Impact of Crude Oil and Dispersant Use on the Baltic Sea Blue Mussel Mytilus trossulus Microbiome
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsIn my opinion, the article is clearly written and ready for publication.
But probably the subsubsection numbers in subsection 3.3 (line 221 and line 240) are missing.
Author Response
In my opinion, the article is clearly written and ready for publication.
But probably the subsubsection numbers in subsection 3.3 (line 221 and line 240) are missing.
Thank you for noticing this, we added the missing subsection numbers.
Reviewer 2 Report
Comments and Suggestions for Authors
The present article, entitled "Impact of crude oil and dispersant use on the Baltic Sea blue mussel Mytilus trossulus microbiome", describes the effects of exposure to water accommodated fraction (WAF) of crude oil and chemically enhanced water accommodated fraction (CEWAF) of crude oil on the mussel microbiome, with a particular focus on the abundance of hydrocarbon degradation-related genes in the bacterial community.
The manuscript is relatively well written (by English standards) and generally easy to understand. The area of study is interesting and the study has produced some interesting results. However, I have some concerns about the methodology used. It may well be that the authors have done the necessary validation work on which the assay is based, which could be included in a revised submission.
The main considerations raised after reading the manuscript are as follow:
Introduction: Please improve references to the paragraph.
Material and Methods: The Materials and Methods section should be enriched:
Line 137: The reference article is not open access, so more information about the primers used should be added.
Line 138: Add the slope values obtained for each standard curve and their corresponding efficiency.
Section 2.4: Add the description of the number of samples treated for analysis and the number of replicates.
Table 1: Add Deviation Standard Values
Line 224: Is it possible to reduce the percentage of "others" by lowering the cutoff to 0.1?
Discussion: Please improve references to the paragraph.
Author Response
The main considerations raised after reading the manuscript are as follow:
Introduction: Please improve references to the paragraph.
We added a recent publication that was missing (Palladino et al. 2023, line 43)
Material and Methods: The Materials and Methods section should be enriched:
Line 137: The reference article is not open access, so more information about the primers used should be added.
We added information about primers and a reference (line 141).
Line 138: Add the slope values obtained for each standard curve and their corresponding efficiency.
We mention the amplification efficiency of the qPCR reactions, which was 100% ±10% for all runs. It is considered to be acceptable since we have the method in routine use. If the qPCR efficiency is within 100% ± 10%, the acceptable slope range for the standard curve is approximately -3.1 to -3.6. These values are dependent of each other so we considered that one of them is sufficient in text.
Section 2.4: Add the description of the number of samples treated for analysis and the number of replicates.
We added “10+10 mussels were pooled from two replicate aquaria per treatment.” to the text. Due to the small size of the Baltic Sea blue mussel pooling is needed for chemical analysis.
Table 1: Add Deviation Standard Values
The table shows results of all replicates, so after careful consideration we decided that standard deviation is not necessary.
Line 224: Is it possible to reduce the percentage of "others" by lowering the cutoff to 0.1?
We tested different values and considered that 0.5 is good for the clarity of the figure.
Discussion: Please improve references to the paragraph.
Thank you for pointing out this. We have now included more relevant citations (lines 327-328, Palladino et al. 2023, Capolupo et al. 2017, Shukla et al. 2007, Gan et al. 2021, Ramachandran et al. 2006, DeLorenzo et al. 2021).
