Illumina sequencing of 16S rRNA genes reveals a unique microbial community in three anaerobic sludge digesters of Dubai

Munawwar Ali Khan; Shams Tabrez Khan; Milred Cedric Sequeira; Sultan Mohammad Faheem; Naushad Rais

doi:10.1371/journal.pone.0249023

Peer Review History

Original SubmissionNovember 26, 2020
29 Jan 2021 Decision Letter - William J Brazelton, Editor PONE-D-20-37224 Illumina sequencing of 16S rRNA genes reveals a unique microbial community in three anaerobic sludge digesters of Dubai. PLOS ONE Dear Dr. Khan, 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 Mar 05 2021 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: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols We look forward to receiving your revised manuscript. Kind regards, William J Brazelton 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. PLOS requires an ORCID iD for the corresponding author in Editorial Manager on papers submitted after December 6th, 2016. 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Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information. 4.Thank you for stating the following in the Funding Section of your manuscript: "This work was supported by Zayed University Microbial Diversity and Ecogenomics Research Cluster Project (Grant code: R16092)." We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." Please include your amended statements within your cover letter; we will change the online submission form on your behalf. Additional Editor Comments: Both reviewers recommend rejection, but one of their key reasons for rejection, "significance", is not a valid reason for rejection from PLoS ONE. Moreover, I think that the reported data could be valuable contributions to the public databases and to the literature if properly discussed in the paper. Therefore, if you can make substantial revisions to the manuscript that address the reviewers' concerns, then I will consider a heavily revised manuscript. The revised manuscript must explain how the current study is related to previous work with the same or similar samples. It must attempt to interpret the results from the perspective of trying to understand the microbial processes in anaerobic digester systems, rather than simply concluding that the microbial communities are "different". All microbial communities are different from each other. All microbial communities are unique. The revised manuscript should explain how the reported microbial diversity data helps to improve our knowledge of anaerobic digesters. Finally, the authors should make their data publicly available. Currently, the provided BioProject accession is not associated with any sequence data that I can see. The issues listed above are the minimum that should be addressed for a revised manuscript to be re-submitted. If the revision does not address all of the concerns from the reviewers, then it may be rejected even after revision. If you require additional time to make all necessary revisions, an extension can be requested. [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: This study investigated the microbial community in three anaerobic digesters of JASTP using the Illumina sequencing. This article simply describes the species and abundance of archaea and bacteria in three anaerobic digesters. The content is relatively simple and has strong regional limitations. In addition, this study is not of great significance. Hence, it is not suggested to be published in PONE. Comments: 1. What is the significance of this study to research the microbial distribution in these three anaerobic digesters? Why did the author choose these three anaerobic digesters of JASTP? Are they representative? What reference can this study provide for sludge disposal in Dubai in the future? 2. “Fig 1” should be corrected as “Fig. 1”, so as the others. 3. there are write mistakes in line 205, 585. 4. there are some format mistakes in the References. Reviewer #2: The authors investigated the microbial community (Bacteria and Archaea) of three anaerobic digesters treating sewage water in Dubai by illumina sequencing. In my opinion, in order to be published in PLOS ONE the paper need a deep revision by the authors and, after that, a re-submission. The biggest issues are related to: Some general mistakes (eg: line 67 Methanogens (methane-producing bacteria) The lack of information about replicates, sampling period etc Some data in the results section that were already present in Khan et al. 2019 (Table 1 and 2). To this regard, I suggest to clarify if the analysed digesters and samples are the same (same extracted DNA?). Accordingly, with this I suggest to report explicitly in the discussion differences or similarities of the two experiments. Moreover, the discussion need to be deepen. As currently, that is not enough. In any case, the bacterial and archaeal populations results should be considered in relation to digester inlet (wastewater) and outlet (digestate and biogas). In my opinion, in the current form, the paper is not enough significant for the current state of the art of anaerobic digestion to be published in PLOS ONE ****** 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. https://doi.org/10.1371/journal.pone.0249023.r001
Revision 1
