Updated Taxonomic Key of European Nycteribiidae (Diptera), with a Host-Parasite Network

Round 1

Reviewer 1 Report

In my opinion the manuscript is very interesting. Follow some suggestions to improve the quality of the paper.

Line 74 and 75. Adrian Marshall has another explanation for the function of ctenidia. See Marshall AG(1980) The function of combs in ectoparsitic insects. In Traub R & Starcke H (eds) Fleas. AA Balkema, Rotterdan. Or other book or chapter book by Marshall. I agree with Marshall.

About the plates. Who made the drawings? You? Oskar Theodor? Explain, please. If you do not cite the authors of the figures, this may be considered plagiarism.

Line 376.  If you explained metastenoxenous, Ithink you have to explain what is mesostenoxenous.

 

Author Response

Respected reviewer,

 

The authors thank you for your comments and suggestions for modification of our manuscript. The manuscript was edited accordingly.

Please find below replies to the comments and suggestions:

 

Main comments:

 

1. Line 74 and 75. Adrian Marshall has another explanation for the function of ctenidia. See Marshall AG(1980) The function of combs in ectoparsitic insects. In Traub R & Starcke H (eds) Fleas. AA Balkema, Rotterdan. Or other book or chapter book by Marshall. I agree with Marshall.

• Response: Mentioned reference and and further explanation of the function of ctenidia have been added.

 

2. About the plates. Who made the drawings? You? Oskar Theodor? Explain, please. If you do not cite the authors of the figures, this may be considered plagiarism.

• Response: Figures are redrawn and modified by first and last authors (L.M. and J.O.) based on the original illustrations by Theodor (1954) [50] (pp. 66 a. Nycteribiidae. Taf. I-IV, VI-VII, IX, XIII-XVI); Theodor (1967) [49] (p. 1 – 506, figures compared with [50]); Hůrka (1970) [45]. (p. 243: fig. 1); Hůrka (1972) [46] (p. 711: fig. 1); Beaucournu & Noblet (1685) [97] (p. 637: fig. 1).

 

 

3. Line 376.  If you explained metastenoxenous, I think you have to explain what mesostenoxenous is.

• Response: The definition of metastenoxenous has been added.

Author Response File: Author Response.docx

Reviewer 2 Report

The study shows relevance and is well structured. It is necessary a minor revision of the language, and better detailing the  analysis data.

Congratulations for research.

Author Response

Respected reviewer,

 

The authors thank you for your comments and suggestions for modification of our manuscript. The manuscript was revised accordingly.

Please find below replies to the comments and suggestions:

 

Main comments:

1. The study shows relevance and is well structured. It is necessary a minor revision of the language, and better detailing the analysis data.

• Response: The manuscript was checked by a native speaker for grammatical and syntax errors.

The data analysis was checked, broadly supplemented and unclear parts were corrected.

Author Response File: Author Response.docx

Reviewer 3 Report

Dear authors,

You provide with your manuscript an updated key to European Nycteribiidae and present an interaction network of host-parasite associations.

I am very intrigued by bat flies and their host specificity or the lack thereof. The key you provide is very useful and allows finally easier identifications of European bat flies. I also like the idea to present host-parasite interactions in form of a network to highlight the presumed low host specificity. However, I have some concerns regarding both, the key as well as the network.

First, in the short time I am given to write a review I was not able to test your identification key for functionality and correctness. Also, the figures you use appear highly familiar. Did you create those by yourself or are they taken from existing sources? If latter, why did you not cite those/get the permission of reuse?

Second, I have questions regarding the origin and treatment of the parasite-host associations. I assume you used presence-absence data? How come that some of the interactions are thicker than others? Or did you count the number of published associations? Therefore, I tried to understand this by revising Szentiványi et al., however I have problems understanding where your numbers come from. As an example: in Szentiványi, Phthiridium biarticulatum has 22 mentions with R. ferrumequinum; 16 mentions with R. hipposideros; 3 with R. mehelyi – in your study none of these interactions are occurring, why? This makes me wonder about the completeness of your whole interaction network and subsequently the correctness of your analyses. Another example: Pe. d. dufourii has 15 interactions with M. schreibersii in Szentiványi et al., and 24 with M. myotis – how come the interaction link in your network is thicker for M. schreibersii compared to M. myotis?
Only based on these observations I must assume the network is flawed. And as you do not provide any raw data, or the interaction matrix, I have no possibility to follow your analysis.

I urge you therefore to revise and explain in more detail your methodology section and how your interaction network was created. This lack of precision also follows through the discussion section. When comparing existing studies to your network, I would recommend you to at least mention that you are talking about different families, in regards of bat flies as well as host bats, and not take these values for granted and applicable to European bat fly – bat networks. I am also wondering why you push so aggressively towards bat flies and zoonotic diseases in your discussion and conclusion section. Sure, as blood-sucking insects bat flies are prone to act as vectors and this is important to keep in mind. However, much still needs to be resolved before you can give bat flies a bad reputation as disease spreader. Also, this part reads a bit clumsy and distracts the reader from your main focus of your manuscript.

More detailed notes below.

 

Further remarks:

