Blood Parasite Diversity and Zoonotic Risk in Captive Sun-Tailed Monkeys from Gabon

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

This study investigates the prevalence and diversity of Plasmodium and 17

Trypanosoma infections in Allochrocebus solatus in Sun-Tailed Monkeys from a Semi-Free-Ranging Colony in Southern Gabon. The findings provide a better understanding of host-parasite dynamics in semi- captivity and their implications for primate biology and conservation. However, there are some points should be addressed.

1. Line 113,122, Unidirectional sequencing or bidirectional sequencing about the the 18S rRNA gene and 5.8S/28S rRNA of trypanosomes parasites and the cyt-b gene of malaria parasites? Is the sequence uploaded to Genbank, If the sequences obtained in this study,please supply the accession numbersof genes obtained in this study deposited in GenBank.
2. The results of the paper lackthe description about the genetic analysis of the obtained sequences, Please supply the relevant informationabout phylogenetic analysis and Figure 2. and Figure 3.
3. The authors mentioned that the χ2test was used to compare the proportions of animals infected by trypanosome and plasmodium species in each sex, age, species, and season. But the results of the paperonly mention the statistical analyses result about age, and have not the statistical analyses result in each sex, species, and season. Please supply these results.
4. Some minor issues

(1) Please use abbreviations for species names, please see line58,77,144,149,176-177,

187,269,275-276,295-296,301,324 and the Figure 2. and Figure 3.

(2) some “spp” should be “spp.”,please see line 28,55.

(3) Please use the italics for the species names,please see line 74,318.

(4) Line 82, “by trained veterinarians who who adhered” should be “by trained veterinarians who adhered”.please check this.

(5) line 151,153, “Solatus” →“A. solatus”

(6)  The scientific name of parasite and host should be italic in the References section.

Author Response

Reviewer 1

This study investigates the prevalence and diversity of Plasmodium and Trypanosoma infections in Allochrocebus solatus in Sun-Tailed Monkeys from a Semi-Free-Ranging Colony in Southern Gabon. The findings provide a better understanding of host-parasite dynamics in semi- captivity and their implications for primate biology and conservation. However, there are some points should be addressed.

1. Line 113,122, Unidirectional sequencing or bidirectional sequencing about the the 18S rRNA gene and 5.8S/28S rRNA of trypanosomes parasites and the cyt-b gene of malaria parasites? Is the sequence uploaded to Genbank, If the sequences obtained in this study,please supply the accession numbersof genes obtained in this study deposited in GenBank.

Response: We thank the reviewer for this important point. All sequences were bidirectionally sequenced to ensure accuracy. The sequences obtained in this study have now been submitted to GenBank with the following accession numbers: PV425447–PV425455 for Trypanosoma spp. and PV454983-PV454997 for Hepatocystis spp.

 

1. The results of the paper lackthe description about the genetic analysis of the obtained sequences, Please supply the relevant informationabout phylogenetic analysis and Figure 2. and Figure 3.

Response: We have now provided a more detailed description of the genetic analysis results, including the topologies and genetic clustering observed in Figures 2 and 3. These figures are now properly cited in Sections 3.1 and 3.2, respectively.

 

1. The authors mentioned that the χ2test was used to compare the proportions of animals infected by trypanosome and plasmodium species in each sex, age, species, and season. But the results of the paperonly mention the statistical analyses result about age, and have not the statistical analyses result in each sex, species, and season. Please supply these results.

Response: We appreciate this suggestion. We have now included the results of the Chi-square analyses by sex and season in the Results section. No significant differences were found for either factor.

1. Some minor issues

• Please use abbreviations for species names, please see line58,77,144,149,176-177,187,269,275-276,295-296,301,324 and the Figure 2. and Figure 3.

Okay, we considered that in the revised manuscript version

 

• Some “spp” should be “spp.”,please see line 28,55.

Thank you for pointing this out. We have corrected “spp” to “spp.” in the specified lines.

(3) Please use the italics for the species names,please see line 74,318.

We have edited the manuscript to ensure that all species names are italicized as requested

(4) Line 82, “by trained veterinarians who who adhered” should be “by trained veterinarians who adhered”.please check this.

We have removed the duplicate "who" in the sentence as suggested.

(5) line 151,153, “Solatus” →“A. solatus”

Okay, we considered this aspect in the new manuscript

(6)  The scientific name of parasite and host should be italic in the References section.

We have revised the References section to ensure that all scientific names are italicized

Reviewer 2 Report

Comments and Suggestions for Authors

This is an interesting article.

Major comments.

1) The authors should clarify whether the novel sequences derived in this manuscript have been submitted to GenBank, and include the accession numbers.

2) Section 3.1. The Figure showing 18S-phylogeny should be cited in this Section as ‘Figure 2’.

3) Section 3.2. The Figure showing Cytb-phylogeny should be cited in this Section as ‘Figure 3’.

