The first complete hand-rearing of two neonatal finless porpoises

Masahiko Kasamatsu; Kazuhiro Hasegawa; Ikuo Wakabayashi; Masami Furuta; Hiroki Inoue; Hidetomo Iwano

doi:10.1371/journal.pone.0288891

Peer Review History

Original SubmissionJuly 9, 2023
13 Dec 2023 Decision Letter - Ulrike Gertrud Munderloh, Editor PONE-D-23-20946The first complete hand-rearing of two neonatal finless porpoisesPLOS ONE Dear Dr. Iwano, 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. First of all, thank you very much for your patience in this lengthy review process. With respect to the comments by Reviewer 1 concerning statistics, I realize that it may not be possible to address all of them by repeating analyses. If you can do so, that would be excellent, but if you are unable to generate new or additional data due to lack of materials or other problems, please explain your inability to follow through with the requested change. Please respond to all comments made by the reviewers, Please submit your revised manuscript by Jan 27 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, Ulrike Gertrud Munderloh, Ph.D. 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. Did you know that depositing data in a repository is associated with up to a 25% citation advantage (https://doi.org/10.1371/journal.pone.0230416)? If you’ve not already done so, consider depositing your raw data in a repository to ensure your work is read, appreciated and cited by the largest possible audience. You’ll also earn an Accessible Data icon on your published paper if you deposit your data in any participating repository (https://plos.org/open-science/open-data/#accessible-data). 3. In your Data Availability statement, you have not specified where the minimal data set underlying the results described in your manuscript can be found. PLOS defines a study's minimal data set as the underlying data used to reach the conclusions drawn in the manuscript and any additional data required to replicate the reported study findings in their entirety. All PLOS journals require that the minimal data set be made fully available. For more information about our data policy, please see http://journals.plos.org/plosone/s/data-availability. "Upon re-submitting your revised manuscript, please upload your study’s minimal underlying data set as either Supporting Information files or to a stable, public repository and include the relevant URLs, DOIs, or accession numbers within your revised cover letter. For a list of acceptable repositories, please see http://journals.plos.org/plosone/s/data-availability#loc-recommended-repositories. Any potentially identifying patient information must be fully anonymized. Important: If there are ethical or legal restrictions to sharing your data publicly, please explain these restrictions in detail. Please see our guidelines for more information on what we consider unacceptable restrictions to publicly sharing data: http://journals.plos.org/plosone/s/data-availability#loc-unacceptable-data-access-restrictions. Note that it is not acceptable for the authors to be the sole named individuals responsible for ensuring data access. We will update your Data Availability statement to reflect the information you provide in your cover letter. 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: No Reviewer #2: Yes ****** 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: This paper reported two cases of successful hand-rearing finless porpoise, which provides invaluable technique, information and data on rescuing the orphans of small cetacean species. This paper mainly provided raw data of their work only with t-test for difference analysis. However, it might be more valubale and helpful for other researchers. So I sugges to further improve the statistical analysis for a better quaility of this work. This paper provided the detailed and supportive data and information on hand-rearing the two finless porpoise babies, which include the formula, the way of hand-rearing, the daily intake, and body weight and blood information of the animals, etc. The process of hand-rearing and nutrition status of the animals were all clearly presented in this paper. But the language needs to be further improved by native speakers. Reviewer #2: This manuscript provides incredibly valuable data about the hand rearing process for porpoise calves, which as the authors say can be extrapolated to endangered species conservation. I commend the authors for sharing as much data and information as they have. Several comments to consider: -Lines 23 and 108: consider clarifying "neglect" with "maternal neglect" -Throughout: consider replacing "force-feeding" with "gavage" or "tube feeding". -Throughout: consider replacing "lactations" with "nursings" or "feedings", as lactations implies the production by the dam as opposed to feeding by calf. -L 44-45: consider adding locations for aquaria (not sure where Point Aquarium is). -L 52: two types of formula are listed, but not discussed or detailed further (the differences between the two). -L 83: This sentence doesn't provide a whole lot of value since all cetaceans are listed. Consider rephrasing as "N asiaorientalis is listed as endangered..." -L 92: Given that a main purpose of the study is to evaluate the nutritional status, consider strengthening conclusions about what the nutritional status of these two animals is based on the data collected. -L 108: consider replacing "breastfeeding" with "nursing". -L 112: I understand that 'neonate A' changes to 'calf A' as the animal ages into that age class, but it may be unclear to readers that this is the same animal; consider changing to 'animal A/B'. -L 130: The fact that the slurry of fish meat was fed separately from formula is an important detail, and Table 1 may benefit from separating the slurry line out for clarity. -Table 1: Consider clarifying "Initial" and "Lactation" as those words don't quite capture the differences between the two (perhaps "Day 1-10" and "Day 11-X", etc.) -L 155: Consider adding total number of days of complete weaning process (I assume 136-88). -L 199: confirming vomiting (versus regurgitation) -L 253-255 and 258-260: listing the mean concentrations of all of these