Line 2: rephrase please “with the host-parasite network”. Perhaps “with a host-parasite network”?
Line 4: for exact numbers on bat species you can also cite batnames.org
Lines 53-54: what is “breadth of food”? Please clarify.
Lines 54-56: Please rephrase, I do not understand what you want to say here.
Line 62: Not sure where in this cited publication the “growing knowledge of the public health importance” is covered. Please address.
Line 82: The paper cited is a network study not focussing on disease transmission. Please cite an appropriate study.
Line 87: replace with “tend to form”, there are quite some species of bats which form mixed species clusters in roosts
Line 88: low abundance of whom? Please clarify.
Lines 91-96: might be better suited in the discussion section
Line 93: What are “normal” ecological conditions?
Line 102: change to “…the study of host-parasite interactions is an…”
Line 107: Please be more precise, what is a “beneficial representation of energy flow”?
Line 115: change to Nycteribiidae
Lines 115-119: This part is really hard to understand, please rephrase
Lines 121-124: necessary?
Line 127: What do you mean with “keys preparing”?
Lines 129-130: Please with a bit more detail. What did you do, how did you do it, and what did you use of these publications? Did you create the key completely new or did you include new species and exclude species not occurring in Europe from an existing key?
Lines 134: How did you treat these host associations? Did you use presence-absence data? Did you weigh them by number of publications/mentions? How did you find all the relevant literature? Did you do a literature search? Please be more detailed. This is crucial for understanding your subsequent steps and your analyses in total.
Line 164: Why the “updated” in parenthesis, or even, why the “updated” at all?
Line 172: Fig. 4 does not refer to the tibia of N. vexata! Please correct!
Line 182: Again, why “updated”? Your title already says that you updated an existing key, no need to write this here.
Page 9-13: Were these figures created by you? They look very similar to those of Theodor (1967), however you do not mention this anywhere.
Lines 375-377: I do not follow here. How come three of the four mesostenoxenous species are also highly specialised, but not the fourth species (P. monoceros)?
Lines 378-279: How many records of B. daganiae did you have in your study?
Line 388: You mention in line 376 that B. italica is highly specialised, however here in this table it is not marked as such. Please address.
Line 398: What do you mean with “relative number of records”? You did not mention anything like this in your methods section.
Figure 40: see comment above. How did you end up with this interaction network? Did you exclude interactions mentioned in the literature? See for example in Szentiványi, Phthiridium biarticulatum has 22 mentions with R. ferrumequinum; 16 mentions with R. hipposideros; 3 with R. mehelyi – in your study none of these interactions are occurring, why? Also: Pe. d. dufourii has 15 interactions with M. schreibersii in Szentiványi et al., and 24 with M. myotis – how come the interaction link in your network is thicker for M. schreibersii compared to M. myotis?
Also, probably the colours you selected are not displayed as supposed to be. I only can distinguish four colours: red for Basilia, purple for Nyteribia, blue for Penicillidia, and green for Phthiridium. You might also want to consider using a colour-blind friendly palette for selecting your colours.
As to the structure of this network, it looks really messy. The r-package bipartite has the option to minimize overlapping interactions, did you try this? If yes, I would recommend for example to order the parasites (or the hosts) alphabetically/phylogenetically to make this figure easier to digest.
Lines 407-410: You create here three types of modules, however, never discuss these findings anywhere. If not important, leave it out – if important, discuss it appropriately!
Figure 41: How did you measure the strength of interaction you show here? Missing information in your methods section.
Line 446: Try to avoid expressions like these.
Line 450-457: Be careful here. First, you have created your network based merely on literature results (some of them quite old) and you have no measure on how exact the identifications are (of bats and bat flies), or how the bats/bat flies were collected in the first place. There are many possible scenarios which can introduce cross-contaminations. This needs to be considered. Second, by generalizing over a large geographic area, you assume that host-parasite associations behave the same over their whole distribution. There is sufficient evidence that host-parasite interactions can change depending on where the study took place, and still remain highly specialised. Further, not all species mentioned in your study have the same distribution. So I wouldn’t be surprised the parasite associations change depending on where the sampling took place. This is from an ecological point of view very logical, however when throwing all records together you dilute these observations.
While it is of course valid to create a species interaction network over all those records, you need to discuss these options accordingly.
Lines 460-461: is this necessary, this appears dismissive. Partial data vs. comprehensive data always depend on the point of view!
Lines 464-465: You base this on only a single record, that has not much merit.
Line 467: which records?
Lines 470-471: higher risks for whom? The bat flies? Are they negatively affected when acting as vector?
Lines 473-474: how did you measure abundance?
Lines 479-483: First, you are comparing your results with results from mainly the Neotropics, where Nycteribiidae are only a small part of the bat fly diversity. Most records are about Streblidae and Phyllostomidae. So be careful when comparing.
Line 481: those cited articles do not support your claims. They show high specificity and a high modularity.
Line 485-486: Modularity basically the same in Duran et al. and your study.
Lines 488-490: Low specificity? Many studies mentioned here have higher specificity than yours. You need to be really careful when comparing your interactions with those from Neotropical regions. As an example, Hiller et al. (https://doi.org/10.1111/icad.12508) showed very high host specificity (H2=0.97) in a network covering over 50 species of each, bats and bat flies. Streblid bat flies tend to be highly specialized.
Line 502: Please change wording, “species complex” is generally used for cryptic or not distinguishable species.
Line 503: change to Rhinolophus
Lines 506-509: Please rephrase, I cannot follow here.
Line 511: Why is this a problem? A problem for whom?
Lines 518-519: How?
Lines 521-522: Are you serious? All sampling of wild animals should follow strict guidelines and laws to guarantee as less impact as possible.
Lines 522-526: Is this necessary here? Your paper is much more than just resolving a complication mentioned in another paper.

Author Response

Respected reviewer,

 

The authors thank you for your comments and suggestions for modification of our manuscript. The manuscript was edited accordingly.

Please find below replies to the comments and suggestions:

 

Main comments:

1. First, in the short time I am given to write a review I was not able to test your identification key for functionality and correctness. Also, the figures you use appear highly familiar. Did you create those by yourself or are they taken from existing sources? If latter, why did you not cite those/get the permission of reuse?

• Response: Response: Figures are redrawn and modified by first and last authors (L.M. and J.O.) based on the original illustrations by Theodor (1954) [50] (pp. 66 a. Nycteribiidae. Taf. I-IV, VI-VII, IX, XIII-XVI); Theodor (1967) [49] (p. 1 – 506, figures compared with [50]); Hůrka (1970) [45]. (p. 243: fig. 1); Hůrka (1972) [46] (p. 711: fig. 1); Beaucournu & Noblet (1685) [97] (p. 637: fig. 1).

