Assessing genotypes of buffel grass (Cenchrus ciliaris) as an alternative to maize silage for sheep nutrition

Sultan Singh; Pushpendra Koli; Tejveer Singh; Madan Mohan Das; Subhendu Bikash Maity; Krishna Kunwar Singh; Rohit Katiyar; Asim Kumar Misra; Sanat Kumar Mahanta; Manoj Kumar Srivastava; Uchenna Young Anele; Olatunde Akeem Oderinwale; YongLin Ren

doi:10.1371/journal.pone.0304328

Peer Review History

Original SubmissionFebruary 27, 2024
3 Apr 2024 Decision Letter - Adham A. Al-Sagheer, Editor PONE-D-24-07834Assessing genotypes of buffel grass (Cenchrus ciliaris) as an alternative to maize silage for sheep nutritionPLOS ONE Dear Dr. Koli, 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 May 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, Adham A. Al-Sagheer 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. We note that your Data Availability Statement is currently as follows: [All relevant data are within the manuscript and its Supporting Information files.] Please confirm at this time whether or not your submission contains all raw data required to replicate the results of your study. Authors must share the “minimal data set” for their submission. PLOS defines the minimal data set to consist of the data required to replicate all study findings reported in the article, as well as related metadata and methods (https://journals.plos.org/plosone/s/data-availability#loc-minimal-data-set-definition). For example, authors should submit the following data: - The values behind the means, standard deviations and other measures reported; - The values used to build graphs; - The points extracted from images for analysis. Authors do not need to submit their entire data set if only a portion of the data was used in the reported study. If your submission does not contain these data, please either upload them as Supporting Information files or deposit them to a stable, public repository and provide us with the relevant URLs, DOIs, or accession numbers. For a list of recommended repositories, please see https://journals.plos.org/plosone/s/recommended-repositories. If there are ethical or legal restrictions on sharing a de-identified data set, please explain them in detail (e.g., data contain potentially sensitive information, data are owned by a third-party organization, etc.) and who has imposed them (e.g., an ethics committee). Please also provide contact information for a data access committee, ethics committee, or other institutional body to which data requests may be sent. If data are owned by a third party, please indicate how others may request data access. 3. Please include your full ethics statement in the ‘Methods’ section of your manuscript file. In your statement, please include the full name of the IRB or ethics committee who approved or waived your study, as well as whether or not you obtained informed written or verbal consent. If consent was waived for your study, please include this information in your statement as well. 4. 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. 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 ******** 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: After reviewing the paper “Assessing genotypes of buffel grass (Cenchrus ciliaris) as an alternative to maize silage for sheep nutrition," The research idea is excellent, particularly because it addresses the issue of feed shortages in animal production. Furthermore, it discusses an important point: which breeding varieties can be ensiled without the use of silage additives. The authors make several interesting points, making it difficult for the reader to follow their main argument. I recommend that the authors identify their main argument and structure their manuscript accordingly. Materials & Methods 1. Material and method are unorganized, author should be organized as follows: 2.1. Experiment 1: Screening assays for different genotypes Cenchrus ciliaris 2.1.1. Fresh raw materials The Experimental location and establishment section is listed 2.1.2. Silage perpration The author should list the method of preparing silage in the laboratory for five types of feed Following that, it should mention how silage is prepared for sheep in vivo digestibility. 2.1.3. Fermentation profiles and chemical analysis • The author should describe the steps taken to prepare the silage extract, which is estimated in terms of silage pH and lactic acid levels. In terms of lactic acid estimation, we must shorten this wholesale preparation method with lactic acid, which was determined using a spectrophotometric method as described in (reference). • The chemical composition of fresh and silage, with a brief mention of sugar contents, should be written as follows: Sugar content was determined using... according to (reference). Additionally, include carbohydrate and protein fractions in this section. 