Residues and Uptake of Soil-Applied Dinotefuran by Lettuce (Lactuca sativa L.) and Celery (Apium graveolens L.)

Round 1

Reviewer 1 Report

This article - in my opinion - is suitable for publication in its current version. It has been prepared neatly and reliably in terms of content, language and editorial. The paper presents interesting results of an experiment that has been properly designed and the results have been processed properly. Related findings have a considerable cognitive and practical value, as they show the risk associated with too early introduction of crops (leafy vegetables: lettuce and celery) after the use of dinetofuran.

Author Response

Dear/ editor-In chief

Subject: Manuscript ID: agriculture-1874126; entitled “Residues and uptake of Soil Applied Dinotefuran by Lettuce (Lactuca sativa L.) and Celery (Apium graveolens L.)”

Dear Sir,

First, we would like to thank you and the reviewers for the valuable comments, which will improve the manuscript. I am pleased to provide the author's responses to the comments raised;

The reviewer's comments and amendments were addressed in the manuscript body in red characters.

Note: Amendments in the manuscript are written in red characters.

Reviewer I:

Comments and Suggestions for Authors

This article - in my opinion - is suitable for publication in its current version. It has been prepared neatly and reliably in terms of content, language and editorial. The paper presents interesting results of an experiment that has been properly designed and the results have been processed properly. Related findings have a considerable cognitive and practical value, as they show the risk associated with too early introduction of crops (leafy vegetables: lettuce and celery) after the use of dinetofuran.

Response: The manuscript was revised carefully for any error.

                                                                                                                            Regards

Author Response File: Author Response.docx

Reviewer 2 Report

The paper examines the fate of dinotefuran in soils at two sites in Korea and consequent residues in plants grown in the soils (Lettuce and celery). The topic is important because pesticides are increasingly used in vegetable production and there are global concerns about possible residues in food. Furthermore, as outlined by the authors, Korea has strict guidelines concerning residue levels foods and there are apparently serious penalties for farmers. The authors conducted very detailed analyses and have carefully explained all of their laboratory protocols. The results are useful in that the explain somewhat how residues are dissipated or assimilated in soils and plants respectively. The authors also explain their results gathering on a vast knowledge of their subject material. The paper therefore has considerable merit. Unfortunately, the paper as it is currently presented is an extremely difficult read and, because of the details and many protocols, it is easy to get lost in the text. So much so, that the main findings become obscured. The authors should therefore considerably improve the presentation of the paper. I would suggest that they include some glossary of terms, or perhaps a model of how dinotefuran is broken down or assimilated by plants in the environment. The authors have overlooked the impact of weather and give no details of rainfall and weathering effects. They might then, using such a diagram, explain where their research fits into the overall model. The authors must also improve their statistical analyses. Currently the appear to have lumped all data into one or two-way ANOVAs, possible treating site as a factor. However, the design is segregated (by sites) and the conditions for an ANOVA are therefore violated. Furthermore, the various analyses are conducted over time in the same plots, so the data should be analysed using repeated measures or some similar MANOVA procedure. The very low residue levels in some samples and higher levels in other, suggest that the data will not have homogenous residuals and the analyses will likely be non-parametric. These statistical problems are apparent in the tables and figures; for example, the figures present error bars that have no meaning (for example, zero values are given error bars). The errors should be recalculated and best presented as standard errors. The tables also demonstrate some errors in analyses, for example all means have different letters to indicate differences, but based on means and errors that are presented - some very similar means are reported as statistically different. This may be correct, but the current handling of statistics erodes the readers' confidence. The authors have also omitted some lables in graphs. Finally, the authors should tidy the paper somewhat, so that readers can follow what is presented without skipping unnecessary details (i.e., some of the calculations that are included). Overall, the study is well conducted, but the presentation needs vast improvement.

Author Response

Dear/ editor-In chief

Subject: Manuscript ID: agriculture-1874126; entitled “Residues and uptake of Soil Applied Dinotefuran by Lettuce (Lactuca sativa L.) and Celery (Apium graveolens L.)”

Dear Sir,

First, we would like to thank you and the reviewers for the valuable comments, which will improve the manuscript. I am pleased to provide the author's responses to the comments raised;

The reviewer's comments and amendments were addressed in the manuscript body in red characters.

Note: Amendments in the manuscript are written in red characters.