Minor comments:

Line 37. Delete the words: ‘, is to’

Line 74. Italic font for: ‘A. solatus’

Line 88. Figure 1 legend. Add the genus name Allochrocebus, and then italic font for genus and species names

Line99. The authors should check the names of the PCR primers here: IST or ITS as in Line 106 & Line 128

Line 149. The authors could state whether there were any mixed infections by more than one Trypanosoma species

Line 151. Should be labelled as Figure 2.

Line 151. Figure title should be ‘…from Allochrocebus solatus,  and then italic font for genus and species names

Line 163. Amend the spelling to ‘Mixed’

Line 171. Should be labelled as Figure 3.

Line 173. Amendment: ‘Red indicates sequences from Allochrocebus solatus’ and then italic font for genus and species names

Line 183/210. Amendment: ‘solatus’ to ‘Allochrocebus solatus’, and then italic font for genus and species names

Line 239/318. ‘A. solatus’ to be in italic font

Line 282. ‘Trypanosoma’ to be in italic font

Line 283. ‘Anopheles’ to be in italic font

Line 361. Abbreviation SOL should be explained as ‘Allochrocebus solatus’, and then italic font for genus and species names

 

Author Response

Reviewer 2

Comments and Suggestions for Authors.

This is an interesting article.

 

Major comments.

• The authors should clarify whether the novel sequences derived in this manuscript have been submitted to GenBank, and include the accession numbers.

Reply: Thank you for your suggestion. We have now included the GenBank accession numbers for each sequence obtained in this study (see the new manuscrit version,). (PV425447-PV425455 for Trypanosomes parasites and PV454983-PV454997 for malaria parasites)

• Section 3.1. The Figure showing 18S-phylogeny should be cited in this Section as ‘Figure 2’.

Reply: We have cited the phylogenetic tree based on 18S as Figure 2 as requested (see the new manuscrit version, section 3.1)

• Section 3.2. The Figure showing Cytb-phylogeny should be cited in this Section as ‘Figure 3’.

Reply: This citation has been updated accordingly (see the new manuscrit version, section 3.2).

Minor comments:

Line 37. Delete the words: ‘, is to’:

Reply: Thank you, i deleted in the new manuscript version

Line 74. Italic font for: ‘A. solatus’:

Reply: Thank you; this has been corrected

Line 88. Figure 1 legend. Add the genus name Allochrocebus, and then italic font for genus and species names

Reply: We have corrected this part to include the genus name in italics

Line99. The authors should check the names of the PCR primers here: IST or ITS as in Line 106 & Line 128

Reply: Thank you, we have edited this for consistency, the good name is ITS.  

Line 149. The authors could state whether there were any mixed infections by more than one Trypanosoma species.

Reply: This information has now been included in the revised manuscript. Howover, in this study, we did not observe a mixed infection between Trypanosoma spp.

Line 151. Should be labelled as Figure 2.

Reply: This labelling has been corrected

Line 151. Figure title should be ‘…from Allochrocebus solatus,  and then italic font for genus and species names

Reply: Okay, we considered that in the manuscript version

Line 163. Amend the spelling to ‘Mixed’

Reply: This correction has been made.

Line 171. Should be labelled as Figure 3.

Reply: thank you, it done now

Line 173. Amendment: ‘Red indicates sequences from Allochrocebus solatus’ and then italic font for genus and species names

Reply: This amendment has been made

Line 183/210. Amendment: ‘solatus’ to ‘Allochrocebus solatus’, and then italic font for genus and species names

Reply: This has been corrected

Line 239/318. ‘A. solatus’ to be in italic font

Reply: This has been updated

Line 282. ‘Trypanosoma’ to be in italic font

Reply: This has been corrected

Line 283. ‘Anopheles’ to be in italic font

Reply: This has been corrected

Line 361. Abbreviation SOL should be explained as ‘Allochrocebus solatus’, and then italic font for genus and species names

Reply: This explanation has been added and formatted accordingly

 

Reviewer 3 Report

Comments and Suggestions for Authors

Dear Authors,

After reviewing the manuscript entitled "Molecular Characterization and Zoonotic Risk Assessment of Blood Parasites in Sun-Tailed Monkeys from a Semi-Free-Ranging Colony in Southern Gabon: Preliminary Insights into Malaria parasite and Trypanosoma Infections "

I see that the article is very important from a phylogenetic point of view as well as Epidemiological point of view and from Infection Prevention and control perspective, in fact this article highlights, using a molecular, qualitative, advanced study, on the Characteristics and the Zoonotic Risk Assessment of different blood borne parasites (namely Malaria Parasites and Trypanosoma Infectious agents) in a specific kind of monkeys mainly SUN-Tailed ones which is frequently present in Southern Gabon..

I have the several comments regarding this work:

01- The text needs English polishing in several of its parts.

02- The Title is too long, I suggest to replace it by a smaller one.

03- You are invited to put the numbers for affiliations in a ascendant order, 1, 2, 3 etc. In your manuscript you used as 1, 3, 2, etc.

04- Since the number of samples is very small (47 samples), I see that this article could not be published as an original research article, I will go more into using it as a Pilot Study.

05- You are invited to put more information in the sub-section related to the statistical analyses, this part should be more clear for readers.

