Effect of orally administered cannabidiol oil on daily tonometric curve in healthy Italian Saddle horses

Marilena Bazzano; Fulvio Laus; Matteo Cerquetella; Andrea Spaterna; Andrea Marchegiani

doi:10.1371/journal.pone.0325191

Peer Review History

Original SubmissionJanuary 27, 2025
27 Jan 2025 Author Response https://doi.org/10.1371/journal.pone.0325191.r001
21 Feb 2025 Decision Letter - Claudia Interlandi, Editor PONE-D-25-04742Effect of orally administered cannabidiol oil on daily tonometric curve in healthy Italian Saddle horsesPLOS ONE Dear Dr. Marchegiani, 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 Apr 04 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. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. 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. 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Journal requirements: When submitting your revision, we need you to address these additional requirements. 1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. To comply with PLOS ONE submissions requirements, in your Methods section, please provide additional information regarding the experiments involving animals and ensure you have included details on (1) methods of anesthesia and/or analgesia, and (2) efforts to alleviate suffering. Additional Editor Comments (if provided): The reviewers agree that many parts of the manuscript are in need of deepening and major revision. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Partly Reviewer #2: No ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: No ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: Please see the attached file. The manuscript entitled “Effect of orally administered cannabidiol oil on daily tonometric curve in healthy Italian Saddle horses.” investigate the use of a natural substance such as cannabis oil to evaluate the effects of such administration on eye pressure trends in the horse. This work is very interesting, in terms of evaluating the health status of the horse's eye but also as it relates to the diagnostic aspect of eye diseases. It is well structured but some parts turn out to be written in too superficial and shallow a manner to be a scientific work. In my opinion this work should be deepened and revised in many parts, and rewritten in the discussion, also following the advice and comments below following a major revision. Reviewer #2: I have read and reviewed the manuscript entitled: Effect of orally administered cannabidiol oil on daily tonometric curve in healthy Italian Saddle horses. Overall, from this reviewer’s perspective, this is an interesting study; however, in its current state, it shows many deficiencies. For example, the methodology section requires an orderly and detailed description to ensure that the study can be replicated. Regarding the statistical analysis, I suggest the authors modify it to a mixed linear model that allows the establishment of statistically significant differences between treatments and the various measurement moments, in addition to correctly analyzing the tonometric curve on the three occasions in which the measurements were repeated. On the other hand, the lack of a control group generates an important bias in the research. Finally, I would like to highlight that the discussion section should be rewritten. This is to explain the neurophysiological events that occur under the effect of CBD to reduce intraocular pressure. Likewise, other observations must be addressed to achieve publication quality. I have left some comments, hoping that they can help the authors. General comments L31-33: Please indicate the IOP values before and after CBD administration, with their respective P values. L66: CBD has also been used in pets for the control of acute pain caused by an ovariohysterectomy and chronic pain caused by osteoarthritis. Please add the following references to your manuscript, which you can also use in the discussion of your manuscript: 10.3389/fvets.2024.1380022 10.3389/fvets.2022.1050884 L69: I suggest that the authors add the following references in this line: 10.3389/fvets.2024.1496473 10.3389/fvets.2024.1341396 10.1016/j.jevs.2019.102880 L73: Please add a hypothesis. L82: What was the statistical method used to determine the sample size? Please clarify. L84: Please indicate which exclusion criteria were considered in your study. L85: What were the analytes or parameters measured in blood before the start of the study? And how long before the administration of CBD were these blood studies performed? L108: Due to the type of experimental design used, I suggest the authors perform a statistical analysis based on a linear mixed model instead of a 2-way ANOVA. This model will allow the analysis of the differences observed between treatments and evaluation times, and even for the analysis of the tonometric curve (comparison of three measurements) with their respective washout periods. L123: Table 1, please indicate the statistical differences observed between treatments and evaluation times with letters and numbers. In this sense, also the results obtained after the administration of CBD at 15 minutes, 30 minutes, 60 minutes, 2 hours, 3 hours, 4 hours, 6 hours, and 8 hours should be integrated into this table; although Figure 1 already shows this information. L141: In general, the discussion is a section that is shown to be deficient. Therefore, I suggest that the authors complement this section. For example, the neurophysiological effects that generate the decrease in intraocular pressure under the effect of CBD should be explained. I suggest consulting and citing the following references: 10.2460/javma.24.06.0360 10.3934/Neuroscience.2024009 10.3390/ph17060748 10.1016/j.ijpharm.2022.121627 10.3390/ijms22073798 10.1016/j.survophthal.2020.07.002 10.1097/01.ijg.0000212260.04488.60 10.3389/fvets.2022.1050884 10.3390/ph16081149 10.1016/j.biopha.2022.112981 10.1055/a-1665-3100 10.1016/j.exer.2020.108266 10.2174/1570159X15666170724104305 10.1016/j.biopha.2016.11.106 10.1155/2016/9364091 10.1016/S0079-6123(08)01131-X https://pubmed.ncbi.nlm.nih.gov/34283478/ L167: I suggest that the authors discuss the limitations and perspectives of their study. L171: Conclusions must be rewritten based on changes arising from modifications made to the statistical analysis. ****** 6. PLOS authors have the option to publish the peer review history of their article (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: No 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". 