Plant Biodiversity Knowledge Varies by Gender in Sustainable Amazonian Agricultural Systems Called Chacras

Round 1

Reviewer 1 Report

 I want to make some appreciations about this paper:

In the Abstract, on line 27 indicate that it refers to "The United Nations Sustainable Development Goal 2 (SGD2)"

I do not see clearly if  "Parcela Borde del río, Parcela Igapo and Parcela colinas" (yellow boxes of Figure 1) are included in this study. I think you should eliminate them.

Place the columns correctly in the paper card design (figure 2).

In figure 4, it is not clear where the values of the table come from.

There is a typographical error on line 294: bites are repeated

Author Response

I want to make some appreciations about this paper:

 

1.      In the Abstract, on line 27 indicate that it refers to "The United Nations Sustainable Development Goal 2 (SGD2)" OK. We have included this.

 

2.      I do not see clearly if  "Parcela Borde del río, Parcela Igapo and Parcela colinas" (yellow boxes of Figure 1) are included in this study. I think you should eliminate them. Ok. We have changed the Figure.

 

3.      Place the columns correctly in the paper card design (figure 2). Ok. We have  changed the figure

 

4.      In figure 4, it is not clear where the values of the table come from. OK The caption of figure 4 has been changed. 

 

5.      There is a typographical error on line 294: bites are repeated. OK.  It has been deleted.

Reviewer 2 Report

In this study, the authors assessed traditional plant knowledge in an isolated community in Amazonian Ecuador. They focus on the difference between males and females in their knowledge on plant diversity and plant uses. This is a beautiful work, with a well-considered and well-prepared methodology. Although I am enthusiastic about the work, I have some concerns that I think should be addressed before the manuscript could be considered for publication.

The introduction starts with the global food- and biodiversity crisis, and then zooms-in on a rotating agricultural system (the Chacras) in a remote area in Ecuador. This structure does not work for me. Such systems may be sustainable in remote areas with low human population densities, but are not a solution for food security on larger scales. In addition, biodiversity is high in such systems because they are relatively small and surrounded by large areas of forest. Thus these fields are continuously invaded by non-agricultural species, and probably half forest / half field for most of the time. On larger scales, that balance, and thus biodiversity, will change. I strongly suggest placing the study in another context, for example to focus only on the importance to consider a gender perspective in ethnobotanical studies.

The manuscript needs to be revised by a native speaker. Grammatical errors are very numerous and in many places unfortunately hinder the interpretation of the text (I have highlighted some errors below, but they are too numerous to point out). In addition, I found the introduction and discussion quite unclear. For many paragraphs, I was left wondering what exactly the authors are trying to point out, or conclude (see some detailed comments below).

 

L20. “potential reservoirs of the traditional biodiversity of chacras plant knowledge”. The word “biodiversity” is not correct here. Suggest using “potential reservoirs of traditional chacras plant knowledge”.

L22. “Uncontacted” is misleading, and not true. Suggest replacing with “isolated” here, and throughout the manuscript.

L41. “forests are bringing down while greenhouse-gas emissions grow up on a large scale” should be something like “deforestation is increasing while greenhouse-gas emissions continuous to increase”.

L47. “up to 40%” in the Amazon?

L52. “identity” should be “identities”.

L59. “interaction between the indigenous woman lifehood” and what other factor?

L61. Suggest using other word for “diagnostic”.

L66. Should “food plains” be “floodplains”?

L81. Suggest using other words for “dominate the forests”.

L96. “communication” should be “connection”.

L97. “originary” should be “original”.

L102. Suggest explaining what is meant with “Pakayaku access is not free.”.

L128. “50x2” add unit (meters?); “method” can be removed here.

L132-133. Suggest removing; this is not relevant information.

L137. What do the authors mean with “cultivated in any region of Ecuador upon de la Torre et al.[29].”?

L87-88. “One attendee..” and L89-90 “One researcher..” Is the same said twice here? If so, I suggest removing the first sentence. If not, what is meant in L87-88?

L198, 201, 209. Should “raw” be “row”?