 

 

2. Second, I have questions regarding the origin and treatment of the parasite-host associations. I assume you used presence-absence data? How come that some of the interactions are thicker than others? Or did you count the number of published associations? Therefore, I tried to understand this by revising Szentiványi et al., however I have problems understanding where your numbers come from. As an example: in Szentiványi, Phthiridium biarticulatum has 22 mentions with R. ferrumequinum; 16 mentions with R. hipposideros; 3 with R. mehelyi – in your study none of these interactions are occurring, why? This makes me wonder about the completeness of your whole interaction network and subsequently the correctness of your analyses. Another example: Pe. d. dufourii has 15 interactions with M. schreibersii in Szentiványi et al., and 24 with M. myotis – how come the interaction link in your network is thicker for M. schreibersii compared to M. myotis?
   Only based on these observations I must assume the network is flawed. And as you do not provide any raw data, or the interaction matrix, I have no possibility to follow your analysis.

• Response: Thank you for the warning - we checked the data and found errors in it - everything is already fixed at this moment.

 

 

3. I urge you therefore to revise and explain in more detail your methodology section and how your interaction network was created. This lack of precision also follows through the discussion section. When comparing existing studies to your network, I would recommend you to at least mention that you are talking about different families, in regards of bat flies as well as host bats, and not take these values for granted and applicable to European bat fly – bat networks. I am also wondering why you push so aggressively towards bat flies and zoonotic diseases in your discussion and conclusion section. Sure, as blood-sucking insects bat flies are prone to act as vectors and this is important to keep in mind. However, much still needs to be resolved before you can give bat flies a bad reputation as disease spreader. Also, this part reads a bit clumsy and distracts the reader from your main focus of your manuscript.

• Response: The methodology has been revised and updated.

 

4. Line 2: rephrase please “with the host-parasite network”. Perhaps “with a host-parasite network”?

• Response: The formulation was rephrased.

 

5. Line 4: for exact numbers on bat species you can also cite batnames.org
   Lines 53-54: what is “breadth of food”? Please clarify.

• Response: The webpage batnames.org was cited. “Breadth of food” was replaced by “diversity of food”.

 

6. Lines 54-56: Please rephrase, I do not understand what you want to say here.
   Line 62: Not sure where in this cited publication the “growing knowledge of the public health importance” is covered. Please address.

• Response: Lines 54-56 have been rephrased. Line 62 – nowhere, the sentence was deleted as irrelevant.

 

7. Line 82: The paper cited is a network study not focussing on disease transmission. Please cite an appropriate study.

• Response: We agree with the reviewer, we have added the relevant sources.

 

8. Line 87: replace with “tend to form”, there are quite some species of bats which form mixed species clusters in roosts

• Response: The formulation was replaced according to the suggestion.

 

9. Line 88: low abundance of whom? Please clarify.

• Response: The original sentence was poorly worded. The sentence was modified.

 

10. Lines 91-96: might be better suited in the discussion section

• Response: The paragraph was moved to Discussion.

 

11. Line 93: What are “normal” ecological conditions?

• Response: The formulation “normal ecological conditions” was removed from the text.

 

12. Line 102: change to “…the study of host-parasite interactions is an…”

• Response: The sentence was corrected according to the reviewer's suggestion.

 

13. Line 107: Please be more precise, what is a “beneficial representation of energy flow”?
    Line 115: change to Nycteribiidae

• Response: The sentence has been modified.

 

Line 115 – a modified term “Nicteribiidae” was inserted to the text

 

14. Lines 115-119: This part is really hard to understand, please rephrase

• Response: The sentence was modified according to the reviewer′s suggestion.

 

15. Lines 121-124: necessary?

• Response: The paragraph was deleted.

 

16. Line 127: What do you mean with “keys preparing”?

• Response: The heading of the chapter has been updated. The meaning is a preparation of the identification key, which consisted of old literature supplemented by new species (more in “Updating of identification key”

 

17. Lines 129-130: Please with a bit more detail. What did you do, how did you do it, and what did you use of these publications? Did you create the key completely new or did you include new species and exclude species not occurring in Europe from an existing key?

• Response: Details were added to the text.

 

18. Lines 134: How did you treat these host associations? Did you use presence-absence data? Did you weigh them by number of publications/mentions? How did you find all the relevant literature? Did you do a literature search? Please be more detailed. This is crucial for understanding your subsequent steps and your analyses in total.

• Response: The chapter was modified and updated.

 

19. Line 164: Why the “updated” in parenthesis, or even, why the “updated” at all?

• Response: We cancelled the “_”, it was necessary to add then unknown species to the original determination keys (Theodor (1954) [50] and Theodor (1967) [49]  were supplemented by works Hůrka (1970) [45] Hůrka (1972) [46] and Beaucournu & Noblet (1685)

 

20. Line 172: Fig. 4 does not refer to the tibia of N. vexata! Please correct!

• Response: Thank you for the warning – the error has been corrected.

 

21. Line 182: Again, why “updated”? Your title already says that you updated an existing key, no need to write this here.

• Response: Thank you for the warning – the error has been corrected.

 

22. Page 9-13: Were these figures created by you? They look very similar to those of Theodor (1967), however you do not mention this anywhere.

• Response: we added the following text - where we explained the origin of the images “Figures are redrawn and modified based on the original illustration by Theodor (1954) [50] (pp. 66 a. Nycteribiidae. Taf. I-IV, VI-VII, IX, XIII-XVI); Theodor (1967) [49] (p. 1 – 506, figures compared with [50]); Hůrka (1970) [45]. (p. 243: fig. 1); Hůrka (1972) [46] (p. 711: fig. 1); Beaucournu & Noblet (1685) [97] (p. 637: fig. 1).”

 

23. Lines 375-377: I do not follow here. How come three of the four mesostenoxenous species are also highly specialised, but not the fourth species (P. monoceros)?

• Response: after reviewing the literature, the data were recalculated and re-evaluated

 

24. Lines 378-279: How many records of B. daganiae did you have in your study?

• Response: We had a total of 5 records in our study (data will be added as an appendix).

 

25. Line 388: You mention in line 376 that B. italica is highly specialised, however here in this table it is not marked as such. Please address.