2.2. Experiment 2: Invivo digestibility by sheep - The corn silage used has no memory of its origins or whether it worked with the experiment treatment. - Rumen metabolites should be referred to as rumen fermentations, and line 224 should be moved to line 227, following line 145. - The method for estimating volatile fatty acids, ammonia, and nitrogen is not mentioned or referenced. For example, when estimating VFA concentration, did you evulate GC? 2. Some errors must be reconsidered; for example, the author estimates dry matter at 100 °C for 72 hours. It is well known that the relative humidity of air-dried materials is estimated at temperatures of 105°C for six hours, 125°C for four hours, or 135°C for three hours, whereas the total humidity of juicy and green materials is estimated at 40-60°C for 24-72 hours. Furthermore, the estimate of total carbohydrates is supposed to be called nitrogen-free extract; it is known that it is estimated at 100- (moisture + protein + fat + fiber + ash), and non-carbohydrate fiber at 100- (moisture + protein + fat + NDF + ash). This is something that the author must correct. 3. Calculations for energy and digestible crude protein values must be combined with statistical analysis. Results 1. Does Table 1 show the chemical composition of a fresh or silage sample? It should be clarified in the title of the table and the text, even if it is a fresh sample, where the chemical analysis of the silage sample is, and vice versa? The same trend with carbohydrates, protein fractions, and energy values in Table 2. Chemical analysis is required for five types of samples, whether silage or fresh. Why does the author review the silage analysis of selected genotype results again in the in vivo assessment? It is a salvation that was discussed during the initial evaluation. I recommend removing it from the in vivo evaluation. Thus, I recommend that the author organize the results as follows: • Chemical composition of fresh material • Fermentation quality and chemical composition of silage • In vivo digestibility by sheep 2. To combine silage fermentation characteristics and chemical analysis, it is advisable to present them in a single table. When writing about materials and methods, arrange them in the order of the table. It is preferable to start with the fermentation characteristics of silage and then include the chemical analysis in a table and write them in a manuscript. This approach will facilitate the interpretation of the results. 3. The results of the evaluation silage of five genotypes show that silage with IG 96-401 had the highest lactic acid, lowest ammonia concentration, and lowest pH. It was preferable to test it in vivo exclusively with maize silage. On what basis did the author select 96-96? Although the silage evaluation indicates that it is the best IG 96-50 after IG 96-401. 4. Why does the author include corn silage in the initial assessment of five types, especially since his main goal is to use it as an alternative feed to corn silage? His genotype selection was supposed to be based on the closest assessment of corn silage as the standard. Discussion 1. In the section of the discussion, the results must be discussed as follows: • Chemical composition of fresh material The author should review studies that agree or disagree with the study of chemical composition of fresh sample. • Fermentation quality and chemical composition of silage - As you discuss the fermentation parameters for silage, you must clarify whether each parameter in the study conforms to the specifications of good silage or not. For example, if the values of the PH of silage, lactic acid, and ammonia for the standared (Maize silage) and genotypes are recommended for the good silage and indicative of its increase or decrease than the values recommended, you must discuss, with references. Furter, auther sould discuss change chemical composition of silage (labotary ensiling). - The author should discuss the reasons for his choice of the two types of genotypes for animal evaluation. • In vivo digestibility by sheep - It should be noted that if there are results that do not have an impact, such as in nutrient digestion, for example, it is essential to refer to references that either agree or disagree with the study and explain the reasons behind this discrepancy. - The author cites plants in the discussion that have nothing to do with the plant under study (underlines 459, 460, 464, and 465); however, he must also cite references to the plant under study. Reviewer #2: Line 96-967. Where all the samples harvested in the same day or in different days? Line 109. Please explain why the authors used these two genotypes (IG96-96 and IG96-401) for the sheep study. Line 229. Please be more specific what kind of experimental design the authors used for both green (fresh) and silage samples, and for the in vivo study. Line 265 Table 2. PB1 of genotype IG96-50= 1310? I think is a typing error. Line 310 Table 6. Please report daily weight gain, if possible, this data will help the readers to visualize the effect of feeding these type of genotypes ****** 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: Yes: 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: cooments PON journal.docx https://doi.org/10.1371/journal.pone.0304328.r001
Revision 1