L206, 213, 216. What do the authors mean with “It is expressed as a both, not in percentage.”?

L261-263. Suggest adding explanation of why management has so much more weight.

L273. “p=0.000” should be “p<0.001”.

L289. Suggest replacing “of the male and female collective” with “of males and females”.

L290-291. “of the categories and subcategories that are related to the divisions of work in society”. This expression is very vague. Do the authors not just mean “of plants”.

L292-293. Suggest replacing “plants, related to the care” with “plants, which is also related to their role in the care”. And add “are” before “used” in the next sentence.

L294. “bites” is mentioned twice here.

L295. What do the authors mean with “by antonomasia”?

L299-302. “Finally we must point that our data suggest that the decline of traditional knowledge related to plant diversity associated with the male population can be related with the fact that hunting is nowadays less practiced than before.” I do not understand what the authors are trying to point out here and how the data would show traditional knowledge is lost.

L303. “quantitative” is mention twice here.

L307-308. “An important aspect to take into account is the valorization of their activities among the population. On the contrary, that traditional knowledge will be lost.” I again do not fully understand what the authors try to point out here, or where the “on the contrary” refers to.

Author Response

In this study, the authors assessed traditional plant knowledge in an isolated community in Amazonian Ecuador. They focus on the difference between males and females in their knowledge on plant diversity and plant uses. This is a beautiful work, with a well-considered and well-prepared methodology. Although I am enthusiastic about the work, I have some concerns that I think should be addressed before the manuscript could be considered for publication. Thank you very much for your appreciations.

 

 

The introduction starts with the global food- and biodiversity crisis, and then zooms-in on a rotating agricultural system (the Chacras) in a remote area in Ecuador. This structure does not work for me. We understand what you mean. You explain perfectly in your position in the following paragraphs.: Such systems may be sustainable in remote areas with low human population densities, but are not a solution for food security on larger scales. In our perspective, global food security has to take in account not only these large scales that you mention but also the scale we are dealing with.  Because, it is also large. Very large. Amazonian populations must be considered because of its social,  ethnobiological, and environmental importance. It is a mistake to forget in our analysis Amazonian reality. 

 In addition, biodiversity is high in such systems because they are relatively small and surrounded by large areas of forest. Thus these fields are continuously invaded by non-agricultural species, and probably half forest / half field for most of the time. On larger scales, that balance, and thus biodiversity, will change.  You are right but we have focused our effort not to the numbers or quantity but the cualitative importance. Our idea is reinforced by the urgencies of the SGD-

I strongly suggest placing the study in another context, for example to focus only on the importance to consider a gender perspective in ethnobotanical studies. We understand what you suggest but we would like to emphasize on the importance of connecting this management practices with food soveringty. This is very important for the local populations. Even though, we have deleted some parts of the text in order to attend your claim as much as possible.

 

The manuscript needs to be revised by a native speaker. Grammatical errors are very numerous and in many places unfortunately hinder the interpretation of the text (I have highlighted some errors below, but they are too numerous to point out). In addition, I found the introduction and discussion quite unclear. For many paragraphs, I was left wondering what exactly the authors are trying to point out, or conclude (see some detailed comments below). Ok. We welcome this criticism in its entirety and to resolve it we have sent the manuscript to the Language Correcting Service of MPDI.  

 

·        L20. “potential reservoirs of the traditional biodiversity of chacras plant knowledge”. The word “biodiversity” is not correct here. Suggest using “potential reservoirs of traditional chacras plant knowledge”. Ok Changed,

 

·        L22. “Uncontacted” is misleading, and not true. Suggest replacing with “isolated” here, and throughout the manuscript. Ok Changed,

 

·        L41. “forests are bringing down while greenhouse-gas emissions grow up on a large scale” should be something like “deforestation is increasing while greenhouse-gas emissions continuous to increase”. Ok Changed,

 

·        L47. “up to 40%” in the Amazon?. Yes.

 

·        L52. “identity” should be “identities”.Ok Changed

 

·        L59. “interaction between the indigenous woman lifehood” and what other factor? Mistake. Changed.

