Inclusion of chia seeds (Salvia hispanica L.) and pumpkin seeds (Cucurbita moschata) in dairy sheep diets

Lizbeth E. Robles Jimenez; Edgar Aranda Aguirre; Maria de los Angeles Colin Cruz; Beatriz Schettino-Bermúdez; Rey Gutiérrez-Tolentino; Alfonso J. Chay-Canul; Ricardo A. Garcia-Herrera; Navid Ghavipanje; Octavio A. Castelan Ortega; Einar Vargas-Bello-Pérez; Manuel Gonzalez-Ronquillo

doi:10.1371/journal.pone.0300864

Peer Review History

Original SubmissionAugust 15, 2023
7 Sep 2023 Decision Letter - Aziz ur Rahman Muhammad, Editor PONE-D-23-22942Inclusion of chia seeds (Salvia hispanica L.) and pumpkin seeds (Cucurbita moschata) in dairy sheep dietsPLOS ONE Dear Dr. Vargas-Bello-Pérez, 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 AuthorsPlease also check the result section carefully, because in result section especially in digestibility some values are under/over estimated. Failing to address the issues or reviewers comments will result in rejection of manuscript. Please submit your revised manuscript by Oct 22 2023 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, Aziz ur Rahman Muhammad Academic Editor PLOS ONE 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. Thank you for stating the following financial disclosure: " The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript " At this time, please address the following queries: a) Please clarify the sources of funding (financial or material support) for your study. List the grants or organizations that supported your study, including funding received from your institution. b) State what role the funders took in the study. If the funders had no role in your study, please state: “The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” c) If any authors received a salary from any of your funders, please state which authors and which funders. d) If you did not receive any funding for this study, please state: “The authors received no specific funding for this work.” Please include your amended statements within your cover letter; we will change the online submission form on your behalf. 3. We note that you have stated that you will provide repository information for your data at acceptance. Should your manuscript be accepted for publication, we will hold it until you provide the relevant accession numbers or DOIs necessary to access your data. If you wish to make changes to your Data Availability statement, please describe these changes in your cover letter and we will update your Data Availability statement to reflect the information you provide. 4. Your ethics statement should only appear in the Methods section of your manuscript. If your ethics statement is written in any section besides the Methods, please delete it from any other section. Additional Editor Comments: Dear Authors Please revise the manuscript as suggested by reviewers. Please also check the result section carefully, because in result section especially in digestibility some values are under/over estimated. [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: Yes Reviewer #3: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes Reviewer #3: N/A ****** 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 Reviewer #3: 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 Reviewer #3: 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: The paper exploring the inclusion of chia seeds (Salvia hispanica L.) and pumpkin seeds (Cucurbita moschata) in dairy sheep diets holds promise for enhancing the nutritional profile and overall health of the animals. Both chia and pumpkin seeds are rich sources of essential nutrients such as omega-3 fatty acids, protein, vitamins, and minerals. Incorporating these seeds into the diet could potentially lead to improved milk quality, enhanced immune function, and better reproductive outcomes for dairy sheep. However, the paper may require corrections to address and establish a clear cause-and-effect relationship between seed inclusion and observed benefits. With these adjustments, the study could contribute valuable insights to optimizing dairy sheep nutrition and potentially benefit both animal health and the dairy industry. The following major points should be addressed: Line 70. I have to disagree with the authors. Not all pumpkin seeds contains that much SFA. The reference no (Yang Li et al., 2023) do not report the FA profile in the pumpkin seed cake, only in the feeds . In a recent study, it was reported that pumpkin seed meal contains up to 51% PUFA, among other nutrients (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8738952/), and because of that the above mentioned information do not stand. Please correct. Line 95. … sample collection instead of harvesting. Table 1. please provide at least for CS and PS the total PUFA and MUFA, the total n-3 and the total n-3, as well as their ratio. This is important