-->PONE-D-25-12452-->-->Nutritional quality and in vitro gas production of Piptocoma discolor (Kunth) Pruski forage across contrasting landscapes in the Colombian Amazon Piedmont-->-->PLOS ONE

Dear Dr. Rios Parra,

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 Oct 18 2025 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,

Vitor Hugo Rodrigues Paiva, 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. In your Methods section, please provide additional information regarding the permits you obtained for the work. Please ensure you have included the full name of the authority that approved the field site access and, if no permits were required, a brief statement explaining why.

3. 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.

4. 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. Please note that, though access restrictions are acceptable now, your entire data will need to be made freely accessible if your manuscript is accepted for publication. This policy applies to all data except where public deposition would breach compliance with the protocol approved by your research ethics board. If you are unable to adhere to our open data policy, please kindly revise your statement to explain your reasoning and we will seek the editor's input on an exemption. Please be assured that, once you have provided your new statement, the assessment of your exemption will not hold up the peer review process.

5. 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.

6. We note that Figure 1 in your submission contain map images which may be copyrighted. All PLOS content is published under the Creative Commons Attribution License (CC BY 4.0), which means that the manuscript, images, and Supporting Information files will be freely available online, and any third party is permitted to access, download, copy, distribute, and use these materials in any way, even commercially, with proper attribution. For these reasons, we cannot publish previously copyrighted maps or satellite images created using proprietary data, such as Google software (Google Maps, Street View, and Earth). For more information, see our copyright guidelines: http://journals.plos.org/plosone/s/licenses-and-copyright.

We require you to either (1) present written permission from the copyright holder to publish these figures specifically under the CC BY 4.0 license, or (2) remove the figures from your submission:

1. You may seek permission from the original copyright holder of Figure 1 to publish the content specifically under the CC BY 4.0 license.

We recommend that you contact the original copyright holder with the Content Permission Form (http://journals.plos.org/plosone/s/file?id=7c09/content-permission-form.pdf) and the following text:

“I request permission for the open-access journal PLOS ONE to publish XXX under the Creative Commons Attribution License (CCAL) CC BY 4.0 (http://creativecommons.org/licenses/by/4.0/). Please be aware that this license allows unrestricted use and distribution, even commercially, by third parties. Please reply and provide explicit written permission to publish XXX under a CC BY license and complete the attached form.”

Please upload the completed Content Permission Form or other proof of granted permissions as an "Other" file with your submission.

In the figure caption of the copyrighted figure, please include the following text: “Reprinted from [ref] under a CC BY license, with permission from [name of publisher], original copyright [original copyright year].”

2. If you are unable to obtain permission from the original copyright holder to publish these figures under the CC BY 4.0 license or if the copyright holder’s requirements are incompatible with the CC BY 4.0 license, please either i) remove the figure or ii) supply a replacement figure that complies with the CC BY 4.0 license. Please check copyright information on all replacement figures and update the figure caption with source information. If applicable, please specify in the figure caption text when a figure is similar but not identical to the original image and is therefore for illustrative purposes only.

The following resources for replacing copyrighted map figures may be helpful:

USGS National Map Viewer (public domain): http://viewer.nationalmap.gov/viewer/

The Gateway to Astronaut Photography of Earth (public domain): http://eol.jsc.nasa.gov/sseop/clickmap/

Maps at the CIA (public domain): https://www.cia.gov/library/publications/the-world-factbook/index.html and https://www.cia.gov/library/publications/cia-maps-publications/index.html

NASA Earth Observatory (public domain): http://earthobservatory.nasa.gov/

Landsat: http://landsat.visibleearth.nasa.gov/

USGS EROS (Earth Resources Observatory and Science (EROS) Center) (public domain): http://eros.usgs.gov/#

Natural Earth (public domain): http://www.naturalearthdata.com/

7. If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise.

Additional Editor Comments:

Along with the comments and suggestions of changes raised by two experts and described below, do not forget to reply to those described on an attached annotated PDF of the manuscript

[Note: HTML markup is below. Please do not edit.]

