https://orcid.org/0000-0002-1983-6022
Peer Review History
| Original SubmissionJanuary 22, 2025 |
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Is the manuscript technically sound, and do the data support the conclusions? Reviewer #1: Partly Reviewer #2: Partly ********** 2. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: I Don't Know Reviewer #2: Yes ********** 3. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes Reviewer #2: Yes ********** 4. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes Reviewer #2: Yes ********** Reviewer #1: I finalized revising the manuscript “Assessing agricultural effects on benthic invertebrate communities in ponds and ditches using δ¹⁵N and δ¹³C isotope niches”. The topic is interesting and attractive to a broad audience. However, major improvements need to be done in the manuscript. The questions and hypotheses, and how the methods were applied to solve these questions, were rather cryptic for me. It was generally hard to understand from the introduction which is the knowledge gap the authors are trying to fulfill. Moreover, without clear questions, assessing the science behind the methods and results was difficult. My major misunderstanding is why the authors expect differences between the two types of ecosystems assessed, and therefore, why this is an interesting question in the framework of agricultural stressors. I think the authors have an interesting and valuable dataset with the potential of answering interesting ecological questions. However, significant work needs to be done. I left detailed comments for each of the sections. Abstract 14: Why do ponds and ditches harbor particularly endangered or scarce taxa? Moreover, why do you mean by scarce, is it low abundant taxa or taxa with low occupancy? I’d recommend removing this last part of the sentence. 16 I disagree. Numerous studies have shown the adverse effects of agricultural land use in biodiversity. Perhaps what you mean is that these habitats are likely to face multiple stressors, all derived from agriculture, which results in a problematic complex to address. More importantly, the reader does not know so far which are these previous approaches and why they were not successful. For example, what do you mean by “classic structural biodiversity metrics”? 19: We analyzed the carbon and nitrogen stable isotope composition of the invertebrates and the pesticides and nutrient residues of water. 21 readers do not know what are Standard ellipse and Layman B community metrics. I recommend leaving technical terms out of the abstract. 22 FFG should be introduced earlier. 28 Which differences? You’re leaving the reader hanging on the unexpected differences but don’t tell them (us) which. 28-32 See my previous comment. These sentences are uninformative for readers. We don’t know why case-by-case patterns emerged, or where the differences lay. What do you mean by the production or consumption level? How was production measured or estimated? We also don’t know which strong fluctuations on parameters did you observed; therefore, we don’t know how did you reach this conclusion. 30 Remove In the end Introduction 40 is it specialization, and use of fertilizer and pesticides homogenizing the landscape? I’d say that the expansion of agricultural land is what homogenizes the landscape. Ref 3 43 What do you mean by semi-natural habitats? Under less intensive agriculture? 45 Starting a paragraph with an example is quite counterintuitive. You haven’t introduced yet non-productive areas, so why are you giving us examples? 49 Same as in the abstract: would you define scarce more precisely. 50 a high richness compared to what? Also, these groups are highly abundant in many aquatic ecosystems; therefore, why are they particularly important in ditches? 51 I think this is your topic sentence for this paragraph; therefore, it should be the first one -or maybe the second one, but at the beginning! 53 but this contradicts your previous sentence in line 48 57 in which sense is challenging? Actually, invertebrates are one of the most used groups for biodiversity assessment because sampling is comparatively easy and cheap to other taxa, and they are very good indicators of ecosystem status and functions. 59 But there is extensive literature showing the effects of agriculture on biodiversity. I think you need to be more specific as to why functional approaches outperform, or complement, taxonomic approaches. For example, why would functional approaches be more sensible to persistent stressors or short gradients? Ref 8 64 why were these other traits not included in your analysis? One important effect of agricultural land use is the change in inputs of organic matter to the water bodies; this justifies the use of feeding groups. However, other effects are channelization or modification of the physical structure, which might produce a response in locomotion; changes in productivity (eutrophication), with effects on body size and respiration; changes in temperature, with effects on thermal tolerance and respiration. In other words, you need either to include more traits, or to justify why feeding groups are the best approach to address the effects of agriculture, including persistent stressors and short gradients. 80 this paragraph is disconnected with the rest of the introduction. While important concepts need to be defined, the reader wants to know how the niche concept(s) is related to your own work and very importantly, to your question and knowledge gap. It is your task, as writers, to tell the reader how this concept can help understanding effects of stressors in biodiversity -not give the reader homework . For example, I’d recommend combining this paragraph with the trait paragraph, since the niche is going to be defined based on those traits. This will provide more coherence between your paragraphs and shorten your introduction. 