and significant information. Table 2. please add the measurement units. Line 141. Please explain in words when using first time FAME (fatty acids methyl esters). Table 3. the live weight, is initial or final? Table 6. Please check the data for C18:3n3, no significant effect? Also for other fatty acids In the discussion part, some parameters deserves more considerations to be discussed, especially Nitrogen balance. The conclusion is not accurate. According to the data presented in table 6, the C18:3n3 is not significantly increased, only a tendency, this do not count as a major result. Reviewer #2: The overall idea of this kind of studies is novel and worth's trying, it helps the livestock producers to find energy sources that may improve the quality of animals products such as milk. There are some issues must be addressed by the authors in order to improve the manuscript. Line 33: delete "DM" Line 34: please delete "LW" Line 35: delete "DMI" Line 36: delete "NDF" Line 37: delete "ADL" and "FCM" Line 38: delete "FE" Line 41: when p value is 0.818, it is not considered tendency, please rewrite "Moreover,..... (p = 0.818). Line 43: according to the results of FA composition in the milk, this statement is not true or at least not enough to draw this conclusion, please rewrite the conclusion statement. Line 61-62: how much Mexico is producing from these two seed per year? Line 67-68: Please add their nutrient content in values - Table 2: please add the units of the ingredients and chemical composition. In the footnote, please define the diets Line 162: don't start any statement with abbreviations -Table 3: the digestibility of ADF and ADL is not accurate please re-visit their actual values in check - Table 4: the values of nitrogen balance are questionable, please re-check them. The N balance is too low - Table 5: please recheck. for example milk composition (g/d) are all wrong Although the discussion section is well written, it has to be rewritten to reflect the previous comments regarding some table (see previous comments) Line 280 and conclusion: the positive results of FA composition in milk may not be related to CS and/or PS, it could be because the control group fed on diet contained butter! please clarify Reviewer #3: When I came across the nutrient consumption table, I noticed a fact that will have to be reviewed for me to continue the review: Let's take the CON treatment as an example: In table 2 the proportion of corn silage was 350 g/kg Table 3 describes: Forage intake: 1078 g Concentrate intake: 1386 g DM intake: 2464g Corn silage ratio: 1078/2464 = 456.14 g/kg (a value well above the 350 g/kg described in table 2) NDF intake: 1061 g DM intake: 2464g NDF proportion in the diet effectively consumed: 1061/2464 = 456.14 g/kg, a value very, very different from that of table 2 (259 g/kg). If we do the same thing for EE consumption, we will see that the diet effectively consumed showed values much higher than those described in Table 2. Crude protein intake was not presented. Digestibility coefficients draw a lot of attention: Digestibility of ADF greater than NDF. ADL digestibility of 430 g/kg, never seen before. So, to follow the correction, I suggest that the calculations, spreadsheets, etc., be reviewed. ******* 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 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. https://doi.org/10.1371/journal.pone.0300864.r001
Revision 1
27 Oct 2023 Author Response Cover letter [14th September 2023] Dear Sirs, On behalf of my coauthors, I would like to thank you for the opportunity to revise and resubmit our manuscript (ID: PONE-D-23-22942), entitled “Inclusion of chia seeds (Salvia hispanica L.) and pumpkin seeds (Cucurbita moschata) in dairy sheep diets”. We found the reviewers’ comments to be helpful in revising the manuscript and have carefully considered and responded to each suggestion. We believe have resulted in an improved revised manuscript. Below we have included a response to each reviewer in which we address each comment the reviewers made. In our response to reviewers, the reviewers’ comments are numbered, and our responses follow below. Please be advised that we uploaded a marked-up copy of manuscript labeled 'Revised Manuscript with Track Changes' and an unmarked version of your revised paper without tracked changes labeled 'Manuscript'. We very much hope the revised manuscript is accepted for publication in PLOS ONE. Sincerely, Einar Vargas-Bello-Perez (corresponding author) School of Agriculture, Policy and Development New Agriculture Building, Earley Gate Whiteknights Road, PO Box 237 Reading RG6 6EU Berkshire UK Response to Reviewer 1 Comments: Reviewer #1: The paper exploring the inclusion of chia seeds (Salvia hispanica L.) and pumpkin seeds (Cucurbita moschata) in dairy sheep diets holds promise for enhancing the nutritional