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. Attachments Attachment Submitted filename: comments for authors.docx https://doi.org/10.1371/journal.pone.0325191.r002
Revision 1
20 Mar 2025 Author Response Below, in bold blue type, the point-to-point reply to reviewers Reviewer #1: Please see the attached file. The manuscript entitled “Effect of orally administered cannabidiol oil on daily tonometric curve in healthy Italian Saddle horses.” investigate the use of a natural substance such as cannabis oil to evaluate the effects of such administration on eye pressure trends in the horse. This work is very interesting, in terms of evaluating the health status of the horse's eye but also as it relates to the diagnostic aspect of eye diseases. It is well structured but some parts turn out to be written in too superficial and shallow a manner to be a scientific work. In my opinion this work should be deepened and revised in many parts, and rewritten in the discussion, also following the advice and comments below following a major revision. The manuscript entitled “Effect of orally administered cannabidiol oil on daily tonometric curve in healthy Italian Saddle horses.” investigate the use of a natural substance such as cannabis oil to evaluate the effects of such administration on eye pressure trends in the horse. This work is very interesting, in terms of evaluating the health status of the horse's eye but also as it relates to the diagnostic aspect of eye diseases. It is well structured but some parts turn out to be written in too superficial and shallow a manner to be a scientific work. In my opinion this work should be deepened and revised in many parts, and rewritten in the discussion, also following the advice and comments below following a major revision. Introduction Line 49 Please unify the number of bibliographical sources and dots through the main text Text has been modified to reflect this comment Line 45-54 As the title of the present article suggest that you were talking about healthy subjects, first of all I suggest to describe the physiological oscillation of the IOP for example after exercise in horses, for example include this source to deep the section: doi: 10.3390/ani12141850. Text has been modified to reflect this comment and reference has been added Line 49-55 this section is described too broadly and in too little depth, so the arguments regarding therapeutic approaches are disconnected with the introductory section Section rephrased to ensure more clarity Line 56-57 some new bibliographic source to accompany the section should be included as: https://doi.org/10.3389/fvets.2024.1496473 https://doi.org/10.3389/fvets.2024.1341396 Text has been modified to reflect this comment and references have been added Materials and methods Line 91-92 if the horses were already stabled in the mentioned way, according to precise routines, what was the purpose of the 5-day acclimatization period? Please be more specific and specify what the horses needed to acclimatize to Lines implemented in the manuscript and a proper reference has been added Line 92-93 With what instrumentation the IOP was measured and by what methodology IOP was evaluated with Tonopen Vet, Reichert Inc., Depew, NY USA. Detailed in the text Line 101-102 The product manufacturer should be reported Results and Discussion Line 116-117 data were normalized results or not? this result should be reported Line 135-136 The mention both eyes and respectively right and left is redundant Line 137 the same mention “No effect of sex was found” is redundant compared to lines 133-134 Line 116-121 In addition, a two-way ANOVA was performed in the statistical analysis. Better clarify which variables were evaluated. Also, in the results section these significant values should be detailed. To establish a significant peak over time, a post hoc analysis should also have been performed, so describe it accurately both in the statistical analysis section and in the results. Results in too little content. According to when IOP measurements over time were assessed before CBD administration and then 3 times every 8 days. Why does it appear in Figure 1 only before and after administration. Please clarify these concepts so that they are easy for a reader to interpret. Table 1 If results were calculated and expressed as mean and standard deviation, this information should be specified in the results rection Line 143-144 Avoid repetitions Line 151 Please mention the examples reported in horses as showed in the previously suggested bibliographical sources Line 155 nothing was previously mentioned about THC use or information in the text. Please explain its importance and usage even in other sections or delete it. The importance of eye pressure changes in both physiological and pathological settings should be emphasized. The discussions are too short and poor in content. there is described only what has been