Reviewer 2:

Comments and Suggestions for Authors

The paper examines the fate of dinotefuran in soils at two sites in Korea and consequent residues in plants grown in the soils (Lettuce and celery). The topic is important because pesticides are increasingly used in vegetable production and there are global concerns about possible residues in food. Furthermore, as outlined by the authors, Korea has strict guidelines concerning residue levels foods and there are apparently serious penalties for farmers. The authors conducted very detailed analyses and have carefully explained all of their laboratory protocols. The results are useful in that the explain somewhat how residues are dissipated or assimilated in soils and plants respectively. The authors also explain their results gathering on a vast knowledge of their subject material. The paper therefore has considerable merit. Unfortunately, the paper as it is currently presented is an extremely difficult read and, because of the details and many protocols, it is easy to get lost in the text. So much so, that the main findings become obscured.

Response: The materials and methods section was revised and summarized by removing the unnecessary details.

The authors should therefore considerably improve the presentation of the paper. I would suggest that they include some glossary of terms, or perhaps a model of how dinotefuran is broken down or assimilated by plants in the environment.

Response: Agreed; glossary of terms and flow chart of metabolites formation were included as suggested. (See supplementary materials, table S4)

The authors have overlooked the impact of weather and give no details of rainfall and weathering effects. They might then, using such a diagram, explain where their research fits into the overall model.

Response; Clarification; the experiments were conducted under the control protected glasshouse condition therefore effects from rainfall and weathering were not expected as mentioned in the manuscript.

The authors must also improve their statistical analyses. Currently the appear to have lumped all data into one or two-way ANOVAs, possible treating site as a factor. However, the design is segregated (by sites) and the conditions for an ANOVA are therefore violated. Furthermore, the various analyses are conducted over time in the same plots, so the data should be analysed using repeated measures or some similar MANOVA procedure.

Response: Agreed; the data were reanalyzed as recommended. (See tables 2 and 4)

The very low residue levels in some samples and higher levels in other, suggest that the data will not have homogenous residuals and the analyses will likely be non-parametric. These statistical problems are apparent in the tables and figures; for example, the figures present error bars that have no meaning (for example, zero values are given error bars). The errors should be recalculated and best presented as standard errors.

Response: Agreed; the error bars were removed from the figures instead the standard error values were provided in the figures as suggested taking into account the comments of reviewers 3 & 4. However in the tables standard deviation were used.

The tables also demonstrate some errors in analyses, for example all means have different letters to indicate differences, but based on means and errors that are presented - some very similar means are reported as statistically different. This may be correct, but the current handling of statistics erodes the readers' confidence. The authors have also omitted some lables in graphs.

Response: The data in tables (2 & 4) were reanalyzed and all (graphs 1-4) were revised taking into account the comments raised by the reviewers 3 & 4.

Finally, the authors should tidy the paper somewhat, so that readers can follow what is presented without skipping unnecessary details (i.e., some of the calculations that are included). Overall, the study is well conducted, but the presentation needs vast improvement.

Response: Agreed; the manuscript was revised and unnecessary details were removed as suggested.

                                                                                                                            Regards

Author Response File: Author Response.docx

Reviewer 3 Report

Comment for authors:

In this study, the dinotefuran and three major metabolites by lettuce and celery from previous season applications and analyze their residues in soil and consumable parts. This study provides a usable method for the detection of dinotefuran and three major metabolites in lettuce and celery for LC-MS/MS, and verified safety interval of dinotefuran for lettuce and celery. This study can provide a chilli production guidance for lettuce and celery planted. However, it needs some corrections and there are some minor queries such as the chart was not standardized and some content is unclear,  the authors should kindly respond to make it good, to the best of my knowledge. In addition, some improvements in English writing are suggested. Finally, I suggested that it can be accepted and publication after revised. I would like to see the authors address the comments before it can be published.

Specific proposals are as follows:

1. Fig1, 2, 3 and 4, Table 2, Lack of significant difference analysis.

2. Please explain the possible causes of “The disappearance of dinotefuran in Chuncheon lettuce soil was faster than that in the Pyeongtaek while in celery soil the vice versa is true as indicated by their half-lives.”

3. Please show the results of “Accuracy Test” in the form of a table in the form of the table in this paper

4. we suggested to modify the manuscript and add the content of dietary risk assessment.

5. We noticed the half-lives of dinotefuran in lettuce, celery and soil were studied in this paper, but in the description of the materials and methods, the plant and soil Samples were collected in 2 time points and the half-life was calculated requires at least 5 time nodes, please check with the author.

6. Discussion: This part needs to be strengthened.

Author Response

Dear/ editor-In chief

Subject: Manuscript ID: agriculture-1874126; entitled “Residues and uptake of Soil Applied Dinotefuran by Lettuce (Lactuca sativa L.) and Celery (Apium graveolens L.)”

Dear Sir,

First, we would like to thank you and the reviewers for the valuable comments, which will improve the manuscript. I am pleased to provide the author's responses to the comments raised;

The reviewer's comments and amendments were addressed in the manuscript body in red characters.

Note: Amendments in the manuscript are written in red characters.

Reviewer 3

Comments and Suggestions for Authors

Comment for authors:

In this study, the dinotefuran and three major metabolites by lettuce and celery from previous season applications and analyze their residues in soil and consumable parts. This study provides a usable method for the detection of dinotefuran and three major metabolites in lettuce and celery for LC-MS/MS, and verified safety interval of dinotefuran for lettuce and celery. This study can provide a chilli production guidance for lettuce and celery planted. However, it needs some corrections and there are some minor queries such as the chart was not standardized and some content is unclear,  the authors should kindly respond to make it good, to the best of my knowledge. In addition, some improvements in English writing are suggested. Finally, I suggested that it can be accepted and publication after revised. I would like to see the authors address the comments before it can be published.

Specific proposals are as follows:

1. Fig1, 2, 3 and 4, Table 2, Lack of significant difference analysis.

Response:  Agreed; figures 1-4 were revised taking into account the comments raised by reviewers 2 and 4. Clarification; The data in table 2 (currently Table 1) contain detected metabolites beside the parent compound, while data in table 3 (currently Table 2) shows the residues of dinotefuran after calculating the levels of detected metabolites (MNG, UF, and DN) and multiplying their levels by conversion factors indicated under the tables and expressed as dinotefuran. However, table 2 (currently Table 1) can be moved to supplementary materials if requested.

2. Please explain the possible causes of “The disappearance of dinotefuran in Chuncheon lettuce soil was faster than that in the Pyeongtaek while in celery soil the vice versa is true as indicated by their half-lives.”

Response: The possible causes of the differences in the disappearance rates of dinotefuran in Chuncheon lettuce and celery soils compared to Pyeongtaek soils was discussed and possible reasons were explained as suggested

3. Please show the results of “Accuracy Test” in the form of a table in the form of the table in this paper

Response: Agreed; the accuracy test was provided in supplementary materials Table S5 as recommended. To remove the confusion the section title was modified to read “Method Validation” instead of “Accuracy Test”.

4. we suggested to modify the manuscript and add the content of dietary risk assessment.

Response: Agreed; the dietary risk assessment was injected into the manuscript as suggested.

5. We noticed the half-lives of dinotefuran in lettuce, celery and soil were studied in this paper, but in the description of the materials and methods, the plant and soil Samples were collected in 2 time points and the half-life was calculated requires at least 5 time nodes, please check with the author.

Response: Clarification; the half-lives were calculated in lettuce soil and celery soil, but not in the plants. See section 2.6 for soil sampling and preparation.

6. Discussion: This part needs to be strengthened.

Response: Agreed; the discussion sections as well as a whole manuscript were revised as recommended.

                                                                                                                            Regards

Author Response File: Author Response.docx

Reviewer 4 Report

The manuscript entitled " Residues and uptake of Soil Applied Dinotefuran by Lettuce (Lactuca sativa L.) and Celery (Apium graveolens L.)" investigated uptake of soil applied dinotefuran and three major metabolites by lettuce and celery from previous season applications and analyze their residues in soil and consumable parts. The paper is well organized, the topic of research is good, but the authors should correct some points and research errors. Hence major revision is needed.

Comments

·       Line 27: keywords are already stated in the title. Please consider changing the keywords list and use synonyms.

·         Line 84: -1 make superscript.

·         Line 115: What does it mean by green house?

·         Lines 179: Replace rpm with “xg”. RCF = (RPM)² × 1.118 × 10^-5 × r Revise through the whole manuscript.

·       The materials and methods contain unnecessary information; the authors should try to shorten them without sacrificing content.

·         Lines 295-289: Delete this paragraph

·     Figure 1: Move this sentence “Vertical bars represented ± SE” to the figure caption. There are no Figure Legends. Statistics, sample sizes, abbreviations, all these should be explained in a legend after the caption of each figure.

·        Why there is no letters of Tukey test above the figures calamus???

·        Lines 299-484The results section lacks state of art and need one sentence at the end of each paragraph to show to readers what happen in the whole paragraph

·       Discussion section is too long repetitive in certain parts of the text as well as in most sections the given information is randomly strung together without a clear structure, and transition between sentences in the discussion is unclear. The authors also did not cite tables and figures in the discussion section.

Kind Regards.

Author Response

Dear/ editor-In chief

Subject: Manuscript ID: agriculture-1874126; entitled “Residues and uptake of Soil Applied Dinotefuran by Lettuce (Lactuca sativa L.) and Celery (Apium graveolens L.)”

Dear Sir,

First, we would like to thank you and the reviewers for the valuable comments, which will improve the manuscript. I am pleased to provide the author's responses to the comments raised;

The reviewer's comments and amendments were addressed in the manuscript body in red characters.

Note: Amendments in the manuscript are written in red characters.

Reviewer 4:

Comments and Suggestions for Authors

The manuscript entitled " Residues and uptake of Soil Applied Dinotefuran by Lettuce (Lactuca sativa L.) and Celery (Apium graveolens L.)" investigated uptake of soil applied dinotefuran and three major metabolites by lettuce and celery from previous season applications and analyze their residues in soil and consumable parts. The paper is well organized, the topic of research is good, but the authors should correct some points and research errors. Hence major revision is needed.

Comments

•       Line 27: keywords are already stated in the title. Please consider changing the keywords list and use synonyms.

Response: editorial change in keywords line 27 was done as suggested.

• Line 84: -1 make superscript.

Response: editorial change in line 84 (currently 86) was done as recommended.

• Line 115: What does it mean by green house?

Response: clarification; the green house mean glass house used for growing of crops under controlled environmental conditions. To remove the confusion the “greenhouse” was changed to read “glasshouse”

• Lines 179: Replace rpm with “xg”. RCF = (RPM)² × 1.118 × 10^-5 × r Revise through the whole manuscript.

Response: Agreed; the editorial changes in line 179 (currently 154) and the whole manuscript were carried out as recommended.

•       The materials and methods contain unnecessary information; the authors should try to shorten them without sacrificing content.

Response: Agreed; this section was shortened by moving table 1 to supplementary materials and removing the unnecessary details as suggested taking into account comments raised by reviewer 2 .

• Lines 295-289: Delete this paragraph

Response: Agreed; the lines 289-295 (subtitle 2.15 Statistical analysis) were removed as suggested.

•     Figure 1: Move this sentence “Vertical bars represented ± SE” to the figure caption. There are no Figure Legends. Statistics, sample sizes, abbreviations, all these should be explained in a legend after the caption of each figure.

Response: Agreed; editorial changes in Figure were done as recommended. Taking account the comment raised by the reviewers 2 & 3 the vertical bars were removed instated the calculated standard error values were provided.

•        Why there is no letters of Tukey test above the figures calamus???

Response: Agreed; the calculated standard error values are provided in the figures columns taking into account the comments raised by the reviewers 2 & 3.

•        Lines 299-484 The results section lacks state of art and need one sentence at the end of each paragraph to show to readers what happen in the whole paragraph

Response: Disagreed; this may increase the length of the manuscript, and lead to many redundancies and contradict with next comment from the same reviewer.

•       Discussion section is too long repetitive in certain parts of the text as well as in most sections the given information is randomly strung together without a clear structure, and transition between sentences in the discussion is unclear. The authors also did not cite tables and figures in the discussion section.

Response: Agreed; the discussion was revised and the tables and figures were cited in the discussion section as recommended. Clarification: Although the  discussion apparently looks too long and repetitive in some parts, however if you carefully review the results you will find that manuscript covered many aspects and therefore the discussion section  explains the possible causes of the major  findings and therefore it is difficult to be shorten further.

                                                                                                                            Regards

Author Response File: Author Response.docx

Round 2

Reviewer 3 Report

The proposed comments have been revised.

Reviewer 4 Report

The authors have addressed my comments, many thanks to their contribution.