Isolation, Identification, and Genetic Evolution Analysis of VP1 Gene of Feline Calicivirus Strain ZZ202306

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

I congratulate the authors on this interesting study. I am sure that it will be interesting for both researchers and practicing veterinarians. I noticed two errors in the text of the manuscript, which I recommend that the authors correct:

1. Marti-noB and colleagues (row 50) – I think this family name has a mistake and probably the right family name is Martinov. These authors are not cited in the Reference list.
2. Grammatical mistake:  3. Detection of FCV byimmunofluorescence assay and electron microscopy.

Additional comments:

This research aims to establish a reference framework for researchers, veterinarians, and cat caregivers to implement effective FCV control strategies and guide future research directions. Ultimately, the goal is to contribute to developing FCV vaccines, thereby reducing the virus's impact on feline populations and their human companions. 

I believe the topic is original because it provides new information about FCV. The authors present new information about FCV by comparing it with other similar viruses. 

Various methods have been used to study and compare FCV with other similar viruses such as, Cell culture and reagents, Sample processing, Specimen testing [primers for FCV, feline parvovirus (FPV), feline herpesvirus (FHV), and feline coronavirus (FCoV), and FCV-specific identification were synthesized], Virus isolation and cultivation, Western blotting, Immunofluorescence assay, TCID50 assay, Viral growth kinetics analysis, and Electron microscopy. This makes the research very rich and original.

The conclusion is based on the following findings: authors successfully isolated and characterized a new FCV strain ZZ202306, providing insights into the genetic variability of the VP1 gene. This research underscores the importance of conducting cross-immunoprotection assays between prevalent local strains and vaccines, emphasizing the critical need for evaluating vaccine efficacy to develop more effective vaccines. This approach is anticipated to reduce FCV infection rates and associated mortality.

The references cited are appropriate for this investigation. 

In this manuscript are presented 2 tables and 7 figures which demonstrate very well the results obtained.

 

 

Author Response

Comments and Suggestions for Authors:I congratulate the authors on this interesting study. I am sure that it will be interesting for both researchers and practicing veterinarians. I noticed two errors in the text of the manuscript, which I recommend that the authors correct:

• Marti-noB and colleagues (row 50) – I think this family name has a mistake and probably the right family name is Martinov. These authors are not cited in the Reference list.

Response: We thank for the reviewer’s suggestion. We have revised the statement in accordance with the reviewer's recommendation. We have thoroughly verified all the references cited in our manuscript, and we are grateful for the reviewer's meticulous evaluation.

• Grammatical mistake

Response: We are grateful for the reviewer's suggestion and have once again checked the writing issues throughout the manuscript, making corrections to the abstract section.

• Detection of FCV byimmunofluorescence assay and electron microscopy.

Response: We appreciate the reviewer's suggestion. We have revised "Detection of FCV by immunofluorescence assay and electron microscopy" to "Detection of FCV using immunofluorescence assay and electron microscopy.

• Additional comments:This research aims to establish a reference framework for researchers, veterinarians, and cat caregivers to implement effective FCV control strategies and guide future research directions. Ultimately, the goal is to contribute to developing FCV vaccines, thereby reducing the virus's impact on feline populations and their human companions.

I believe the topic is original because it provides new information about FCV. The authors present new information about FCV by comparing it with other similar viruses. Various methods have been used to study and compare FCV with other similar viruses such as, Cell culture and reagents, Sample processing, Specimen testing [primers for FCV, feline parvovirus (FPV), feline herpesvirus (FHV), and feline coronavirus (FCoV), and FCV-specific identification were synthesized], Virus isolation and cultivation, Western blotting, Immunofluorescence assay, TCID50 assay, Viral growth kinetics analysis, and Electron microscopy. This makes the research very rich and original.

The conclusion is based on the following findings: authors successfully isolated and characterized a new FCV strain ZZ202306, providing insights into the genetic variability of the VP1 gene. This research underscores the importance of conducting cross-immunoprotection assays between prevalent local strains and vaccines, emphasizing the critical need for evaluating vaccine efficacy to develop more effective vaccines. This approach is anticipated to reduce FCV infection rates and associated mortality.

