Microfluidically-Assisted Isolation and Characterization of Achromobacter spanius from Soils for Microbial Degradation of Synthetic Polymers and Organic Solvents

Round 1

Reviewer 1 Report

Well done. The methods are well explained and presented data of sample characteristics and dose-response curves are of high quality.

I would have liked to have seen statistical analysis if any were performed.

Minor comments: Fig 4: has error bars. Could you elaborate if analysis were performed in multiples ( ie replicates).

Line 333..TX060 shows a significant increase in optical density... What is the p value?

Dose -response curves: were they perform3ed in replicates?

Author Response

Please see the attachment

Author Response File: Author Response.docx

Reviewer 2 Report

Authors present data on the application of a micro segmented-flow approach for the isolation soil bacteria that can degrade selected polymers. The presented work shows signs of the original study and demonstrates a novelty. The experimental setup seems adequate for the field. Most of the tools and methods both experimental and bioinformatical employed are well documented. Appropriate figures demonstrating the main achievements are offered, however, several issues should be addressed to accept the manuscript for publishing.

Major issues

1.      A language of the manuscript should be extensively improved.

2.      A rationale to test “places with ancient human activities” is not appropriately addressed.

3.      A clearer description of microfluidics should be presented for readers from related fields. For example, it is not clear where and how the microorganisms in microdroplets are kept during a prolonged incubation.

4.      A lack of discussion why only Achromobacter spanius strains have been isolated. Is it a bias due to an experimental setup? For example, it is not clear under which conditions (anaerobic, aerobic, microaerophilic etc) microorganisms have been cultivated in microdroplets. That should be critical especially for growth of bacteria during prolonged incubations.

5.      The paragraphs (lines 201-210; 292-298) should be transferred to introduction or discussion part.

Minor issues

1.      Lines 420,421. Authors state “The five genetically closely related isolates…”, however, such statement is too strong, since authors analysed the 16S rRNA encoding sequences only. There are no data on genome sequences presented.

2.      Lines 18,19. The statement “Furthermore, all strains showed similar characteristics in 1,4-dioxane and diglyme” is unclear.

3.      Line 60. “Rhodococcus erythroplis” should be changed to “Rhodococcus erythropolis”

4.      Line 101 “Units” = “units”

5.      Line 182 “0,5” = “0.5”

6.      Line 205 “nutrient cyclin” (?)

7.      Line 274 “PAM-metabolized A. spanius strains” = “PAM-metabolizing A. spanius strains”   

Author Response

Please see the attachment

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

Authors addressed all the raised points.