Algae Diversity and Ecology during a Summer Assessment of Water Quality in the Abraham Lincoln Birthplace National Historical Park, USA
Round 1
Reviewer 1 Report
Algae diversity and ecology in…. by Barinova and Smith.
The authors have studied microalgal diversity in a new location (not studied and reported so far as claimed by the authors), a National Historical Park in USA. The study even though is a one-time study, having taken samples just one day, is more of a survey type investigation. The sampling areas are located in a protected area but with varying physiogeography. That by itself is interesting and provides the readers an opportunity to learn about what is out there in such an environment. The data is quite diverse, the analysis were bit complicated still trends are discernible and interpretation and conclusions are quite in an expected line. The results demonstrated the usefulness of micro algal flora in determining water quality. I like the study and would recommend publication, but have the following points to get addressed before it can go any further.
The description of the study areas is quite confusing, the numbering of study sites are not clear to me. The diversity measurements are bit too much with lots of basic data represented in too many tables and figures. Why don’t the authors reduce the numbers so that the readers get a clearer picture of the data distribution and the trends. The data analysis is not described properly, may be due to the too many tests being carried out and tried to incorporate all the results in just one manuscript. This looks like an undergraduate thesis or report not as a journal paper. The discussion is quite weak, as I would have liked to have it more compared and contrasted with studies from other locations either from USA or other parts. The English used needs modification I think… seems to me as so casual in their approach in many places. All in all, a good work. We don’t generally have opportunities to read manuscripts such as this now a days and reminds me of old school approach and an effective use of statistical analysis.Author Response
Dear Reviewer 1,
Thank you for the review of our ms, your comments and notes.
Our responses are in the attached file.
With best regards,
Prof Sophia Barinova,
corresponding author
Author Response File: 
Author Response.docx
Reviewer 2 Report
The manuscript presented for review is well written, the results are properly documented and discussed. The conclusions are correct. I propose to transfer some results in the form of tables to supplementary materials. Excessive information impairs readability of results. I suggest moving tables 3, 4 and 6. Text editing needs some improvement. Figures showing the location of positions need to be improved (Figs. 1 and 3). In Figure 13, the axis descriptions are too small.
Author Response
Dear Reviewer 2,
Thank you for the review of our ms, your comments and notes.
Our responses are in the attached file.
With best regards,
Prof Sophia Barinova,
corresponding author
Author Response File: Author Response.docx
Reviewer 3 Report
Revision of the manuscript “Algae diversity and ecology in assessment of water quality in the Abraham Lincoln Birthplace National 3 Historical Park, U.S.A.” by Barinova and Smith.
General comments
Introduction. Add some info on which regulations apply in your country, and how/where algae fits into them. This will increase the relevance of your study
Information provided in M&M is not enough: no mention on how the indexes are calculated, or any of the statistical analyses performed. Remember that someone might want to use your methodology in another park in the USA or elsewhere in the world and you should provide enough information to do so.
The fact that the sampling was only made once, and not per season (which might drastically change the species composition and abundance) reduces the impact of the work. Which season is July in the states? If winter, likely less species??
The text must be check for English usage. Prepositions are missing all over the text, wrong words, time verbs, etc.
Discussion can be improved.
Abstract
L18-23: sentences of 7 lines are extremely hard to follow.
Introduction
L36-37: there’s a typo there. Correct.
L43-45. Same as above. And quote missing.
L46-49. Quote missing
Materials and Methods
How many replicates per site? Not specified
Description of study sites- hard to follow convert to table of rephrase. Prepositions are missing here and there which makes reading difficult.
No information of data analysis provided, but then given in results.
No information of the indexes applied, or the reason for choosing them. How were these indexes calculated? Not specified either
L63: What kind of equipment is Shimadzu ICPE-9000 ICP? I had to look for it in internet.... please add what it is and what for did you use it.
L65: “widely used in algology” (this term also refers to the study of pain, please use Phycology) but no quote provided?
L65. Preposition missing.
L66: light microscope used twice in the same sentence.
L70: Delete “of”
L71. Why did you chose to use canonical correspondence analysis? Why is this important? (Same for other indexes/analysis calculated/performed)
L73: “Bioindication” is not a term. Please rephrase.
L78: word missing.
L81: why is this interesting?
Results
L108. Word missing.
L113 “ware”?
L112-113: “Total 112 phosphorus (TP) and nitrate values” how did you measure this? Not mentioned in M&M.
L114. Growth. Not grows
L114: you measured metal as well? Not mentioned in M&M.
L124: word missing.
L127: revise sentence
L133. Use past tense.
L135. Revise use of English
L152: “Bioindication” is not a term. Please rephrase.
L243. I see here some info that can be moved to M&M. Again, you have to say how you calculated this index.
Discussion
L320-322. Comment here that you only sampled one, so you only captured geographical variability and not seasonal.
L332: “very individual”? Rephrase. And change Y axes labelling according to previous comments.
L360: use past tense.
Figs and Tables
How is table 2 and figures 7 to 10 different? One in presented in numbers and other in percentages? This is pretty redundant. Choose one way of presenting your data or justify why is important to represent the same data twice.
Figure 2. So only 2 locations with 7 sampling points? Then how many water bodies in the park in total were sampled? This gives an idea of the sampling effort.
Figs 1 to 3 can be merged into one.
Fig 7 and 8. Rename Y axis as Species (%) and Indicators (%) and remove the % sign from the axis next to the numbers. Decrease the amount of numbers to improve data presentation (i.e. every 20 instead of 10).
Table 1. Please check text.
Table 3: why is there a code provided? it’s not used in other part of the text. Add headings of the table in pages 8 to 10 to facilitate the reading of the table. As it is, you have to go back to the page to see what each column is (this also for the reminder of the tables).
Table 6. Similarity matrix for species richness…this is shown in other Fig/Table.
Figure 12 and 13a are redundant.
Table 4: if abbreviations are the same as table 1, no need to repeat.
Author Response
Dear Reviewer 3,
Thank you for the review of our ms, your comments and notes.
Our responses are in the attached file.
With best regards,
Prof Sophia Barinova,
corresponding author
Author Response File: Author Response.docx
Round 2
Reviewer 3 Report
Please make an effort in elaborating the better rebuttal letter. Changes must be indicated with line numbers, AT LEAST.
"Fixed" is not an anwser and no comments on the general concerns were made.
Author Response
Dear Reviewer,
Thank you very much for your comments. Responses to your comments and notes are in the attached file.
With best regards,
Prof Sophia Barinova,
corresponding author
Author Response File: Author Response.docx
Round 3
Reviewer 3 Report
Revision of the ms “Algae diversity and ecology in assessment of water quality in the Abraham Lincoln Birthplace National Historical Park, U.S.A” by Barinova and Smith.
General comments.
NONE of my General comments were addressed in the ms and/or rebuttal letter, and I think they highlight important issues in the ms. Specially, these:
“Introduction. Add some info on which regulations apply in your country, and how/where algae fits into them. This will increase the relevance of your study”
Intro can be extended including information that will increase the impact of your work.
“The fact that the sampling was only made once, and not per season (which might drastically change the species composition and abundance) reduces the impact of the work. Which season is July in the states? If winter, likely less species??”
This information must be included somewhere or change the title. Diversity assessment in one season will underestimate total (annual) diversity.
“The text must be check for English usage. Prepositions are missing all over the text, wrong words, time verbs, etc”
Did you have the ms checked by a native speaker? It does not seem that way. Some parts are still hard to follow.
How is table 4 and figures 7 to 10 different? One in presented in numbers and other in percentages? This is pretty redundant. Choose one way of presenting your data or justify why is important to represent the same data twice. Use a program to modify the figures. The fact that in the Y axes says “percentage” and the the number have the % symbol is just incorrect. The M&M section has been improved. 6 figures and 1 table for describing the sampling sites still seems excessive.
Specific comments
L18. L159. L325. Remove “The”.
From my previous revision: “Abstract: L18-23: sentences of 7 lines are extremely hard to follow.”
Now you have an equally 6 lines sentence (19-23) that is still hard to follow. Rephrase.
L43-49: I asked for quotations here, not to change the sentence. Quotations have not been added.
L55: what does “original material” mean?
L56. Rephrase.
L64. Specify the number of water samples.
L65-66 This does not give an idea on how you collected the samples. Please rephrase indicating how you collected them.
L358-360. Unclear what you are saying here.
Comments for author File: 
Comments.pdf
Author Response
Dear Editor,
Thank you and the Reviewer for your comments. Please find below the responses to the Reviewer comments point by point. Changes in ms were made in respect to your comments and highlighted by yellow or red font in the text. Many small corrections has been done in English editing that not highlighted. Figures 3-7 and 15 were replaced.
With best regards,
Prof Sophia Barinova,
Corresponding author
COMMENTS AND RESPONSES
NONE of my General comments were addressed in the ms and/or rebuttal letter, and I think they highlight important issues in the ms. Specially, these:
“Introduction. Add some info on which regulations apply in your country, and how/where algae fits into them. This will increase the relevance of your study”
Intro can be extended including information that will increase the impact of your work.