06- The Figures (second and third ones) used in the present manuscript are clear to understand but I think that you should work on the quality of the presentation, for example the font used must be common in all figures, you can change the color of the font but not the font itself.

07- The second and third figure missed a Figure number, It should be Figure 2 and Figure 3. 

08- You Should remove the Point (06- Patent) after the conclusion.

09- The conclusion is not very rich in information, you just re-mentioned the prevalence of each infection and that of the co-infection, then you talked about the limitation! Try to be more persuasive and make your conclusion more rich in information.

BR,

 

Comments on the Quality of English Language

The English Language in this manuscript is average it must be improved by an English native speaker.

Author Response

Reviewer 3

Dear Authors,

After reviewing the manuscript entitled "Molecular Characterization and Zoonotic Risk Assessment of Blood Parasites in Sun-Tailed Monkeys from a Semi-Free-Ranging Colony in Southern Gabon: Preliminary Insights into Malaria parasite and Trypanosoma Infections "

I see that the article is very important from a phylogenetic point of view as well as Epidemiological point of view and from Infection Prevention and control perspective, in fact this article highlights, using a molecular, qualitative, advanced study, on the Characteristics and the Zoonotic Risk Assessment of different blood borne parasites (namely Malaria Parasites and Trypanosoma Infectious agents) in a specific kind of monkeys mainly SUN-Tailed ones which is frequently present in Southern Gabon..

I have the several comments regarding this work:

• The text needs English polishing in several of its parts.

Reply: Thank you, we have decided to engage an English editor for MDPI to ensure the manuscript meets high language standards

• The Title is too long, I suggest to replace it by a smaller one.

Reply: We edited the title to “Blood Parasite Diversity and Zoonotic Risk in Captive Sun-Tailed Monkeys from Gabon”

• You are invited to put the numbers for affiliations in a ascendant order, 1, 2, 3 etc. In your manuscript you used as 1, 3, 2, etc.

Reply: We have corrected the affiliations in the manuscript to follow the ascending order as recommended.

• Since the number of samples is very small (47 samples), I see that this article could not be published as an original research article, I will go more into using it as a Pilot Study.

Reply: We have modified the title and abstract to reflect that this is a preliminary (pilot) study, acknowledging the sample size.

• You are invited to put more information in the sub-section related to the statistical analyses, this part should be more clear for readers.

Reply: We have edited the statistical analyses section to provide clearer and more detailed information for readers.

• The Figures (second and third ones) used in the present manuscript are clear to understand but I think that you should work on the quality of the presentation, for example the font used must be common in all figures, you can change the color of the font but not the font itself.

Reply: We have harmonized Figures 2 and 3, ensuring consistency in font style and formatting

• The second and third figure missed a Figure number, It should be Figure 2 and Figure 3

Reply: Figures 2 and 3 have been correctly numbered and labeled in the revised manuscript.

08- You Should remove the Point (06- Patent) after the conclusion.

Reply: We understand this requirement is per journal guidelines and have removed that section.

09- The conclusion is not very rich in information, you just re-mentioned the prevalence of each infection and that of the co-infection, then you talked about the limitation! Try to be more persuasive and make your conclusion more rich in information.

Reply: We have enriched the conclusion to emphasize the importance of monitoring the health of captive primates, the significance of a One Health perspective, and the potential role of A. solatus as reservoirs.

Reviewer 4 Report

Comments and Suggestions for Authors

This is an Epidemiological Survey Articles. The discussion part is described in detail which a plus of this manuscript.

I have only minor comments as indicated below.

1. Line 151 and 171：The author should notify the number of the Figure: Figure 2 and Figure3.
2. The species name of the parasite shows in the phylogenetic tree showed be italic font.

Author Response

Reviewer 4

 

Comments and Suggestions for Authors

This is an Epidemiological Survey Articles. The discussion part is described in detail which a plus of this manuscript.

I have only minor comments as indicated below.

1. Line 151 and 171：The author should notify the number of the Figure: Figure 2 and Figure3.

Reply: Thank you for your observation. We have ensured that the figures are correctly labeled as Figure 2 and Figure 3 in the revised manuscript

 

1. The species name of the parasite shows in the phylogenetic tree showed be italic font.

Reply: The species names in the phylogenetic tree have been updated to ensure they are in italic font.

 

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The author has made good revisions according to the reviewer's comments.

Reviewer 2 Report

Comments and Suggestions for Authors

Thank you for making the changes. Congratulations on the good work in this article.

Reviewer 3 Report

Comments and Suggestions for Authors

Dear Authors,

After reviewing the revised version of the manuscript entitled "Molecular Characterization and Zoonotic Risk Assessment of Blood Parasites in Sun-Tailed Monkeys from a Semi-Free-Ranging Colony in Southern Gabon: Preliminary Insights into Malaria parasite and Trypanosoma Infections "

I see that the article is more suitable for publication in its present form,

Best Regards,

Reviewer 4 Report

Comments and Suggestions for Authors

The corrected article turned out to be clear and easy to understand, and the author answered every question reasonably well