How Can Nanoplastics Affect the Survival, Reproduction, and Behaviour of the Soil Model Enchytraeus crypticus?
Round 1
Reviewer 1 Report
Review of the research paper on “How can nanoplastics affect the survival, reproduction, and behavior of the soil model Enchytraeus crypticus? by Angela Barreto, Joana Santos, Mónica J. B. Amorim, Vera L. Maria
My overall impression of the research paper is very positive. It considers a very important and urgent problem related to the assessment of micro- and nanoplastics effects on soil organisms. Authors also indicate the methodological problems which may lead to the misinterpretation of the results, especially due to toxic effects of accompanying substances like NaN3 indicating bactericide effects. The presented research paper is very interesting and very well designed. The statistical methods are appropriately applied and the results are clearly presented.
The only I recommend is the spelling correction of some words but it does not influence the overall positive impression.
Summarizing the review, I recommend publishing the paper in the present version.
Author Response
Manuscript ID: applsci-976266
Title: How can nanoplastics affect the survival, reproduction, and behaviour of the soil model Enchytraeus crypticus?______________________________________________________________
Comments of Reviewer 1
My overall impression of the research paper is very positive. It considers a very important and urgent problem related to the assessment of micro- and nanoplastics effects on soil organisms. Authors also indicate the methodological problems which may lead to the misinterpretation of the results, especially due to toxic effects of accompanying substances like NaN3 indicating bactericide effects. The presented research paper is very interesting and very well designed. The statistical methods are appropriately applied and the results are clearly presented.
The only I recommend is the spelling correction of some words but it does not influence the overall positive impression.
Summarizing the review, I recommend publishing the paper in the present version.
Answer: Thank you very much for the review and your positive considerations. We have reviewed the text language and style.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
I would recommend this article for publishing after a minor revision which should include a moderate English language and style editing. I think that the experiments and obtained results are well described. I do have several questions:
- " NPls concentrations <15 μg.L−1 have been predicted to be environmentally relevant concentrations [7]." In which environment? If in soil that concentrations in your experiments are very high.
- Why did you use standard error instead of standard deviation in your data analysis?
- "Would had been interesting to assess the avoidance for 900, 1200, 1500 like in the ERT range." I do not understand this sentence. You deliberately decided to use a different range of concentrations due to the results of ERT and then this sentence?
Author Response
Manuscript ID: applsci-976266
Title: How can nanoplastics affect the survival, reproduction, and behaviour of the soil model Enchytraeus crypticus?_________________________________________________________________
Comments of Reviewer 2
I would recommend this article for publishing after a minor revision which should include a moderate English language and style editing. I think that the experiments and obtained results are well described.
Answer: Thank you for the care that you have put in the review process. We did a careful spell check, which we believe improved the quality of the manuscript. We have answered all the comments below.
I do have several questions:
- " NPls concentrations <15 μg.L−1 have been predicted to be environmentally relevant concentrations [7]." In which environment? If in soil that concentrations in your experiments are very high.
Answer 1: The mentioned concentration refers to seawater. Despite having tested (also) higher NPls concentrations (>15 µg/kg soil), we included the nominal concentration of 0.015 mg NPls / kg soil (i.e., 15 µg NPls / kg soil) when we performed enchytraeid reproduction test (ERT). We intended to do a dose -response assessment for hazard characterization. Afterward, the estimation of the effect concentrations (ECx) based on reproduction or mortality was possible (Table S3-supplementary data).
2. Why did you use standard error instead of standard deviation in your data analysis?
Answer 2: Because we used a small sample of the whole population, i.e., n=4 replicates for each treatment analyzed.
3. "Would had been interesting to assess the avoidance for 900, 1200, 1500 like in the ERT range." I do not understand this sentence. You deliberately decided to use a different range of concentrations due to the results of ERT and then this sentence?
Answer 3: You are right. This sentence is out of context and it was removed from the manuscript (Results). We deliberately decided on the tested concentrations for avoidance assay based on the ERT data.
Author Response File: Author Response.pdf
