Molecular Detection of Acarapis woodi Using Hive Debris as Innovative and Non-Invasive Matrix

Round 1

Reviewer 1 Report

The present study indicates the detection of Acarapis woodi in Italy, a widely distributed parasite. The number of samples analyzed were low compared to similar previous studies.
The article could improve for example:
- Material and methods: include a map with the location of the sampling areas and the positives.
- Results: this section should be expanded and provide more information on the number of specimens sampled, location, etc.

Author Response

Dear Reviewer,

Thank you for your comments and suggestions to improve the manuscript.

As the sampling plan was on voluntary basis, avoiding the management of personal data (except from the farm/beekeeper name), it was not asked to participants to record GPS data or location of the sampling. For this reason, it is not possible to provide an accurate map with the location of the sampling areas and positives.

Reviewer 2 Report

The study presented by authors Pietropaoli et al. is interesting and addresses the need to screen for all potential pests and parasites in honey bee colonies. I only have one minor comment. Please state the exact N. The numbers are confusing in section 2.3 in the methods.

Author Response

Dear Reviewer,

Thank you for your comment. We corrected the number of sections.

Reviewer 3 Report

The authors have tried to introduce the simple P C R method as an early detection technique for the acarine mite and have actually confirmed a high infection rate from honey bee rearing colonies in Italy. I judge that the paper has important technical implications for the future quarantine of acarine mites.

 

On the other hand, I consider it problematic that the paper does not provide any physical evidence in the form of figures. Detailed data, such as information on the location of the colonies, electrophoretic images of the PCR amplified products detected in each colony, and their nucleotide sequence information, should be disclosed first.

 

The manuscript in its current form would be difficult to publish in our journal.

Author Response

Dear Reviewer,

Thank you for your comments and suggestions to improve the manuscript.

As the sampling plan was on voluntary basis, avoiding the management of personal data (except from the farm/beekeeper name), it was not asked to participants to record GPS data or location of the sampling. For this reason, it is not possible to provide an accurate map with the location of the sampling areas and positives.

We added an electrophoretic images of the PCR products, and the nucleotide sequences obtained in our study have been added as supplementary material.

Reviewer 4 Report

The manuscript “Molecular detection of Acarapis woodi using hive debris as innovative and non-invasive matrix” by Pietropaoli et al developed a method for detecting the pathogen of honey bees. Through PCR, the results in this brief report demonstrated the potential underestimation of the pathogen A. woodi occurrence in Italian apiaries. Overall, the method is useful and well established in this manuscript. Accordingly, this reviewer recommends publication.

Author Response

Dear Reviewer,

Thank you for the positive comments on the manuscript.

Best regards

Round 2

Reviewer 1 Report

No comments

Author Response

Dear Reviewer,

Thank you for reading the manuscript and your interest for it.

Best regards

Reviewer 3 Report

Dear authors,

I understand that the author cannot specify the point of origin information for privacy reasons. The author should rather state the reason for this to reduce stress for the reader.

 

Thank you also for the electrophoresis photos.

However, in my opinion, the photographs are also inadequate in the following respects.

Firstly, there are no positive and negative control lanes.

Secondly, you did not indicate which band represents the tick DNA fragment.

I would like to see these points improved.

 

Also, for Figure S1, please add an explanation of which sequence lane is the sample obtained and which lane is the reference data.

Author Response

Dear Reviewer,

Thank you for your comments and suggestions to improve the manuscript.

We added a new electrophoretic images of the PCR products,  in which more explanations and negative and positive control have been added. In brackets the number of base pairs obtained close to the tick DNA fragments.

Moreover we improved the figure S1, with a reference sequence and the explanation of sequences lane by lane.

King regards, 

Dr. Silvia Tofani