14 Feb 2021 Author Response Response to Reviewer 1 Comments Comment-1: This study investigated the microbial community in three anaerobic digesters of JASTP using the Illumina sequencing. This article simply describes the species and abundance of archaea and bacteria in three anaerobic digesters. The content is relatively simple and has strong regional limitations. In addition, this study is not of great significance. Hence, it is not suggested to be published in PONE. Response: JASTP is one of the two main municipal sewage treatment plants operating in Dubai. The successful operation of JASTP is of critical importance to Dubai since both wastewater and digested sludge is reused for further application. The biogas produced by anaerobic digesters of the JASTP is partly utilized to maintain digester temperature and run the boiler systems. However, there are plans in Dubai for the future to capture and use of biogas produced during the treatment of domestic organic waste to energy. However, understanding microbial community structure and functions of anaerobic digester systems in this part of the world is lacking. The results reported in our manuscript not only describe the microbial community in these digesters but also helps us to predict the roles of various microorganisms detected in the treatment of sludge during the anaerobic digester stage of the wastewater treatment process. This knowledge will be beneficial for devising appropriate control methods, leading to more efficient operation of anaerobic digester systems of wastewater treatment plants in the UAE. Furthermore, as reported in the manuscript, microorganisms involved in the process seem to be influenced by the local environmental condition and the fact that most of the water used in the country is desalinated water from the sea. This finding has much greater implications as in most Gulf countries; the conditions are almost similar. Furthermore, so far, no such study to our knowledge is available on this aspect from the region. In light of the above discussion, the study is not merely an isolated study of some digesters in Dubai, but it will provide a basis for the whole region. Comment-2: What is the significance of this study to research the microbial distribution in these three anaerobic digesters? Why did the author choose these three anaerobic digesters of JASTP? Are they representative? What reference can this study provide for sludge disposal in Dubai in the future? Response: As discussed above JASTP is one of the two main municipal sewage treatment plants in Dubai. The three AD systems chosen for this study are operating for a long time and are considered representative digesters of JASTP and Dubai. The significance of the study is manifold, as discussed above. It is the first study of this type, which may have greater implications for the ADs of the gulf region. The climatic condition and the type of water used may influence the process of anaerobic digestion. Therefore, this study's findings not only provide a reference to the sludge disposal in Dubai but also to other gulf countries sharing similar climatic condition and water quality. Since the sludge produced during anaerobic digestion is used as a biofertilizer in public parks and local agricultural farms in Dubai. Which may further affect the soil microbial communities and soil fertility. Ensuring stable operation of the digesters is of critical importance to the city of Dubai making it necessary to understand the microbial community involved. Comment-3: “Fig 1” should be corrected as “Fig. 1”, so as the others. Response: Suggested style for citing figures for PLOS ONE journal is “Fig 1”. We have followed journal formatting style for the figures. Comment-4: there are write mistakes in line 205, 585. Response: We have corrected mistakes in line 205 and 585. Comment-5: there are some format mistakes in the References. Response: The reference format is corrected as per the PLOS One formatting style using EndNote software. Response to Reviewer 2 Comments Comment-1: The authors investigated the microbial community (Bacteria and Archaea) of three anaerobic digesters treating sewage water in Dubai by illumina sequencing. In my opinion, in order to be published in PLOS ONE the paper need a deep revision by the authors and, after that, a re-submission. Response: We have thoroughly revised the manuscript as per the reviewer's suggestion. Comment-2: The biggest issues are related to: Some general mistakes (eg: line 67 Methanogens (methane-producing bacteria) Response: This error has been corrected. Comment-3: The lack of information about replicates, sampling period etc Response: We have included this missing information in the methodology section between lines 88 to 90. Comment-4: Some data in the results section that were already present in Khan et al. 2019 (Table 1 and 2). To this regard, I suggest to clarify if the analysed digesters and samples are the same (same extracted DNA?). Accordingly, with this I suggest to report explicitly in the discussion differences or similarities of the two experiments. Response: The samples used for extraction of genomic DNA for Illumina sequencing were different but collected simultaneously with samples used in our previous published studies involving Fluorescence in situ hybridization FISH and real-time PCR assay. JASTP laboratory has a weekly or biweekly schedule for analysis of digester operational parameters. JASTP officials provided the reported digesters parameters for that sampling period. As suggested, we have reported it clearly in the introduction section (lines 78-84). Comment-5: Moreover, the discussion need to be deepen. As currently, that is not enough. In any case, the bacterial