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: 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 study on the nutritional quality and in vitro gas production of Piptocoma discolor across contrasting landscapes in the Colombian Amazon Piedmont provides valuable insights into its potential as a forage resource for ruminant supplementation in silvopastoral systems. Its importance lies in addressing sustainable livestock production and soil fertility challenges in a deforestation-impacted region. With the minor revisions suggested—enhancing clarity, scientific precision, and ecological context through suggested citations- the manuscript can be well-positioned for acceptance, offering a robust contribution to tropical agroforestry research.

Comments and Suggestions

1. Line 27-40 (Abstract):

� Comment: The abstract is concise but could benefit from a brief mention of the study’s broader implications (e.g., sustainability or livestock productivity).

� Suggestion: Add after line 40: "These results highlight the potential of P. discolor to enhance ruminant nutrition and support sustainable silvopastoral practices in the region."

2. Line 46-47 (Introduction):

� Comment: The phrase "rainforest and tropical forest fragmentation" is slightly redundant since rainforests are a type of tropical forest.

� Suggestion: Revise to: "Livestock production is a major driver of tropical rainforest fragmentation worldwide, altering the composition, configuration, and ecological functions of these ecosystems."

3. Line 61-62 (Introduction):

� Comment: The nutritional values (6.6% CP, 75% NDF, 65% digestibility) lack a clear source or context, which might confuse readers.

� Suggestion: Add qualifier: "The predominant forage base consists of grasses with low nutritional value, e.g., 6.6% CP, 75% NDF, and 65% digestibility, as reported in regional studies [13]."

4. Line 83 (Introduction):

� Comment: The benefits of native trees in silvopastoral systems are mentioned, but soil biota’s role in sustainability could be emphasized.

� Suggestion: After line 83, add: "Soil organisms like earthworms and fungi enhance nutrient availability and combat land degradation, supporting sustainability, cite https://doi.org/10.1002/ldr.5446 as it link soil health to forage quality, reinforcing the study’s ecological context.

5. Line 104 (Materials and Methods):

� Comment: The transition to "Materials and Methods" feels abrupt without a closing statement for the Introduction.

� Suggestion: Add before- Suggestion: Before line 104, add: "To address these gaps, this study evaluated the nutritional quality and rumen fermentation potential of P. discolor across two distinct landscapes."

6. Line 111 (Study Area):

� Comment: "Köppen climate type A (Tropical Rainforest - Equatorial - Af)" could be simplified for readability.

� Suggestion: Revise to: "classifying it as a Köppen Af (tropical rainforest) climate in Colombia [27]."

7. Line 119 (Materials and Methods - Study Area):

� Comment: The study area description could suggest future research methods like remote sensing for landscape analysis.

� Suggestion: After line 119, add and cite https://doi.org/10.1016/j.pce.2024.103667 , https://doi.org/10.1007/s11356-022-20305-y, https://doi.org/10.1080/10807039.2015.1122509. "Future studies could use remote sensing, as in to map forage quality variability across Amazonian landscapes” as it offers a methodological tool for scaling up landscape-scale forage studies.

8. Line 129 (Sampling Design):

� Comment: The sample size calculation lacks detail on how 36 and 23 trees were derived.

� Suggestion: Add: "The sample size was calculated using Cochran’s formula [30], yielding 36 trees in the hill and 23 in the alluvial plain, totaling 59 (Fig 1)."

9. Line 147 (Nutritional Composition):

� Comment: "Dry matter (DM) content" is introduced abruptly without linking to the drying process.

� Suggestion: Revise to: "Samples were dried… to determine dry matter (DM) content, calculated as the percentage of weight remaining [AOAC, 2005; Method 950.46]."

10. Line 171 (In Vitro Gas Production):

� Comment: "2.5 ± 0.5 years old" is vague and could be more precise.

� Suggestion: Replace with: "approximately 2.5 years old (range: 2–3 years)."