profile and overall health of the animals. Both chia and pumpkin seeds are rich sources of essential nutrients such as omega-3 fatty acids, protein, vitamins, and minerals. Incorporating these seeds into the diet could potentially lead to improved milk quality, enhanced immune function, and better reproductive outcomes for dairy sheep. However, the paper may require corrections to address and establish a clear cause-and-effect relationship between seed inclusion and observed benefits. With these adjustments, the study could contribute valuable insights to optimizing dairy sheep nutrition and potentially benefit both animal health and the dairy industry. The following major points should be addressed: � Authors: Thank you for your valuable feedback on our manuscript. We appreciate your comments and suggestions, and we have carefully considered your feedback in preparing our revised manuscript. Line 70: I have to disagree with the authors. Not all pumpkin seeds contains that much SFA. The reference no (Yang Li et al., 2023) do not report the FA profile in the pumpkin seed cake, only in the feeds. In a recent study, it was reported that pumpkin seed meal contains up to 51% PUFA, among other nutrients (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8738952/), and because of that the above mentioned information do not stand. Please correct � Authors: We acknowledge that there is conflicting results in literature for fatty acid composition of pumpkin seed oil which is probably caused by the different variety, climatic conditions, cultivation practices, soil, and etc. Although, we would like to drew your attention on a recent review (10.21608/zvjz.2020.22530.1097) which reports that the pumpkin seed possesses a comparatively modest composition of fatty acids, predominantly the essential fatty acids: linoleic, stearic, oleic and palmitic acids, those four fatty acids estimate almost (98 ± 0.13%) of the total amount of fatty acids. However, the text were revised according to you suggestion, please see line 72. Line 95: … sample collection instead of harvesting. � Authors: This has been done, Please see lines 95. Table 1: please provide at least for CS and PS the total PUFA and MUFA, the total n-3 and the total n-3, as well as their ratio. This is important and significant information. � Authors: This has been done, Please see Table 1. Table 2: please add the measurement units. � Authors: This has been done, Please see Table 2. Line 141: Please explain in words when using first time FAME (fatty acids methyl esters). � Authors: This has been corrected throughout the text. Table 3: the live weight, is initial or final? � Authors: This has been done, Please see Table 3. Table 6: Please check the data for C18:3n3, no significant effect? Also for other fatty acids � Authors: All data were re-checked the data for significance. We would like to clarify that all p-values were within the specified thresholds for declaring significance and tendency. As mentioned in lines 170-171, significance was declared at p ≤ 0.05, and tendency was declared at 0.05< p <0.10. We hope this explanation adequately addresses the reviewer's concern. Please let us know if there or clarifications needed. In the discussion part, some parameters deserve more considerations to be discussed, especially Nitrogen balance. The conclusion is not accurate. According to the data presented in table 6, the C18:3n3 is not significantly increased, only a tendency, this do not count as a major result. � Authors: Thank you for your feedback. We acknowledge that there is a room for more discussion for nitrogen balance. We gave more consideration to this section. Please see lines 233-250. Additionally, we made note of your concerns in conclusion. Thanks for bringing this to my attention. Please see lines 362-363. Response to Reviewer 2 Comments: Reviewer #2: The overall idea of this kind of studies is novel and worth's trying, it helps the livestock producers to find energy sources that may improve the quality of animals products such as milk. There are some issues must be addressed by the authors in order to improve the manuscript. � Authors: Thank you for your valuable feedback on our manuscript. We appreciate your comments and suggestions, and we have carefully considered your feedback in preparing our revised manuscript. Line 33: delete "DM" � Authors: This has been removed as suggested. Line 34: please delete "LW" � Authors: This has been removed as suggested. Line 35: delete "DMI" � Authors: This has been removed as suggested. Line 36: delete "NDF". � Authors: This has been removed as suggested. Line 37: delete "ADL" and "FCM". � Authors: This has been removed as suggested. Line 38: delete "FE" � Authors: This has been removed as suggested. Line 41: when p value is 0.818, it is not considered tendency, please rewrite "Moreover,..... (p = 