found in the literature on the topic. The discussion section should be a discussion regarding the results obtained, so one should first describe what was obtained, comment on its possible meanings, and discuss comparison with other studies that found similar or opposite results based on the variables analyzed, so the section is all in need of rewriting Conclusions should also more emphasize the results obtained and indicate in what ways these results can be useful in the scientific community Statistical analysis has been run again, applying both a mixed effect model analysis and post hoc analysis to reflect your recommendations, so the paragraph has been rewritten accordingly. Results, Discussion and Conclusion sections have been completely rewritten to be more specific and consistent, incorporating all your suggestion Reviewer #2: I have read and reviewed the manuscript entitled: Effect of orally administered cannabidiol oil on daily tonometric curve in healthy Italian Saddle horses. Overall, from this reviewer’s perspective, this is an interesting study; however, in its current state, it shows many deficiencies. For example, the methodology section requires an orderly and detailed description to ensure that the study can be replicated. Regarding the statistical analysis, I suggest the authors modify it to a mixed linear model that allows the establishment of statistically significant differences between treatments and the various measurement moments, in addition to correctly analyzing the tonometric curve on the three occasions in which the measurements were repeated. On the other hand, the lack of a control group generates an important bias in the research. Finally, I would like to highlight that the discussion section should be rewritten. This is to explain the neurophysiological events that occur under the effect of CBD to reduce intraocular pressure.Likewise, other observations must be addressed to achieve publication quality. I have left some comments, hoping that they can help the authors. Thanks for your insights and comments. We have amended the text in the proper parts to reflect your suggestions. General comments L31-33: Please indicate the IOP values before and after CBD administration, with their respective P values. Text modified to reflect this comment L66: CBD has also been used in pets for the control of acute pain caused by an ovariohysterectomy and chronic pain caused by osteoarthritis. Please add the following references to your manuscript, which you can also use in the discussion of your manuscript: 10.3389/fvets.2024.1380022 10.3389/fvets.2022.1050884 Text modified to reflect this comment L69: I suggest that the authors add the following references in this line: 10.3389/fvets.2024.1496473 10.3389/fvets.2024.1341396 10.1016/j.jevs.2019.102880 Text modified to reflect this comment L73: Please add a hypothesis. Text modified to reflect this comment L82: What was the statistical method used to determine the sample size? Please clarify. The sample size needed was calculated with Sealed Envelope™ calculator, setting significance level at 5% and power at 95%, detailed in the text L84: Please indicate which exclusion criteria were considered in your study. Text modified to reflect this comment L85: What were the analytes or parameters measured in blood before the start of the study? And how long before the administration of CBD were these blood studies performed? Text modified to reflect this comment L108: Due to the type of experimental design used, I suggest the authors perform a statistical analysis based on a linear mixed model instead of a 2-way ANOVA. This model will allow the analysis of the differences observed between treatments and evaluation times, and even for the analysis of the tonometric curve (comparison of three measurements) with their respective washout periods. Statistical analysis has been run according to your suggestion and text modified to reflect this comment L123: Table 1, please indicate the statistical differences observed between treatments and evaluation times with letters and numbers. In this sense, also the results obtained after the administration of CBD at 15 minutes, 30 minutes, 60 minutes, 2 hours, 3 hours, 4 hours, 6 hours, and 8 hours should be integrated into this table; although Figure 1 already shows this information. Table 1 has been updated to reflect your comment L141: In general, the discussion is a section that is shown to be deficient. Therefore, I suggest that the authors complement this section. For example, the neurophysiological effects that generate the decrease in intraocular pressure under the effect of CBD should be explained. I suggest consulting and citing the following references: 10.2460/javma.24.06.0360 10.3934/Neuroscience.2024009 10.3390/ph17060748 10.1016/j.ijpharm.2022.121627 10.3390/ijms22073798 10.1016/j.survophthal.2020.07.002 10.1097/01.ijg.0000212260.04488.60 10.3389/fvets.2022.1050884 10.3390/ph16081149 10.1016/j.biopha.2022.112981 10.1055/a-1665-3100 10.1016/j.exer.2020.108266 10.2174/1570159X15666170724104305 10.1016/j.biopha.2016.11.106 10.1155/2016/9364091 10.1016/S0079-6123(08)01131-X https://pubmed.ncbi.nlm.nih.gov/34283478/ L167: I suggest that the authors discuss the limitations and perspectives of their study. L171: Conclusions must be rewritten based on changes arising from modifications made to the statistical analysis. Discussion and Conclusion sections have been completely rewritten to be more specific and consistent, incorporating all your suggestion Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0325191.r003