 

·        L61. Suggest using other word for “diagnostic”. Ok. Description has been used.

 

·        L66. Should “food plains” be “floodplains”? Ok. Yes.

 

·        L81. Suggest using other words for “dominate the forests” Ok, Changed by “be specialized in”

 

·        L96. “communication” should be “connection”.Ok. Changed.

 

·        L97. “originary” should be “original”. Ok. Changed.

 

·        L102. Suggest explaining what is meant with “Pakayaku access is not free.”It means that you cannot get into if you are not allowed by the kuraka or the President of Pakayaku.  If you do it, you can be reduced or attacked.

 

·        L128. “50x2” add unit (meters?); “method” can be removed here. OK .

 

·        L132-133. Suggest removing; this is not relevant information. We prefer to maintain it because it is usually asked by Botanical Referees.

 

·        L137. What do the authors mean with “cultivated in any region of Ecuador upon de la Torre et al.[29].”? Cultivated in Ecuador, as it is said in the Enciclopedia de las Plantas Utiles de Ecuador, a book that has been written by those authors.

 

·        L87-88. “One attendee..” and L89-90 “One researcher..” Is the same said twice here? If so, I suggest removing the first sentence. If not, what is meant in L87-88? Ok. Changed.

 

·        L198, 201, 209. Should “raw” be “row”? Ok. Changed.

 

·        L206, 213, 216. What do the authors mean with “It is expressed as a both, not in percentage.”? Ok, -changed. Both per one

 

·        L261-263. Suggest adding explanation of why management has so much more weight.Ok. included.

 

·        L273. “p=0.000” should be “p<0.001”. Yes. Corrected.

 

·        L289. Suggest replacing “of the male and female collective” with “of males and females”. Ok. Changed.

 

·        L290-291. “of the categories and subcategories that are related to the divisions of work in society”. This expression is very vague. Do the authors not just mean “of plants”. Corrected. In means “of plant uses”.

 

·        L292-293. Suggest replacing “plants, related to the care” with “plants, which is also related to their role in the care”. And add “are” before “used” in the next sentence. Corrected.

 

·        L294. “bites” is mentioned twice here. Corrected.

 

·        L295. What do the authors mean with “by antonomasia”? Corrected, changed.

 

·        L299-302. “Finally we must point that our data suggest that the decline of traditional knowledge related to plant diversity associated with the male population can be related with the fact that hunting is nowadays less practiced than before.” I do not understand what the authors are trying to point out here and how the data would show traditional knowledge is lost. Better explanation included.

 

·        L303. “quantitative” is mention twice here. Corrected.

 

L307-308. “An important aspect to take into account is the valorization of their activities among the population. On the contrary, that traditional knowledge will be lost.” I again do not fully understand what the authors try to point out here, or where the “on the contrary” refers to. Ok, -sentence modified.

Reviewer 3 Report

1. The Enlgish should be improved, e.g. amazonic, kitchwa, latin – with capital

2. lie, rub and burn – sound strange – is it not better to write: slash-and-burn?

3. The paper lacks information on what are the specific uses of the studied plants.

4. There is a large literature on gender differences in knowledge od plants, including South America. It is not discussed here.

5. Voucher numbers in the herbarium are missing.

6. A map would be useful.

7. Some diagram showing the chakras system could help readers to imagine it.

8. There is no need to directly show every step of data input in Excel. It is usually not done in papers.

Author Response

1. The Enlgish should be improved, e.g. amazonic, kitchwa, latin – with capital. Ok Done The manuscript has been sent to MPDI Language Service..

 

2. lie, rub and burn – sound strange – is it not better to write: slash-and-burn? Ok. Changed.

 

3. The paper lacks information on what are the specific uses of the studied plants. Sorry but this is not the objective of the paper. We have included just the uses mentioned by the informants.

 

4. There is a large literature on gender differences in knowledge of plants, including South America. It is not discussed here. We have tried to improve bibliography on this topic and we have included it in this new version of the paper.