and archaeal populations results should be considered in relation to digester inlet (wastewater) and outlet (digestate and biogas). Response: We have improved the discussion section. The collected digester samples are composite samples. There was only one accessible sampling port used for digester parameters analysis. Comment-6: In my opinion, in the current form, the paper is not enough significant for the current state of the art of anaerobic digestion to be published in PLOS ONE. Response: We have improved the manuscript considerably. We have already discussed the manifold significance of the study. The authors hope that the revised manuscript will be of great interest to its readers and attract readership, especially in the gulf region. This study will also attract readers elsewhere as there is no report to our knowledge on the Gulf region's ADs with particular regard to its climatic condition. Response to Additional Editor Comments 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 Response 1: We have reformatted the manuscript to meet PLOS ONE's style requirements. 2. PLOS requires an ORCID iD for the corresponding author in Editorial Manager on papers submitted after December 6, 2016. Please ensure that you have an ORCID iD and that it is validated in Editorial Manager. To do this, go to ‘Update my Information’ (in the upper left-hand corner of the main menu), and click on the Fetch/Validate link next to the ORCID field. This will take you to the ORCID site and allow you to create a new iD or authenticate a pre-existing iD in Editorial Manager. Please see the following video for instructions on linking an ORCID iD to your Editorial Manager account: https://www.youtube.com/watch?v=_xcclfuvtxQ Response 2: The corresponding author ORCID iD has been updated in the Editorial Manager. 3. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information. Response 3: We have added supporting table information on the last page of the manuscript and amended in-text citations of supporting information according to the journal's guidelines. 4.Thank you for stating the following in the Funding Section of your manuscript: "This work was supported by Zayed University Microbial Diversity and Ecogenomics Research Cluster Project (Grant code: R16092)." We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." Please include your amended statements within your cover letter; we will change the online submission form on your behalf. Response 4: This is true that the funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. However, the work would not have been possible without the generous financial support from the funding agency, that is why we want to acknowledge the funding agency. We have moved funding-related information to the acknowledgement section in the revised manuscript. Additional Editor Comments: Point 1: Both reviewers recommend rejection, but one of their key reasons for rejection, "significance", is not a valid reason for rejection from PLoS ONE. Moreover, I think that the reported data could be valuable contributions to the public databases and to the literature if properly discussed in the paper. Therefore, if you can make substantial revisions to the manuscript that address the reviewers' concerns, then I will consider a heavily revised manuscript. Response: We appreciate the opportunity provided by the editor to considerably improve our manuscript for possible publication in PlosOne. Keeping in view the comments from the editor and reviewers the manuscript has been improved significantly. I hope the revised manuscript meets the standard of PlosOne and will be published in the journal. Point 2: The revised manuscript must explain how the current study is related to previous work with the same or similar samples. It must attempt to interpret the results from the perspective of trying to understand the microbial processes in anaerobic digester systems, rather than simply concluding that the microbial communities are "different". All microbial communities are different from each other. All microbial communities are unique. The revised manuscript should explain how the reported microbial diversity data helps to improve our knowledge of anaerobic digesters. Response: The sludge samples for the next-generation sequencing analysis, FISH and qPCR analysis were collected simultaneously from the digesters. This information has been added to the revised manuscript. We agree to the editors comment, and we have elaborated the possible role of various microorganisms in the process of anaerobic digestion as also summarised in figure 4. Further information on the role of these microorganisms in the process of anaerobic digestion is also added to the revised manuscript, especially in the discussion section. We have concluded in the manuscript that the processes are mediated by the halophilic and/or thermophilic microorganisms which may be due to the influence of the local climatic condition. This study, therefore, is a primer for the possible role of microorganisms in the process of anaerobic digestion in gulf region. In future studies, we can try to isolate such halophilic or thermophilic bacteria from local environment that can help to improve the process of anaerobic digestion. In future we will do metagenome based study of the same digesters to understand the functional aspects more clearly. Point 3: Finally, the authors should make their data publicly available. Currently, the provided BioProject accession