11. Line 200 (Data Analysis):

� Comment: "Cdb and Cwb" appear in correlation analysis (line 231) but lack prior definition.

� Suggestion: Add at line 151: "Organic carbon was measured on a dry basis (Cdb) and wet basis (Cwb) using the Walkley and Black method [33]."

12. Line 214 (Results - Forage Nutritional Quality):

� Comment: The sentence "The DM content was higher in the alluvial plain…" is clear but could be more engaging.

� Suggestion: Revise to: "Forage from the alluvial plain showed significantly higher DM content than the hill landscape (36.1% vs. 30.9%, P = 0.0458)."

13. Line 231-234 (Results - Pearson’s Correlation):

� Comment: Abbreviations "Cdb," "Cwb," "M," and "OC" are used without prior definition, potentially confusing readers.

� Suggestion: Define at line 151 and simplify: "Pearson’s analysis showed strong positive correlations between CP and N (r > 0.6, P < 0.05) and Mg with Ca, Mn (r > 0.6, P < 0.05), and negative correlations between NDF and DM, Cwb (r < -0.6, P < 0.05) (Fig 2)."

14. Line 259-260 (Results - In Vitro Gas Production):

� Comment: "4.5 ml/g IDM less" is awkwardly phrased.

� Suggestion: Revise to: "whereas the hill landscape produced 19.9 ml/g IDM, 4.5 ml/g IDM less than the alluvial plain (P < 0.05)."

15. Line 291-292 (Discussion):

� Comment: The comparison to Riascos et al. and Guayara lacks context for CP content differences.

� Suggestion: Clarify: "CP content (9.48% hill vs. 14.62% alluvial plain) was lower than 21% [20] and 20.4% [23], likely due to regrowth age and environmental factors."

16. Line 335-336 (Discussion):

� Comment: "Soil-type effect" is vague and could be more specific.

� Suggestion: Revise to: "Differences in DM, ash, NDF, CP, N, and P reflect soil fertility, likely enhanced by soil biota like earthworms and fungi in the alluvial plain” cite https://doi.org/10.1002/ldr.5446, as it explains soil biota’s role in nutrient retention, supporting the study’s findings.

17. Line 363-364 (Conclusions):

� Comment: "Bromatological analysis" may not be clear to all readers.

� Suggestion: Replace with: "Nutritional analysis of P. discolor from the alluvial plain suggests favorable properties for ruminant supplementation…"

18. Line 370 (Conclusions):

� Comment: The call for further research could include specific methods for broader application.

� Suggestion: After line 370, add: "Remote sensing, as in [Bhattacharjee et al., 2024], could assess landscape-scale adaptability of P. discolor in silvopastoral systems." https://doi.org/10.1016/j.pce.2024.103667 provides a method to expand the study’s scope, enhancing future research.

19. Line 374-378 (Acknowledgments):

� Comment: The acknowledgment is brief and could be more formal.

� Suggestion: Revise to: "The authors thank the Universidad de la Amazonia and COFEMA S.A., especially Mr. Milton Chávez López, for their valuable support."

Reviewer #2: The paper, titled "Nutritional quality and in vitro gas production of Piptocoma discolor (Kunth) Pruski forage across contrasting landscapes in the Colombian Amazon Piedmont", addresses an important and timely topic. I found the subject matter of the article fascinating and read the manuscript with great interest. The paper aligns well with the scope of the journal.

Main Questions and Relevance

The main question this research tackles is how the nutritional quality and in vitro total gas production of Piptocoma discolor forage differs when collected from two very distinct landscapes—alluvial plain and hill—in the Colombian Amazon piedmont. They're basically trying to see if where this plant grows affects how good it is as feed for ruminants and how much gas (like methane, which is a big deal for climate change) it produces.