The references cited are appropriate for this investigation.

In this manuscript are presented 2 tables and 7 figures which demonstrate very well the results obtained

Response: We appreciate the recognition and valuable feedback provided by the reviewers on our research. The reviewers' professional and insightful summary of our study has been incorporated into Section 5 (Conclusions) of the manuscript. We extend our gratitude for the endorsement of our research and the successful publication of the article.

Reviewer 2 Report

Comments and Suggestions for Authors

This study characterizes the FCV strain ZZ202306 isolated from Zhengzhou, employing RT-PCR, CRFK cell culture, and immunofluorescence techniques. Phylogenetic analysis revealed its genetic proximity to global FCV strains. While the methods employed are robust, further investigation into clinical epidemiology and in vivo pathogenicity is essential to enhance translational relevance. The manuscript is suitable for publication pending revisions addressing these gaps.

 

Minor Comments:

1. The title should be revised to “Isolation, Identification, and Genetic Evolution Analysis of the VP1 Gene of Feline Calicivirus Strain ZZ202306” for greater precision and clarity.
2. Line 70: The sentence “Vaccination remains the most effective preventive measure, with prevalent strains such as 255 and F9” should be rephrased as “Vaccination, notably targeting prevalent strains such as 255 and F9, remains the primary preventive measure against FCV.”
3. Line 140: The description “Samples were centrifuged at 12,000 × g for 5 minutes at 4°C, and the supernatant was collected” should be revised to “Following centrifugation at 12,000 g for 5 minutes at 4°C, the supernatant was harvested.”
4. Line 177: The sentence “Virus samples were collected at 6, 12, 18, 24, 30, 36, 42, and 48 hours post-infection, and viral nucleic acids were extracted for real-time fluorescent quantitative analysis” should be adjusted to “At 6, 12, 18, 24, 30, 36, 42, and 48 hours post-infection, virus samples were gathered and subjected to extraction of viral nucleic acids for real-time fluorescent quantitative analysis.”

Author Response

This study characterizes the FCV strain ZZ202306 isolated from Zhengzhou, employing RT-PCR, CRFK cell culture, and immunofluorescence techniques. Phylogenetic analysis revealed its genetic proximity to global FCV strains. While the methods employed are robust, further investigation into clinical epidemiology and in vivo pathogenicity is essential to enhance translational relevance. The manuscript is suitable for publication pending revisions addressing these gaps.

Minor Comments:

(1) The title should be revised to “Isolation, Identification, and Genetic Evolution Analysis of the VP1 Gene of Feline Calicivirus Strain ZZ202306” for greater precision and clarity.

Response: We acknowledge the reviewer's recommendation and have accordingly revised the title.

(2) Line 70: The sentence “Vaccination remains the most effective preventive measure, with prevalent strains such as 255 and F9” should be rephrased as “Vaccination, notably targeting prevalent strains such as 255 and F9, remains the primary preventive measure against FCV.”

Response: Following the reviewers' recommendations, we have made corrections to the grammar errors in this section and have conducted a review of the remaining sections of the manuscript.

(3) Line 140: The description “Samples were centrifuged at 12,000 × g for 5 minutes at 4°C, and the supernatant was collected” should be revised to “Following centrifugation at 12,000 g for 5 minutes at 4°C, the supernatant was harvested.”

Response: As per the recommendations of the reviewers, we have corrected the grammar errors in this section and have reviewed the other sections of the manuscript.

(4) Line 177: The sentence “Virus samples were collected at 6, 12, 18, 24, 30, 36, 42, and 48 hours post-infection, and viral nucleic acids were extracted for real-time fluorescent quantitative analysis” should be adjusted to “At 6, 12, 18, 24, 30, 36, 42, and 48 hours post-infection, virus samples were gathered and subjected to extraction of viral nucleic acids for real-time fluorescent quantitative analysis.

Response: We appreciate the support and feedback provided by the reviewers, and have made the necessary corrections accordingly.