0.818). � Authors: This is a Typo, we apologize for this oversight. Please see line 41. Line 43: according to the results of FA composition in the milk, this statement is not true or at least not enough to draw this conclusion, please rewrite the conclusion statement. � Authors: This has been corrected, Please see line 43. Line 61-62: how much Mexico is producing from these two seed per year? � Authors: This has been done, Please see lines 62-64. Line 67-68: Please add their nutrient content in values � Authors: This has been done, Please see lines 67-69. Table 2: please add the units of the ingredients and chemical composition. In the footnote, please define the diets � Authors: This has been done, Please see Table 2. Line 162: don't start any statement with abbreviations � Authors: This has been done see line 162. Table 3: the digestibility of ADF and ADL is not accurate please re-visit their actual values in check � Authors: We appreciate your attention to detail and the opportunity to address these issues. We have thoroughly reviewed the data, calculations, and spreadsheets. the potential errors should be related to the FAD and measurement techniques. Considering these concerns, we have made the decision to remove the affected data from our analysis. Unfortunately, this means that we no longer have any remaining residue to further analyze. Table 4: the values of nitrogen balance are questionable, please re-check them. The N balance is too low � Authors: This has been done, the data and calculations were re-checked and the table corrected accordingly, Please see Table 4. Table 5: please recheck. for example milk composition (g/d) are all wrong � Authors: This has been done, the data and calculations were re-checked and the table corrected accordingly, Please see Table 5. Although the discussion section is well written, it has to be rewritten to reflect the previous comments regarding some table (see previous comments). � Authors: This has been done, please see lines 178-180, 184-186, and 233-243. Response to Reviewer 3 Comments: Reviewer #3: When I came across the nutrient consumption table, I noticed a fact that will have to be reviewed for me to continue the review: Let's take the CON treatment as an example: Table 3 describes: Forage intake: 1078 g Concentrate intake: 1386 g DM intake: 2464g Corn silage ratio: 1078/2464 = 456.14 g/kg (a value well above the 350 g/kg described in table 2) NDF intake: 1061 g DM intake: 2464g NDF proportion in the diet effectively consumed: 1061/2464 = 456.14 g/kg, a value very, very different from that of table 2 (259 g/kg). If we do the same thing for EE consumption, we will see that the diet effectively consumed showed values much higher than those described in Table 2. Crude protein intake was not presented. Digestibility coefficients draw a lot of attention: Digestibility of ADF greater than NDF. ADL digestibility of 430 g/kg, never seen before. So, to follow the correction, I suggest that the calculations, spreadsheets, etc., be reviewed. � Authors: We sincerely appreciate your valuable input and the opportunity to rectify these issues. We have carefully re-evaluated the data and calculations as per your suggestion. Upon review, we have identified an error in the calculations for ADF and ADL. Considering these concerns in data, we have removed these affected data from our analysis. Furthermore, we acknowledge your observation regarding silage consumption. We have thoroughly verified the data and recalculated the consumption of both silage and concentrate based on the revised information. Thank you once again for your time and attention. Attachments Attachment Submitted filename: editorial observations evbp.docx* https://doi.org/10.1371/journal.pone.0300864.r002
4 Jan 2024 Decision Letter - Aziz ur Rahman Muhammad, Editor PONE-D-23-22942R1Inclusion of chia seeds (Salvia hispanica L.) and pumpkin seeds (Cucurbita moschata) in dairy sheep dietsPLOS ONE Dear Dr. Vargas-Bello-Pérez, 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 Feb 18 2024 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, Aziz ur Rahman Muhammad 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. [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 #1: All comments have been addressed Reviewer #2: All comments have been addressed ******** 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 #1: Yes Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 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 #1: Yes Reviewer #2: 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 #1: Yes Reviewer #2: 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 #1: The authors addressed all the questions and observations. The paper has been reviewed in a nice manner and the corrections are highlighted accordingly. No further observations. Reviewer #2: Authors have addressed all comments