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Thank you for addressing the last comments/suggestions.
[# PeerJ Staff Note - this decision was reviewed and approved by Xavier Pochon, a PeerJ Section Editor covering this Section #]
There a few minor issues to consider.
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Dear Editor and authors,
The manuscript has much improved since the first submission, I wish to thank the authors for taking the suggestions in good spirit and adapting the manuscript. I understand now more the reasoning behind the study design, methodology, and mostly the choice of temperatures in the experiment. It is also clear why my previous suggestion of incorporating the heatwave aspect is not possible in this case.
One remark which now remains is to look at instances where there is speculated about increasing temperatures (e.g. L123 (simple markup): “could affect”, L647: “would increase”, L657: “can be”), and rewrite these in the present tense, since the 20°C temperatures used in the experiment as “future increasing temperatures” are apparently already occurring frequently in summer.
Also, please check the last edited sections of the manuscript, these contain several instances of missing or double spaces and punctuation marks.
The authors have properly addressed all my comments.
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The authors have made a good kob replying to all the comments. I just include a few comments here. Lines refer to new ms:
L125. I still think that he natural conditions comment has to be removed as it has to be made clear that these conditions do not include tides or light.
L194. Why this measuring on one single day is representative, still has to be explained, even if previous experiments have done experiments ranging from a few days to several weeks or months.
L220:
What does "settled" mean? In any case, in order for diffusive oxygen fluxes to be affected (and as a consequence other anaerobic processes), it is not necessary to have a stratified column.
L233. Authors mention that they covered the cores during dark incubations. Did they allow for the system to adjust to these new conditions? If so, how long?
L260. The AFDW of benthic fauna is always considered after removing shells, on the flesh of the mollusks. It should be made clear if the entire animal including the shell was muffled. Please also mention the conditions for obtaining the AFDW.
L542. I apologise for the wrong reference. I was referring to this one doi:10.3354/meps192203 Christensen, Bjarne, Anders Vedel, and Erik Kristensen. “Carbon and Nitrogen Fluxes in Sediment Inhabited by Suspension-Feeding (Nereis Diversicolor) and Non-Suspension-Feeding (N. Virens) Polychaetes.” Marine Ecology Progress Series 192 (2000): 203–17
L669. I understand your point of maintaining this section but it should be made clear that predictions about the effects of climate change on the invasive algae are unknown or not taken into account when you are looking into the additional effect of A.m.
See above
See above
See above
Please provide a point-by-point rebuttal to all of the reviewers' comments along with your revised manuscript.
The article meets the basic reporting standards. Please see the PDF review for more extended comments.
The research question is relevant, but the experimental design does not fully match the original research question. In my opinion, the design better matches a heatwave experiment, which is why I suggest rewriting several sections. Please see the PDF review for more extended comments.
Experimental design does not allow for some of the conclusions being drawn. Please see the PDF review for more extended comments.
Please see the attached PDF review for additional comments.
Dear Authors,
I thoroughly read your manuscript and I find it clear and well-structured. The aim of your study was to describe the effect of temperature rise on the bioturbating activity of 4 common macrofaunal species in the Wadden Sea and on the nutrient fluxes and oxygen consumption. The manuscript is well-written, the language is clear, and in most cases, professional English was used. Methodology, figures, and tables are well-presented and it is easy for the readers to follow.
The objectives of the study and the hypothesis tested are well-defined and of great interest. The experimental design was appropriate for the confirmation or rejection of the hypothesis tested. Methods were described sufficiently, although in a few points extra info is needed.
The findings of the study are well-presented, although once again there is a part of the text that needs clarification. Moreover, post hoc comparisons are needed in cases where there is an interaction between the factors. Conclusions are well stated and supported by the results.
Specific comments can be found in the attached file.
The article is properly structured, contains appropriate figures and tables. Minor English corrections of some expressions required. Results are relevant to hypothesis.
The experimental design needs to be clarified further with potential problems in terms of the water column mixing and light/dark conditions. Other than that, although is is basic, only one day of fluxes, one temperature change non gradual change, setup seems correct. Interpretation should reflect the shortcomings.
Some corrections in all the aspects are needed.
There is one key point that the authors do make very clear though out the ms. Any increase in the fluxes is due to animal activities in addition to, usually, a stimulation of the sediment microbial activity. The somehow refer to it in L92 but do not discuss this in the results and discussion.
All parts of the manuscript can de reduced. There are repetition of concepts throughout (e.g. L66, L71, L102 all refer to ecosystem functions and bioturbation).
The selection of the temperature increase is not justified in the introduction.
The experimental setup has one serious flaw. Stirring of the cores is not continuous but only mixed at the beginning, middle and end, we assume which means that the water column become stratified. Also, was the water replaced when the nutrients sample was obtained? How was the calibration of the oxygen sensor made?
L231. Oxygen concentration should not go below 80% saturation because the oxygen penetration depth changes and alters the desnitrification-nitrification depth and associated fluxes.
Also an 80% saturation definitely does not make macrofauna enter anaerobic respiration, whatever this means, as they only respire oxygen. Nereis and Arenicola are known to tolerate very low oxygen and high H2S concentrations.
Animals collected in situ were found at a temperature of? Then the animals added in the "hot" temperature. This rather simulated a heatwave rather than a progressive temperature increase over decades. Animals will have more time to adapt. Authors should comment on this factor.
Fluxes and all calculated ratios etc should use the absolute values and not relative to the controls. Not only this allows for comparisons with other studies but also because subtraction changes the ratios. Also we should observed whether in controls fluxes increase or decrease with temperature.
To follow the tests made I think it would help if they report the factor, F value, DF. Also on some occasions they report exact values (preferable) and in others not (p<xxx).
One concept that is very little explored here is the effect on the microbial community itself. It is well known that bioturbation increases fluxes and metabolic rates not only by the animal's own metabolism and activities but also by enhancing microbial community metabolism.
Figure captions should be made more details. Figures 3 and 4 can be combined in two panels. We are unaware what the boxplots and whiskers exactly represent
No dotted line observed in Fig 6.
Fig 7 I guess the Note is wrong as it mentions "positive" into the sediment whereas in the entire article it is mentioned a that an efflux is observed.Please clarify this.
Table 1 and 3 no errors are given.
Figure 8. It is very hard to differentiate between treatments. Maybe authors should split temperatures in two plots per treatment and add at least an spline.
More comments is attached pdf
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