is not associated with any sequence data that I can see. The issues listed above are the minimum that should be addressed for a revised manuscript to be re-submitted. If the revision does not address all of the concerns from the reviewers, then it may be rejected even after revision. If you require additional time to make all necessary revisions, an extension can be requested. Response: We have updated the deposited Nucleotide sequences files in the National Center for Biotechnology Information (Accession no. SRX10004771-SRX10004780) under the Bio-Project ID: PRJNA602372 on February 3, 2021. The sequence data records are now open to the public and accessible with the following link: https://www.ncbi.nlm.nih.gov/sra/PRJNA602372 Attachments Attachment Submitted filename: Authors response to Academic editor.docx https://doi.org/10.1371/journal.pone.0249023.r002
22 Feb 2021 Decision Letter - William J Brazelton, Editor PONE-D-20-37224R1 Illumina sequencing of 16S rRNA genes reveals a unique microbial community in three anaerobic sludge digesters of Dubai. PLOS ONE Dear Dr. Khan, 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. Thank you for revising the manuscript. The revision addresses some but not all of the reviewer concerns. The responses to the reviewer's comments (e.g. the general importance of this study for effective wastewater management in Dubai and why these digesters were chosen) are not fully reflected in the text. Please ensure that the points made in your responses to the reviewers are also in the manuscript text. Also, Figure 4 contains a nice summary of some key results, but this figure is only briefly cited in the text. Please incorporate this figure more fully into the Results and Discussion. Finally, please check the entire manuscript for English grammar. For example, multiple sentences beginning with "Although" and "But" are not complete sentences. Please submit your revised manuscript by Apr 08 2021 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: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols We look forward to receiving your revised manuscript. Kind regards, William J Brazelton Academic Editor PLOS ONE [Note: HTML markup is below. Please do not edit.] [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. https://doi.org/10.1371/journal.pone.0249023.r003
Revision 2
28 Feb 2021 Author Response Response to Reviewer’s /Additional Editor Comments Comment: 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. Response: Authors thank the reviewers for providing an opportunity to revise the manuscript. The manuscript is thoroughly revised keeping in view the suggestions of reviewers and academics editor. Manuscript is also considerably improved to avoid grammatical, language and editing mistakes. Comment: Thank you for revising the manuscript. The revision addresses some but not all of the reviewer concerns. The responses to the reviewer's comments (e.g. the general importance of this study for effective wastewater management in Dubai and why these digesters were chosen) are not fully reflected in the text. Please ensure that the points made in your responses to the reviewers are also in the manuscript text. Response: The manuscript is revised to include the importance of the study in the manuscript text. Furthermore, we have also modified the manuscript to include the changes suggested by reviewers earlier. Comment: Also, Figure 4 contains a nice summary of some key results, but this figure is only briefly cited in the text. Please incorporate this figure more fully into the Results and Discussion. Response: A detailed and duly discussion on figure 4 (now figure 2) is now included in the revised manuscript. Figure 4 is renamed as figure 2 to follow the in-text figure sequence style of the journal. Comment: Finally, please check the entire manuscript for English grammar. For example, multiple sentences beginning with "Although" and "But" are not complete sentences. Response: The manuscript is thoroughly checked for grammatical and editing mistakes by a native speaker. The authors hope that the language is significantly improved in the revised manuscript. Attachments Attachment Submitted filename: Author response to reviewers-2nd-revision.docx https://doi.org/10.1371/journal.pone.0249023.r004
10 Mar 2021 Decision Letter - William J Brazelton, Editor Illumina sequencing of 16S rRNA genes reveals a unique microbial community in three anaerobic sludge digesters of Dubai. PONE-D-20-37224R2 Dear Dr. Khan, 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 for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, 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, William J Brazelton Academic Editor PLOS ONE Additional Editor Comments (optional): Thank you very much for responding to all of the comments and concerns. I think the manuscript is much improved, and I hope you agree. Best wishes. https://doi.org/10.1371/journal.pone.0249023.r005
Formally Accepted
23 Mar 2021 Acceptance Letter - William J Brazelton, Editor PONE-D-20-37224R2 Illumina sequencing of 16S rRNA genes reveals a unique microbial community in three anaerobic sludge digesters of Dubai. Dear Dr. Khan: I'm 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 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 plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr. William J Brazelton Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0249023.r006

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