Supplementary Material
ESPR (Ref.: Ms. No. ESPR-D-12-01482). Reviewers’ comments
Thank you very much for the opportunity to revise the manuscript. We studied the comments carefully and tried our
best to revise and improve the manuscript according to the suggestions. A detailed list of changes and responses are
given below.
Dear Catarina Mansilha,
Reviewers' comments on your work have now been received. You will see that they are advising against publication
of your work.
Therefore I must reject the manuscript as is, but I encourage you to resubmit as a new manuscript after the
appropriate changes or research have been completed.
The reviewers' comments can be found at the end of this email or can be accessed by following the provided link.
Thank you for giving us the opportunity to consider your work.
Yours sincerely,
Dr. Henner Hollert
Editor
Environmental Science and Pollution Research
08/12/2012
Reviewers' comments:
Reviewer #1: General comments:
The MS submitted by Catarina Mansilha addresses a topic that has found considerable interest both, in the scientific
community and in the lean public. The results as such are convincing and the MS sets it to a round figure but only as
long as the phenomenological level is considered. The problem begins when taking mechanistical aspects in
consideration. This and further topics explained below deserve attention by the authors when revising the MS. I
personally have little doubts that is will be a valuable contribution to the field in a revised form and an asset for
ESPR. General remarks:
*
The choice of daphnids as test organisms is poorly motivated. The existing information from chemical testing,
endocrinology and genome analyses indicate that (1) Daphnia does not specifically respond to estrogenic compounds
in general and BPA in particular (cf. Caspers (1998): No estrogenic effects ?), (2) estrogens do not have a functional
role in crustaceans and (3) there is no estrogen receptor orthologue in the genome or transcriptome of any crustacean
species analysed so far (cf. reviews on this topic, for example Oetken et al. (2004): International Review of Cytology
236, 1-44). Despite the lack of molecular reasoning for the choice of the test species there are few studies indicating
that crustaceans may respond to estrogens for example with an increased reproduction (cf. Andersen et al. (1999):
Ecotoxicol. Environ. Saf. 44, 56-61). I miss any of this information in the discussion of the MS which should be
amended accordingly.*
In line with this statement, the literature cited in the current version of the MS is rather
biased. There is an apparent emphasis on technical papers such as guidelines while original papers or reviews are
under-represented.
It is true that we can found in literature some controversy about the use of daphnids as test organisms. However, as
stated in introduction section, the newly revised OECD Conceptual Framework adopted Daphnia tests as guideline
methods - level 5 - for testing and evaluation of adverse effects of EDCs).
A revision was done according to reviewer´s instruction and some references, as Oetken et al. 2004, were included in
the text, as suggested.
*
Furthermore, I do not see any justification for the acute tests within this study. I propose to delete them
completely and to use the free space for a more in-depth and detailed mode-of-action-based discussion of the results
from the chronic testing.
We evaluated carefully the possibility of withdrawing the acute tests, but we consider that they are very important as
a basis of comparison with previous studies, and to validate the remaining ecotoxicological tests’ results
*
Three key words are already part of the title and can therefore be deleted/substituted by other phrases.
Two of the key words that were already part of the title were deleted/substituted, however we decided to maintain
BPA
*
Starting from page 6, line 23 all BPA test concentrations are provided in µ/L. Given that you lowest
concentration is 1000 µ/L it would be more straight forward to provide all test concentrations as mg/L.
The correction was done as requested
*
The language needs substantial corrections. The authors should seek advice from a native speaker.
As suggested, English was improved according reviewers` instructions.
Specific comments (by page and line):
Page 1:
Abstract:
l. 27: replace "exposition" by exposure" here and throughout the MS (e.g. p. 7, l. 26)
The correction was done as suggested
l. 29: what do you mean with "reproductive burden"? Is it rather "impact" or "effects" you are referring to?
Yes, it was our intention to refer the impact of BPA
l. 31: replace "specie" with "species"
The correction was done as suggested
Page 2:
Introduction:
l. 3f.: shorten the first sentence to "Plastic materials are considered extremely convenient for food storage."
The correction was done as suggested
l. 5: "polymers made from plasticizers' compounds can be leached" - This sentence is a complete misunderstanding.
Plasticizers are either the monomers for the synthesis of plastics (such as BPA in the case of PC) or additives (like
phthalates, flame retardants, etc.). The migrating or leachable part of plastics are not the polymers but typically
monomers or the short short-chained polymers.
The correction was done as suggested
l. 8-12: I doubt whether you can address the "real overall food contamination and integrated cumulative effects" in
your study using Daphnia as a model organism because the protection target when testing plastic materials for food
stuff are humans. Be modest in this your ambition.
It was not our intention evaluate the “real overall food contamination and integrated cumulative effects". It was just a
statement. The text was changed.
Page 3:
l. 9: replace "as a biomonitoring species" with "for standardised toxicity tests"
The paragraph was changed
l. 12: replace "reach" with "reaches" and "of leaving" with "after leaving"
The correction was done as suggested
Material and methods:
Page 6:
l. 46: replace "mater" with "the parent"
The correction was done as suggested
Results and discussion:
Page 8:
l. 43f.: replace "it was not necessary any procedural review" with "no procedural review was necessary"
The correction was done as suggested
l. 52ff. join the first two sentences of the paragraph: "Table 3 summarizes the results of the high temperature
migration tests [not "testes"], showing unequivocally ?"
The two sentences were joined as suggested
Page 9:
l. 2: replace "no-reactive" with "non-reactive"
The correction was done as suggested
l. 37f: replace "There were obtained the followed values:" with "The following values were obtained:"
The correction was done as suggested
Page 10:
l. 21: replace "valid" with "significant"
The correction was done as suggested
Page 11:
l. 2f.: what do you mean with "offspring in plastic containers belong to the same population as control."? Do you
mean: "offspring numbers in plastic containers is significantly different from the control."?
As suggested, the phrase was rewritten.
l. 3: delete "Results suggested two subpopulations."
The correction was done as suggested
l. 3f.: replace "It was observed a significantly higher offspring in ? and C-3 when compared with control." with "A
significantly higher offspring number in ? and C-3 was observed when compared with control."
The correction was done as suggested
l. 20: replace "There were used immature juveniles" with "Immature juveniles were used"
The correction was done as suggested
l. 46: replace "all over" with "over all"
The correction was done as suggested
Page 12:
l. 6: add "et al." after "Kortenkamp" here and also on p. 12, l. 18; p. 13, l. 23 and 29.
The correction was done as suggested
l. 7: replace "remain" with "remained"
The correction was done as suggested
Conclusions:
l. 28: replace "A method" with "An analytical method"
The correction was done as suggested
Page 13:
l. 1-16: I propose to delete the first two paragraphs on this page completely. Due to the lack of any mechanistical data
this is pure speculation which is not appropriate for the conclusion chapter.
As suggested, the two paragraphs were deleted.
Page 17:
Figure captions:
l. 21: delete "all" after "brood size"
The correction was done as suggested
Figure 3: Please add information which correlation is shown specifically ba the two lines in the inset: multigeneration
study in the upper part, single species in the lower part.
As suggested, information was added.