arisen in the previous revision. However,, a few comments should be addressed in this version of the paper. Line 66: replace "30-40 g" with "30-40 g/100g" line 67: replace "30-34 g" with "30-34 g/100g" Line 59-70: please add some information about one or two references about the use of chia seeds in the livestock diets Lines 66-67 or 71 or elsewhere: please be consistent , you either use g/100 g or % Lines 74-75: move it to the previous section of chia seeds Lines 107, 125 or elsewhere in the manuscript: don't start any sentence with abbreviations Line 131: please provide full information about the mikoscan analyzer such as the version number, company name, country...... In Table 3: where is the nutrient digestibility for the other nutrients? In Table 4: still believe that the N lost in the feces and urine is too high and the retained N is too low, please clarify Line 368: 7%!!! I believe it is 6.1% ****** 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 #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. https://doi.org/10.1371/journal.pone.0300864.r003
Revision 2
22 Jan 2024 Author Response Response to Reviewers Authors: Thank you for your valuable feedback on our manuscript. We appreciate your comments and suggestions, and we have carefully considered your feedback in preparing our revised manuscript. Reviewer #2: Authors have addressed all comments arisen in the previous revision. However, a few comments should be addressed in this version of the paper. Line 66: replace "30-40 g" with "30-40 g/100g" Authors: This has been done, Please see line 79 line 67: replace "30-34 g" with "30-34 g/100g" Authors: This has been done, Please see line 80 Line 59-70: please add some information about one or two references about the use of chia seeds in the livestock diets Authors: This has been done, Please see lines 66 - 78 Lines 66-67 or 71 or elsewhere: please be consistent, you either use g/100 g or % Authors: This has been done, Please see line s79-80, 84 L84 seed has a crude protein content of approximately 35 g/100g and from 30 to 50 g/100g of oil [2]. Changed accordingly Lines 74-75: move it to the previous section of chia seeds Authors: This has been done, Please see lines 66 -78 Lines 107, 125 or elsewhere in the manuscript: don't start any sentence with abbreviations Authors: This has been done, Please see linen 119 Line 131: please provide full information about the mikoscan analyzer such as the version number, company name, country...... R Authors: This has been done, Please see line 143 SL60. Milkotronic LTD. Nova Zagora, Bulgaria In Table 3: where is the nutrient digestibility for the other nutrients? Authors: In the past revision we explained the following regarding missing nutrient digestibility data: We sincerely appreciate your valuable input and the opportunity to rectify these issues. We have carefully re-evaluated the data and calculations as suggested. Upon review, we have identified an error in the calculations for ADF and ADL. Considering these concerns regarding the data, we have removed all affected data from our analysis. In Table 4: still believe that the N lost in the feces and urine is too high and the retained N is too low, please clarify Authors: We have reviewed the data and they are correct, especially because they are producing milk and in this case the greatest loss of N was in urine, we reviewed the data and they are correct, and when we make the correction of N in milk is when we get negative. That would be normal for a cow or ewe at the beginning of lactation in many cases. Line 368: 7%!!! I believe it is 6.1% Authors: This has been done, Please see line 380 You are right, this data was corrected in the previous revision, and we did not adjust it in the conclusions, thank you for your observation. https://doi.org/10.1371/journal.pone.0300864.r004
1 Feb 2024 Decision Letter - Aziz ur Rahman Muhammad, Editor PONE-D-23-22942R2Inclusion of chia seeds (Salvia hispanica L.) and pumpkin seeds (Cucurbita moschata) in dairy sheep dietsPLOS ONE Dear Dr. Vargas-Bello-Pérez, 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. Although you have addressed major issues of the current manuscript, however, there are few changes especially grammar and some numerical values need to be addressed. Authors need these to be addressed before publication. Authors are advised to respond and address the said comments Please submit your revised manuscript by Mar 17 2024 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, Aziz ur Rahman Muhammad Academic Editor PLOS ONE Additional Editor Comments : Dear Authors Although you have addressed major issues of the current manuscript, however, there are few changes especially grammar and some numerical values need to be addressed. Authors need these to be addressed before publication. Authors are advised to respond and address the said comments [Note: HTML markup is below. Please do not edit.] [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. https://doi.org/10.1371/journal.pone.0300864.r005
