Genotoxicity Set Up in Artemia franciscana Nauplii and Adults Exposed to Phenanthrene, Naphthalene, Fluoranthene, and Benzo(k)fluoranthene

Round 1

Reviewer 1 Report

I recommend the acceptance of the manuscript.

Author Response

Dear Reviewer 1,

 I want to thank you for review.

Author Response File: Author Response.pdf

Reviewer 2 Report

Dear authors,

Congratulation for your work. This manuscript is an interesting study that provides information on the large-scale genotoxicity of PAHs in Artemia Franciscana nauplii and adults. It is very detailed research with many results presented. However, some aspects must be clarified. Specifically, my indications/comments are:

1.         The abbreviation should be used consistently (i.e. benzo(k)fluoranthene is B(k)F  or BkF)-  lines 23, 25, 75, 153, 258, etc.). Please check the entire document. 

2. Can you mention, please, what represents the abbreviation IC in Supplementary Table S3. Are the confidence intervals? Please mention the name of this abbreviation.

3. Line 158: what represents centrifugation at 4000g; probably you should express this in revolutions per minute (rpm).

4. Line 179: Please delete the point before “using the following…”

5. Equation 1: it was impossible to see the entire equation. Maybe it is better to use also the equation such as Ratio= …..

6. Line 159: Please delete “.6,” Correct this phrase.

7. The quality of the figures must be improved (high resolution needed).

8. Lines 326–327: please correct this phrase (Both contaminants, NAP, PHE and BkF- you have mentioned 3 contaminants not two).

9. Line 347: please correct dawn-regulated with down-regulated.

10. The conclusion part should be improved by introducing a more pertinent conclusion to the work.

Thus, my decision is a MINOR REVISION.

Author Response

Dear Reviewer 2, 

I want to thank you for review and the usefull comments.

Author Response File: Author Response.pdf

Reviewer 3 Report

Title: Genotoxicity in Artemia franciscana nauplii and adults exposed to phenanthrene, naphthalene, fluoranthene and benzo(k)fluoranthene

General Comments:

This manuscript explores the toxicity of PAHs to two lifestages of Artemia franciscana using both mortality and a genotoxicity assay. The data presented in this paper is highly suspect as the exposure concentrations for a lot of these PACs exceed the maximum solubility of these compounds in water by almost 100 fold! In table one they state they measured phenanthrene in water at a concentration of 223.4 mg/L when the maximum experimental solubility of phenanthrene in water is 1.15mg/L. For fluoranthene, they said their maximum exposure concentration was measured to be 325 mg/L when the maximum solubility is 0.26 mg/L!! This may be a unit error, and if it is the authors need to completely re-write and re-graph the manuscript before considering resubmission.

Specific Comments:

Line 18: Actually factually incorrect. There have been many PAH toxicity tests conducted with artemia, see Rojo-nieto et al. 2012, Cong et al. 2021, and Colvin et al. 2021 just to name a select few. This statement is repeated in the intro and then followed with citations for the Rojo-Nieto paper…. So this needs to be revised.

Line 42: Depends on number of benzene rings or Kow value? Cite source.

Line 45: Citation needed.

Line 156: How were you able to expose the Artemia to phenanthrene at concentrations as high as 84.6 mg/L when the maximum solubility of phenanthrene in water is 1.15mg/L??

Author Response

Dear Reviewer 3,

I want to thank you for your review and the usefull comments.

Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

The authors cite papers that generally describe the increase in solubility of the PAHs used with co-solvents, but no where do these papers support a 100 fold increase with only a 1% solvent ratio. Also, the majority of the effects observed in the study occur at concentrations that far exceed the maximum solubility of the compounds in seawater.. so is the assay actually sensitive and useful?

If this manuscript is going to be published the fact that these exposures exceed maximum solubility by 100 fold should be clear in the abstract, methods, and discussion of the manuscript. The authors also need to discuss the sensitivity of their assay in more detail in the discussion and relate their results more clearly to environmental exposure concentrations.

Author Response

Dear Reviewer,

I want to thank you for review and comments

Author Response File: Author Response.pdf

Round 3

Reviewer 3 Report

The authors have not addressed any of the comments I made on the 2nd version of the manuscript. They need to be forthcoming in the abstract, intro and discussion that the exposure concentrations used to generate the data far exceed the solubility of these compounds in water, their assay is mechanistic (which is good!) but not very sensitive which limits its application.  

Author Response

Dear Reviewer,

We are sorry, but last time we uploaded the wrong file.

Author Response File: Author Response.pdf

Round 4

Reviewer 3 Report

Thank you for addressing my questions, I have no further comments to make on the manuscript.