I definitely consider this topic original and highly relevant to the field of animal nutrition and sustainable agriculture, especially in tropical regions. It directly addresses a pretty significant gap: while we know a lot about common forage species, there's less research on the potential of native trees and shrubs, particularly how their nutritional value might vary based on local environmental conditions like soil type. This is crucial for developing sustainable silvopastoral systems. It’s not just about finding new feed, but understanding how to use existing, native resources more effectively in specific ecological contexts. Many studies just look at one area, but here, they’ve thought about the different landscapes.

Contribution to the Subject Area

What this paper adds, really, is a specific and detailed look at Piptocoma discolor in a region where such data is pretty scarce. Compared to other published material that might focus on more common forage species or broad comparisons, this study provides empirical data on a promising native species, highlighting how landscape-level differences can impact key nutritional parameters like crude protein, fiber, and mineral content, as well as gas production. The finding that P. discolor's degradability is moderately acceptable regardless of landscape is quite useful, and the differences in cumulative gas production are certainly noteworthy. It really helps to fill in the picture for those looking to implement sustainable livestock practices in the Colombian Amazon.

However, I believe that in its current form, it has several shortcomings.

Specific comments

Introduction

The introduction does a good job of setting the stage, emphasizing the importance of silvopastoral systems in the Amazonian context. However, it feels like it could be a tad more concise in certain parts. For example, some of the initial sentences on deforestation and climate change, while true, are quite broad and could be streamlined to get to the core problem statement a bit faster. Also, when discussing previous work on P. discolor, perhaps a slightly more critical look at why this study is needed, beyond just stating its promising nature, would be beneficial. What specific questions remain unanswered by earlier research that this paper uniquely addresses? The hypothesis is clearly stated, which is a big plus, but a stronger link between the literature review and the novelty of their specific landscape comparison would be good.

In the Introduction, line 45, it says "fragmentation worldwide, altering the composition, configuration, and ecological functions of these ecosystems." The phrase "these ecosystems" is a bit vague, could perhaps specify "rainforest and tropical forest ecosystems" again for clarity.

Still in the Introduction, line 55 talks about the "hilly landscape, which dominates the region (67.9%) has been disproportionately affected (70%)". The percentages here are very close but refer to different things; maybe a quick rephrasing for precision would help avoid a misreading, like "the hilly landscape, comprising 67.9% of the region, has been disproportionately affected (70% of the conversion)."

In the Introduction, when discussing previous work on P. discolor and its promising nature, there might be room to add more citations if there's research that specifically details why this plant is so "promising." It’s kinda generally stated.

In line 73, the secondary compounds must be listed, maybe they are tannins, please see andi cite: 10.1016/j.vas.2025.100434 to support your statement.

Materials and Methods

The methodology is generally well-described, but I have a few specific points that need attention. First, on the sampling design: While the sample size calculation is mentioned as per Cochran [30], it would be helpful to briefly explain how that calculation was applied, perhaps noting the variance estimates used, especially given the differing number of trees sampled in each landscape (36 vs. 23). This just adds to the transparency. Second, when describing the rumen fluid collection, mentioning that cattle had "free access to fresh water" is good, but any information on their feed prior to slaughter would be immensely valuable. The diet of the donor animals can significantly influence the rumen microbial population, which, in turn, impacts in vitro gas production results. This is a pretty important detail for replicability and interpretation. Finally, for the gas production calculations, Equation (1) is provided, but it's not clear what "x" represents in that specific equation. Is it pressure? Is it time? The text says "per unit of pressure x," which suggests pressure, but clarification would remove any ambiguity. It just seems a little bit confusing for the reader.

In the Materials and Methods, line 107, there's a typo in "Municipality of Florencia (Fig 1), covering an area of 107 2,262 km2." The "107" seems like a stray line number or something that got included by mistake.

Also in Methods, line 131, it mentions "height ranging from 16 to 20 m". Is this referring to the height of the trees or the sampling height on the trees? I'm assuming trees, but clarifying would be good.

Supporting your in vitro methods I suggest citing 10.1080/1828051X.2021.1899063.