25 Apr 2024 Author Response Dear Editor, We would like to thank you for all reviewers for their highly insightful comments, which significantly helped me in the improvisation of the quality of our article. In the following pages, I have written the responses point by point for each comment of reviewers. Hopefully, I have incorporated reviewers all comments and advice. Thanks, and kind regards, Pushpendra Response to Reviewers Reviewer# 1 After reviewing the paper “Assessing genotypes of buffel grass (Cenchrus ciliaris) as an alternative to maize silage for sheep nutrition," The research idea is excellent, particularly because it addresses the issue of feed shortages in animal production. Furthermore, it discusses an important point: which breeding varieties can be ensiled without the use of silage additives. The authors make several interesting points, making it difficult for the reader to follow their main argument. I recommend that the authors identify their main argument and structure their manuscript accordingly. Response: Thanks for your in-depth review and critical suggestions to improve our manuscript. Materials & Methods 1. Material and method are unorganized, author should be organized as follows: 2.1. Experiment 1: Screening assays for different genotypes Cenchrus ciliaris 2.1.1. Fresh raw materials The Experimental location and establishment section is listed 2.1.2. Silage perpration The author should list the method of preparing silage in the laboratory for five types of feed Following that, it should mention how silage is prepared for sheep in vivo digestibility. Response: The above suggestions are incorporated in the manuscript. Outline and subheadings are formed as suggested. 2.1.3. Fermentation profiles and chemical analysis • The author should describe the steps taken to prepare the silage extract, which is estimated in terms of silage pH and lactic acid levels. In terms of lactic acid estimation, we must shorten this wholesale preparation method with lactic acid, which was determined using a spectrophotometric method as described in (reference). Response: The above suggestions are incorporated in the manuscript and reference also incorporated in the text. • The chemical composition of fresh and silage, with a brief mention of sugar contents, should be written as follows: Sugar content was determined using... according to (reference). Additionally, include carbohydrate and protein fractions in this section. Response: The above suggestions are incorporated with modification of text. The suitable reference also incorporated in the text. 2.2. Experiment 2: In vivo digestibility by sheep - The corn silage used has no memory of its origins or whether it worked with the experiment treatment. Response: Yes, maize is considered as one of the treatments along with Cenchrus genotypes during the silage feeding experiments. The values are given in table 4 and accordingly reported and discussed in respective sections. - Rumen metabolites should be referred to as rumen fermentations, and line 224 should be moved to line 227, following line 145. Response: The word rumen metabolites replaced by rumen fermentation. Sentence rearranged as per the suggestions. - The method for estimating volatile fatty acids, ammonia, and nitrogen is not mentioned or referenced. For example, when estimating VFA concentration, did you evulate GC? Response: Thanks for notifying this, we did not use GC here because we have not assessed individual component of volatile fatty acids, we focused on total VFAs. They are mentioned with the reference 26, 27 and 28 are described volatile fatty acids, ammonia, and total nitrogen respectively. The sentence also rephrases by incorporation important information. 2. Some errors must be reconsidered; for example, the author estimates dry matter at 100 °C for 72 hours. It is well known that the relative humidity of air-dried materials is estimated at temperatures of 105°C for six hours, 125°C for four hours, or 135°C for three hours, whereas the total humidity of juicy and green materials is estimated at 40-60°C for 24-72 hours. Furthermore, the estimate of total carbohydrates is supposed to be called nitrogen-free extract; it is known that it is estimated at 100- (moisture + protein + fat + fiber + ash), and non-carbohydrate fiber at 100- (moisture + protein + fat + NDF + ash). This is something that the author must correct. Response: Thank you for your input and for providing the information. We would like to emphasize that all calculations are conducted on a dry matter basis, and we adhered to the standard protocol for drying the samples until no further reduction in weight was observed. This typically involves drying the sample until it reaches a constant weight, meaning that there is no further reduction in weight observed over successive drying periods. We were not focused on the relative humidity which is bit different concept. 3. Calculations for energy and digestible crude protein values must be combined with statistical analysis. Response: Modified as per the suggestions. Results: 1. Does Table 1 show the chemical composition of a fresh or silage sample? It should be clarified in the title of the table and the text, even if it is a fresh sample, where the chemical analysis of the silage sample is, and vice versa? Response: Thanks for pointing out, yes this is for fresh samples and changes have been made in table and within text as well. The same trend with carbohydrates, protein fractions, and energy values in Table 2. Chemical analysis is required for five types of samples, whether silage or fresh. Why does the author review the silage analysis of selected genotype results again in the in vivo assessment? It is a salvation that was discussed during the initial evaluation. I recommend removing it from the in vivo evaluation. Thus, I recommend that the author organize the results as follows: • Chemical composition of fresh material • Fermentation quality and chemical composition of silage • In vivo digestibility by sheep Response: The table 2 also for the fresh samples and changes have been made accordingly. To review the silage analysis for the selective genotype because, our investigation, we found two lines (IG96-96 and IG96-401) exhibiting nutritional profiles comparable to maize (reported in our previous studies). Therefore, we selected only these two genotypes out of the initial five used for fresh sample analysis for further consideration. The primary objective in this study was to identify Cenchrus grass genotypes that could serve as substitutes or alternatives to maize, commonly used for silage preparation. 