Inclusion of 8% alfalfa silage advances feeding time and improves late-finishing feed efficiency in pigs

Yu Chen; Xiaogang Zhao; Xionggui Hu; Ji Zhu; Huibo Ren; Huali Li; Lihua Cao; Qingming Cui; Yuan Deng; Zhicai Li; Zhongshan Wei; Weimin Jiang; Yingying Liu; Yinglin Peng; Chen Chen

doi:10.1371/journal.pone.0341429

Peer Review History

Original SubmissionNovember 12, 2025
3 Dec 2025 Decision Letter - Andrey Nagdalian, Editor Dear Dr. Chen, 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 Jan 17 2026 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. 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'. 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We look forward to receiving your revised manuscript. Kind regards, Andrey Nagdalian 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. When completing the data availability statement of the submission form, you indicated that you will make your data available on acceptance. We strongly recommend all authors decide on a data sharing plan before acceptance, as the process can be lengthy and hold up publication timelines. 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If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references.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? Reviewer #1: Yes Reviewer #2: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: Yes Reviewer #2: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes Reviewer #2: Yes ****** Reviewer #1: All key results are correctly described. Perhaps, for completeness, it would be worthwhile to add explicit numerical totals of the final mass and total gain by group, so that the reader can directly assess the effect. However, the lack of such a proposal is not critical, since the figures and models overlap this information. The authors conscientiously interpret the absence of significant differences as the absence of a negative effect of alfalfa on productivity, and pay attention to the minor differences in time dynamics – this is a balanced approach without exaggerating the results. The statistical analysis is beyond doubt. The only thing that can be recommended is to check the wording in the text again for typos related to statistics, for example, in line 1425, the phrase "time period were significantly different" should be corrected to "significantly different". Such small things do not affect the results, but it is important to eliminate them. In general, statistical data processing was carried out carefully and in full compliance with modern standards; the results reliably confirm the conclusions. In the discussion section, it would be worth adding a couple of suggestions about the limits of the application of the results. For example, to emphasize the limited sample (24 animals of a single breed) and the fact that the results were obtained under controlled conditions with adlibitum feeding. This will help the reader understand that the conclusions are correct for these conditions, but may require testing on larger livestock or other breeds. It may also make sense to mention that the economic effect is implied, but was not directly calculated – such a remark will save from overloading conclusions with practical significance. Otherwise, the conclusions fully correspond to the data obtained and answer the questions posed in the introduction. There are only minor typos and stylistic flaws in the text. The suffixes in adverbs were skipped several times – it should be "significantly different" instead of "significantly different" (line 1422) and "significantly lower" instead of "significantly lower" (line 1439). Also in line 1729, the word "In" is capitalized inside the sentence. These little things are easy to fix when editing. In general, the language and structure of the manuscript comply with the standards of scientific publications and, after minimal proofreading, will be understandable to a wide range of specialists. The final recommendation is Minor revisions. The article is a technically high-quality study that meets the PLOS ONE criteria. Minor edits are needed to improve clarity – to clarify some details of the methods (feeding time), make minor language corrections, and discuss the limitations of the experiment. After these edits, the manuscript, in my opinion, will be ready for publication. There are no grounds for rejection or global recycling – the work fulfills its goals and provides valuable results for the pig feeding industry. The manuscript deserves high praise for the novelty of the question, its rigorous experimental approach and the depth of statistical analysis. The score was lowered only due to a limited sample and some clarifications that need to be worked out. Otherwise, the research was carried out at a decent level. Reviewer #2: The text should be carefully checked for typos and grammatical errors. The work is formatted using data indicating the relevance of the problem. The manuscript is technically sound, has sufficient sample sizes, controls and replicas that have undergone statistical processing. The data obtained may become outdated, since the relevance of this problem remains for a long time. It is advisable to refer to modern works touching on similar topics for the corresponding conclusions that are given. ****** 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.] To ensure your figures meet our technical requirements, please review our figure guidelines: https://journals.plos.org/plosone/s/figures You may also use PLOS’s free figure tool, NAAS, to help you prepare publication quality figures: https://journals.plos.org/plosone/s/figures#loc-tools-for-figure-preparation. NAAS will assess whether your figures meet our technical requirements by comparing each figure against our figure specifications. https://doi.org/10.1371/journal.pone.0341429.r001