Revision 3
12 Feb 2024 Author Response Dear Authors Although you have addressed major issues of the current manuscript, however, there are few changes especially grammar and some numerical values need to be addressed. Authors need these to be addressed before publication. Authors are advised to respond and address the said comments AUTHORS We have made English style corrections and double-checked numbers. All changes have been done in red colour. Attachments Attachment Submitted filename: REBUTTAL LETTER PlosOne.docx https://doi.org/10.1371/journal.pone.0300864.r006
19 Feb 2024 Decision Letter - Aziz ur Rahman Muhammad, Editor PONE-D-23-22942R3Inclusion of chia seeds (Salvia hispanica L.) and pumpkin seeds (Cucurbita moschata) in dairy sheep dietsPLOS ONE Dear Dr. Vargas-Bello-Pérez, 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 I am unable to find detailed author response on the reviewer comments. Furthermore, I am also unable to find color changes in the manuscript as authors claimed. Therefore, I would like to send it back to author for revision. Please submit your revised manuscript by Apr 04 2024 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, Aziz ur Rahman Muhammad Academic Editor PLOS ONE Dear Editor I am unable to find author response on the reviewer comments. Furthermore, i am also unable to find color changes in the manuscript as authors claimed. Therefore, i would like to send it back to author for revision. [Note: HTML markup is below. Please do not edit.] [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. https://doi.org/10.1371/journal.pone.0300864.r007
Revision 4
19 Feb 2024 Author Response REBUTTAL LETTER Dear Authors Although you have addressed major issues of the current manuscript, however, there are few changes especially grammar and some numerical values need to be addressed. Authors need these to be addressed before publication. Authors are advised to respond and address the said comments AUTHORS We have made English style corrections and double-checked numbers. All changes have been done in red colour. Attachments Attachment Submitted filename: REBUTTAL LETTER PlosOne.docx https://doi.org/10.1371/journal.pone.0300864.r008
7 Mar 2024 Decision Letter - Aziz ur Rahman Muhammad, Editor Inclusion of chia seeds (Salvia hispanica L.) and pumpkin seeds (Cucurbita moschata) in dairy sheep diets PONE-D-23-22942R4 Dear Dr. Vargas-Bello-Pérez, 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 http://www.editorialmanager.com/pone/ 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, Aziz ur Rahman Muhammad Academic Editor PLOS ONE Additional Editor Comments (optional): Dear Authors Thanks for addressing the comments of reviewers. congrats 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 #2: All comments have been addressed ******** 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 #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: Yes ****** 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 #2: 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 #2: Yes ****** 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 #2: No more comments, the authors addressed all comments arisen in the previous version. I recommend accepting the manuscript in the current form ****** 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 #2: No ******** https://doi.org/10.1371/journal.pone.0300864.r009
Formally Accepted
27 Mar 2024 Acceptance Letter - Aziz ur Rahman Muhammad, Editor PONE-D-23-22942R4 PLOS ONE Dear Dr. Vargas-Bello-Pérez, 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 If revisions are needed, the production department will contact you directly to resolve them. If no revisions are needed, you will receive an email when the publication date has been set. At this time, we do not offer pre-publication proofs to authors during production of the accepted work. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few weeks 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 Dr. Aziz ur Rahman Muhammad Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0300864.r010

Open letter on the publication of peer review reports

PLOS recognizes the benefits of transparency in the peer review process. Therefore, we enable the publication of all of the content of peer review and author responses alongside final, published articles. Reviewers remain anonymous, unless they choose to reveal their names.

We encourage other journals to join us in this initiative. We hope that our action inspires the community, including researchers, research funders, and research institutions, to recognize the benefits of published peer review reports for all parts of the research system.

Learn more at ASAPbio .