2. To combine silage fermentation characteristics and chemical analysis, it is advisable to present them in a single table. When writing about materials and methods, arrange them in the order of the table. It is preferable to start with the fermentation characteristics of silage and then include the chemical analysis in a table and write them in a manuscript. This approach will facilitate the interpretation of the results. Response: We thank for your directions but regarding the fermentation quality we assessed all the five genotypes of C. ciliaris and then further we selected two genotypes (IG96-401 and IG96-96) along with maize for large scale ensiling in pits for the animal feeding. Therefore, we kept these two tables separately. 3. The results of the evaluation silage of five genotypes show that silage with IG 96-401 had the highest lactic acid, lowest ammonia concentration, and lowest pH. It was preferable to test it in vivo exclusively with maize silage. On what basis did the author select 96-96? Although the silage evaluation indicates that it is the best IG 96-50 after IG 96-401. Response: That’s correct one. In general, all the three (IG 96-50, IG 96-401 and IG 96-96) genotypes were found suitable for the ensiling characteristics. But due to the availability of material we selected two (IG 96-401 and IG 96-96) genotypes. This work was the extension of our previous work where we screened almost 79 accessions belonging to 6 species of Cenchrus germplasm for key nutritional and silage quality traits. (Singh, S., Singh, T., Singh, K.K., Srivastava, M.K., Das, M.M., Mahanta, S.K., Kumar, N., Katiyar, R., Ghosh, P.K. and Misra, A.K., 2023. Evaluation of global Cenchrus germplasm for key nutritional and silage quality traits. Frontiers in Nutrition, 9, p.1094763.) 4. Why does the author include corn silage in the initial assessment of five types, especially since his main goal is to use it as an alternative feed to corn silage? His genotype selection was supposed to be based on the closest assessment of corn silage as the standard. Response: As mentioned in the response above, our main goal was to use this grass for silage without any additives. After studying for 5-6 years, we found a few genotypes with sugar content over 6-7%, making them suitable for silage. That's why we didn't initially consider maize, but we included it in the feeding trial as it's a well-known standard for comparison. Discussion 1. In the section of the discussion, the results must be discussed as follows: • Chemical composition of fresh material The author should review studies that agree or disagree with the study of chemical composition of fresh sample. Response: Suggestion is accepted, and changes have been made accordingly. • Fermentation quality and chemical composition of silage - As you discuss the fermentation parameters for silage, you must clarify whether each parameter in the study conforms to the specifications of good silage or not. For example, if the values of the PH of silage, lactic acid, and ammonia for the standared (Maize silage) and genotypes are recommended for the good silage and indicative of its increase or decrease than the values recommended, you must discuss, with references. Furter, auther sould discuss change chemical composition of silage (labotary ensiling). Response: The information is incorporated into the text. The reference 54, 55 ,56. The Cenchrus genotypes are found a good tropical grasses for ensiling purpose. - The author should discuss the reasons for his choice of the two types of genotypes for animal evaluation. Response: Following the fermentation quality assessment, three out of the five genotypes (IG 96-50, IG 96-401, and IG 96-96) were deemed suitable for ensiling. However, due to the unavailability of material for IG 96-50, we proceeded with only these two genotypes (IG 96-401 and IG 96-96) for further chemical analysis and in vivo feeding trials. The same is incorporated in the discussion. • In vivo digestibility by sheep - It should be noted that if there are results that do not have an impact, such as in nutrient digestion, for example, it is essential to refer to references that either agree or disagree with the study and explain the reasons behind this discrepancy. Response: We already have reference in relation to the digestibility correlated with the nutritional and chemical composition of the feed. - The author cites plants in the discussion that have nothing to do with the plant under study (underlines 459, 460, 464, and 465); however, he must also cite references to the plant under study. Response: The unrelate content is removed, and text is modified. Reviewer #2: Line 96-97. Where all the samples harvested in the same day or in