Review Reports

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The present manuscript entitled “Genotypes of Bovine Viral Diarrhea Virus Infecting Vaccinated     

and Nonvaccinated Cattle in Thrace District, Türkiye” aims to detect BVDV associated with respiratory and diarrhea in cattle.

To this end, the authors:

-collected 533 nasal and rectal swabs from animals showing clinical respiratory disease or diarrhea in 26 farms in the Thrace district between the years 2021 and 2023.

-95 samples were PCR-positive for BVDV genomic material.

-The Pestivirus brazilense subgenotype was reported for the first time in the Thrace district.

-Provide in general valuable information for BVDV control programs in the region.

The manuscript is well written and the data are presented clearly and support the conclusions. Therefore, I only have the following minor points that might improve the manuscript.

Minor points

-More explanation should be added to table 2.

-Line 54: cytopathic effects

Change to: cytopathogenicity

-Line 68: to virus

Change to: the virus

-Line 69: that not

Change to: that are not

-Line 76-77: A very short description, in two or three sentences should be provided to understand the differences between the production systems.

-Line 203: Reference for the Fisher´s exact test should be cited (Fisher, R. A. (1934). Statistical methods for research workers (5th ed.). Oliver & Boyd.)

-Line 214-215: Is it possible to provide some information of the vaccine, e.g. genotype ?.

-Lines 245-246: the ages of higher positivity rates coincide with the time at which the maternal antibodies of the calf decline. This aspect should be considered.

-In the supplementary table 4 the subgenotype 1f showed the lowest Ct value. Did this correlate with impaired clinical condition ?.

-Lines 290 to 298: This is more introductory information and should not be part of the discussion section. It should removed from the manuscript.

-Line 303-304: What is the impact of the introduction of the  Pestivirus brazilense genotype ? Is it more pathogenic ? How divergent is the amino acid sequence of the E2 protein compared to E2 of the vaccines used in Türkiye ? 

-Line 313-314: Which seroprevalence was reported in the district of this study ?

-Line 317: What was the association of BVDV in animals with diarrhea and or respiratory disease ?.

-Line 341: The prevalence is actually underestimated.

-Line 342: Do animals from small family owned farms share pastures with conventional farms ?

-Line 354: In order to assess the effectivity of the vaccination to prevent clinical infections, the percent of infected animals at herd level should be compared. In this case, only clinically affected animals were compared. Maybe at herd level, more animals are protected in herds with vaccination than in herds without it.

-Line 361: It can be that the maternal antibodies are declining, which typically occur in calves between 2 to 6 months. In the case of herds without vaccination, the natural „booster“ by infections might stimulate higher antibody titers than the ones provided by the vaccine before/around the calving time.

-Line 363: Very often inactivated vaccines do not provide sterile immunity (block viral infection/replication).

-Line 383: Which E gene ? The E2 protein encoding gene ?

-Line 391: Check for consistency on the genotypes names written in cursive.

-Line 395: What is it meant with no amplification. A clearer sentence should be added highlighting the step that seemed to be critical for not obtaining sequencing data.

-Lines 400 to 405: Globally…..and 1i.

This part can be removed to keep the discussion more concise and concentrate on the situation/reports in Türkiye.

-Lines 409-414: In contrast…and subgenotypes.

This information is more introductory and do not add much in the discussion.

-Lines 424-426: Could you provide any situation by wich this strain might have reached the Turkish farms ?

-Lines 433 and 436: Virus shedding instead of shedding.

-Line 439: The serological surveillance might be useful and amenable since antibodies against the non-structural protein 3 are barely detectable in animals vaccinated with inactivated BVDV vaccines (Makoschey et al., 2007. Vaccine 25: 6140-6145).

-Line 450: Could you give an example on the implications for disease control on the region ?

-Line 455: The term prevalence should be avoided in this study as it refers to the association of BVDV with respiratory and gastrointestinal symptoms in cattle in this region.

-Line 457 to 461: This sentence prompts me as reviewer to request further sequencing data with the mentioned genes. Nevertheless, as the aim of this study is to determine the presence of BVDV in cattle with clinical disease, this might not be relevant.

-Lines 463-464: In this case it should be at least possible to know the vaccines that are approved to vaccinate cattle in the region. With this information the sequence of the E2 protein of the vaccine and the circulating strains could be compared.

-The title of supplementary table 2 should be: Primers and probe used for detection and sequencing of BVDV genome.

-The title of supplementary table 3 should be: List of reference strains of pestiviruses for phylogenetic comparison with isolates.

Author Response

Dear Reviewer

Thank you very much for your thorough and constructive comments on our manuscript. We greatly appreciate your time and expertise in evaluating our work. Please find below our detailed point-by-point responses to your valuable suggestions. We have carefully considered each comment and have revised the manuscript accordingly to address all concerns raised (see revised manuscript and response letter).

Best regards

Corresponding author

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

All comments are included in the attached file

Comments for author File: Comments.pdf

Author Response

Dear Reviewer

Thank you very much for your thorough and constructive comments on our manuscript. We greatly appreciate your time and expertise in evaluating our work. Please find below our detailed point-by-point responses to your valuable suggestions. We have carefully considered each comment and have revised the manuscript accordingly to address all concerns raised (see revised manuscript and response letter).

Best regards

Corresponding author

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

The authors addressed the reviewer's comments and suggestions. The corrections significantly increased the scientific value of the manuscript. I fully support the publication of this paper.