• Response: After reviewing the literature, the data were recalculated and re-evaluated.

 

26. Line 398: What do you mean with “relative number of records”? You did not mention anything like this in your methods section.

• Response: We supplemented the methodology and tried to explain it better - the data consists of qualitative data, not quantitative data (that is, one record was added to the table for one literary source, regardless of the quantity of the parasite).

 

27. Figure 40: see comment above. How did you end up with this interaction network? Did you exclude interactions mentioned in the literature? See for example in Szentiványi, Phthiridium biarticulatum has 22 mentions with R. ferrumequinum; 16 mentions with R. hipposideros; 3 with R. mehelyi – in your study none of these interactions are occurring, why? Also: Pe. d. dufourii has 15 interactions with M. schreibersii in Szentiványi et al., and 24 with M. myotis – how come the interaction link in your network is thicker for M. schreibersii compared to M. myotis?

• Response: Thank you for the warning - we checked the data and found errors in it - everything is already fixed at this moment.

 

28. Also, probably the colours you selected are not displayed as supposed to be. I only can distinguish four colours: red for Basilia, purple for Nyteribia, blue for Penicillidia, and green for Phthiridium. You might also want to consider using a colour-blind friendly palette for selecting your colours.

• Response: Thank you for the warning, we accept the comment. The colors have been changed and selected from the color-blind friendly palette.

 

29. As to the structure of this network, it looks really messy. The r-package bipartite has the option to minimize overlapping interactions, did you try this? If yes, I would recommend for example to order the parasites (or the hosts) alphabetically/phylogenetically to make this figure easier to digest.

• Response: By default, plotweb minimises overlap of lines and therefore we consider it the best solution to leave it that way. In our opinion, this is the most transparent way of displaying, given the large number of interactions. The parasite species are logically and intuitively (digestible for the reader) ordered so that the closest are those that have the most common hosts, and on the other hand the parasites are also ordered by similarity in terms of common hosts.

 

30. Lines 407-410: You create here three types of modules, however, never discuss these findings anywhere. If not important, leave it out – if important, discuss it appropriately!

• Response: The adjacency matrix and identified modules have been removed from the manuscript. Because the network of relationships is extremely complex, repeated calculations resulted to different modules that were difficult to interpret biologically, parasitologically, or ecologically. We could not resolve this even using the function computeMetaModules, which re-runs the algorithm several times, returning the most modular result, to stabilize modularity computation. With repeated computations, the resulting meta-modules were different and difficult to interpret, too.

 

31. Figure 41: How did you measure the strength of interaction you show here? Missing information in your methods section.

• Response: The adjacency matrix and identified modules have been removed from the manuscript. See response above.

 

32. Line 446: Try to avoid expressions like these.

• Response: The expression has been deleted.

 

33. Line 450-457: Be careful here. First, you have created your network based merely on literature results (some of them quite old) and you have no measure on how exact the identifications are (of bats and bat flies), or how the bats/bat flies were collected in the first place. There are many possible scenarios which can introduce cross-contaminations. This needs to be considered. Second, by generalizing over a large geographic area, you assume that host-parasite associations behave the same over their whole distribution. There is sufficient evidence that host-parasite interactions can change depending on where the study took place, and still remain highly specialised. Further, not all species mentioned in your study have the same distribution. So I wouldn’t be surprised the parasite associations change depending on where the sampling took place. This is from an ecological point of view very logical, however when throwing all records together you dilute these observations.While it is of course valid to create a species interaction network over all those records, you need to discuss these options accordingly.

• Response: We are aware that such an approach may have weaknesses, but we tried to avoid duplicate data and also repetitive data. the discussion was supplemented.

 

34. Lines 460-461: is this necessary, this appears dismissive. Partial data vs. comprehensive data always depend on the point of view!

• Response: The sentence was deleted.

 

35. Lines 464-465: You base this on only a single record, that has not much merit.

• Response: The sentence has been deleted.

 

36. Line 467: which records?

• Response: A reference Szentivanyi et al. [23] has been added.

 

37. Lines 470-471: higher risks for whom? The bat flies? Are they negatively affected when acting as vector?

• Response: The unclear formulation has been modified and corrected.

 

38. Lines 473-474: how did you measure abundance?

• Response: We did not measure abundance. The formulation has been deleted.

 

39. Lines 479-483: First, you are comparing your results with results from mainly the Neotropics, where Nycteribiidae are only a small part of the bat fly diversity. Most records are about Streblidae and Phyllostomidae. So be careful when comparing.

• Response: Theparagraph has been modified.

 

40. Line 481: those cited articles do not support your claims. They show high specificity and a high modularity.

• Response: Accepted. The text has been edited and citations removed.

 

41. Line 485-486: Modularity basically the same in Duran et al. and your study.

• Response: the modularity index Q = 0.57 in Duran et al is much higher than Q = 0.37 in our study. The number of modules was the same but we removed the modules and their interpretation from the MS.

 

42. Lines 488-490: Low specificity? Many studies mentioned here have higher specificity than yours. You need to be really careful when comparing your interactions with those from Neotropical regions. As an example, Hiller et al. (https://doi.org/10.1111/icad.12508) showed very high host specificity (H2=0.97) in a network covering over 50 species of each, bats and bat flies. Streblid bat flies tend to be highly specialized.

• Response: We accept the comment. We did not write this clear. The text has been edited for clarity (we were referring to the low specificity indexes H2' and d' in many bat-flies species from our results) and in accordance with the reviewer's comment.
•  

 

43. Line 502: Please change wording, “species complex” is generally used for cryptic or not distinguishable species.

• Response: “species complex” has been replaced by “taxons”.

 

44. Line 503: change to Rhinolophus

• Response: A grammatical mistake was corrected.

 

45. Lines 506-509: Please rephrase, I cannot follow here.

• Response: The text was rephrased.

 

46. Line 511: Why is this a problem? A problem for whom?

• Response: “Problem” replaced by “problematics”.

