New horizons in live and dehydrated black soldier fly larvae usage: Behavioral and welfare implications in “Bianca di Saluzzo” cockerels

Valentina Bongiorno; Edoardo Fiorilla; Marta Gariglio; Valeria Zambotto; Eleonora Erika Cappone; Stefania Bergagna; Isabella Manenti; Elisabetta Macchi; Francesco Gai; Achille Schiavone

doi:10.1371/journal.pone.0318793

Peer Review History

Original SubmissionJune 29, 2024
29 Jun 2024 Author Response https://doi.org/10.1371/journal.pone.0318793.r001
9 Oct 2024 Decision Letter - Mohamed Mahrous Amer, Editor PONE-D-24-26609New horizons in live and dehydrated black soldier fly larvae usage: behavioral and welfare implications in “Bianca di Saluzzo” cockerelsPLOS ONE Dear Dr. Fiorilla, 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 Nov 23 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. 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The PLOS ONE style templates can be found at https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf 2. To comply with PLOS ONE submissions requirements, in your Methods section, please provide additional information regarding the experiments involving animals and ensure you have included details on (1) methods of sacrifice, (2) methods of anesthesia and/or analgesia, and (3) efforts to alleviate suffering. Additional Editor Comments: Reviewer 1 recommended: Minor Revision The study examines the effects of live and dehydrated black soldier fly larvae on behavioral and welfare implications in “Bianca di Saluzzo” cockerels. The paper is well-written and provides valuable insights into the efficiency of environmental enrichment with black soldier fly larvae in slow-growing male chickens. However, there are some points that need further attention. - Why only male birds were used in this study? L 25: “excreta corticosterone metabolites," which metabolites do the authors mean? L 36-37: “However, the effects of live and dehydrated larvae (LL and DL, respectively) supplementation on slow-growing male chicken welfare have never been formerly investigated.”. Add this sentence to the final paragraph of the introduction to highlight the research gap and to express the innovation of the work. L 114-118: Have the authors analyzed the chemical composition of black soldier fly larvae? For the basal diet, what were the feed ingredients and chemical composition? 218: “2 birds/pen (48 birds in total)” With 6 replicates and 2 birds sampled from each replicated pen, there will be 36 birds in total not 48. L 374-375: What were the reasons behind the better performance in BSFL-fed birds compared with control, despite the higher H/L ratio? L 539-558: The conclusion should be more concise, emphasizing the practical outcomes of the study. Reviewer 2 recommended mijor revision Strictly address the following points: 1. in the introduction section, need of the study should be ideally raised followed by the hypothesis of the study. 2. in materials and methods section, bird's husbandry should be more elaborated 3. in statistical analysis section, add mathematical model for better understanding of the data analysis. 4. in results section, add actual p-value of each result. 5. in discussion section, add logical reasoning of each result before discussing with the previous studies. 6. in conclusion section, add limitations and implication of the study. [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: Partly ******** 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: The study examines the effects of live and dehydrated black soldier fly larvae on behavioral and welfare implications in “Bianca di Saluzzo” cockerels. The paper is well-written and provides valuable insights into the efficiency of environmental enrichment with black soldier fly larvae in slow-growing male chickens. However, there are some points that need further attention. - Why only male birds were used in this study? L 25: “excreta corticosterone metabolites," which metabolites do the authors mean? L 36-37: “However, the effects of live and dehydrated larvae (LL and DL, respectively) supplementation on slow-growing male chicken welfare have never been formerly investigated.”. Add this sentence to the final paragraph of the introduction to highlight the research gap and to express the innovation of the work. L 114-118: Have the authors analyzed the chemical composition of black soldier fly larvae? For the basal diet, what were the feed ingredients and chemical composition? 218: “2 birds/pen (48 birds in total)” With 6 replicates and 2 birds sampled from each replicated pen, there will be 36 birds in total not 48. L 374-375: What were the reasons behind the better performance in BSFL-fed birds compared with control, despite the higher H/L ratio? L 539-558: The conclusion should be more concise, emphasizing the practical outcomes of the study. Reviewer #2: Dear Authors, Strictly address the following points: 1. in the introduction section, need of the study should be ideally raised followed by the hypothesis of the study. 2. in materials and methods section, bird's husbandry should be more elaborated 3. in statistical analysis section, add mathematical model for better understanding of the data analysis. 4. in results section, add actual p-value of each result. 5. in discussion section, add logical reasoning of each result before discussing with the previous studies. 6. in conclusion section, add limitations and implication of the study. Thank You! ****** 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: Hossein Ali Ghasemi 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: Revewer comments.docx https://doi.org/10.1371/journal.pone.0318793.r002
Revision 1
11 Dec 2024 Author Response Dear Editor, We have addressed all the suggestions raised by the two reviewers and have incorporated their feedback into our revised manuscript. We hope that the revisions meet the expectations of both the reviewers and the journal, and we believe that these changes have strengthened our work. Thank you for the opportunity to improve our paper, and we look forward to your positive consideration. Reviewer 1: Comment: The study examines the effects of live and dehydrated black soldier fly larvae on behavioral and welfare implications in “Bianca di Saluzzo” cockerels. The paper is well-written and provides valuable insights into the efficiency of environmental enrichment with black soldier fly larvae in slow-growing male chickens. However, there are some points that need further attention. Response: Thank you for your positive feedback on the study. We appreciate your recognition of the value of our research on the effects of live and dehydrated black soldier fly larvae (BSFL) on the behavioral and welfare aspects of "Bianca di Saluzzo" cockerels. We are grateful for your thoughtful comments and are happy to address the points you have raised for further attention. We will carefully consider your suggestions and make the necessary revisions to enhance the clarity and rigor of the manuscript. Comment: Why only male birds were used in this study? Response: We chose to focus on male birds for this study because they are typically more relevant for meat production. However, we are currently planning a follow-up trial to investigate the effects of BSFL on laying hens, to address the potential impacts on egg production and overall health. Comment: L 25: “excreta corticosterone metabolites," which metabolites do the authors mean? Response: Thank you for your comment. By "excreta corticosterone metabolites," we are referring specifically to the primary metabolites of corticosterone excreted in the feces. These metabolites are commonly used as biomarkers to assess stress in avian species. Comment: L 36-37: “However, the effects of live and dehydrated larvae (LL and DL, respectively) supplementation on slow-growing male chicken welfare have never been formerly investigated.”