Suitable Mouse Model to Study Dynamics of West Nile Virus Infection in Culex quinquefasciatus Mosquitoes
, Silvana de Mendonça 1, Ellen Santos 2, Bruno Marçal 1, Amanda Cupertino de Freitas 1, Fernanda Rezende 1, Rafaela Moreira 1,3, Viviane Sousa 1, Sara Comini 1, Mariana Lima 1, Flávia Ferreira 2, João Paulo de Almeida 2, Emanuele Silva 2, Siad Amadou 2, Marcele Rocha 1, Thiago Leite 2, Yaovi Todjro 2, Camila de Carvalho 4, Viviane Santos 5, Marta Giovanetti 1,6, add
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Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsEstimated Editors of TropicalMed,
Estimated Authors,
I've been invited to review this original study on a mouse model for studying the dynamics of the WNV in C quinquefasciatus mosquitors. Due to the ever increasing impact of arbovirus on the public health, the present study is both interesting and potentially of high significance. In fact, in this study, Authors provide to the basic research two very important input: 1) the mouse model they suggest could be suitable for the study of arbovirus in vertebrates, providing an effective trade-off between consistence with mammalian biology and laboratory costs; 2) the laboratory approach for collecting salivary samples could be used even in different laboratory models.
According to the hereby reported results, following the intraperitoneal injection, AG129 mice exhibited a transient viremia that lasted several days (peak in 2nd to 3rd day), and this timeframe is reasonably sufficient for allowing C quinquefasciatus to take its blood meal, that resulted in the invasion of arthropod saliva within the following 16 days.
From my point of view, the study was properly designed and performed.
My only recommendation is to remove the Figure 1, that could be provided as supplementary / appendix document. Similarly, introduction could be simplified as some information (more precisely rows 58 to 84) provide information that are not strictly necessary for the understanding of the present paper.
Author Response
Dear Reviewer,
Thank you for your valuable feedback and for taking the time to review our manuscript. Please find below our detailed responses, with corresponding revisions/corrections highlighted in the re-submitted files.
Point-by-Point Response to Comments and Suggestions for Authors
Comment 1:
My only recommendation is to remove Figure 1, which could be provided as a supplementary/appendix document.
Response 1:
We agree with your suggestion and have moved Figure 1 to a supplementary appendix document to streamline the main text.
Comment 2:
Similarly, the introduction could be simplified, as some information (specifically, lines 58 to 84) is not strictly necessary for understanding the present paper.
Response 2:
As recommended, we have simplified the introduction by removing the non-essential information between lines 58 and 84, focusing on key content that enhances the manuscript's clarity and focus. We appreciate your valuable insights and believe these changes will improve the quality of our paper.
Reviewer 2 Report
Comments and Suggestions for Authors1. Please add the detection limit of the viral load detection of WNV and cut-off point by RT-qPCR method. This is very important for your system.
2. Regarding the mosquitoes, which generation was used in this experiment such as (F1, F2 etc) Please add at the revised manuscript.
3. In this study, the authors described the gene bank accession number of the WNV. Did the authors check the any amino acids or nucleotide mutation before using the virus by sequencing? If the authors perform this, please add at the revised manuscript.
4. In materials and methods section, the authors used arbovirus “ 4.5. Mosquito Infection with Arbovirus”. In fact, the authors tested WNV only, one of flaviviruses . There are many arboviruses eg not only flaviviruses but also alpha virus. So the authors used specific virus only at the revised manuscript.
5. The authors tested infection of WNV to mice by means of infected mosquitoes after 16dpi. Did the authors check again mortality of mice after getting infection from mosquitoes biting? If the authors check, please describe at the revised one.
Author Response
Dear Reviewer,
Thank you for your valuable feedback and for taking the time to review our manuscript. Below, you will find our detailed responses, with the corresponding revisions/corrections highlighted in the re-submitted files.
Comment 1:
Please add the detection limit of the viral load detection of WNV and the cut-off point by the RT-qPCR method. This is very important for your system.
Response 1:
We appreciate your suggestion. The detection limit and cut-off point for the RT-qPCR method used to detect WNV viral load have been added to the revised manuscript in the "Methods" section. These details are crucial for interpreting the sensitivity and reliability of our detection system (line 245-246)
Comment 2:
Regarding the mosquitoes, which generation was used in this experiment (e.g., F1, F2)? Please add this information in the revised manuscript.
Response 2:
Thank you for pointing this out. The generation of mosquitoes used in the experiments has been clarified and included in the revised manuscript. Specifically, this information has been added to the "Materials and Methods" section (line 135 -136).
Comment 3:
In this study, the authors described the gene bank accession number of the WNV. Did the authors check for any amino acid or nucleotide mutations before using the virus by sequencing? If the authors performed this, please add it to the revised manuscript.
Response 3:
We have not sequenced the virus to check for any amino acid or nucleotide mutations before using it in the experiments. While we understand the importance of this suggestion, due to technical limitations, we were unable to perform the sequencing of the virus stock used in this study.
Comment 4:
In the Materials and Methods section, the authors used the term "arbovirus" in section "4.5. Mosquito Infection with Arbovirus." In fact, the authors tested WNV only, which is one of the flaviviruses. There are many arboviruses, e.g., not only flaviviruses but also alphaviruses. So the authors should specify the virus used in the revised manuscript.
Response 4:
We agree with your suggestion. The term "arbovirus" has been replaced with "West Nile Virus (WNV)" in the revised manuscript to accurately reflect the specific virus tested in this study (line 175).
Comment 5:
The authors tested the infection of WNV in mice by means of infected mosquitoes after 16 dpi. Did the authors check again the mortality of mice after getting infected from mosquito bites? If the authors checked, please describe this in the revised manuscript.
Response 5:
We did not test the infection of mice by allowing infected mosquitoes to take a blood meal, although we consider this an important experiment to ultimately validate that the mosquitoes are capable of transmitting WNV to a vertebrate host. We have discussed the significance of this experiment in the discussion section, highlighting its importance for future studies (line 436 - 439). However, this experiment was beyond the scope of the current study.