 

 

47. Lines 518-519: How?

• Response: It is important to collect data on the circulation of pathogens in vector populations. Good quality and up-to-date identification keys allow vectors to be correctly identified.

 

48. Lines 521-522: Are you serious? All sampling of wild animals should follow strict guidelines and laws to guarantee as less impact as possible.

• Response: The co-authors' idea was not properly formulated. Several belong to a bat conservation group. The paragraph has been modified.

 

49. Lines 522-526: Is this necessary here? Your paper is much more than just resolving a complication mentioned in another paper.

• Response: The paragraph was deleted according to the suggestion.

Author Response File: Author Response.docx

Reviewer 4 Report

The authors gathered all existent identification keys of European nycteribiids into this manuscript. Additionally, they have completed a specificity degree estimation based on previously published host-parasite occurrences. I have some comments, which I strongly suggest the authors to consider.

L14-16: I suggest deleting the first sentence. Going from taxonomical keys to the sustainability of human society is a little bit far-fetched in my opinion. However I do agree taxonomical keys are important.

L18: I generally do not agree with the whole zoonotic disease and bats as carriers of diseases tone in this work. As you clarify later in the work, bat flies probably do not contribute to spreading pathogens to humans or any other mammals besides bats, and therefore their role in pathogen spillovers is extremely unlikely. I suggest mentioning once, but removing these paragraphs as it quite out of context, considering the aims of this work.

L25-26: The terminology is incorrect throughout the manuscript. The usage of metastenoxenous and mesostenoxenous is not correct (see below), and I personally have never seen these terminology used in works related to bat parasites.

 metastenoxenous meaning: utilizing hosts of more than one genus but still within a single host family

 mesostenoxenous: utilizing different host species from a single genus.

I suggest using the conservative terminology, which is applied throughout the literature, such as Monoxenous, Stenoxenous, Oligoxenous, polyxenous (see Dick, C. W., & Dittmar, K. (2014). Parasitic bat flies (Diptera: Streblidae and Nycteribiidae): host specificity and potential as vectors. Bats (Chiroptera) as vectors of diseases and parasites: Facts and myths, 131-155.)

L26: I strongly suggest elaborating in the discussion about why Basilia daganiae seems to be monoxenous. Partially, because there are few reports and research in the area where this species occurs. It could potentially parasitize other host species, however we do not know as we don’t have data in the area.

 

L33: again, saying bats are source of zoonoses is correct, however I do not see why emphasizing so strong on this point throughout this work. Please delete.

L42-44: same comment as above. Why starting the introduction with this. You work is not focusing on pathogens or vectorial potential in any way.

L54-55: same comment.

L60-61: please add references.

L65: ectoparasites as ecological importance, please elaborate.

L80-81: reference is incorrect. please add references that actually shows bat flies transmit pathogens between bats.

L86-87: untrue, bats occasionally form species-specific groups in colonies, however they very often form mixed species colonies. Please correct.

L91-93: disturbance may in turn cause an increase in accidental or transient associations of bats and their parasites with non-primary hosts. Please add reference. Otherwise speculative.

L98-100: this sentence does not make sense, also I suggest again deleting and not focusing on the whole zoonotic and spillover perspective of bats as this work is not about that. -àHowever, correct identification and information on distribution, host species and parasite-host networks are critical for knowledge, prevention and rapid response to potential new animal-born epidemics and epizootics [29].

L115: nycteribiids as in plural

L122: as much as I like that the authors gathered al existent taxonomical keys for the occurring 16 species, it is not new. The authors did not describe new species, nor added new morphological characters to identify existent ones, therefore this identification key has already existed since the 60s, but in several publications. I suggest correcting this.

Additionally, I miss basic description of the ecology and biology of bat flies form the introduction. I think you should give much more details about their reproduction, behaviour, ecology etc. instead of focusing the zoonotic disease parts.

L133-134: multiple works have been dismissed during data collection, which contains more bat-bat fly association data. Why were these dismissed? Without these this work cannot be published. I only mentione here a few, but please do a more throughout literature search.

Fort instance:

Haelewaters, D., Pfliegler, W. P., Szentiványi, T., Földvári, M., Sándor, A. D., Barti, L., ... & Pfister, D. H. (2017). Parasites of parasites of bats: Laboulbeniales (Fungi: Ascomycota) on bat flies (Diptera: Nycteribiidae) in central Europe. Parasites & Vectors, 10(1), 1-14.

Szentiványi, T., Haelewaters, D., Pfliegler, W. P., Clément, L., Christe, P., & Glaizot, O. (2018). Laboulbeniales (Fungi: Ascomycota) infection of bat flies (Diptera: Nycteribiidae) from Miniopterus schreibersii across Europe. Parasites & vectors, 11, 1-7.

Pejić, B., Budinski, I., & Blagojević, J. (2021). Ectoparasite bat flies (Diptera: Nycteribiidae) of Schreiber's bent-winged bat and their fungus parasite. Programme & Abstract book: 13th European Multicolloquium of Parasitology; 2021 Oct 12-16; Belgrade, Serbia, 81.

Sándor, A. D., Péter, Á., Corduneanu, A., Barti, L., Csősz, I., Kalmár, Z., ... & Mihalca, A. D. (2021). Wide distribution and diversity of malaria-related haemosporidian parasites (Polychromophilus spp.) in bats and their ectoparasites in Eastern Europe. Microorganisms, 9(2), 230.

Sándor, A. D., Földvári, M., Krawczyk, A. I., Sprong, H., Corduneanu, A., Barti, L., ... & Földvári, G. (2018). Eco-epidemiology of novel Bartonella genotypes from parasitic flies of insectivorous bats. Microbial Ecology, 76, 1076-1088.

Kemenesi, G., Tóth, G. E., Mayora-Neto, M., Scott, S., Temperton, N., Wright, E., ... & Jakab, F. (2022). Isolation of infectious Lloviu virus from Schreiber’s bats in Hungary. nature communications, 13(1), 1706.

