Peer Review History

Original SubmissionMarch 24, 2020
Decision Letter - Leonidas Matsakas, Editor

PONE-D-20-08433

Morphological and physiological characterization of filamentous Lentzea aerocolonigenes: Comparison of biopellets by microscopy and flow cytometry

PLOS ONE

Dear Prof. Dr. Krull,

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Kind regards,

Leonidas Matsakas

Academic Editor

PLOS ONE

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3. Thank you for providing the following Funding Statement: 

"The authors gratefully acknowledge financial support by the German Research Foundation (DFG) in the Priority Programme 1934 DiSPBiotech – Dispersity, structural and phase modifications of proteins and biological agglomerates in biotechnological processes (SPP 1934 DiSPBiotech – 315384307 (HB) and 315457657 (RK/AK)). The work of LV and CH was supported by the Austrian Research Promotion Agency (FFG) (Grant number: 844608) and within the framework of the Competence Center CHASE GmbH, funded by the Austrian Research Promotion Agency (grant number 868615) as part of the COMET program-Competence Centers for Excellent Technologies by BMVIT, BMDW, the Federal Provinces of Upper Austria and Vienna.

The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript."

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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: Partly

Reviewer #2: Yes

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2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: Yes

Reviewer #2: No

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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: No

Reviewer #2: Yes

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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

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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: Manuscript Number PONE-D-20-08433

The present manuscript studied L. aerocolonigenes cultures, comparing two different methods for morphological and physiological characterization.

Generally, the manuscript is well written and structured. However, there are a few aspects that should be improved in order to make the work clearer.

General comments:

Legends should be inserted close to the Figures, in order to make easy the reading of the paper.

Figures are not presented sequentially; Figure 4 is not referred in the text.

Specific comments:

Line 116 – Authors should specify if glucose was separately sterilized;

Line 177 – Authors referred that they have used Syto9 and PI dyes for CLSM analysis; however, they did not explain how these dyes work, as they did - and well done – for the Flow cytometry protocol, in which they explained how PI and FDA work (Line 221).

Line 223 – Please indicate the number of events/s, threshold and compensation settings for FC analysis;

Line 276 – “The transition from growth phase to stationary phase occurs between day 1 and 2 (Fig 2b)”. This is wrong, since the growth phases are identified in natural logarithmic (Ln) biomass versus time plots, not in biomass versus time plots, as Fig. 2b) shows. Please correct the sentence.

Line 338 – “Due to the high manual effort of pellet slicing only one pellet per sample point was examined.” Do Authors think that this procedure is statistically relevant? Please comment.

-It would be interesting to see the cytograms with FDA/PI results

Figures:

Figure 2 a) –Some experimental points show an error > 20%, which is unacceptable; The same for the biomass curve depicted in Figure 2 b)

More experimental points should be presented during the first 24 h, in order to have a more accurate biomass profile.

Figures 3) and b) should present error bars.

Reviewer #2: I have read throughout the manuscript; it’s a valuable and comprehensive research showing metabolic activity in macro morphology of filamentous bacteria producing rebeccamycin. However, in several instances, the presentation and the discussion of the achieved results and the comparison with the relevant literature are rather limited.

Minor Comments: _

1. Provide a suitable reference for sentence mentioned in line no 405 to 406 for metabolic activity.

2. Is there any evidence that rebeccamycin has fluorescence properties (Line no 427-429).

3. Any suitable experiment/reference showing that the smaller and uniform morphology support more substrate uptake and oxygen into pellet. However, results are contradictory, where all glucose are utilized in 2 days under both condition (With glass beads and without glass beads. Explain this.

4. Did you optimize the substrate concentration during experiment (>3.5 g/L).

5. Why authors used only 100 g/L of beads or with same diameter, did they check if more or less quantity of beads have different observation than the current study?

6. Please provide statistical analysis of data.

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Reviewer #1: No

Reviewer #2: Yes: ALOK PATEL

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Revision 1

Thank you very much for your letter with the reviewer assessment. In the name of all authors I would like to express our gratitude for the fast processing of our manuscript. We appreciate the comments on our paper. The suggestions given were processed. The revised manuscript was resubmitted as instructed.

We include in our revised manuscript:

- A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). This letter was uploaded as separate file and labeled 'Response to Reviewers'.

- A marked-up copy of your manuscript that highlights changes made to the original version. This file was uploaded as separate file and labeled 'Revised Manuscript with Track Changes'.

- An unmarked version of your revised paper without tracked changes. This file was uploaded as separate file and labeled 'Manuscript'.

Kind regards,

Rainer Krull (for all authors)

Attachments
Attachment
Submitted filename: Response to Reviewers.docx
Decision Letter - Leonidas Matsakas, Editor

Morphological and physiological characterization of filamentous Lentzea aerocolonigenes: Comparison of biopellets by microscopy and flow cytometry

PONE-D-20-08433R1

Dear Dr. Krull,

We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements.

Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication.

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If your institution or institutions have a press office, please notify them about your upcoming paper to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and 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.

With kind regards,

Leonidas Matsakas

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

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 #2: All comments have been addressed

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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 #2: Yes

**********

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

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 #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 #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 #2: Authors responded to all queries raised by reviewer satisfactorily, so I recommend acceptance for this article.

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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 #2: Yes: ALOK PATEL

Formally Accepted
Acceptance Letter - Leonidas Matsakas, Editor

PONE-D-20-08433R1

Morphological and physiological characterization of filamentous Lentzea aerocolonigenes: Comparison of biopellets by microscopy and flow cytometry

Dear Dr. Krull:

I am pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please notify them about your upcoming paper at this point, to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, 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.

For any other questions or concerns, please email plosone@plos.org.

Thank you for submitting your work to PLOS ONE.

With kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Leonidas Matsakas

Academic Editor

PLOS ONE

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