parameters is dense and confusing; consider adding column to Table 2 listing mean values there. -L 301: unsure why 'grow in the first week' is there; this information appears valuable throughout the entire hand-rearing process; recommend clarification. -L 310 and 312: "13 and 17 weeks" and "2 months and 4-7 months" - consider standardizing units (weeks or months) for clarity. -L 352: would argue that this is not just necessary for zoo animals, but all animals (including in situ conservation). -L 355: the authors have not adequately described why higher plasma amino acid concentrations are associated with hypoproteinemia. Also, is there information to cite that this is an unexpected finding for neonates versus adults (e.g. not formula-raised; is this pathologic, or expected)? -L 362: Apologies, as above, it is not clear why reduced protein synthesis would occur if amino acid levels were elevated. -L 382: "bases" - I believe this might be "basis". -L 388: as above, are there citations that suggest that this is a pathologic metabolism disorder based on what is known in other neonate species? E.g. that hypertriglyceridemia is not expected in nursing calves of the same age? -L 391: citation needed for "fatty liver changes and lipid metabolism disorders in neonatal porpoises" -L 399: same as with L 355 above: please explain why hypoproteinemia was associated with elevated amino acid levels in the porpoises. ****** 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: Yes: Claire A Simeone ******** [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: PONE-D-23-20946-Reviewer1.docx https://doi.org/10.1371/journal.pone.0288891.r001
Revision 1
10 Mar 2024 Author Response Reviewer #1 This paper reported two cases of successful hand-rearing finless porpoise, which provides invaluable technique, information and data on rescuing the orphans of small cetacean species. This paper mainly provided raw data of their work only with t-test for difference analysis. However, it might be more valubale and helpful for other researchers. So I sugges to further improve the statistical analysis for a better quaility of this work. This paper provided the detailed and supportive data and information on hand-rearing the two finless porpoise babies, which include the formula, the way of hand-rearing, the daily intake, and body weight and blood information of the animals, etc. The process of hand-rearing and nutrition status of the animals were all clearly presented in this paper. But the language needs to be further improved by native speakers. In response to your kind comments and constructive suggestions, we would like to express our sincere appreciation. Your thorough examination of our research and your valuable suggestions are highly appreciated. I understand and acknowledge the significance of your suggestion regarding the statistical analysis of our research. But it is difficult to obtain additional hand-rearing data sets from neonatal porpoises because the breeding and hand-rearing of porpoises are extremely rare. We would like to increase the number of cases in the future and verify the data as statistically meaningful. Even though the number of cases in this study is small, we consider it important that this report will be valuable in the conservation of endangered animals, and we would like to deliver it to researchers around the world as soon as possible. Our manuscript has been edited in English by native speakers again. Thank you again to the reviewers for their comments and constructive advice. We hope that the changes that have been made to the manuscript meet to your satisfaction. We wish to take this opportunity to thank you for your consideration of our paper for publication in your journal, PLOS ONE. Reviewer #2 In response to your kind comments and constructive suggestions, we would like to express our sincere appreciation. Your thorough examination of our research and your valuable suggestions are highly appreciated. Lines 23 and 108: consider clarifying "neglect" with "maternal neglect". →"neglect" have been changed to "maternal neglect" as suggested (revised manuscript (Rev. M.), line 23 and line 111). Throughout: consider replacing "force-feeding" with "gavage" or "tube feeding". →"force-feeding" has been changed to "tube feeding" as suggested (Rev. M., throughout the manuscript). Throughout: consider replacing "lactations" with "nursings" or "feedings", as lactations implies the production by the dam as opposed to feeding by calf. →"lactations" have been changed to "nursings" as suggested (Rev. M., throughout the manuscript). L 44–45: consider adding locations for aquaria (not sure where Point Aquarium is). →Locations for aquaria have been added as suggested (Rev. M., line 44–45). L 52: two types of formula are listed, but not discussed or detailed further (the differences between the two). →The additional information regarding two types of formula has been added as follow: “Safflower oil in the April formula and salmon oil in the Sea World formula have been used for the hand-rearing of dolphins, but whipping cream has only been used in the Sea World formula [1,2]. (Rev. M., line 54). L 83: This sentence doesn't provide a whole lot of value since all cetaceans are listed. Consider rephrasing as "N asiaorientalis is listed as endangered...". →This sentence has been rephrased as “is listed as endangered species along with all other cetaceans” as suggested (Rev. M., line 87). L 92: Given that a main purpose of the study is to evaluate the nutritional status, consider strengthening conclusions about what the nutritional status of these two animals is based on the data collected. →The sentence strengthening conclusions has been added as suggested (Rev. M., line 411). As follow: “Given the observed hyperlipidemia in the early stages of porpoise hand-rearing, it is possible that there is not a high demand for lipid supplementation in the form of fish oil. The results of this study suggest that neonatal porpoises have a high demand for amino acids, and we found that we needed to add BCAAs to artificial formula for the neonatal porpoises.” L 108: consider replacing "breastfeeding" with "nursing". →"breastfeeding" has been changed to "nursing" as suggested (Rev. M., line 112 and 119). L 112: I understand that 'neonate A' changes to 'calf A' as the animal ages into that age class, but it may be unclear to readers that this is the same animal; consider changing to 'animal A/B'. →“neonate A”and “neonateB” have been changed to “animal A” and “animal B”, respectively as suggested (Rev. M., throughout the manuscript). L 130: The fact that the slurry of fish meat was fed separately from formula is an important detail, and Table 1 may benefit from separating the slurry line out for clarity. →Table 1 has been rearranged as suggested (Rev. M., line 145). Table 1: Consider clarifying "Initial" and "Lactation" as those words don't quite capture the differences between the two (perhaps "Day 1-10" and "Day 11-X", etc.). →The specific number of days have been added in Table 1 as suggested (Rev. M., line 145). L 155: Consider adding total number of days of complete weaning process (I assume 136-88). →Total number of days of the complete weaning process were have been added as suggested (Rev. M., line 158). L 199: confirming vomiting (versus regurgitation). →“vomiting” has been changed to “regurgitation” as suggested (Rev. M., line 202, line 234 and line 247). L 253-255 and 258-260: listing the mean concentrations of all of these parameters is dense and confusing; consider adding column to Table 2 listing mean values there. →The mean concentrations of all of these parameters have been deleted and the mean values have been listed in Table 2 (Rev. M., line 261). L 301: unsure why 'grow in the first week' is there; this information appears valuable throughout the entire hand-rearing process; recommend clarification. →We agree with the comments from Reviewer #2. The sentences have been added as follows: “The highest mortality rate in ex situ neonatal finless porpoises typically occurs within the first 15 days following birth. The data obtained in this study are indispensable for determining the conditions under which artificial hand-rearing enables newborn porpoises to grow throughout the hand-rearing period.” (Rev. M., line 300). L 310 and 312: "13 and 17 weeks" and "2 months and 4-7 months" - consider standardizing units (weeks or months) for clarity. →"13 and 17 weeks" have been changed to "3 and 4 months" as suggested (Rev. M., line 312). L 352: would argue that this is not just necessary for zoo animals, but all animals (including in situ conservation). →“of zoo animals” has been changed to “of animals both in situ and ex situ” as suggested (Rev. M., line 355). L 355, L 362: the authors have not adequately described why higher plasma amino acid concentrations are associated with hypoproteinemia. Also, is there information to cite that this is an unexpected finding for neonates versus adults (e.g. not formula-raised; is this pathologic, or expected)? Apologies, as above, it is not clear why reduced protein synthesis would occur if amino acid levels were elevated. →We would like to discuss hypoproteinemia and plasma amino acid profiles in neonatal porpoises separately. The sentences have been added as follows: “The temporary hypoproteinemia observed early in the hand-rearing of neonatal porpoises that was not accompanied by significant clinical symptoms is interesting. Serum total protein consists mainly of albumin synthesized in the liver. In rapidly growing newborns, liver function might be overloaded, which in turn can affect protein synthesis. Neonates require a specific balance of amino acids based on growth stage, and amino acid requirements differ between neonatal and adult pigs, for instance [29,30]. Protein synthesis and muscle growth can be stimulated in neonatal pigs by providing amino acids with anabolic properties [29]. These results suggest that neonatal porpoises require protein and supplementation with certain appropriate amino acids in the early stages of hand-rearing. Supporting early muscle growth with an adequate supply of amino acids may be essential for proper growth of neonatal porpoises. We believe that this is an important finding for future case studies and could lead to the development of an ideal artificial milk. (Rev. M., line 364). L 382: "bases" - I believe this might be "basis". →"bases" has been revised to “basis” following your comments (Rev. M., line 393). L 388: as above, are there citations that suggest that this is a pathologic metabolism disorder based on what is known in other neonate species? E.g. that hypertriglyceridemia is not expected in nursing calves of the same age? →Yes, the hypertriglyceridemia has not been found in sera from nursing calves of dolphins. Hypertriglyceridemia has been reported to occur in human neonatal diabetes and in equine neonates (Rev. M., line 398). L 391: citation needed for "fatty liver changes and lipid metabolism disorders in neonatal porpoises". →The "lipid metabolism disorders" and related citation as suggested have been additionally described from L392 to L403. L 399: same as with L 355 above: please explain why hypoproteinemia was associated with elevated amino acid levels in the porpoises. →“Conclusion” section has been revised as suggested and the sentence has been added as follows: “The results of this study suggest that neonatal porpoises have a high demand for amino acids, and we found that we needed to add BCAAs to artificial formula for the neonatal porpoises.” (Rev. M., line 413). Thank you again to the reviewers for their comments and constructive advice. We hope that the changes that have been made to the manuscript meet to your satisfaction. We wish to take this opportunity to thank you for your consideration of our paper for publication in your journal, PLOS ONE. Attachments Attachment Submitted filename: Response to Reviewers_Kasamatsu.pdf https://doi.org/10.1371/journal.pone.0288891.r002
28 Mar 2024 Decision Letter - Ulrike Gertrud Munderloh, Editor The first complete hand-rearing of two neonatal finless porpoises PONE-D-23-20946R1 Dear Dr. Iwano, 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, Ulrike Gertrud Munderloh, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: https://doi.org/10.1371/journal.pone.0288891.r003

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 .