Szentiványi, T., Haelewaters, D., Rádai, Z., Mizsei, E., Pfliegler, W. P., Báthori, F., ... & Glaizot, O. (2019). Climatic effects on the distribution of ant-and bat fly-associated fungal ectoparasites (Ascomycota, Laboulbeniales). Fungal ecology, 39, 371-379.

Dondini, G., Vanin, S., Vergari, S., & Vergari, S. (2017). First record of Basilia mediterranea Hůrka, 1970 from Italy (Diptera: Nycteribiidae). Onychium, 13, 139-142.

Szentiványi, T., Vincze, O., & Estók, P. (2017). Density-dependent sex ratio and sex-specific preference for host traits in parasitic bat flies. Parasites & vectors, 10(1), 1-9.

Also, you need to explain how you exclude accidental host occurrences. These are crucial to determine if the parasite species truly polyxenous for example, or just occasional “accidental” host use is recorded.

 

L373: delete ; after 15

Table 1: use correct terminology, as mentioned above

Discussion:

L427-428: One of the most interesting characteristics of bat 427 flies is that they are obligate parasites of bats [68]. This should go to introduction.

L433: again, not a new key.

L437: which species did they not cover? All existent species have been covered previously, the authors did not add any new species description. I suggest deleting this.

L440: they are not recently known from Europe, but for decades.

L447-449: the authors did not do any analysis regarding the effect of colony size or individuality and specificity level between nycteribiids, or did I miss it? I suggest deleting it.

L450-454: again where is the data and analysis for these conclusions? The authors absolutely did not cover any of this to make these conclusion.

464: B. dagania is incorrect, as several species name across the manuscript. Pleaser carefully check both bat fly and host names.

L4674: you cannot make conclusion that this species is highly specialized based on one paper reporting data (as I mentioned above).

L467: do you mean biotic and abiotic? If not please add explanation to this terminology.

L469-470: same comment as above: conspecifics or 469 heterospecifics

L470: increased competition or higher pathogen infection 470 risks in bats --- again speculative, no data to support this. Delete.

L479: are also very interesting – please delete and let the reader decide about this.

L493: B. m. nudir --- incorrect

L501-502: Also, the 501 species complex comprising N. (N) schmidlii, P. conspicua and Ph. biatriculatum --- this is absolutely cannot be called species complex.

L502: Ph. biatriculatum – incorrect

L503: genus Rhynolophus --- incorrect. Please check all names very carefully throughout the manuscript.

 

L505-508: again absolutely speculative, no data to support any of these claims. Please delete.

L511: pathogen without plural

 

Additionally, several very important and relevant papers weren’t cited, which you should definitely include (besides the ones I already mentioned above):

McKee, C. D., Krawczyk, A. I., Sándor, A. D., Görföl, T., Földvári, M., Földvári, G., ... & Sprong, H. (2019). Host phylogeny, geographic overlap, and roost sharing shape parasite communities in European bats. Frontiers in Ecology and Evolution, 7, 69.

Szentiványi, T., Estók, P., Pigeault, R., Christe, P., & Glaizot, O. (2020). Effects of fungal infection on the survival of parasitic bat flies. Parasites & vectors, 13(1), 1-8.

Sándor, A. D., Péter, Á., Corduneanu, A., Barti, L., Csősz, I., Kalmár, Z., ... & Mihalca, A. D. (2021). Wide distribution and diversity of malaria-related haemosporidian parasites (Polychromophilus spp.) in bats and their ectoparasites in Eastern Europe. Microorganisms, 9(2), 230.

Kwak, M. L., Gorecki, V., & Markowsky, G. (2022). Parasites in peril: abundance of batflies (Diptera: Nycteribiidae) declines along an urbanisation gradient. Journal of Insect Conservation, 26(4), 627-638.

Verrett, T. B., Webala, P. W., Patterson, B. D., & Dick, C. W. (2022). Remarkably low host specificity in the bat fly Penicillidia fulvida (Diptera: Nycteribiidae) as assessed by mitochondrial COI and nuclear 28S sequence data. Parasites & Vectors, 15(1), 1-16.

Rivera, C. L., Padilla, J. F., Ospina Pérez, E. M., Urbano, F. M., Guarín, D. V., Mejía Fontecha, I. Y., ... & Ramírez-Chaves, H. E. (2022). Interaction networks between bats (Mammalia: Chiroptera) and ectoparasitic flies (Diptera: Hippoboscoidea) in the Colombian Orinoquia Region. Acta Chiropterologica, 24(2), 379-394.

Author Response

Respected reviewer,

 

The authors thank you for your comments and suggestions for modification of our manuscript. The manuscript was edited accordingly.

Please find below replies to the comments and suggestions:

 

Main comments:

1. L14-16: I suggest deleting the first sentence. Going from taxonomical keys to the sustainability of human society is a little bit far-fetched in my opinion. However I do agree taxonomical keys are important.

• Response: The link between taxonomic keys and the sustainability of human society has been broken. Only the mention of the importance of keys for a better understanding of biodiversity was retained.

 

2. L18: I generally do not agree with the whole zoonotic disease and bats as carriers of diseases tone in this work. As you clarify later in the work, bat flies probably do not contribute to spreading pathogens to humans or any other mammals besides bats, and therefore their role in pathogen spillovers is extremely unlikely. I suggest mentioning once, but removing these paragraphs as it quite out of context, considering the aims of this work.

• Response: The comment has been strictly taken into account and the text has been modified.

 

3. L25-26: The terminology is incorrect throughout the manuscript. The usage of metastenoxenous and mesostenoxenous is not correct (see below), and I personally have never seen these terminology used in works related to bat parasites. metastenoxenous meaning: utilizing hosts of more than one genus but still within a single host family mesostenoxenous: utilizing different host species from a single genus. I suggest using the conservative terminology, which is applied throughout the literature, such as Monoxenous, Stenoxenous, Oligoxenous, polyxenous (see Dick, C. W., & Dittmar, K. (2014). Parasitic bat flies (Diptera: Streblidae and Nycteribiidae): host specificity and potential as vectors. Bats (Chiroptera) as vectors of diseases and parasites: Facts and myths, 131-155.)

• Response: Response: Accepted and corrected.

 

 

4. L26: I strongly suggest elaborating in the discussion about why Basilia daganiae seems to be monoxenous. Partially, because there are few reports and research in the area where this species occurs. It could potentially parasitize other host species, however we do not know as we don’t have data in the area.

• Response: The claim has been deleted and the text rephrased according to the reviewer’s comment.

 

 

5. L33: again, saying bats are source of zoonoses is correct, however I do not see why emphasizing so strong on this point throughout this work. Please delete.

• Response: It was deleted according to the suggestion.

 

6. L42-44: same comment as above. Why starting the introduction with this. You work is not focusing on pathogens or vectorial potential in any way.

• Response: The paragraph has been deleted according to the suggestion.

 

7. L54-55: same comment.

• Response: Mentioned sentences have been deleted.

 

8. L60-61: please add references.

• Response: Sentences under these lines have been deleted in the text.

 

9. L65: ectoparasites as ecological importance, please elaborate.

• Response: The sentence has been deleted.

 

10. L80-81: reference is incorrect. Please add references that actually shows bat flies transmit pathogens between bats.

• Response: Relevant references have been added.

 

11. L86-87: untrue, bats occasionally form species-specific groups in colonies, however they very often form mixed species colonies. Please correct.

• Response: Thank you for the comment. We added it to the text and modified this part of the text.

 

12. L91-93: disturbance may in turn cause an increase in accidental or transient associations of bats and their parasites with non-primary hosts. Please add reference. Otherwise speculative.

• Response: The sentence was deleted.

 

13. L98-100: this sentence does not make sense, also I suggest again deleting and not focusing on the whole zoonotic and spillover perspective of bats as this work is not about that. -àHowever, correct identification and information on distribution, host species and parasite-host networks are critical for knowledge, prevention and rapid response to potential new animal-born epidemics and epizootics [29].

• Response: The sentence was deleted.

 

14. L115: nycteribiids as in plural

• Response: The grammatical mistake was corrected.

 

15. L122: as much as I like that the authors gathered al existent taxonomical keys for the occurring 16 species, it is not new. The authors did not describe new species, nor added new morphological characters to identify existent ones, therefore this identification key has already existed since the 60s, but in several publications. I suggest correcting this.

• Response: We agree, it has been corrected.

 

16. Additionally, I miss basic description of the ecology and biology of bat flies form the introduction. I think you should give much more details about their reproduction, behaviour, ecology etc. instead of focusing the zoonotic disease parts.

• Response: added

 

17. L133-134: multiple works have been dismissed during data collection, which contains more bat-bat fly association data. Why were these dismissed? Without these this work cannot be published. I only mentione here a few, but please do a more throughout literature search.

Fort instance:

Haelewaters, D., Pfliegler, W. P., Szentiványi, T., Földvári, M., Sándor, A. D., Barti, L., ... & Pfister, D. H. (2017). Parasites of parasites of bats: Laboulbeniales (Fungi: Ascomycota) on bat flies (Diptera: Nycteribiidae) in central Europe. Parasites & Vectors, 10(1), 1-14.

Szentiványi, T., Haelewaters, D., Pfliegler, W. P., Clément, L., Christe, P., & Glaizot, O. (2018). Laboulbeniales (Fungi: Ascomycota) infection of bat flies (Diptera: Nycteribiidae) from Miniopterus schreibersii across Europe. Parasites & vectors, 11, 1-7.

Pejić, B., Budinski, I., & Blagojević, J. (2021). Ectoparasite bat flies (Diptera: Nycteribiidae) of Schreiber's bent-winged bat and their fungus parasite. Programme & Abstract book: 13th European Multicolloquium of Parasitology; 2021 Oct 12-16; Belgrade, Serbia, 81.

Sándor, A. D., Péter, Á., Corduneanu, A., Barti, L., Csősz, I., Kalmár, Z., ... & Mihalca, A. D. (2021). Wide distribution and diversity of malaria-related haemosporidian parasites (Polychromophilus spp.) in bats and their ectoparasites in Eastern Europe. Microorganisms, 9(2), 230.

Sándor, A. D., Földvári, M., Krawczyk, A. I., Sprong, H., Corduneanu, A., Barti, L., ... & Földvári, G. (2018). Eco-epidemiology of novel Bartonella genotypes from parasitic flies of insectivorous bats. Microbial Ecology, 76, 1076-1088.

Kemenesi, G., Tóth, G. E., Mayora-Neto, M., Scott, S., Temperton, N., Wright, E., ... & Jakab, F. (2022). Isolation of infectious Lloviu virus from Schreiber’s bats in Hungary. nature communications, 13(1), 1706.

Szentiványi, T., Haelewaters, D., Rádai, Z., Mizsei, E., Pfliegler, W. P., Báthori, F., ... & Glaizot, O. (2019). Climatic effects on the distribution of ant-and bat fly-associated fungal ectoparasites (Ascomycota, Laboulbeniales). Fungal ecology, 39, 371-379.

Dondini, G., Vanin, S., Vergari, S., & Vergari, S. (2017). First record of Basilia mediterranea Hůrka, 1970 from Italy (Diptera: Nycteribiidae). Onychium, 13, 139-142.

Szentiványi, T., Vincze, O., & Estók, P. (2017). Density-dependent sex ratio and sex-specific preference for host traits in parasitic bat flies. Parasites & vectors, 10(1), 1-9.

• Response: Using the records in Szentivanyi et al. [23] as starting point, we checked the literature on the host associations of European bat louse for new, overlooked or misattributed records [46-49; 87-174]. In addition, we conducted a literature search using the keywords ‘Nycteribiidae*’, ‘European bat flies*’, ‘host associations *’, ‘bats *and ‘Europe*’ in Scopus and Google Scholar [14,43-44; 175-188].

 

18. Also, you need to explain how you exclude accidental host occurrences. These are crucial to determine if the parasite species truly polyxenous for example, or just occasional “accidental” host use is recorded.