The authors should consider including references to support the statistical methods used in the analysis. Some key areas to address include: 10.3390/ani13061107 for pearson correlation; 10.3389/fvets.2024.1332207 for PCA; 10.29261/pakvetj/2020.067 for Shapiro wilk; 10.1080/01652176.2024.2347928 for tukey; 10.1186/s12917-023-03823-w for GLM.

Results

The results section is largely clear, but the presentation could be improved, particularly in the text versus table consistency. For instance, in Table 1, the "I" for Iron seems like a typo and should probably be "Fe" for consistency with other mineral abbreviations and standard chemical symbols. Just a small thing, but those sorts of details can distract a reader. When discussing the PCA, Figure 3 is very illustrative, but the explanation in the text about PC1 and PC2 could be slightly more explicit about which variables are loading strongly in which direction along those axes, beyond just listing them. For example, stating that "higher carbon content on a dry basis (Cdb) observed in the alluvial plain landscape" is a bit vague – is Cdb associated with the alluvial plain on PC1? Making that more explicit helps the reader interpret the biplot. Also, the description of Figure 2's chord diagram notes "Coloration and width of the ribbons indicate the direction and magnitude of the correlation, with blue representing positive values (r > 0.4 ) and red representing negative values (r < -0.3)." This is a bit of a broad range for "strong" correlations. Clarifying what the bands represent within that range (e.g., darker blue for stronger positive, etc.) would be helpful for full interpretation.

In Results, line 215, "1.54 times more CP" is clear, but "P < 0.05" is typically presented in tables. If it's in the text, usually it’s fine, but just be consistent whether you always mention p-values in the text for significant differences or only in the tables.

Table 1 has "Iron (I)" which should be "Iron (Fe)" for standard chemical notation. Just a little correction.

Discussion

The discussion does a commendable job of comparing the findings with existing literature, which is a major strength. However, sometimes the connections between the study's specific findings and the broader implications could be drawn more strongly. For example, while the differing CP and NDF levels are discussed in comparison to other studies, there could be more in-depth speculation on why these differences occur beyond just soil type – perhaps linking it to specific plant physiology responses to nutrient availability in those contrasting landscapes. There's a slight disconnect when you say "Differences in DM, ash, NDF, CP, nitrogen, and phosphorus content indicate a soil-type effect on forage quality," then directly after, "This continuous supply of nutrients through sediment deposition contrasts with the highland zone, where nutrient loss is more pronounced due to leaching and suboptimal soil management." This is good, but it could be woven more deeply into the prior comparisons of CP and NDF. Also, the statement that "CGP per degraded organic matter (DOM) was significantly higher in the hill landscape compared to the alluvial plain (102.7 vs. 78.99 ml/g)" is a very interesting result, but the discussion doesn't fully delve into the implications of this difference. Why might the hill landscape forage produce more gas per unit of degraded organic matter? This is a key finding that warrants more detailed interpretation in the discussion.

In the Discussion, line 290, "The CP content observed in P. discolor trees in this study (9.48% in the hill landscape vs. 14.62% in the alluvial plain) was lower than the values reported by Riascos et al. [20], who found 21% CP in trees with 60-day regrowth, and by Guayara [23] who reported 20.4% CP in trees with 45-day regrowth." There's a slight awkwardness in flow here, perhaps "The CP content observed in P. discolor trees in this study (9.48% vs. 14.62% for hill and alluvial plain landscapes, respectively) was lower than..." might read a bit better.

Generally, throughout the text, ensure that species names like Piptocoma discolor are consistently italicized wherever they appear. I think they mostly are, but it's always worth a double-check.

More concretely, in the Discussion section, around line 310, where it talks about ADF values as high as "27.25% have been documented in other regions, such as the Putumayo Department in Colombia." This specific claim about Putumayo needs a citation. It sounds like a direct piece of data from another study, and if it's not cited, it could look like unbacked information, which is a big no-no.