different days? Response: All the samples were collected on the same day and they were processed together and then stored at 4°C for further chemical and other studies. Line 109. Please explain why the authors used these two genotypes (IG96-96 and IG96-401) for the sheep study. Response: Our primary objective in this study was to identify Cenchrus grass genotype that could serve as substitutes or alternatives to maize, commonly used for silage preparation. Through our investigation, we discovered two lines (IG96-96 and IG96-401) exhibiting nutritional profiles comparable to maize. Therefore, we selected only these two lines out of the initial five for further consideration." Line 229. Please be more specific what kind of experimental design the authors used for both green (fresh) and silage samples, and for the in vivo study. Response: In our study, we employed a Completely Randomized Design for both the green (fresh) and silage samples, ensuring unbiased treatment allocation. For the in vivo animal study, we utilized a Randomized Block Design to enhance precision by accounting for variation among animals. The same is incorporated into the text. Line 265 Table 2. PB1 of genotype IG96-50= 1310? I think is a typing error. Response: Thanks for notifying yes it was typo error, now it is rectified. Line 310 Table 6. Please report daily weight gain, if possible, this data will help the readers to visualize the effect of feeding these type of genotypes Response: Thank you for the notification. Our primary objective was to assess the feeding value in respect of intake, nutrients digestibility and rumen fermentation. We did not specifically focus on studying animal production purposes, which is why we did not calculate body weight parameters. Attachments Attachment Submitted filename: Response to Reviewers.pdf https://doi.org/10.1371/journal.pone.0304328.r002
8 May 2024 Decision Letter - Adham A. Al-Sagheer, Editor PONE-D-24-07834R1Assessing genotypes of buffel grass (Cenchrus ciliaris) as an alternative to maize silage for sheep nutritionPLOS ONE Dear Dr. Koli, 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 22 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, Adham A. Al-Sagheer 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 thorough English language review. [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: (No Response) 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: After reviewing the paper "Assessing genotypes of buffel grass (Cenchrus ciliaris) as an alternative to maize silage for sheep nutrition," the author followed all existing comments with one minor change: the paragraph for Rumen fermentations and microbial counts must be moved from line 165 to 170, after line 210. Reviewer #2: Line 114 rephrase the sentence… mixture samples were analyzed as described (14) Line 129 correct the sentence…Total carbohydrate (g/kg of dry matter) of green (fresh) and silage samples were determined Lines 167-168 correct the sentence… entodiniomorphs were identified by the method of Ogimoto and Imai (25) Line 222- 224: correct the reference style…. Fonnesbeck et al. (1984); Khalil et al. (1986). Table 1. revise the hemicellulose value… 2158 Line 360. Correct English…An another ****** 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: Yes: 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.0304328.r003
Revision 2
9 May 2024 Author Response Response to Reviewers Reviewer# 1 After reviewing the paper "Assessing genotypes of buffel grass (Cenchrus ciliaris) as an alternative to maize silage for sheep nutrition," the author followed all existing comments with one minor change: the paragraph for Rumen fermentations and microbial counts must be moved from line 165 to 170, after line 210. Response: Changes are accepted and modified into the manuscript. Reviewer #2: Line 114 rephrase the sentence… mixture samples were analyzed as described (14) Response: Sentence is rephrased as per the suggestion. Line 129 correct the sentence…Total carbohydrate (g/kg of dry matter) of green (fresh) and silage samples were determined Response: Sentence is corrected and rephrased. Lines 167-168 correct the sentence… entodiniomorphs were identified by the method of Ogimoto and Imai (25) Response: Sentence is corrected. Line 222- 224: correct the reference style…. Fonnesbeck et al. (1984); Khalil et al. (1986). Response: the reference style corrected for both. Table 1. revise the hemicellulose value… 2158 Response: Oh! thanks for notifying. that was typo error, now rectified. Line 360. Correct English…An another Response: Checked for English language. Attachments Attachment Submitted filename: Response to Reviewers-Final.docx https://doi.org/10.1371/journal.pone.0304328.r004
10 May 2024 Decision Letter - Adham A. Al-Sagheer, Editor Assessing genotypes of buffel grass (Cenchrus ciliaris) as an alternative to maize silage for sheep nutrition PONE-D-24-07834R2 Dear Dr. Koli, 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, Adham A. Al-Sagheer Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0304328.r005
Formally Accepted
14 May 2024 Acceptance Letter - Adham A. Al-Sagheer, Editor PONE-D-24-07834R2 PLOS ONE Dear Dr. Koli, 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. Adham A. Al-Sagheer Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0304328.r006

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 .