88 we used. You haven’t defined isotopic niche nor realized niche. Feeding resources? And which habitats? Do you mean between ponds and ditches? If so, why feeding resources and, therefore, isotopic niches would differ between them? 89 we compared 90 which groups? 92 are you assessing more than one stressor? Or is it pesticides the only metric you are using? 93 we asked. can 94 I don’t fully understand your questions. Are you assessing isotopic niches in a gradient of agricultural stress, as measured by pesticides? Why do you expect differences, or not, between ponds and ditches? 96 to answer these questions… 98 what is the difference between niche differentiation and niche specialization? And why reduced niche specialization would result in less overlap? I’d expect the opposite. How about the expectations for ponds? Do ponds have different effects of agriculture? It is not clear to me which questions are being answered and how. Is it the question related to differences in niche width due to different levels of agricultural stress? Then, would these effects differ between ponds and ditches? Why? What do you mean by habitat variability? Is this variability inside each water body, or among types? Methods 103 I think that saying During the 1980s, the wet lowlands… is enough. Moreover, I’d turn the sentence around: The wet lowlands in the Havelländisches region were drained … during the 1980s. 105: What do you mean by traditional grasslands? As in, they were already agricultural lands but transformed into monoculture crops? Or were they natural grasslands converted to agriculture? 107 What about ditches? 113 describe the site’s characteristics in order, from Pond 1 forward, and ditch 1 on. Table 1: How were these habitats characterized? Do they correspond to the littoral zone, the whole water body? Which area of the water body was estimated or quantified? Same for the riparian zone: which distance from the shore was considered riparian, and how these percentages were estimated/quantified? You could highlight the dominant type of riparian vegetation to help readers focus. 124 Benthic invertebrates were sampled using hand netting… 126 meaning that samples were composed by combining all habitats? 126 how were feeding groups distinguished in the field? Based on morphology? 129 how is dispatch with boiling water humanely? To what end were they kept alive for 24 hours? And why did you use this boiling water method, instead of preservatives like alcohol or formol. This is related to the isotopic signal, but the reader does not know this. 131 Explain why their shells were remove. 149 I don’t see how conductivity and pH can affect the food web structure or isotopic niche. Oxygen and temperature, indirectly, if they reflect productivity? 175 Why was toxicity for macrophytes and algae estimate? 187 So far, FFGs have not being defined or described. What are they, what do they represent, how do you assign an organism to a FFG, or which categories exist. This table should be probably moved to the Results. 188 family determination based on which literature? FFGs can vary in different regions because organisms are flexible in their diets, even given certain morphology. Moreover, they can usually be assigned to more than one category. Even in the book of Merrit, Cummins, and Berg (the “inventors” of FFGs), taxa are usually categorized in more than one group. Table 3. CPOM and FPOM are in the legend but not the table. Columns after the TU are not explained. How were all these metrics quantified and to what aim? Move to Results. 200 was observed 201 of all invertebrates or for primary consumers? This does not seem to be an appropriate justification, from my point of view, precisely because you are looking for potential differences between water body types. To my knowledge, there are two ways of correcting dN: first, by using a basal resource dN value as a baseline; this might have the problem of having different values among sites, if they differ in the amount of fertilizer. Second, using the dN value of the primary consumer with the lowest dN to obtain comparable trophic positions across sites. 204 difference between what and what? 209 what difference? 210 an alternative is centering or scaling the deltas of all sites to obtain comparable values, but I don’t understand why to alter the variability of the sites showing effects of agriculture, if that’s your question? Moreover, it seems odd and a bit arbitrary to transform values of one site. 212 Explain what these analyses are and why were they performed 233 what information 235 to what end? What is the information provided by these metrics? 237 I don’t understand why do you want to correct for the effects of agriculture? Also, is this another correction or the same one mentioned in the previous subsections? Results 265 it hasn’t been explained or justified why these two types of habitats would differ in the isotopic composition Figure 4 the resolution of the figure is very poor, please improve it. 271 replace clearly by a more appropriate term, like statistically 274 … for the MNND (Fig 5c)… 275 evenness is Fig 5f, not e 284 Which clear differences, between what and what? Please, try to avoid using words like “clear” or “clearly” since patterns might not be as clear or obvious for the readers. Figure 6: the boxes correspond to SEA.B or SEAc? 284 this paragraph needs to be more organized. It is mixing the isotope values with abundances and diversity for one FFG. 306-309 these sentences are quite hard to read; please, revise the grammar. 321 were the explanatory variables scaled before running the analysis? Discussion 327 but how was this tested? 339 why did you expect more abundance and diversity in ponds? 341 was any of this measured? Is the relative abundance of herbivores significatively higher in ditches? This all seems very speculative 346 homogenous in which sense? How was homogeneity measured? 347 then, why citing 46? 349 I don’t think secondary production is an appropriate term here, since this was not measured. Secondary production is a complex metric requiring, at the very least, biomass estimation. 