Revision 1
5 Dec 2025 Author Response Reviewer #1: All key results are correctly described. Perhaps, for completeness, it would be worthwhile to add explicit numerical totals of the final mass and total gain by group, so that the reader can directly assess the effect. However, the lack of such a proposal is not critical, since the figures and models overlap this information. The authors conscientiously interpret the absence of significant differences as the absence of a negative effect of alfalfa on productivity, and pay attention to the minor differences in time dynamics – this is a balanced approach without exaggerating the results. The statistical analysis is beyond doubt. The only thing that can be recommended is to check the wording in the text again for typos related to statistics, for example, in line 1425, the phrase "time period were significantly different" should be corrected to "significantly different". Such small things do not affect the results, but it is important to eliminate them. In general, statistical data processing was carried out carefully and in full compliance with modern standards; the results reliably confirm the conclusions. In the discussion section, it would be worth adding a couple of suggestions about the limits of the application of the results. For example, to emphasize the limited sample (24 animals of a single breed) and the fact that the results were obtained under controlled conditions with adlibitum feeding. This will help the reader understand that the conclusions are correct for these conditions, but may require testing on larger livestock or other breeds. It may also make sense to mention that the economic effect is implied, but was not directly calculated – such a remark will save from overloading conclusions with practical significance. Otherwise, the conclusions fully correspond to the data obtained and answer the questions posed in the introduction. There are only minor typos and stylistic flaws in the text. The suffixes in adverbs were skipped several times – it should be "significantly different" instead of "significantly different" (line 1422) and "significantly lower" instead of "significantly lower" (line 1439). Also in line 1729, the word "In" is capitalized inside the sentence. These little things are easy to fix when editing. In general, the language and structure of the manuscript comply with the standards of scientific publications and, after minimal proofreading, will be understandable to a wide range of specialists. The final recommendation is Minor revisions. The article is a technically high-quality study that meets the PLOS ONE criteria. Minor edits are needed to improve clarity – to clarify some details of the methods (feeding time), make minor language corrections, and discuss the limitations of the experiment. After these edits, the manuscript, in my opinion, will be ready for publication. There are no grounds for rejection or global recycling – the work fulfills its goals and provides valuable results for the pig feeding industry. The manuscript deserves high praise for the novelty of the question, its rigorous experimental approach and the depth of statistical analysis. The score was lowered only due to a limited sample and some clarifications that need to be worked out. Otherwise, the research was carried out at a decent level. Response: Thank you for your detailed and constructive comments. We are grateful for the positive feedback on the novelty, experimental approach, and depth of statistical analysis in our study. We have addressed all minor revisions as per the reviewer’s suggestions. We have made the necessary clarifications to improve the clarity of the methods, corrected the language issues, and incorporated the recommended discussion on limitations. We have addressed the points as follows: 1. Typo Corrections: Thank you for pointing out the typographical errors. We have carefully reviewed the entire manuscript and corrected the spelling as suggested. 2. Method section: We have made revisions to the method section (feeding time) to enhance clarity, as follows: “The linear analysis allowed for the evaluation of overall feeding trends over time, capturing changes in feeding behavior along a continuous temporal scale. In contrast, the circular analysis addressed the periodic nature of feeding patterns, where the 24-hour cycle repeats, enabling the detection of daily rhythms and shifts in behavior that may not be apparent in a linear framework”. 3. Results section: We acknowledge the reviewer’s suggestion to include explicit numerical totals of final mass and total gain by group. In response, we have added the detail in the result section as follow “The final mass and weight gain for pigs with feed type A were 131.2 ± 10.8 kg and 66.4 ± 5.7 kg, respectively. Pigs on feed type B exhibited a final mass of 125.3 ± 11.5 kg and a weight gain of 63.9 ± 10.8 kg. Pigs receiving feed type C showed a final mass of 134.2 ± 7.3 kg and a weight gain of 72.0 ± 6.6 kg”. 