. Add this sentence to the final paragraph of the introduction to highlight the research gap and to express the innovation of the work. Response: Thank you for your suggestion. The sentence highlighting the research gap and the innovation of the work has been incorporated in lines 94-102, rather than in lines 36-37, in order to better align with the overall flow of the introduction. We believe this placement improves the clarity and coherence of the manuscript. Comment: L 114-118: Have the authors analyzed the chemical composition of black soldier fly larvae? For the basal diet, what were the feed ingredients and chemical composition? Response: We have added further details in the manuscript regarding the chemical composition of black soldier fly larvae and the ingredients used in the basal diet. These are now provided in lines 113-115 and 121-125. Comment: 218: “2 birds/pen (48 birds in total)” With 6 replicates and 2 birds sampled from each replicated pen, there will be 36 birds in total not 48. Response: Thank you for pointing that out. You are correct; with 6 replicates and 2 birds sampled from each pen, the total number of birds should be 36, not 48. We will correct this discrepancy in the manuscript to reflect the accurate total (line 228). Comment: L 374-375: What were the reasons behind the better performance in BSFL-fed birds compared with the control, despite the higher H/L ratio? Response: Thank you for your interesting question. The enhancement of birds’ performance in relation to BSFL administration may not necessarily be linked with the H/L ratio findings, especially when a distinction between eustress and distress is not possible, as in this case. However, since we found similar results in a previous experiment, we hypothesized that the difference between supplemented and control groups may be attributed to the excitement of the birds in receiving the larvae, which would not have had negative implications on birds’ performance (line 555-557). Another option is related to the microbial load of BSFL which could have modulated H/L ratio levels (line 549-551). Further evaluations, such as the use of a thermal camera, would be helpful in better understanding the origin and the meaning of such stress. Comment: L 539-558: The conclusion should be more concise, emphasizing the practical outcomes of the study. Response: The conclusion has been thoroughly revised based on the insightful comments from both reviewers. Reviewer 2 recommended minor revision Strictly address the following points: 1. in the introduction section, need of the study should be ideally raised followed by the hypothesis of the study 2. in materials and methods section, bird's husbandry should be more elaborated 3. in statistical analysis section, add mathematical model for better understanding of the data analysis. 4. in results section, add actual p-value of each result. 5. in discussion section, add logical reasoning of each result before discussing with the previous studies. 6. in conclusion section, add limitations and implication of the study. Response: 1. Thank you for your valuable suggestions! We incorporated them to revise the introduction, specifically in lines 40–49 and 94–102. 2. Accordingly, to your comment we added information and references in lines 113-115 and 121-125 3. Thank you for your feedback. We would like to clarify that the statistical analysis was performed using IBM SPSS Statistics software, which constructs the mathematical models based on the specific input parameters provided for each analysis. The methods reported, including GLMMs and ANOVA, were implemented in SPSS to ensure robust statistical power and appropriate model fit. Each analysis was carefully selected to match the data distribution and research design, thereby optimizing the reliability of the results. We believe that the use of SPSS to generate these models provides the necessary rigor for our analysis and ensures that the statistical assumptions are appropriately met. We added the mathematical formula in the text (line 255-262). 4. Thank you for your feedback. We report p-values only when between 0.05 and 0.10, to guarantee better readability of the text. For p-values smaller than 0.05, we use the notation "P<0.05, P<0.01, P<0.001" to maintain consistency with the journal's formatting requirements. We hope this clarification addresses your concern. 5. Thank you for your suggestion. We have decided to follow the current approach in the discussion section, where we begin by providing the context before discussing the results, as we believe this approach allows for a clearer understanding of our findings within the theoretical framework, before comparing them with previous studies. We hope that this structure is still satisfactory and meets the journal's expectations. 6. Thank you for your suggestion. The conclusion section has been carefully and completely revised in response to the valuable feedback provided by the reviewers. We believe these changes have strengthened the manuscript, and we appreciate your input in improving our work. Attachments Attachment Submitted filename: review_R1.docx https://doi.org/10.1371/journal.pone.0318793.r003
22 Jan 2025 Decision Letter - Mohamed Mahrous Amer, Editor New horizons in live and dehydrated black soldier fly larvae usage: behavioral and welfare implications in “Bianca di Saluzzo” cockerels PONE-D-24-26609R1 Dear Dr. Fiorilla, 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, Mohamed Mahrous Amer, PhD Academic Editor PLOS ONE Additional Editor Comments (optional): article is now accepted Reviewers' comments: https://doi.org/10.1371/journal.pone.0318793.r004
Formally Accepted
Acceptance Letter - Mohamed Mahrous Amer, Editor PONE-D-24-26609R1 PLOS ONE Dear Dr. Fiorilla, 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 Professor Mohamed Mahrous Amer Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0318793.r005

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