22 Apr 2025 Decision Letter - Claudia Interlandi, Editor PONE-D-25-04742R1Effect of orally administered cannabidiol oil on daily tonometric curve in healthy Italian Saddle horsesPLOS ONE Dear Dr. Marchegiani, 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 Jun 06 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. Please include the following items when submitting your revised manuscript: A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'. A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'. An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'. 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, Claudia Interlandi, Ph.D Academic Editor PLOS ONE Journal Requirements: Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice. Additional Editor Comments: The manuscript needs further revision before it can be considered for publication. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #3: (No Response) ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #3: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #3: I Don't Know ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #3: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #3: No ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #3: I reviewed the edited version of the manuscript and provide the following comments to the authors. The majority of my comments are intended to correct the manuscript for grammatical errors and clarity. Line 52/53 – suggest changing to “…common and represents a major cause of retinal…. Line 65 – suggest rewording the section “determine a lowering in IOP” to be more clear and understandable. Line 74 – 76 – suggested edit “In the last few years…investigated in pet medication, …” Line 116 – how was the length of the washout period determined? This information should be added to the manuscript. Line 165 – suggested edit “….after one hour post CBD administration…left eyes) and maintaining….” Line 178 – 180 – this sentence should be reworded to expand and provide clarity on the intended information this sentence is adding to the manuscript. Line 188 – suggested edit “…was started to be studied for its…” Line 190 – edit to “…the insight into the mechanism….” Line 201 – 207 – this section needs to be reworded to ensure clarity for the reader. Line 231 – 233 – please expand this section so that the point you are making is clear to the reader. ****** 7. PLOS authors have the option to publish the peer review history of their article (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 #3: 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". 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. Attachments Attachment Submitted filename: Reviewer 3.docx https://doi.org/10.1371/journal.pone.0325191.r004
Revision 2
5 May 2025 Author Response Dear Editor and Reviewer, on behalf of all other Authors, I would thank for the time and efforts spent in reviewing the manuscript and for the points raised, which allow to improve the overall quality of the paper. We have carefully considered your suggestions and addressed them throughout thew text, using the revision function of Microsoft Word to highlight changes in the text. In addition, English language has been reviewed by a native speaker. Below, in bold blue type, the point-to-point reply to reviewers I reviewed the edited version of the manuscript and provide the following comments to the authors. The majority of my comments are intended to correct the manuscript for grammatical errors and clarity. Line 52/53 – suggest changing to “…common and represents a major cause of retinal…. Text has been modified to reflect this comment Line 65 – suggest rewording the section “determine a lowering in IOP” to be more clear and understandable. Text has been modified to reflect this comment Line 74 – 76 – suggested edit “In the last few years…investigated in pet medication, …” Text has been modified to reflect this comment Line 116 – how was the length of the washout period determined? This information should be added to the manuscript. Detailed in the text Line 165 – suggested edit “….after one hour post CBD administration…left eyes) and maintaining….” Text has been modified to reflect this comment Line 178 – 180 – this sentence should be reworded to expand and provide clarity on the intended information this sentence is adding to the manuscript. Text has been modified to reflect this comment Line 188 – suggested edit “…was started to be studied for its…” Text has been modified to reflect this comment Line 190 – edit to “…the insight into the mechanism….” Text has been modified to reflect this comment Line 201 – 207 – this section needs to be reworded to ensure clarity for the reader. Text has been modified to reflect this comment Line 231 – 233 – please expand this section so that the point you are making is clear to the reader. Text has been modified to reflect this comment https://doi.org/10.1371/journal.pone.0325191.r005
9 May 2025 Decision Letter - Claudia Interlandi, Editor Effect of orally administered cannabidiol oil on daily tonometric curve in healthy Italian Saddle horses PONE-D-25-04742R2 Dear Dr. Andrea Marchegiani, 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. If you have any questions relating to publication charges, please contact our Author Billing department directly at authorbilling@plos.org. 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, Claudia Interlandi, Ph.D Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0325191.r006
Formally Accepted
Acceptance Letter - Claudia Interlandi, Editor PONE-D-25-04742R2 PLOS ONE Dear Dr. Marchegiani, 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. 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 Professor Claudia Interlandi Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0325191.r007

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