• Response:we proceeded data according to the following methodology: Using the records in Szentivanyi et al. [23] as starting point, we checked the literature on the host associations of European bat louse for new, overlooked or misattributed records [46-49; 87-174]. In addition, we conducted a literature search using the keywords ‘Nycteribiidae*’, ‘European bat flies*’, ‘host associations *’, ‘bats *and ‘Europe*’ in Scopus and Google Scholar [14,43-44; 175-188]. We are aware that such an approach may have weaknesses, but we tried to avoid duplicate data and also repetitive data. the discussion was supplemented.
•  

 

 

19. L373: delete ; after 15

Table 1: use correct terminology, as mentioned above

• Response: Accepted, deleted. The terminology was corrected in the text and table.

 

Discussion:

20. L427-428: One of the most interesting characteristics of bat 427 flies is that they are obligate parasites of bats [68]. This should go to introduction.

• Response: The sentence has been moved to introduction.

 

21. L433: again, not a new key.

• Response: We agree. It has been corrected.

 

22. L437: which species did they not cover? All existent species have been covered previously, the authors did not add any new species description. I suggest deleting this.

• Response: The sentence was corrected.

 

23. L440: they are not recently known from Europe, but for decades.

• Response: “recently was replaced by “currently. This is what we meant.

 

24. L447-449: the authors did not do any analysis regarding the effect of colony size or individuality and specificity level between nycteribiids, or did I miss it? I suggest deleting it.

• Response: We have not written it clearly enough. The text has been edited and the mentioned problematic part deleted.

 

25. L450-454: again where is the data and analysis for these conclusions? The authors absolutely did not cover any of this to make these conclusion.

• Response: We have not written it clearly enough. The text has been edited. We have relied on results published by other cited authors

 

26. L464:  daganiais incorrect, as several species name across the manuscript. Pleaser carefully check both bat fly and host names.

• Response: The fly and host names were corrected.

 

27. L4674: you cannot make conclusion that this species is highly specialized based on one paper reporting data (as I mentioned above).

• Response: The conclusion was deleted in the text.

 

28. L467: do you mean biotic and abiotic? If not please add explanation to this terminology.

• Response: Yes, biotic and abiotic factors. Information has been added to the text for better clarity.

 

29. L469-470: same comment as above: conspecifics or 469 heterospecifics

• Response: It was replaced by “bats” which were primarily meant.

 

30. L470: increased competition or higher pathogen infection 470 risks in bats --- again speculative, no data to support this. Delete.

• Response: it was deleted.

 

31. L479: are also very interesting – please delete and let the reader decide about this.

• Response: It was deleted according to the suggestion.

 

32. L493: B. m. nudir --- incorrect

• Response: The name was corrected.

 

33. L501-502: Also, the 501 species complex comprising (N) schmidlii, P. conspicua and Ph. biatriculatum --- this is absolutely cannot be called species complex.

• Response: “species complex” has been replaced by “taxons”.

 

34. L502: Ph. biatriculatum – incorrect

• Response: The grammatical mistake was corrected.

 

35. L503: genus Rhynolophus --- incorrect. Please check all names very carefully throughout the manuscript.

• Response: The particular mistake was corrected and names were checked.

 

 

36. L505-508: again absolutely speculative, no data to support any of these claims. Please delete.

• Response: We agree with the reviewer. The speculative claim was deleted in the text.

 

37. L511: pathogen without plural

• Response: The grammatical mistake was corrected.

 

 

38. Additionally, several very important and relevant papers weren’t cited, which you should definitely include (besides the ones I already mentioned above):

McKee, C. D., Krawczyk, A. I., Sándor, A. D., Görföl, T., Földvári, M., Földvári, G., ... & Sprong, H. (2019). Host phylogeny, geographic overlap, and roost sharing shape parasite communities in European bats. Frontiers in Ecology and Evolution, 7, 69.

 

Szentiványi, T., Estók, P., Pigeault, R., Christe, P., & Glaizot, O. (2020). Effects of fungal infection on the survival of parasitic bat flies. Parasites & vectors, 13(1), 1-8.

 

Sándor, A. D., Péter, Á., Corduneanu, A., Barti, L., Csősz, I., Kalmár, Z., ... & Mihalca, A. D. (2021). Wide distribution and diversity of malaria-related haemosporidian parasites (Polychromophilus spp.) in bats and their ectoparasites in Eastern Europe. Microorganisms, 9(2), 230.

 

Kwak, M. L., Gorecki, V., & Markowsky, G. (2022). Parasites in peril: abundance of batflies (Diptera: Nycteribiidae) declines along an urbanisation gradient. Journal of Insect Conservation, 26(4), 627-638.

 

Verrett, T. B., Webala, P. W., Patterson, B. D., & Dick, C. W. (2022). Remarkably low host specificity in the bat fly Penicillidia fulvida (Diptera: Nycteribiidae) as assessed by mitochondrial COI and nuclear 28S sequence data. Parasites & Vectors, 15(1), 1-16.

 

Rivera, C. L., Padilla, J. F., Ospina Pérez, E. M., Urbano, F. M., Guarín, D. V., Mejía Fontecha, I. Y., ... & Ramírez-Chaves, H. E. (2022). Interaction networks between bats (Mammalia: Chiroptera) and ectoparasitic flies (Diptera: Hippoboscoidea) in the Colombian Orinoquia Region. Acta Chiropterologica, 24(2), 379-394

• Response: Response: Using the records in Szentivanyi et al. [23] as starting point, we checked the literature on the host associations of European bat louse for new, overlooked or misattributed records [46-49; 87-174]. In addition, we conducted a literature search using the keywords ‘Nycteribiidae*’, ‘European bat flies*’, ‘host associations *’, ‘bats *and ‘Europe*’ in Scopus and Google Scholar [14,43-44; 175-188].

Author Response File: Author Response.docx

Round 2

Reviewer 4 Report

Dear Authors,

Thank you for considering my comments and improving the text accordingly. I have no further comments.