what the text is describing or the figures. I think Hviz_HV4 should actually be Lmex_HV4 and Hviz_DV1 should be Lmex_DV1.
My other comments and notes are in the pdf of the document.
Overall, I found this to be a clearly written, concise study that makes a significant contribution to the study of leech anticoagulant diversity.
Comments for author File: 
Comments.pdf
Author Response
Comments and Suggestions for Authors
This manuscript presents a reanalysis of publicly available datasets to identify and characterize the molecular and functional profiles of anticoagulant transcriptomes of two species of American leeches, Limnobdella mexicana andHaementeria vizottoi. The paper is well-organized, concise, and well-written. Overall, my comments and suggestions are minor and I expect this will not need extensive revisions to be ready for publication.
One reoccurring point that needs to be reconsidered is that Limnobdella mexicana is considered a North American species since it is distributed broadly across Mexico (although the species needs to be revised this will ultimately be refined). I don’t think this will change the ideas presented in the paper, but Limnobdella species are not only a Middle American or South American species.
Answer:
We follow the suggestion of the reviewer and have deleted the terms "middle and south" across the manuscript.
In the Introduction (pg 2, line 91), it would be helpful to provide a citation for the feeding behavior and host preferences of Limnobdella species. I am not aware of records of Limnobdella species parasitizing birds. They are most commonly found on mammals (e.g. cattle) and amphibians, usually frogs.
Answer:
Many thanks for the very helpful hint! In their paper on the salivary transcriptome of Limnobdella mexicana Iwama et al. (2019) stated that "little is known about the host diversity and bloodfeeding behavior of this species" and that most members of the Praobdellidae (like Limnobdella mexicana) "have been found to feed indiscriminately on mammals (including humans) and birds". We drew the conclusion that this feeding behavior likely also applies to Limnobdella mexicana. However, a careful check of the available literature on Limnobdella mexicana revealed that there is indeed no direct evidence for parasitism of Limnomdella mexicana also on birds. For that reason we have skipped the term "birds" from the respective sentence in the Introduction section.
In the abstract, it says “The leech species represent the two basal orders of leeches,”. The orders Rhynchobdellida and Arhynchobdellida are not basal in any way. If Acanthobdellida is recognized as the third order of leeches (this is a relatively minor point of taxonomic debate), it would be basal to Rhychobdellida and Arhynchobdellida. If Acanthobdellida is not considered a separate order within Hirudinida, then there are only two orders and one is not basal to the other. It is best in this case to remove the word “basal” from the abstract.
Answer:
To avoid any misunderstanding we follow the suggestion of the reviewer and deleted the term "basal".
A very important point comes up at the end of section 2.3 Functional characterization I: thrombin inhibition. In the final paragraph, the names of the factors do not match what the text is describing or the figures. I think Hviz_HV4 should actually be Lmex_HV4 and Hviz_DV1 should be Lmex_DV1.
Answer:
The reviewer is absolutely right and we deeply apologize for our mistake. We have corrected the respective notations.
My other comments and notes are in the pdf of the document.
Answer:
Unfortunately we did not have access to that particular pdf file. Neither the downloadable pdf file nor the doc file of the manuscript contained the mentioned comments and notes. We will ask the editor to get access.
Overall, I found this to be a clearly written, concise study that makes a significant contribution to the study of leech anticoagulant diversity.
Answer:
Thank you very much for the kind statement and for the very careful reading of our manuscript.
Author Response File: 
Author Response.docx