4. Discussion Section: We appreciate the reviewer’s suggestion to discuss the limitations of the results in the context of the sample size and controlled conditions. In response, we have expanded the discussion as follows: “The generalizability of the findings is constrained by the relatively small sample size (24 animals of a single breed) used in this study. Furthermore, the experiment was carried out under controlled conditions with ad libitum feeding, which may not accurately replicate typical field conditions. To enhance the applicability of the results, further research involving larger sample sizes or different breeds would be necessary. While the potential economic implications of incorporating alfalfa were acknowledged, a detailed economic analysis was not performed, as it fell outside the scope of the current investigation.” Reviewer #2: The text should be carefully checked for typos and grammatical errors. The work is formatted using data indicating the relevance of the problem. The manuscript is technically sound, has sufficient sample sizes, controls and replicas that have undergone statistical processing. The data obtained may become outdated, since the relevance of this problem remains for a long time. It is advisable to refer to modern works touching on similar topics for the corresponding conclusions that are given. Response: We greatly appreciate the constructive comments provided. In response, we have carefully reviewed the entire manuscript and made corrections to the typos and grammatical errors, as also suggested by Reviewer 1. Regarding the suggestion to “refer to modern works touching on similar topics,” we would like to clarify that our discussion and reference list already include several recent and relevant studies on alfalfa, dietary fiber, and feeding behavior in pigs (e.g., Gao et al., 2024; Luo et al., 2021; Piles et al., 2025; Huangfu et al., 2024). These citations effectively connect our findings to contemporary research on the role of alfalfa in swine growth and precision feeding. Our study specifically addresses a gap in the current literature by investigating the stage-specific effects of alfalfa substitution during the fattening period. While the long-term relevance of feed efficiency is acknowledged, our novel contribution lies in demonstrating that: 1. Overall performance is not compromised with 8% alfalfa inclusion. 2. Phase-dependent effects are observed (e.g., improved FCR in the late finishing phase for alfalfa groups), supporting modern precision feeding strategies. 3. Alfalfa silage significantly alters feeding behavior, particularly in terms of temporal patterns, with potential practical implications for farm management. We believe that the manuscript already engages with the current scientific literature, and we hope this explanation effectively addresses the reviewer’s concerns. Attachments Attachment Submitted filename: Response to Reviewers.docx https://doi.org/10.1371/journal.pone.0341429.r002
7 Jan 2026 Decision Letter - Andrey Nagdalian, Editor Inclusion of 8% alfalfa silage advances feeding time and improves late-finishing feed efficiency in pigs PONE-D-25-60915R1 Dear Dr. Chen, 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. For questions related to billing, please contact billing support . 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, Andrey Nagdalian Academic Editor PLOS One Reviewers' comments: Reviewer's Responses to Questions Comments to the Author Reviewer #1: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions??> Reviewer #1: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? -->?> Reviewer #1: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available??> The PLOS Data policy Reviewer #1: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English??> Reviewer #1: Yes ****** Reviewer #1: It is recommended to carefully proofread the text for individual stylistic details before the final publication, although in general the language of the article already meets high standards and does not contain the errors indicated by the reviewer. These remarks are advice for the final polishing of the text. ****** 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 ******** https://doi.org/10.1371/journal.pone.0341429.r003
Formally Accepted
Acceptance Letter - Andrey Nagdalian, Editor PONE-D-25-60915R1 PLOS One Dear Dr. Chen, I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS One. Congratulations! Your manuscript is now being handed over to our production team. At this stage, our production department will prepare your paper for publication. This includes ensuring the following: * All references, tables, and figures are properly cited * All relevant supporting information is included in the manuscript submission, * There are no issues that prevent the paper from being properly typeset You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps. Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing. 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. Andrey Nagdalian Academic Editor PLOS One https://doi.org/10.1371/journal.pone.0341429.r004

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