A Reliable and Cost-Efficient PCR-RFLP Tool for the Rapid Identification of Cetaceans in the Mediterranean Sea

Round 1

Reviewer 1 Report

  Dear Authors, congratulation on this work which is very well-designed and developed.

Strandings are a big challenge in term of species population status, health monitoring and management. Any new framework such as protocols or new tools is an essential support for the research and conservation on these species. This paper offer an overview of the Italian situation of the presence of cetaceans and the logistic difficulties to manage strandings specimens, especially when the condition code of the carcass doesn't allow to conduct complete post mortem investigation. I strongly believe that the new cost-effective tool proposed can support research on this field, particularly in areas where there is still an important knowledge gap or lack of resources. This is very common in many areas in the Mediterranean Sea and worldwide. It is very common indeed, that, especially within the climate change concerns, non-regular species enter into the Mediterranean basin without any chance to be detected by e-DNA studies or at sea monitoring. In this context, strandings represents the most useful opportunity to assess many aspects regarding the life of marine mammal species, one of these is represented by the identification of the species which is usually based on subjective external observation without any objective scientific support.    Regarding some small reviews:   > Introduction section: I would add the effectiveness of your protocol also for sunk carcasses that sometimes are found with no chance to be collected. Moreover, this protocol could also be adapted to other marine animals difficult to monitor, such as sea turtles and monk seals. It is very frequent that small cetaceans are confused with monk seals and sea turtles species are confused each other. A mention of this aspect could be very useful for the readers and future research perspectives highiting the importance of the application of this protocol for the objective identification of stranded cetaceans.
> Fig 2. I would avoid reporting "Stranded" several times for each species, as it is quite obvious by the images and the description.
> I strongly believe that this approach could be a valuable contribution to conservation In the Discussion, it would be very useful if it could be expanded the conservation aspect of your tool. For example, describing the conservation applications highlighting the importance of the detection of non-regular species in the climate change or environment change should be addressed with more attention.

Author Response

Dear Reviewer, thank you very much for the appreciation of our work and for the useful suggestions. For a point-by point reply,  please see the attachment.

With our best regards, 

Stefania Chiesa and co-Authors.

Author Response File: Author Response.docx

Reviewer 2 Report

The paper entitled "A reliable and cost-efficient....." is focused on the use of molecular tools to provide taxonomic identification, especially useful for cetacean species in the Mediterranean sea. 

The strength of the paper consiste on cost-efficient PCR-REFLP tool for rapid identification for cetacean species occurring in the Mediterranean sea opening  a window on the applicability of the tool for identification of other species.

The results show that the tool used is accessible and allows to obtain results in a short time on a large number of specimens

My suggestions is to add to molecular data where possible given the conditions of the species considered an adequate morphological description.

Author Response

Dear Reviewer, thank you very much for the appreciation of our work and for the useful suggestions. For a point-by point reply,  please see the attachment.

With our best regards, 

Stefania Chiesa and co-Authors.

Author Response File: Author Response.docx