 

5. Voucher numbers in the herbarium are missing. We can include it in an Annex. They appear in the Ph Tesis of one of the autors C.X. Luzuriaga Quichimbo (2017). This bibliographic cite is included in the reference list. We did not include each voucher number in order to facilitate the reading. But the addition in an Annex is available upon request.

 

6. A map would be useful. OK. It has been included.

 

7. Some diagram showing the chakras system could help readers to imagine it. Ok It has been included.

 

8. There is no need to directly show every step of data input in Excel. It is usually not done in papers. We think it is necessary because the process is somehow tedious and it requires to be explained in detail as to favour the paper can be replicated anywhere.

Round 2

Reviewer 2 Report

The authors have properly addressed most previous comments. Although they give valid arguments for their choices, none of the main concerns raised previously have been addressed in the revision. I recommend that some of the arguments given in the reply to justify this are included in the introduction/discussion. In addition, in some cases (e.g. “Pakayaku access is not free.”), the unclarity remains in the text, and the authors only explain what they mean in the reply. This is of course not useful for other readers (who may also wonder what is meant with “free” in this context). I recommend to edit the text where unclarities were previously indicated, if not yet done so. The grammar also still needs to be revised. Lastly, I have a few minor suggestions left: (i) remove “perfect” in the first line of the abstract (this relates again to my previous comment on small-scale versus large-scale; on large scales such systems are unlikely sustainable), (ii) “primary forest” should be “secondary forest in figure 1, (iii) Figure 6 and 7 (and maybe all figures except figure 1 and 2) are in fact tables, and should thus be named "Table 1”, etc.

Author Response

The authors have properly addressed most previous comments. Although they give valid arguments for their choices, none of the main concerns raised previously have been addressed in the revision. I recommend that some of the arguments given in the reply to justify this are included in the introduction/discussion. [Ok. The aforementioned arguments have been included in the Introduction, and this Part has been partially modified in order to attend better the concerns] In addition, in some cases (e.g. “Pakayaku access is not free.”), the unclarity remains in the text, and the authors only explain what they mean in the reply. This is of course not useful for other readers (who may also wonder what is meant with “free” in this context). I recommend to edit the text where unclarities were previously indicated, if not yet done so. [Yes, these explanations improve the previous draft, so we have included now, as you suggest ] The grammar also still needs to be revised.  [We have sent it to the MPDI Services, and we agree if the editor includes grammatical modifications]. Lastly, I have a few minor suggestions left: (i) remove “perfect” in the first line of the abstract (this relates again to my previous comment on small-scale versus large-scale; on large scales such systems are unlikely sustainable),[ Ok. Removed ] (ii) “primary forest” should be “secondary forest in figure 1, ,[ Sorry, but I cannot find these words, but tropical forest ]  (iii) Figure 6 and 7 (and maybe all figures except figure 1 and 2) are in fact tables, and should thus be named "Table 1”, etc .

 

This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.

Round 1

Reviewer 1 Report

The authors evaluated "Revalorization of a phytogenetic resource: the Western Mediterranean chamomile [Chamaemelum fuscatum (Brot.) Vasc.]".

From my point of view, the topic is interesting and the authors did a great work. English is readable and the background is established. So, I recommend the MS for publication after some revisions.

I have some comments:

TITLE:

I am not sure if the word "revalorization" is correct. What did the authors mean with "revalorization". Did they want to use "valorization". In this case, how?

ABSTRACT:

OK

INTRODUCTION:

As far as I understood the authors combined a research work with a review. Taking into account the structure, I would suggest not including in Introduction figures to clarify the overall idea.

In Introduction, I would suggest the authors focusing on the main aim lines 58-61 and summarizing lines 37-42.

Please, check if Table 1 can be in one page.

Authors should put in context "valorization"

M&M:

OK

CONCLUSIONS:

Taking into account this paper could be used as a reference for the Western Mediterranean chamomile [Chamaemelum fuscatum (Brot.) Vasc. conclusions section should be expanded. What about the origin influence, method of extraction, how to obtain valuable compounds (if the aim is the valorization)

Line 234: Please, check grammar.