351 again, secondary production was not measured. 355 this paragraph is generally confusing. I’d recommend you explain your results at the beginning, as a topic sentence, and then develop the ideas that support your findings. In the current version, sentences supporting and contradicting your findings are mixed making it hard for readers to get the main message you want to transmit. 356 is there a better term than indifferent? I think generalist could be more appropriate 357 meaning enriched values either due to high levels of omnivory or secondary predators. 360 herbivorous feeding preference 364 but you measured pollution. This is something you should be able to test. 368 why is this related to wider habitats? 370 lower diversity of littoral microhabitats in ditches? You started your paragraph talking about ponds. 373 a well-documented effect of eutrophication is the development of cyanobacteria, that outcompete other primary producers, but are commonly toxic for animals, explaining why eutrophication could actually reduce grazers diversity. 374 it is not clear what do you mean by wider habitat? Is the width of the water body, or the number of microhabitats available? 374 higher taxa richness where? These results were not presented before 375 what is a taxa group? 381 could pesticides have important direct effects on the invertebrates, leading to higher mortality rates, in addition to the indirect effects through primary production? 386 it makes sense for me to have higher variability in these groups related to their diets; they are after all generalists 393 Why? 405 I am surprised to read here that ponds were supposed to be a reference community. This kind of statement should be mentioned very early in the Methods or Introduction. Moreover, I don’t think ponds embedded in an agricultural landscape could be used as a reference, less impacted community by agriculture. I’d suggest using the water bodies as a gradient of agricultural effects, including nutrient enrichment, pesticides, and the proportion of surrounding crops to test your hypotheses. Reviewer #2: Manuscript: Assessing agricultural effects on benthic invertebrate communities in ponds and ditches using δ¹⁵N and δ¹³C isotope niches Authors: Fee Nanett Trau, Kathrin Fisch, Stefan Lorenz Overall, the manuscript presents interesting and valuable research, but several key areas require further clarification to improve its structure, transparency and readability. The study's aim is currently too vague and should be made more specific, particularly in regard to which aspects of benthic invertebrates are being investigated. The introduction would benefit from a clearer distinction between ponds and ditches, specifying their purpose, shape, and typical formation or usage. In the methods section, there is a lack of clarity surrounding the toxicity calculations mentioned. It is important to explicitly connect these methods to the study's objectives so the reader can follow the logic of your approach. At the moment, the description of toxicity tests, seem disconnected from the rest of the paper and need to be more clearly integrated into the study's goals. If toxicity testing was conducted, please describe the specific methods used, including whether test organisms or particular pesticides were involved. Further, the process for sorting organisms into taxonomic and functional feeding groups (FFGs) should be described more clearly. The use of boiling water as a method needs more explanation- if this method is employed instead of freeze-drying, please justify why it was chosen and explain how it might affect the isotopic analysis. It would be valuable to emphasize whether the observed differences between ponds and ditches are statistically significant. Discussing the statistical significance of these differences would provide a clearer understanding of their reliability and help contextualize the results within the broader field. Additionally, providing information about the statistical tests used, the p-values, and any relevant confidence intervals could strengthen the interpretation of the findings and give readers more confidence in the conclusions drawn. Some aspects of the figures and tables require additional detail. For example, Figure 1 should have a more informative caption, specifying what the red labels represent and distinguishing between the different symbols. Some Tables also need clearer explanations. In addition, there are a number of minor issues that need addressing. It would be useful to include more details on the taxonomic composition of riparian vegetation, ideally at the family level. The citation format should be checked according to the authors' guidelines to ensure consistency. Also, please specify the number of replicates used at each location and for each taxonomic or functional feeding group, as well as the amount of material weighed for each replicate sample. More specific comments and suggestions are provided in the attached PDF. Addressing these points will strengthen the manuscript, making it clearer, more precise, and easier for readers to follow. ********** what does this mean? ). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy Reviewer #1: Yes: Daniela Cortes Guzman Reviewer #2: No ********** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". 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Dear Trau, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. ============================== ACADEMIC EDITOR: Dear authors, many thanks for revising your manuscript. Two referees have evaluated it again and despite some improvements were made, the manuscript still needs significant work before its consideration for publication, particularly in terms of writing and interpretation. I invite you to go in detail through both reviewers’ comments and suggestions and modify the manuscript accordingly. Please submit your revised manuscript by Jul 12 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.