Conclusions

The conclusions are consistent with the evidence presented and largely address the main question posed. They effectively summarize the key findings regarding nutritional properties and degradability. The statement that P. discolor "possesses favorable nutritional properties for ruminant supplementation" is well-supported. However, the conclusion that it's "directly associated with gas production at the rumen level" could be expanded slightly to clarify how this association is demonstrated by their data – i.e., whether higher nutrient content led to lower or higher gas production, linking back to the differing CGP results. It’s a bit vague at the end. Perhaps also, reiterate the implications for climate change, given it's mentioned in the introduction.

References

The references appear mostly appropriate and cover relevant literature. The formatting seems consistent with the journal's requirements. No major issues noted here, but I didn't do a deep dive into every single one. Just make sure all linked URLs are still active.

Tables and Figures

Overall, the tables and figures are clear and help in understanding the data. Table 1 is very comprehensive, but as noted, the "I" for Iron should probably be "Fe". A quick check on units is always good too, just to be extra sure everything is clear. Figure 1 is a fantastic addition, clearly showing the sampling locations, which is vital for understanding the study's geographical context. Figure 2, the chord diagram, is visually appealing and effectively conveys the correlations, though the clarification on the interpretation of ribbon width/color within the defined ranges would be good. Figure 3, the PCA biplot, is also very well done and effectively visualizes the relationships between variables and landscapes. My only thought there is, perhaps a slightly larger font for the variable labels within the plot would improve readability for those of us with older eyes!

Figure legends: For Figure 3, in the legend for the PCA, "CO" is listed for organic carbon, but in the Methods section, it was "OC." Consistency here would be good, sticking to one abbreviation. Just a very small thing.

**********

-->6. PLOS authors have the option to publish the peer review history of their article (what does this mean? ). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy .-->

Reviewer #1: No

Reviewer #2: No

**********

[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: PLOS_one.pdf

-->PONE-D-25-12452R1-->-->Nutritional quality and in vitro gas production of Piptocoma discolor (Kunth) Pruski forage across contrasting landscapes in the Colombian Amazon Piedmont-->-->PLOS One

Dear Dr. Rios Parra,

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 Apr 12 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.

Please include the following items when submitting your revised manuscript:-->

• A 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,

Vitor Hugo Rodrigues Paiva, Ph.D.

Academic Editor

PLOS One

Journal Requirements:

If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise.

Please review your reference list to ensure that it is complete and correct. If you have cited papers that have been retracted, please include the rationale for doing so in the manuscript text, or remove these references and replace them with relevant current references. Any changes to the reference list should be mentioned in the rebuttal letter that accompanies your revised manuscript. If you need to cite a retracted article, indicate the article’s retracted status in the References list and also include a citation and full reference for the retraction notice.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

-->Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.-->

Reviewer #1: (No Response)

Reviewer #2: All comments have been addressed

**********

-->2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. -->

Reviewer #1: Yes

Reviewer #2: Yes

**********

-->3. Has the statistical analysis been performed appropriately and rigorously? -->

Reviewer #1: Yes

Reviewer #2: Yes

**********

-->4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.-->

Reviewer #1: Yes

Reviewer #2: Yes

**********

-->5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.-->

Reviewer #1: Yes

Reviewer #2: Yes

**********

-->6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)-->

Reviewer #1:The manuscript is scientifically sound and significantly improved. If the authors confirm the inclusion of the minimal raw data set and add the specific map attribution, it is ready for publication .

Reviewer #2: well done! no additional comments from my side, all the points were properly addressed by the authors

**********

-->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: SADASHIV CHATURVEDI

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.

-->

Nutritional quality and in vitro gas production of Piptocoma discolor (Kunth) Pruski forage across contrasting landscapes in the Colombian Amazon Piedmont

PONE-D-25-12452R2

Dear Dr. Rios Parra,

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,

Vitor Hugo Rodrigues Paiva, Ph.D.

Academic Editor

PLOS One

Additional Editor Comments (optional):

Reviewers' comments: