Effects of Glyphosate or Glyphosate-Based Herbicide during the Zebrafish Life Cycle: A Review Addressing the Mechanisms of Toxicity
, Luís M. Félix 1,2, António Fontaínhas-Fernandes 1,2,3, Sandra Mariza Monteiro 1,2,3,* and Carlos Venâncio 1,2,4Round 1
Reviewer 1 Report
Line 16-17: The sentence is confusing, please rephrase “Consequently, and due to its inappropriate use, 16 its residues have often been found in aquatic ecosystems.”
Line 47: “… used in over 140 countries [6].” Please mention the year.
Line 68: Figure 1: The result for what year to what year?
Figure 1: Residual concentrations of glyphosate in the aquatic environment, food, and 90 humans. Adapted from [18,19]. References (Water [26,108-112], Food [13,113-115] and Hu- 91 mans [116-118]). References are jump from 28to 108, 113 and 116. Please refer to MDPI guidelines. thank you.
Line 95: Suggestion: Please add on 1 figure that summaries the main topic.”Toxicological effects of GLY/GBHs on zebrafish.”. The figure includes 2.1 Effects on lethality/mortality, 2.2 Effects on the hatching, 2.3 Malformations, 2.4 Cardiotoxicity, 2.5 Neurotoxicity and behavioural changes, 2.6 Oxidative stress, 2.7 Genotoxicity and 2.8 Apoptosis.
Line 112-114: “For this critical review, we reviewed and synthesized publications from the NCBI Pub- Med and Web of Science databases in March/2023, using the keywords “glyphosate, toxicity, zebrafish”. What are the duration year?
Line 116 and 119: Please rephrase. avoid to used “we” “us”.
Line 508: “2.7.1 New targets of GLY in zebrafish through insights from computational biology “ no need sub-topic if only one topic.
In my opinion, this subtopic “Proposal for future research on this subject” is not suitable for review paper. May be the authors should write and discuss more on “The limitation of the study”
Comments for author File: 
Comments.pdf
Author Response
REVIEWER 1
Comments and Suggestions for Authors
Line 16-17: The sentence is confusing, please rephrase “Consequently, and due to its inappropriate use, 16 its residues have often been found in aquatic ecosystems.”
Response: We thank the editor for the suggestion. Accordingly, it was rephrased to:
“Consequently, its residues have often been found in aquatic ecosystems.”
Line 47: “… used in over 140 countries [6].” Please mention the year.
Response: We thank the editor for the suggestion. As requested, the year was mentioned:
“used in over 140 countries in 2014”.
Line 68: Figure 1: The result for what year to what year?
Response: We thank the editor for the suggestion, Figure 1 shows data from 1991 to 2022.
Figure 1: Residual concentrations of glyphosate in the aquatic environment, food, and 90 humans. Adapted from [18,19]. References (Water [26,108-112], Food [13,113-115] and Hu- 91 mans [116-118]). References are jump from 28 to 108, 113 and 116. Please refer to MDPI guidelines. thank you.
Response: We thank the editor for the indication, the references were correct according to MDPI guidelines.
Line 95: Suggestion: Please add on 1 figure that summaries the main topic.”Toxicological effects of GLY/GBHs on zebrafish.”. The figure includes 2.1 Effects on lethality/mortality, 2.2 Effects on the hatching, 2.3 Malformations, 2.4 Cardiotoxicity, 2.5 Neurotoxicity and behavioural changes, 2.6 Oxidative stress, 2.7 Genotoxicity and 2.8 Apoptosis.
Response: We thank the editor for the suggestion that improved the manuscript quality. Figure 2 was added.
Line 112-114: “For this critical review, we reviewed and synthesized publications from the NCBI Pub- Med and Web of Science databases in March/2023, using the keywords “glyphosate, toxicity, zebrafish”. What are the duration year?
Response: We thank the editor for the note. The time interval was comprised between 2011 to 2023. This information was added in the text.
Line 116 and 119: Please rephrase. avoid to used “we” “us”.
Response: We thank the editor for the suggestion, changes were made in accordance.
Line 508: “2.7.1 New targets of GLY in zebrafish through insights from computational biology “ no need sub-topic if only one topic.
Response: We thank the editor for the suggestion. Accordingly, the sub-topic was removed and the text improved.
In my opinion, this subtopic “Proposal for future research on this subject” is not suitable for review paper. May be the authors should write and discuss more on “The limitation of the study”
Response: We thank the editor for the suggestion that clearly contributed to improving the focus of the manuscript. Accordingly, to conclude the manuscript, a brief conclusion was elaborated highlighting the questions raised by the literature review and pointing to some directions that may contribute to improving the current knowledge on the topic.
Reviewer 2 Report
Review Report
General Comments
The manuscript entitled “A comprehensive review of glyphosate and glyphosate-based herbicides toxicity in zebrafish (Danio rerio)” contributes the knowledge in the area that how chemicals in one ecosystem pollute the other environmental component. Glyphosate herbicide is widely used in worldwide crop production.
Consequently, its active ingredient, surfactants, and adjuvants commonly reach the aquatic ecosystem, thereby harming the biota. Authors have tried to investigate how this herbicide affects aquatic species especially fish, as they have the ability to absorb and concentrate toxins.
Specific Comments
To improve the quality of manuscript, I have suggested the following points:
Title of the review paper should be broader. Similar work has already been reported titled as "Glyphosate vs. Glyphosate-Based Herbicides Exposure: A Review on Their Toxicity".
The novelty of this review is the addition of toxicity in zebrafish. Please rewrite the title more comprehensively.
Please add more relevant keywords.
Please explain the novelty of this review paper to clear how this review is different from other review papers on the similar topic.
The heading environmental levels is not clear and appropriate, please replace with more suitable heading.
Figure title should be mentioned below the figure not above.
Heading "Effects on lethality/mortality" is not clear, please rewrite.
In the manuscript, 11 Tales are given. Information given in 11 Tales can be summarized in 4-5 Tales.
In the whole manuscript most of the headings are not clear and incomplete. Please rewrite.
In review paper, continuity of text should break by adding flow charts, tales and diagrams.
In this manuscript, authors should add mechanism-based diagram/diagrams.
Conclusion section is missing.
More proofreading of the manuscript is advised to remove typographical mistakes.
The minor editing of English language is required to improve the quality of manuscript for publication in such a high impact factor journal.
Author Response
General Comments
The manuscript entitled “A comprehensive review of glyphosate and glyphosate-based herbicides toxicity in zebrafish (Danio rerio)” contributes the knowledge in the area that how chemicals in one ecosystem pollute the other environmental component. Glyphosate herbicide is widely used in worldwide crop production.
Consequently, its active ingredient, surfactants, and adjuvants commonly reach the aquatic ecosystem, thereby harming the biota. Authors have tried to investigate how this herbicide affects aquatic species especially fish, as they have the ability to absorb and concentrate toxins.
Specific Comments
To improve the quality of manuscript, I have suggested the following points:
Title of the review paper should be broader. Similar work has already been reported titled as "Glyphosate vs. Glyphosate-Based Herbicides Exposure: A Review on Their Toxicity".
The novelty of this review is the addition of toxicity in zebrafish. Please rewrite the title more comprehensively.
Response: We thank the editor for the suggestion, the title was changed to …
“A comprehensive review of glyphosate and glyphosate-based herbicides toxicity in zebrafish: perspective on effects on the zebrafish life cycle.”
Please add more relevant keywords.
Response: We thank the reviewer for the suggestion and the keyword Environmental Pollutants was added.
Please explain the novelty of this review paper to clear how this review is different from other review papers on the similar topic.
Response: We consider that this review presents updated content on the toxic effects of glyphosate and GBHs in multiple systems at the molecular, cellular and organic level. The scope of the studies included, and the effects evaluated make this review one of the most complete and comprehensive to our knowledge. This information was added to manuscript (line 122).
The heading environmental levels is not clear and appropriate, please replace with more suitable heading.
Response: We thank the reviewer for the suggestion, the heading was changed to 1.2 Glyphosate and glyphosate-based herbicides in the environment.
Figure title should be mentioned below the figure not above.
Response: We thank the reviewer for the suggestion, the figure title position was corrected accordingly.
Heading "Effects on lethality/mortality" is not clear, please rewrite.
Response: We thank the reviewer for the suggestion, the heading was changed to “2.1 Lethality and mortality caused by exposure to GLY or GBHs on zebrafish”.
In the manuscript, 11 Tales are given. Information given in 11 Tales can be summarized in 4-5 Tales.
Response: We thank the reviewer for the suggestion. However, for the sake of organization and accessible reading, we believe it is an asset to keep the current number of tables, since they present the information in a synthetic and systematized way. We kindly request the maintenance of the tables presented.
In the whole manuscript most of the headings are not clear and incomplete. Please rewrite.
Response: We thank the reviewer for the suggestion, all the headings were improved.
In review paper, continuity of text should break by adding flow charts, tales and diagrams.
In this manuscript, authors should add mechanism-based diagram/diagrams.
Response: We thank the reviewer for the suggestion, for this purpose the Figure 2 was added aiming to summarize the main topic 2. Toxicological effects of GLY/GBHs on zebrafish.
Conclusion section is missing.
Response: We thank the reviewer for the suggestion, a conclusion section alongside proposals for future research was included.
More proofreading of the manuscript is advised to remove typographical mistakes.
Response: We thank the reviewer for the suggestion, a revision of typographical errors was conducted.
Reviewer 3 Report
In this review, the findings of toxicological studies on glyphosate and glyphosate-based herbicides were summarized, and the structure of this article was designed and organized very well. The review seemed quite instructive and helpful to people participating in not only the toxicological research but also the activity of protecting the environment. But it was to be regretted that the presentation of previous findings seemed complicated a little or rather troublesome to grasp whole picture. So, it would be desirable to improve the disclosure manner of previous findings, or would be better to switch them over any other expression, if possible. It could be expected to devise the presentation method simple and easy to grasp whole picture of herbicide toxicities.
Regarding English writing, there were only minor points to be reconsidered.
1) Line 246: The phrase “Na+/K+-ATPase e” seemed strange, and a letter “e” was unnatural. It would probably be a simply mistake, and might be possible to guess that this phrase was supposed to be “Na+/K+-ATPase and”.
2) Line 300: The phrase “an increase” seemed a mistake, and it might be “an increase in” or “an increased”.
3) Lines 300-303: The sentence had the words “increase” and “decrease”, and the usage of these words seemed incomprehensible. It would be question whether they might be verb or noun.
4) Line 423: The word “formulations” might be misused, and it would be possible to guess that “formula” could be correct.
5) Line 519: The word “compromised” seemed unnatural, or rather inadequate. This word was usually used in the case of giving a damage to one’s honor and reputation, but not to say biological and physical damage. Therefore, the phrase “in the removal of harmful or compromised cells” at lines 518-519 might be better to say “in the removal of harmful or invasive cells”.
6) Line 577: The word “compromising” also seemed inadequate, as with the comment 5).
7) Line 579: The preposition “from” could probably be considered a mistake, and it might be possible to guess “in” instead of “from”.
Through the whole manuscript, it should be a little more careful to put a comma and to avoid the frequent use of inverted sentences.
Manuscript had no serious problem, and easy to read and understand. But it might be still necessary to do very minor revision.
Author Response
In this review, the findings of toxicological studies on glyphosate and glyphosate-based herbicides were summarized, and the structure of this article was designed and organized very well. The review seemed quite instructive and helpful to people participating in not only the toxicological research but also the activity of protecting the environment. But it was to be regretted that the presentation of previous findings seemed complicated a little or rather troublesome to grasp whole picture. So, it would be desirable to improve the disclosure manner of previous findings, or would be better to switch them over any other expression, if possible. It could be expected to devise the presentation method simple and easy to grasp whole picture of herbicide toxicities.
We thank the reviewer for the pertinent comments. To be illustrative of all the existing information it is difficult to summarize more. Even so, some changes were made in the description of the results throughout the text and new figures were included (also responding to reviewer 1's request) to try to systematize the results more clearly. The conclusion was also improved, trying to extract the main features of the current knowledge about the research topic.
Regarding English writing, there were only minor points to be reconsidered.
1) Line 246: The phrase “Na+/K+-ATPase e” seemed strange, and a letter “e” was unnatural. It would probably be a simply mistake, and might be possible to guess that this phrase was supposed to be “Na+/K+-ATPase and”.
Response: We thank the reviewer for the note, the error was corrected.
2) Line 300: The phrase “an increase” seemed a mistake, and it might be “an increase in” or “an increased”.
Response: We thank the reviewer for the note, the error was corrected.
3) Lines 300-303: The sentence had the words “increase” and “decrease”, and the usage of these words seemed incomprehensible. It would be question whether they might be verb or noun.
Response: We thank the reviewer for the note, the text was adjusted as suggested.
4) Line 423: The word “formulations” might be misused, and it would be possible to guess that “formula” could be correct.
Response: We thank the reviewer for the note, the error was corrected.
5) Line 519: The word “compromised” seemed unnatural, or rather inadequate. This word was usually used in the case of giving a damage to one’s honor and reputation, but not to say biological and physical damage. Therefore, the phrase “in the removal of harmful or compromised cells” at lines 518-519 might be better to say “in the removal of harmful or invasive cells”.
Response: We thank the reviewer for the note, the text was adjusted as suggested.
6) Line 577: The word “compromising” also seemed inadequate, as with the comment 5).
Response: We thank the reviewer for the note, the error was corrected.
7) Line 579: The preposition “from” could probably be considered a mistake, and it might be possible to guess “in” instead of “from”.
Response: We thank the reviewer for the note, the error was corrected.
Through the whole manuscript, it should be a little more careful to put a comma and to avoid the frequent use of inverted sentences.
Response: We thank the reviewer for the suggestion. A textual revision was conducted.
Round 2
Reviewer 1 Report
Dear authors,
I accept all the corrections. Well done. Thank you.
Reviewer 2 Report
Please add the revised title "A comprehensive review of glyphosate and glyphosate-based herbicides toxicity in zebrafish: perspective on effects on the zebrafish life cycle" in manuscript file.
