Composition and Concentration of the Biogenic Components of the Aerosols Collected over Vasyugan Marshes and Karakan Pine Forest at Altitudes from 500 to 7000 m

Round 1

Reviewer 1 Report

In general, the article is interesting, original in idea and a rare method of sampling aerosol at different heights. The authors have obtained new experimental results that are thought out and laid out in a reasonable scheme for interpretation.

Remarks:

1. The title of the article is too verbose. It seems to me that it can be shortened to, for example, this: Composition and concentration of biogenic components of the aerosols collected over Vasyugan marshes and Karakan pine forest at altitudes from 500 to 7000 m

2. Line 53: The coordinates of one point located on the territory of the Vasyugan marshes are indicated. But the sampling was not carried out at this point, it is better to specify the coordinates of the point for which the back-trajectories of air mass transport were calculated.

3. Line 92 et seq.: For the Karakan forest, no coordinates are given at all. It would also be possible to specify the point of calculation of back-trajectories. You can also put these two points on the map in Figure 1, which would allow you to visualize everything.

4. In several places of the article, starting with the abstract, it is said about flight. But nowhere is it described what kind of flight (special?) it was, on what plane, according to what program. This is important and interesting to readers.

5. Line 130: it says about sampling up to heights of 5500 m, and then the analysis goes up to 7000 m. Probably a typo?

6. Lines 160-166. The points where aerosol samples were taken and for which air mass transport trajectories were calculated are located close to each other (approximately two degrees of latitude along the meridian), at a distance of about 200 km. This is a small distance from the point of view of the horizontal scales of atmospheric vortices (cyclones and anticyclones) that determine the ways of air transport. This is noticeable by the trajectories of air mass transport to these points, given by the authors in Supplementary, calculated for two points for the same day. Trajectories for the same heights are fundamentally different from trajectories for other heights, but they are similar and sometimes very close for different points. However, the statement that they were all over the northern territories 10 days ago is not entirely true. The trajectories for heights of 1500 and 2000 m (Figures S3 and S7) do not go beyond the mainland and are located mainly to the west or southwest of the aerosol sampling points, being located for some time over the desert and steppe regions of Kazakhstan and over the center of the European part of Russia. The authors' opinion is interesting as to how this could affect and whether it affected the composition of the studied aerosol biocomponents in Western Siberia.

7. To be honest, I am not an expert in chemistry and biochemistry, so I cannot correctly assess the methodology for analyzing the biological components of the aerosol, the reliability and adequacy of the indicators used. I think there will be a reviewer who is well familiar with this topic.

8. Regarding terminology, I would like to note that when describing the movement of air masses and atmospheric impurities, the verb “transport” is more often used, and not “transfer” and especially not “introduction” (line 210). However, in general, I do not undertake to judge the quality of the English language.

Author Response

Dear reviewer 1. Here below is our response to your remarks. All corrections are introduced into the paper’s text using blue font.

1. The title of the article is too verbose. It seems to me that it can be shortened to, for example, this: Composition and concentration of biogenic components of the aerosols collected over Vasyugan marshes and Karakan pine forest at altitudes from 500 to 7000 m

The Title is changed according to Reviewer’s remark.

2. Line 53: The coordinates of one point located on the territory of the Vasyugan marshes are indicated. But the sampling was not carried out at this point, it is better to specify the coordinates of the point for which the back-trajectories of air mass transport were calculated.

Point coordinate of Vasyugan marshes is omitted from the text. Coordinates of backward trajectories initial points for Vasyugan marshes and Karakan pine forest are included in figures S1 – S8.

3. Line 92 et seq.: For the Karakan forest, no coordinates are given at all. It would also be possible to specify the point of calculation of back-trajectories. You can also put these two points on the map in Figure 1, which would allow you to visualize everything.

The coordinates are added to Fig. 1 capture.

4. In several places of the article, starting with the abstract, it is said about flight. But nowhere is it described what kind of flight (special?) it was, on what plane, according to what program. This is important and interesting to readers.

The flight was organized in the frame of France-Russian project YAK-AEROSIB (CNRS, France) "Large-scale studies of greenhouse and oxidizing gases in Siberia and the Arctic".

5. Line 130: it says about sampling up to heights of 5500 m, and then the analysis goes up to 7000 m. Probably a typo?

Yes, it is a misprint. Corrected.

6. Lines 160-166. The points where aerosol samples were taken and for which air mass transport trajectories were calculated are located close to each other (approximately two degrees of latitude along the meridian), at a distance of about 200 km. This is a small distance from the point of view of the horizontal scales of atmospheric vortices (cyclones and anticyclones) that determine the ways of air transport. This is noticeable by the trajectories of air mass transport to these points, given by the authors in Supplementary, calculated for two points for the same day. Trajectories for the same heights are fundamentally different from trajectories for other heights, but they are similar and sometimes very close for different points. However, the statement that they were all over the northern territories 10 days ago is not entirely true. The trajectories for heights of 1500 and 2000 m (Figures S3 and S7) do not go beyond the mainland and are located mainly to the west or southwest of the aerosol sampling points, being located for some time over the desert and steppe regions of Kazakhstan and over the center of the European part of Russia. The authors' opinion is interesting as to how this could affect and whether it affected the composition of the studied aerosol biocomponents in Western Siberia.

Lines 160-166: An analysis of the backward trajectories of the movement of air masses from which samples were taken shows that mainly all of them were over the northern territories/water areas 10 days ago and then moved to the sampling points without descending far to the south (Figures S1 - S8, except the trajectories for heights of 1500 and 2000 m (Figures S3 and S7) ). Thus, it can be assumed that the air masses that contribute to the observed concentrations of total protein and culturable microorganisms in the studied samples of atmospheric aerosols were mainly enriched with biocomponents from sources located in the northern regions.

7. To be honest, I am not an expert in chemistry and biochemistry, so I cannot correctly assess the methodology for analyzing the biological components of the aerosol, the reliability and adequacy of the indicators used. I think there will be are viewer who is well familiar with this topic.

No answer.

8. Regarding terminology, I would like to note that when describing the movement of air masses and atmospheric impurities, the verb “transport” is more often used, and not “transfer” and especially not “introduction” (line 210). However, in general, I do not undertake to judge the quality of the English language.

The verb “introduction” is replaced in line 210: pine forest in autumn 2016 as a result of the air masses with dust transport from Central. The verb “transfer” is replaced with the verb “transport” throughout the text where it is possible.

Author Response File: Author Response.pdf

Reviewer 2 Report

The manuscript presents the results of a study of biological compounds present in the aerosol from several locations in northern Siberia using samples collected from one flight.

The authors should point out the limitations of the study in that it is a single sampling even though it was carried out at several different points and that its extrapolation to other dates and conditions may not be valid due to the seasonality of living beings.

The importance of the place of sampling for the study should also be emphasized in the introduction; in its current wording, the feeling the reader has is that it is an area that is passed through to see what is there and that it is good for me to take samples here and not elsewhere, and then it is justified with casual arguments that the work is carried out in that place.

In addition, the work is too similar to one published by Belan et al. in 2015 at the same location, but now they have modernized the measurements by employing the measurement of other variables instead of those from the 2012 flight. Therefore, the authors should highlight what improvements and new contributions this manuscript has over previous work done by themselves.

In the abstract it is not necessary to indicate the parts of the abstract, the words methods, results and conclusions should not be put; it should be constructed as a continuous paragraph.

In line 119 the data of the software used and its location should be indicated.

The methodology does not describe how the statistical treatment of the data was carried out to obtain the results presented.

The set of results and discussion is well presented and discussed, although some works mentioned are published in Russian language and are difficult for readers to use. A review of the English grammar would also be desirable, as expressions and turns of phrase appear.

The manuscript does not present conclusions of the work that relate to the objectives formulated in the introduction. It should at least be concluded whether the presence detected is high or low and whether it is different from that of other places with similar characteristics or whether it is similar. In addition, it should be pointed out whether the procedure used on this occasion improves the results obtained in previous works.

Regarding the bibliography, it is observed that many of the references indicated are self-cited in a percentage that could be considered high, being advisable that the number of self-citations does not exceed 15%, either by substituting different ones or by trying to reduce the percentage present.

Table S2 should indicate the meaning of the presence of dashes and crosses.

Table S3 should indicate that the letters indicate the name of the corresponding amino acid.

Author Response

Dear reviewer 2. Here below is our response to your remarks. All corrections are introduced into the paper’s text using blue font.

The manuscript presents the results of a study of biological compounds present in the aerosol from several locations in northern Siberia using samples collected from one flight.

1. The authors should point out the limitations of the study in that it is a single sampling even though it was carried out at several different points and that its extrapolation to other dates and conditions may not be valid due to the seasonality of living beings.

Within the framework of this program, monthly flights were planned over the Karakansky pine forest as a relatively clean region to study changes in the concentrations of aerosol and its components and greenhouse gases. It became possible in 2018, within the framework of one flight, to compare the entire complex of the studied characteristics over the Vasyugan marshes. Undoubtedly, single measurements do not give a complete picture of the differences in the observed characteristics of aerosol over the Karakan pine forest and over the unique powerful source of greenhouse gases, aerosol, etc. - the Vasyugan marshes.

2. The importance of the place of sampling for the study should also be emphasized in the introduction; in its current wording, the feeling the reader has is that it is an area that is passed through to see what is there and that it is good for me to take samples here and not elsewhere, and then it is justified with casual arguments that the work is carried out in that place.

However, the autumn measurements carried out almost simultaneously (when the bioaerosol emission is still not limited by snow cover) make it possible to reveal some differences in the concentrations and composition of the biogenic components of atmospheric aerosol over these regions.

3. In addition, the work is too similar to one published by Belan et al. in 2015 at the same location, but now they have modernized the measurements by employing the measurement of other variables instead of those from the 2012 flight. Therefore, the authors should highlight what improvements and new contributions this manuscript has over previous work done by themselves.

Previously, a similar study over these regions was carried out in winter, when the surface of swamps is covered with snow and aerosol emission from them is highly limited (Belan, et al., 2016). In addition, the biogenic components of atmospheric aerosol did not study at all [Belan, et al., 2016].

4. In the abstract it is not necessary to indicate the parts of the abstract, the words methods, results and conclusions should not be put; it should be constructed as a continuous paragraph.

Indication of the abstract’s parts are excluded.

5. In line 119 the data of the software used and its location should be indicated.

Information was insert into the text: (web version, https://www.ready.noaa.gov/HYSPLIT.php)

6. The methodology does not describe how the statistical treatment of the data was carried out to obtain the results presented.

To estimate the concentration of total protein in atmospheric aerosol, samples were taken at altitudes up to 7000 m on AFA-XA-20 fibrous filters [35], approximately 2 m³ of air per filter. When determining the concentration of total protein in samples, a fluorometric method was used, based on the acquisition of intense fluorescence by a protein after its modification with a fluorogenic reagent. As modifying reagents, 3-4-carboxybenzoyl quinoline-2-carboxyaldehyde (CBQCA) was used, a reagent that forms fluorescent derivatives with a higher quantum yield than other dyes upon interaction with proteins [36]. Proteins were determined in the presence of lipids, detergents, and surfactants. The limit of detection of total protein on a Shimadzu RF-520 (Japan) spectrofluorimeter using CBQCA was 0.5 ng/ml of a concentrated sample, and the error in determining its concentration did not exceed 20%. Bovine serum albumin standard solutions were used to calibrate the Shimadzu RF-520 spectrofluorimeter.

The inaccuracy of determining the concentration of microorganisms in a sample did not exceed 30% [41].

7. The set of results and discussion is well presented and discussed, although some works mentioned are published in Russian language and are difficult for readers to use. A review of the English grammar would also be desirable, as expression sand turns of phrase appear.

Unfortunately, the studies of the Vasyugan marshes were mainly carried out by Russian scientists, who published their results in Russian, there are practically no English-language papers.

If the Publisher deems it necessary, we are ready to pay for the correction of the English language by the Publisher’s service.

8. The manuscript does not present conclusions of the work that relate to the objectives formulated in the introduction. It should at least be concluded whether the presence detected is high or low and whether it is different from that of other places with similar characteristics or whether it is similar. In addition, it should be pointed out whether the procedure used on this occasion improves the results obtained in previous works.

It should be noted that the recorded values of the concentration of total protein in the atmosphere at different heights above the Karakan forest are significantly lower than the average annual concentration of total protein for this region in 2018 [We 2022]. Despite the fact that, as a rule, this concentration in the warm season is higher than the average annual value. The recorded concentrations of total protein in the atmosphere over the Vasyugan marshes were even lower.

The measured values of the concentration of culturable microorganisms in the atmosphere at low altitudes above the Karakan pine forest, on the contrary, are significantly higher than the average annual concentration in 2018 at these altitudes. The measured values of the concentration of culturable microorganisms in the atmosphere above the Vasyugan marshes at the same heights are close to those for the Karakan pine forest. At the same time, no culturable microorganisms were found at altitudes of 4000 m and above over the Karakan pine forest, in contrast to the same heights above the Vasyugan marshes. Such a unique situation over the Karakan pine forest was revealed for the first time in more than 20 years of observations. Also, the data on the diversity of culturable microorganisms in the atmosphere above the Vasyugan marshes were obtained for the first time.

Thus, the data obtained in this work require further research both over the Karakan pine forest and over the Vasyugan marshes.

9. Regarding the bibliography, it is observed that many of the references indicated are self-cited in a percentage that could be considered high, being advisable that the number of self-citations does not exceed 15%, either by substituting different ones or by trying to reduce the percentage present.

Many self-citing references are excluded or replaced by references of another authors. See new list of references.

10. Table S2 should indicate the meaning of the presence of dashes and crosses.

The footnote of Table S2 is added:

“+++” means very good microorganism culture growth, “++” - good microorganism culture growth, “+” - moderate microorganism culture growth, “±”- weak microorganism culture growth, and “-“ – no growth at all.

11. Table S3 should indicate that the letters indicate the name of the corresponding amino acid.

The footnote of Table S3 is added:

Four types of bases are found in a DNA molecule usually: letter “A” stands for adenine, letter “C” stands for cytosine, letter “G” stands for guanine, and letter “T” stands for thymine.

Author Response File: Author Response.pdf

Reviewer 3 Report

Although the subject of the article is interesting, there are some problems in conveying its focus, interpreting it, and highlighting its novelty. The article should be modified for the following reasons:

1. The authors need to revise the abstract by including the removing the title of background, methods, results and conclusions, which is not common in the atmosphere journal. Also, main quantitative findings are suggested.

2. Please provide a flow chart in the introduction or in the method section so that reviewers and readers can easily understand this study.

3. The supplementary documents were sufficient, however, the section of the “2. Materials and Methods” were not documented in a well-written way.

4. For the results, the subsection is required to indicate which finding was presented, and the structure organization was not clear.

5. The conclusions was missing, which should be appropriately simplified and highlight the conclusions and insightful findings of the study.

6. The authors should realized that the limitations and future consideration are not included in the present content.

Author Response

Dear reviewer 3. Here below is our response to your remarks. All corrections are introduced into the paper’s text using blue font.

Although the subject of the article is interesting, there are some problems in conveying its focus, interpreting it, and highlighting its novelty. The article should be modified for the following reasons:

1.The authors need to revise the abstract by including the removing the title of background, methods, results and conclusions, which is not common in the atmosphere journal. Also, main quantitative findings are suggested.

Indication of the abstract’s parts are excluded. Quantitative information is added into the abstract.

2. Please provide a flow chart in the introduction or in the method section so that reviewers and readers can easily understand this study.

The flow chart was prepared and introduced into the text as Fig. 2.

3. The supplementary documents were sufficient, however, the section of the “2. Materials and Methods” were not documented in a well-written way.

The section of the “2. Materials and Methods” was partly rewritten. Some information was added into it.

4. For the results, the subsection is required to indicate which finding was presented, and the structure organization was not clear.

“3. Results and Discussion” section was rewritten with subsections.

5. The conclusions was missing, which should be appropriately simplified and highlight the conclusions and insightful findings of the study.

The conclusions were added to the text.

6. The authors should realized that the limitations and future consideration are not included in the present content.

The description of some limitations of the study is insert into “Introduction”. Future consideration of the work is included into new section “4 Conclusions”.

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

The authors have made the modifications proposed in my first revision and the content is correct.

Only one detail that could be improved is that the results presented in Table 2 and Table 3 do not have a coherent presentation, since in the first one they are presented in integer notation and in the second one in exponential scientific notation. It would be desirable for the two tables to be presented in exponential notation, including incorporating the information from table 3 in table 2, as this will reduce the overall width when making this change.

Author Response

Thank you for your remark!

We changed data format in the Table 2 to exponential notation.

Author Response File: Author Response.pdf

Reviewer 3 Report

The current manuscript can be accepted after the significant revisions.

Author Response

Dear reviewer 3! You gave us no suggestions on what should be improved.

Author Response File: Author Response.pdf