Response: Inserted the paragraph: Algae indicators are used in the National Aquatic Resource Surveys [49,50] as a part of the rivers and stream water quality assessment program are based on density and abundance values for both soft-bodied algae (non-diatom) and diatoms [20]. Diatom Bioassessment Index (DBI) is multimetric index, developed in Kentucky based on a calculation of Bacillariophyta algae only (diatom richness, Shannon index, PTI, Siltation index, Fragilaria richness, and Cymbella richness) and cannot help in the assessment of contaminated waters [20,51]. However, in many cases, data from diatoms only are not enough for assessing aquatic habitat condition because can have mostly non-diatom algal community. However, algae species autecology is likely correlated with nutrient concentrations. Therefore, bioindicator methods, which included a full species list of community in assessment, can help in the multi-parametric indication of aquatic habitats.
“The fact that the sampling was only made once, and not per season (which might drastically change the species composition and abundance) reduces the impact of the work. Which season is July in the states? If winter, likely less species??”
This information must be included somewhere or change the title. Diversity assessment in one season will underestimate total (annual) diversity.
Response:
Added to L13 and L56 - during the summer
Changed title to include season –
Algae diversity and ecology summer assessment of water quality in the Abraham Lincoln Birthplace National Historical Park, U.S.A.
“The text must be check for English usage. Prepositions are missing all over the text, wrong words, time verbs, etc”
Did you have the ms checked by a native speaker? It does not seem that way. Some parts are still hard to follow.
Response: native speaker that is one of the authors corrects English across ms.
How is table 4 and figures 7 to 10 different? One in presented in numbers and other in percentages? This is pretty redundant. Choose one way of presenting your data or justify why is important to represent the same data twice. Use a program to modify the figures. The fact that in the Y axes says “percentage” and the the number have the % symbol is just incorrect. The M&M section has been improved. 6 figures and 1 table for describing the sampling sites still seems excessive.
Response: Table 4 presents the number of species of each taxonomic or environmental category that represents the result of our research. However, it is possible to compare the share of species in each category in percentage graphs 7-10, while using a tabular version of the pattern is extremely difficult to see. Axis Y name has been changed in Figures 7-10, 15, and figures are replaced in ms.
Specific comments
L18. L159. L325. Remove “The”.
Response:
Removed all 3 “The” from L18. L159. L325.
From my previous revision: “Abstract: L18-23: sentences of 7 lines are extremely hard to follow.”
Now you have an equally 6 lines sentence (19-23) that is still hard to follow. Rephrase.
Response:
Changed the text to the following:
This method was used for the first time in the U.S.A. during this study. This demonstrated that benthic and planktonic-benthic algae preferred temperate temperatures, middle-oxygenated mesotrophic waters, low-to-middle enriched by chlorides, well oxygenated, sometimes saturated by sulfides, low-alkaline, low-to-middle organic enriched, and of Class 1-3 of Water Quality with high self-purification capacity. This is very important for habitat protection and cannot be easily accomplished strictly thought chemical analysis.
L43-49: I asked for quotations here, not to change the sentence. Quotations have not been added.
Response: corrected as: Floristic studies in aquatic systems are very important because the flora is an environmental indicator [20] and does infer environmental conditions on the natural, climatic and economic from the Knob Region.
Studies of algae are of particular interest since the formation of their flora and algal composition is strongly influenced by land-water interactions [20]. Thus, the algal list represents the combined result of natural and anthropogenic processes in the entire catchment area for many years.
L55: what does “original material” mean?
Response:
Removed the term “original”
L56. Rephrase.
Response: added –
This was a one-time sampling collection. The samples collected came from 7 sites and 4 different waterbodies with a total of 21 samples (3 replicate samples per site).
L64. Specify the number of water samples.
Response:
Added – L67 There were 21
L65-66 This does not give an idea on how you collected the samples. Please rephrase indicating how you collected them.
Response:
Rephrased –
The epilithic rock samples were cleaned with a soft toothbrush by lightly scrubbing in a circular motion, then rinsed with distilled water. The toothbrush was cleaned with distilled water after each sample. This rinse water was collected in a sterile Whirl-pak®. Phytobenthos samples were collected with a large nylon liquid baster and placed in a sterile Whirl-pak®. Algal and water samples were transported to the lab in an ice cooler at 0 °C until they could be taken to the lab for processing.
L358-360. Unclear what you are saying here.
Response:
Rephrased:
Nevertheless, the indication of eutrophication and algae productivity with chlorophyll has been documented in U.S.A. [58]. Biondicator methods of water quality assessment with the use of algae and Cyanobacteria species’ autoecology were first used in the U.S.A. on aquatic habitats during this study. The bioindicator results demonstrated the algal preferences
139-142 red text. Corrected as: Diatoms prevailed in all studied communities but in site 7 it was accompanied by twenty Cyanobacteria species. Figure 7a demonstrates that in sites 3-7, cyanobacteria species also played an important role in algal communities where it contains up to 10-15 percent of total species richness in the site's community.
Author Response File: Author Response.docx
