The relationship between microbial population ATP and quantitative PCR bioburdens in diesel fuel microcosms
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Historically, fuel microbiology studies have relied on culture data. Potentially relevant but unculturable bacteria were not detected. Although ATP can quantify total microbial bioburdens in fuels, it cannot differentiate among the taxa present. Quantitative PCR (qPCR) testing promises to fill this gap by quantifying targeted amplicon sequences thereby detecting both culturable and non-culturable taxa and quantifying specifically targeted taxa. In this study, fluid samples drawn from the fuel, interface and water phases of fuel over water microcosms were tested for cellular ATP concentration ([cATP]) and qPCR bioburdens. Additionally, surface swab samples from steel corrosion coupon surfaces exposed to each of these three phases were collected and tested for total ATP concentration ([tATP]) and qPCR bioburdens. Statistical relationships between ATP and qPCR bioburdens were examined. Correlation coefficients between the two variables were matrix dependent and ranged from negligible (| r |=0.2) to strong (| r |=0.7). When results were categorized into negligible, moderate and heavy bioburdens, parameter agreement was again matrix dependent. Percentage agreement between [ATP] and qPCR gene copies ranged from 11 % to 89 % – with qPCR-bioburden ratings typically being greater than ATP-bioburden ratings.
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Thank you for addressing the remaining minor concerns in your revised manuscripts. Prior to publication I suggest amending the first two sentences of the abstract which currently read "Historically, fuel microbiology studies have relied on culture data. Potentially relevant but unculturable were not detected." I suggest rewriting it to something like: Historically, fuel microbiology studies have relied on live culture data, however this approach cannot detect unculturable microorganisms and is biased by the cultivation technique."
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The abstract only reports the findings, there is no introductory sentence where the context of this study is explained and it also lacks a conclusion of how the findings are important in this area of research. The comment from reviewer 2 regarding the clarification of ASTM D7687 testing would aid a non-expert reader immensely, I do not see that this has been fully addressed yet- I suggest the authors expand this with a brief explanation on the assay principle in the method section: Probably 1-2 sentences explaining what this assay measures and how this is done, in addition to the reference provided. Avoid single sentence paragraphs (line 90-91). It makes it really hard to follow the rationale of the authors. An overall conclusion is also lacking at the very end, how does this improve practice and helps this area of research in the …
The abstract only reports the findings, there is no introductory sentence where the context of this study is explained and it also lacks a conclusion of how the findings are important in this area of research. The comment from reviewer 2 regarding the clarification of ASTM D7687 testing would aid a non-expert reader immensely, I do not see that this has been fully addressed yet- I suggest the authors expand this with a brief explanation on the assay principle in the method section: Probably 1-2 sentences explaining what this assay measures and how this is done, in addition to the reference provided. Avoid single sentence paragraphs (line 90-91). It makes it really hard to follow the rationale of the authors. An overall conclusion is also lacking at the very end, how does this improve practice and helps this area of research in the future, the only indication of this is given that further research is needed.
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This is a study that would be of interest to the field and community. The reviewers have highlighted minor concerns with the work presented. Please ensure that you address their comments. The language used is poor, which can cause ambiguity at times. Please carefully rewrite it. We offer a discounted translation service, Editage (https://www.editage.com/; see https://www.microbiologyresearch.org/prepare-an-article#13 for more information).
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Comments to Author
The article is very well written, complete, clear and with excellent statistical analysis. I leave just two suggestions and congratulate the authors for their work. Line 103: Please write the definition of the ASTM D7687 test in full, even if it has a cited reference. Line 109: Add to the reference any paper that uses the commercial method.
Please rate the manuscript for methodological rigour
Very good
Please rate the quality of the presentation and structure of the manuscript
Very good
To what extent are the conclusions supported by the data?
Strongly support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other …
Comments to Author
The article is very well written, complete, clear and with excellent statistical analysis. I leave just two suggestions and congratulate the authors for their work. Line 103: Please write the definition of the ASTM D7687 test in full, even if it has a cited reference. Line 109: Add to the reference any paper that uses the commercial method.
Please rate the manuscript for methodological rigour
Very good
Please rate the quality of the presentation and structure of the manuscript
Very good
To what extent are the conclusions supported by the data?
Strongly support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
Yes
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Comments to Author
The authors declared, "The Relationship between microbial population adenosine triphosphate and quantitative polymerase chain reaction bioburdens in diesel fuel microcosms ". Despite the importance of the study, the article lacks a good presentation. It has many grammar and language mistakes. The order of event writing should be the same in the abstract, introduction, material, ……….so on. The title of the study needs to be more appropriate English language should be revised before publishing. The following minor points must be taken into consideration: Abstract: - The authors may improve the abstract and make it concise Introduction: -The introduction needs to be more informative. The introduction should be improved(illustrating the aim of the work): * Line 60 long term should be changed to …
Comments to Author
The authors declared, "The Relationship between microbial population adenosine triphosphate and quantitative polymerase chain reaction bioburdens in diesel fuel microcosms ". Despite the importance of the study, the article lacks a good presentation. It has many grammar and language mistakes. The order of event writing should be the same in the abstract, introduction, material, ……….so on. The title of the study needs to be more appropriate English language should be revised before publishing. The following minor points must be taken into consideration: Abstract: - The authors may improve the abstract and make it concise Introduction: -The introduction needs to be more informative. The introduction should be improved(illustrating the aim of the work): * Line 60 long term should be changed to long-term * Line 68 between change to for * Line 84 next generation change to next-generation * Line 91 invert change to inverted * The objective of this study needs to be rewritten Methods -Statistical analysis should be mentioned in brief and explain the program and its version Discussion and conclusion -Authors need to improve discussion and explore the significance of the study compared to other studies. -Line 342,343 tables change to table
Please rate the manuscript for methodological rigour
Very good
Please rate the quality of the presentation and structure of the manuscript
Very good
To what extent are the conclusions supported by the data?
Partially support
Do you have any concerns of possible image manipulation, plagiarism or any other unethical practices?
No
Is there a potential financial or other conflict of interest between yourself and the author(s)?
No
If this manuscript involves human and/or animal work, have the subjects been treated in an ethical manner and the authors complied with the appropriate guidelines?
No: this manuscript not involves human or animal work
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