If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols . We look forward to receiving your revised manuscript. Kind regards, Marina Vilenica Academic Editor PLOS ONE Reviewers' comments: Reviewer's Responses to Questions Comments to the Author Reviewer #1: (No Response) Reviewer #2: (No Response) ********** 2. Is the manuscript technically sound, and do the data support the conclusions??> Reviewer #1: Partly Reviewer #2: Yes ********** 3. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: No Reviewer #2: Yes ********** 4. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes Reviewer #2: Yes ********** 5. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: No Reviewer #2: Yes ********** Reviewer #1: The manuscript has improved. The authors addressed some of the concerns; however, considerable work is still needed in order to have a manuscript ready for publication. It seems to me that some changes in the manuscript were made to please the reviewers, but without giving deeper thought about why these requests were made. In that sense, changes lack attention to detail. For example, Table 3 was moved to results, as suggested, but it is not described, and Figure 4 is missing. Furthermore, I disagree with the interpretation of results the authors made. They found statistical differences in several metrics yet concluded that there are no clear patterns. I have several major and minor comments across all sections that I detailed here: Abstract 15: the last part of the sentence, after landscape is a bit redundant and unnecessary; it could be removed. 15-20 I still find the wording here difficult to justify the use of functional approaches (or not use of taxonomic ones). Might I suggest a phrasing like: Biodiversity metrics based on taxonomy, such as richness or evenness indices, might fail to describe changes in ecosystem functions that result from the effects of agriculture. In contrast, functional approaches, such as the use of functional feeding groups and isotopic composition, better describe the use of resources by the invertebrates, improving our understanding of the influence of agricultural stressors on aquatic communities (just a suggestion, please use your own words) 29 we still don’t know why these differences are unexpected. I suggest removing that word. 33-34 this seems to be part of your results, rather than a general statement. I would write it in past tense. More importantly, this might be, in effect, a consequence of your small number of water bodies. Therefore, I don’t think your sentence at line 31 is justified: communities respond in complex ways to stressors… since this is, again, an effect of the sample size. I suggest re-ordering this last section of the abstract to provide more accurate conclusions. For example: The strong fluctuations in biotic and abiotic parameters resulted in pronounced differences among water bodies, but no clear patterns between water body types. However, larger datasets… However, after reading your main text, I disagree with this conclusion. Introduction 51 water bodies. Suggested phrasing: Although these small water bodies are usually not included in monitoring programs (5,7), their importance for benthic macroinvertebrate biodiversity and ecosystem services is increasingly gaining recognition in research activities (5,8–13). 53-56 These last sentences are a bit repetitive. It was already stated that these ecosystems are valuable habitats for invertebrates. Moreover, the sentence structure is a bit confusing. I think mentioning the orders at the end of the sentence is unnecessary since, as I have suggested before, these are abundant and common groups in many aquatic ecosystems. My previous comment was not meant to have an aesthetic change in the wording, but to justify the inclusion of such a sentence in your introduction. I recommend closing the paragraph with a statement highlighting the knowledge gap that the manuscript is addressing. 58-59 I still disagree on that it is challenging being one of the most common bioindicators. I think you could instead jump directly into the previous findings and avoid this sentence. 62-63 long-term persistent stressors would be likely reflected in taxonomic diversity or EPT, whereas short stressor gradients might result in stochasticity, for which functional approaches consistently reflecting habitat types and resources might be a solution 69 the acronym FFG has not been defined in the introduction. Please do so 69-71 this sentence does not belong here since you haven’t told us yet anything about the water bodies under study. The justification for the use of FFG, from an ecological point of view, is your next sentence. 73 explain why they reflect the effects of agriculture. Is it because of the change in quality, quantity, type of feeding resources? 74-78 this definition of niche could be better described in terms of the feeding/trophic niche, given that it is the topic of the manuscript and to better connect with the previous section of the paragraph. 91 please define realized ecological niche 92 previous sentence was spelled realized... please, confirm with the journal British or American spelling 93-94 not necessary if the niche is explained in that context in the previous sentences 95-98 this paragraph could be written more concisely. Suggestion: We aimed to assess the influence of agricultural stressors, measured as water toxicity from pesticide residues and eutrophication, on the isotopic niche of FFGs in pond and ditch communities. 106-108 this phrasing is confusing. In my understanding, is not the body type, pond or ditch, what is used as a gradient (since this a categorical factor with only two levels; if so, which is higher and which lower?). It is the continuous values of nutrient enrichment, pesticides, and the proportion of surrounding crops. 111 I am not sure I understand your second question. Do you mean that, on one side, you expect differing effect of agriculture between ditches and ponds on the food web structure (Q1 and H1), but similarities in the isotopic niches of specific FFGs (Q2 and H?)? 114 ... and presence of generalists compared to ponds. To keep parallelism and include the ponds in your hypothesis. Methods 131 and ditch 4? 149 specimens 150 otherwise? You collected as many as possible or discarded samples that did not reach 10 individuals? 159, 161 grounded 163 10-15 samples, even if they belonged to different genera? 164 isotopic... were carried 177 that can explain... I still don’t understand this. Why do you expect that conductivity and pH explain the food web structure? Your answer was unfortunately not sufficient. I know these are standard variables typically measured during sampling, but that does not justify their inclusion in your statistical analysis. I could understand that you included them to control for intrinsic differences among water bodies; therefore, they are not explanatory but control variables. 253 explain why these metrics were calculated, or what information did you get from them. 269 check parallelism of the sentence 289 why do you have small samples sizes in EC and oxygen if they were measured with a probe? Results 293 You have to describe the table, not just reference it. For example, what common patterns did you observe across ponds, and in comparison to ditches? Which type of water body tended to have higher nutrient concentrations, toxicity, or pesticides? Table 3. For the sake of completeness, describe all acronyms. Sample was used before to describe the individuals analyzed for isotopic composition. I recommend using here Site or water body. C° are the units of temperature. To keep it consistent with your other columns, use T [°C] or something similar. Fig 4 is missing. Is it maybe Fig S2? If so, could you show the biplot after correction? Maybe this will distinguish 313 dC13 range is 5d not b 316 whereas 324 is this Figure S3? What different information is provided between Fig 5 and S3? It seems that many of the differences shown in Fig 5 disappeared in S3. This should be important to explain. In a more general comment, all tables and figures in the text are meant to be described and explained, not just mentioned. 326 since Fig. 6 is comparing FFG among wb types, I suggest placing each FFG in pond and ditches next to each other, and distinguish them with colors, as you have done. This way is easier for the reader to notice the differences between, e.g., predators between water body types. You also need less labels in the x axis this way. 327 I disagree with your initial statement. I think there are differences between ponds and ditches, as you mentioned in the following sentences. 332 what about the diversity of the other groups? Also, you didn’t really quantify abundance, since you stopped gathering individuals when you collected 10. 342 for all FFG, overlap was greater when corrected for both C and N. Moreover, why this comparison between corrections is important? Is one correction providing different information than the other? Why not simply correct by both and compare overlaps? 348 a lower ... in ponds compared to ditches, and higher... in ditches... 351 higher 354 this is the description of table 3 that should be with the table, not in a different section. Discussion 366 what exactly do you mean by comparable? The isotopic niche width? However, you found marked differences in Fig 5 369 I don’t agree with this interpretation. Again, you found statistical differences in some niche metrics and in some FFGs. 376 since the isotopic niche is indirectly measuring feeding resources, I’d recommend sticking to that word and not use it together with habitats. 381 what patterns did you expect to find? Since your main message seems to be that there were no differences among habitats, despite several metrics in your results showing differences, I wonder which were the patterns you expected to observe? 382 however, in your introduction you stated that ditches could also harbour high diversity. Therefore, why is this contrary to your expectations? 384 but is this true? Did you observe more plants or algae in ditches? Did you observe higher diversity of primary consumers? Moreover, this is assuming that the food web is based on primary production, but disregarding the role of detritus to sustain aquatic food webs, this is, all allochthonous inputs. 393 but you didn’t compare species richness 396 what do you mean by more homogeneous communities for predators? This term could be confusing, and it is not to be compared to having lower diversity. 400 this paragraph is describing the results from the figure, not discussing the further implications of such results. It should be moved to the Results section and discussed in the Discussion. Moreover, I failed to see why the ample preference of omnivorous is restricted to polluted waters, since they are, by definition, feeding on multiple types of resources. 410 same, these are Results. Please reference the figure or table that they refer to. 414 as mentioned before, I recommend you keep your interpretation in terms of the feeding resources, not the habitats or microhabitats 415 but you found higher diversity in ponds. Where is the link with the lake’s reference? 418 what do you mean by primary producer-rich? As in having higher primary production? Measured how? Or as in having higher diversity of primary producers? Measured how? 427 see my previous comments on this conclusion 436-439 these sentences are disconnected to the starting sentence of the paragraph. 443 explain why their food resources would be low in November 444 in deed, organisms need current to filter; therefore, it is expected to have low densities of filterers in water bodies without current. 448 explain why sampling in autumn affects the detectability of invertebrates and pesticides 456 explain why is this recommended Conclusions If the other sections are revised, this section would also require modifications accordingly. Reviewer #2: Dear Authors, Thank you for your thorough revision of the manuscript. It is evident that you have made significant efforts to address the reviewers' comments, and many aspects of the manuscript have improved as a result. That said, there are still several points that require further attention to enhance the clarity, coherence, and scientific rigor of the work. Most notably, improvements to the language are recommended to increase readability and avoid unnecessary or unclear repetitions. In particular: _Some sentences are redundant, as the information they convey has already been clearly stated in preceding text. _Paragraph structure can be improved by shifting certain sentences to new paragraphs for better flow and thematic clarity. _Several sentences would benefit from rephrasing to improve clarity and avoid grammatical issues such as the repetitive use of conjunctions (e.g., multiple occurrences of “but” in the same sentence). _In a few instances, sentences are overly long and difficult to follow, and should be restructured into more concise and digestible forms. _All abbreviations should be defined at their first mention in the text to ensure clarity for all readers. _Certain terminology (e.g., "realized niche") may be unclear in this context. More precise phrasing is suggested to better convey the intended meaning. _Some key methodological choices would benefit from the inclusion of appropriate references to justify their use. _As a general comment, enhancing the narrative structure and clarity of expression will improve the overall impact of the manuscript and make it more accessible to a broader readership. A careful final language edit is strongly recommended. For more detailed suggestions and specific examples, please refer to the attached PDF version of the comments. ********** what does this mean? ). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? 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| Revision 2 |
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Dear Dr. Trau, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. ============================== Please submit your revised manuscript by Aug 23 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.
If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols . We look forward to receiving your revised manuscript. Kind regards, Marina Vilenica Academic Editor PLOS ONE Reviewers' comments: Reviewer's Responses to Questions Comments to the Author Reviewer #1: (No Response) ********** 2. Is the manuscript technically sound, and do the data support the conclusions??> Reviewer #1: Partly ********** 3. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: No ********** 4. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes ********** 5. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes ********** Reviewer #1: I have finished reviewing the manuscript for the third time. The manuscript has improved compared to the previous versions. However, I still have significant comments that, in my opinion, must be addressed before publication. I regret to say, once more, that the authors’ responses are occasionally not satisfactory. Moreover, while I appreciate their responses and explanations directed to me, my request and questions are finally made to improve understanding from the readers. In that sense, replying to me, but not fully addressing the issues in the text is not helpful for future readers of their work. Some explanations and changes made from the previous version to this one turned out to be more confusing to me. Specifically, the maize corrections for C. Some analyses and figures are presented with the correction, which leads to the conclusion that there are not differences between water body types, whereas others are presented without the correction, in which case significant differences emerged. In my previous comments, I asked about the different results when correcting versus not correcting, and the answer was that it was only intended as an additional visualization. However, the conclusions are dramatically different between the two methods. Additional comments are detailed below. Abstract 15 delete only, as they are not only based on taxonomy, they also account for abundance. 30 again, you didn’t measure abundance because you stopped counting when reaching 10 individuals. They were more diverse because you identified more taxa. Introduction 54 for measuring biodiversity in ponds and ditches affected by agricultural activities… or something similar that provides context for the sentence. 86 taxonomic group since taxa is plural 87 for resource use or consumption 91 the habitat use is not reflected in isotopic compositions because it is not incorporated in the tissue. This applies for the rest of your ms, when you mention habitat use or habitat diversity. For this reason, I recommended in my previous comments to keep the terminology as feeding resources and remove the habitat/microhabitats. 96 connect this sentence with the following paragraph 100 morphological characteristics 101 in addition, ditches are probably more contaminated… 103 protected by a riparian buffer? 104 act as a source of pesticides? How? 109 I personally find the word comparable vague because I don’t know if you mean they can be compared with one being larger than the other (which, of course, they can, using isotopic compositions!) or they are similar. Methods 150 necessary for the isotopic analyses. Moreover, how did you estimated the weight in the field? 148-149 however, in your response to my previous comment, you said that taxa were actually sorted more than at the family level (i.e., you did not gather individuals from different genera in the same group). 179 that could explain 225 by lack of information or because the family includes species from different feeding groups? 262 move this paragraph up to line 258 269 what is the difference from TA from the previous paragraph? Were the metrics in the previous paragraph not calculated as well for FFG in ponds and ditches? 272 meaning that in practical terms, TA, SEA.B and SEA.c provide the same information: niche width. Any reason for including the three of them? 281 I do not follow this new justification. Both C- and N-corrections were made because one water body showed enrichment of either C or N. Then, why now is only one being considered? Moreover, if the C-correction was not used in the analysis, remove the section where it is explained. However, in your Fig. 4 there is a clear difference in the isotopic space with and without correction, so it does have an effect. 294 my previous question was not answered. Why EC and O2 have small sample size but temperature and pH don’t? they were all measured with a probe. Additionally, in the text and in your answer, it was stated they were measured once, then why there is variability during the day? 305 this are the number of samples, correct? Then, I recommend not using N since it might be confusing with nitrogen. Results 310 then again, you said that the maize correction was not necessary. 317 significantly distinct 319 closing dash is missing (and they should be m-dashes)… are significantly larger… Why is Fig. 5 again only corrected by N? It is very confusing how the authors are mixing results regarding the corrections. This makes very hard to assess the outcomes of these analyses. 331 …for primary consumers, a mean value 26.5 N for omnivores, and a mean value of 42.25 N for predators… See my previous comment about the use of N for sample size Moreover, I assume primary consumers refer to grazers, collectors, and shredders? Please, stick to the same terms throughout the manuscript, in this case, terms of table 2. Otherwise, explain why this grouping was made. 337 groups 344 I see how my previous comment was not addressed. Your method does not estimate abundance, because you stopped counting until reaching a certain number of individuals. Therefore, you cannot argue that they were observed in larger numbers. Could taxa group be replaced simply by order? 365 is M the mean? This is not an standard acronym. Do these values correspond to the column Pesticide detections in Table 4? The column does not have units. Discussion 383 but you did find differences in the C range, TA, distance to centroid, MNND (Fig. 5). However, this is because this figure was not corrected by C. The fact that some figures/analyses are corrected and some others are not makes the interpretation difficult. Which results are to be used, with or without correction? And based on that, are or are there not differences? This applies to the whole discussion since some paragraphs seem to be based on corrected values (e.g., paragraph 2) and others not (paragraph 1). 391 expected 395 is higher in ponds 395 led to enriched values 401 these are not diversity values. These are the number of samples that you used, which should not be used as a diversity metric. A diversity metric is, for example, the number of families. 402 but again, this is highly speculative since you didn’t measure primary production. More prey could also come from filterers, gatherers, or shredders consuming FPOM or CPOM. 407 ok but highest densities are not to be compared with higher diversity. They could have found higher densities of a few (or even one) taxa, while you found higher diversity (more families) or predators. 408-411 but these two sentences are completely contradicting each other! 412 but nutrient concentration was higher in ditches. In general, this paragraph is very confusing and contradictory. 414 it seems to be a typo: more less 427 I partly disagree. The shift could occur in polluted water bodies, or in any water body for multiple reasons. Mainly, resource availability, which changes seasonally, according to the longitudinal position, riparian structure, etc… that is why they are omnivores. 434 how do you conclude they are highly abundant? 434 see my comment in the intro about why isotopic compositions are not meant to be used for habitat use, but for feeding resources, which are not the same. Also, keep consistency between habitat and microhabitat-unless you mean something different-, although my suggestion is to remove the habitat part. 466 do you mean autumn? ********** what does this mean? ). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy Reviewer #1: Yes: Daniela Cortes ********** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step. |
| Revision 3 |
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Dear Dr. Trau, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Dear authors, the reviewer has completed the review of your manuscript once again and acknowledged its substantial improvement. However, two major points related to data analysis and discussion are still to be addressed before your manuscript can be accepted for publication. I invite you to take this final step and consider the suggestions thoroughly. Please submit your revised manuscript by Oct 17 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.
If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter. If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols . We look forward to receiving your revised manuscript. Kind regards, Marina Vilenica Academic Editor PLOS ONE Journal Requirements: If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author Reviewer #1: (No Response) ********** 2. Is the manuscript technically sound, and do the data support the conclusions??> Reviewer #1: Yes ********** 3. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: Yes ********** 4. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes ********** 5. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes ********** Reviewer #1: I have finalized revising the manuscript again. I acknowledge the effort of the authors to address all previous comments, which is reflected in the improvement of the manuscript. I still have one general comment and two major issues. My general comment is about the effects of the N and C corrections. I think that the authors could emphasize the reasoning of the corrections in the discussion, to clarify and remind readers of why some figures and results are based on one/both corrections and others are not. My major issues are: 1. The assignment of taxonomic groups without FFG information to omnivores. This is not trivial, as you are arbitrary assigning NAs to one functional group, which could have an important effect on your results. For example, this could explain the clusters you observed in your analysis. Dealing with NA is an extensively discussed topic in Ecology, and among the multiple approaches to address it, arbitrary assigning values is not a recommended one. My suggestions are as follow: you can remove these taxa from your FFG analysis, and justify this because of the lack of information. You can extend your literature sources to find the appropriate FFG category for such taxa; given the generally coarse taxonomic level you managed (order, family, or genus), I am convinced that this information is available for European taxa. Finally, you can repeat your analysis excluding these taxa and compare to your current results to discard (or not) the influence of your approach. 2. Your discussion about the predator diversity explained by increased primary production. Once more, I consider that adding “However we have to be careful with this conclusions as we did not measure primary production.” is not enough to address the comment. As I explained before: first, this is especulative as you didn't measure PP; however, you did measure FFG, and you can quantify the number of potential preys available and easily check/discuss if they are more preys in ditches than in ponds. Second, more importantly, one would expect that increasing PP—although only hypothetical—leads to a higher number of primary consumers, particularly grazers. But this is not what you observed. An alternative, which is not discussed in your text, is that predators are not being sustained by a green food web, but by a brown one. This is, instead of PP, there are more detritus sustaining preys (but this also needs to be supported by your own results) and, therefore, sustaining predators. As I mentioned in my previous comment, more available prey could come from filterers, gatherers, or shredders consuming FPOM or CPOM. ********** what does this mean? ). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy Reviewer #1: Yes: Daniela Cortes ********** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/ . PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email PLOS at figures@plos.org . Please note that Supporting Information files do not need this step. |
| Revision 4 |
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Dear Dr. Trau, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. ============================== ACADEMIC EDITOR: Dear authors, many thanks for thorough revision, the Reviewer has no additional requests. However, I have made several minor comments and suggestions in the word document of your manuscript that need to be considered before your manuscript can be accepted for publication. ============================== Please submit your revised manuscript by Dec 07 2025 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org . When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.
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| Revision 5 |
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Assessing agricultural effects on benthic invertebrate communities in ponds and ditches using δ15N and δ13C isotope niches PONE-D-25-03786R5 Dear Mrs. Trau, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact billing support . If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Marina Vilenica Academic Editor PLOS ONE |
| Formally Accepted |
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PONE-D-25-03786R5 PLOS ONE Dear Dr. Trau, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. If we can help with anything else, please email us at customercare@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. Marina Vilenica Academic Editor PLOS ONE |
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