REFERENCES:

Lines 293-295. Please, be consistent with the use of uppercase and lowercase letters. Check other references in the same situation.

Author Response

The authors evaluated "Revalorization of a phytogenetic resource: the Western Mediterranean chamomile [Chamaemelum fuscatum (Brot.) Vasc.]".

From my point of view, the topic is interesting and the authors did a great work. English is readable and the background is established. So, I recommend the MS for publication after some revisions. Thank you very much.

I have some comments:

TITLE:

I am not sure if the word "revalorization" is correct. Yes, it is a mistake. It must be changed. What did the authors mean with "revalorization". Did they want to use "valorization". In this case, how? The title has been changed.

ABSTRACT:

OK. Thanks

INTRODUCTION:

As far as I understood the authors combined a research work with a review. Taking into account the structure, I would suggest not including in Introduction figures to clarify the overall idea. Authors consider that this figure can be useful for differentiation of this species from other chamomiles. However, we let the editor make the decision on this matter.

In Introduction, I would suggest the authors focusing on the main aim lines 58-61 and summarizing lines 37-42. We have introduced all these data because they are the ones that make different this chamomile from others that are more wellknown and very often available in the markets.

Please, check if Table 1 can be in one page. Done.

Authors should put in context "valorization".  

The term “valorization” in the previous version of the manuscript was used to explain our intention to confer more value to C. fuscatum as a natural source of bioactives and to contribute this way to reinforce its use and its preservation. This word has been replaced in the new version of the manuscript and text in Abstract and Introduction has been conveniently changed to clarify this point.

M&M:

OK. Thanks.

CONCLUSIONS:

Taking into account this paper could be used as a reference for the Western Mediterranean chamomile [Chamaemelum fuscatum (Brot.) Vasc. conclusions section should be expanded. What about the origin influence, method of extraction, how to obtain valuable compounds (if the aim is the valorization)

Conclusions have been rewritten according to the referee’s suggestions.

Line 234: Please, check grammar. Ok. Changed.

REFERENCES:

Lines 293-295. Please, be consistent with the use of uppercase and lowercase letters. Check other references in the same situation. References have been conveniently formatted.

Reviewer 2 Report

The authors presented the chemical composition and bioactivity of different parts of the western Mediterranean chamomile (Chamaemelum fuscatum) in a very satisfactory way. The work is interesting, informative and well-organized and I enjoyed reading it.  I only have some minor observations that are listed in the following lines.

1.      Minor English revision is required.

2.      Figure 1: Add a scale to pictures b and c.

3.      Line 78: only 71.42% of the components were identified. Have you tried using HPLC to identify the unknowns?

4.      Table 2: Add the unknown compounds and their percentages.

5.      Lines 202-203: “The extraction yield (%) was calculated as the amount of essence (in mL) extracted by hydrodistillation from 100 grams of dry plant”. Provide more details on the calculation.

 

In summary, I think this manuscript needs minor revisions prior to acceptance.

Author Response

The authors presented the chemical composition and bioactivity of different parts of the western Mediterranean chamomile (Chamaemelum fuscatum) in a very satisfactory way. The work is interesting, informative and well-organized and I enjoyed reading it.  I only have some minor observations that are listed in the following lines.

1.      Minor English revision is required. MPDI Language Editinig consulted

2.      Figure 1: Add a scale to pictures b and c. Done

3.      Line 78: only 71.42% of the components were identified. Have you tried using HPLC to identify the unknowns? Here we have focused on the most volatile fraction, since it comprises a huge percentage of the essential oils of both the flowers and the whole plant, and these parts are the most frequently used in popular medicine (the preferred according to IECTB). However, HPLC methods might help as well for the study of the less volatile fraction and we probably consider for our next project.

4.      Table 2: Add the unknown compounds and their percentages. The unknown compounds >0.5% were added to take into account the referee’s suggestion. The several unknown compounds of <0.5% were not included.

5.      Lines 202-203: “The extraction yield (%) was calculated as the amount of essence (in mL) extracted by hydrodistillation from 100 grams of dry plant”. Provide more details on the calculation. It is a simple relation mL/g of dry plant expressed in %. This form of expression of results is commonly used in EO research. Text has been slightly changed to clarify this point.

In summary, I think this manuscript needs minor revisions prior to acceptance. Authors thank the referee for his/her comments on our manuscript.

Reviewer 3 Report

General comments:

In the manuscript entitled “Revalorization of a phytogenetic resource: the Western Mediterranean   chamomile [Chamaemelum fuscatum (Brot.) Vasc.]”, the authors propose as main objective of this study to undertake a detailed characterization of Chamaemelum fuscatum essential oils from flowers, steam and whole plant by GC-MS analysis to be correlated with the ethnobotanical information currently available in Spain. In general, the manuscript is well organized. However, this study is very preliminary, and some major issues have to be addressed before publication.

1) The article title is not clear. It is not representing the content of the article.

2) Line 45. Figure 1 is not a major contribution to the study.

3) Line 49, In table 1. The author should indicate the IECTB meaning (Inventario Español de los Conocimientos Tradicionales relativos a la Biodiversidad).

4) In Table 1, First row, fourth column, please delete comma after Province.

5) Line 57. The authors should add what part of the plant was used to identified sesquiterpene lactones and aliphatic esters. In addition, they should add what extract was obtained to carry out this objective.

6) Line 61. The IECTB meaning should be before the table 1.

7) Lines 58-68 are redundant. Lines 58-64 mean the same thing as line 66-68.

8) Lines 74-76 and lines 78-79 does not mean the same. In the first statement, the authors say that only 71,42% of components identified in the steam EO are present in other plant part, but in the second statement (It is right according to the table 2), the authors say that 71.42 % of components in the steam EO was identified.

9) Line 80 and 117. When you say "important" and "some", respectively. Do you refer to the most abundant compounds"? Please, clarify it.

10) In Table 2. The author should add a column with Linear retention index (I) from literature.

11) Line 91. "...(10.39-14.97%)..." it should be (14.97-10.39%).

12) Lines 92 and whole manuscript. When the authors say, "other compounds" or "Others" (in Table 2), they should indicate that long chains fatty acids are the major compounds classified in "other compounds"

13) Lines 94, 96, 99, 100. The author use "concentration" in a wrong way (check it in whole manuscript). The results should be presented as relative abundance (%). It indicates the amount of a compound about the total IDENTIFIED compounds. However, in the present study, the total identified compounds were different in each EO. Then, compare these values is not correct, but compare concentrations (e.g. mg/L) is possible to do.

14) In Figure 2. The authors should number the chemical structures according to the Table 1 and delete "acid" in the second row. Additionally, they should draw the aliphatic esters with the organic acid part on the same side, in order to make a better structural comparison.

15) In Table 3. The data in the second column is redundant. Moreover, Why the authors chose only these compounds to report its biological activity? Considering that these compounds are not the most abundant in the Chamomile EOs.

16) Lines 124-126 is not clear. Please clarify it.

17) Lines 134-140. This statement was not based on properly reference. Additionally, what is the mechanism of this physiological role? Is it related with lipophilicity of the compounds? Please, improve this assumption.

18) Lines 142-143. The authors say that information related with essential oils of related species to C. fuscatum is scarse. However, more information has been reported in literature related with it.

19) Lines 143-153. The cited studies were done using different extraction methodologies. For example, in [10] the plant material was extracted with n-hexane. In this study the EOs were obtained by hydrodistillation. A non-polar solvent like n-hexane is capable to extract non-polar compounds, leaving the more polar compounds. Non-polar compound and more polar compounds can be extracted by hydrodistillation. So, this comparison is unsuitable.

20) Lines 154-155. This statement has not any relation with the aim of this study

21) Lines 156-160. The hydrodistillation involve heated the plant material and water until 100 °C approximately (during 8h in this case). So, if hydrolysis of esters occurs probably you results show it.

22) Lines 194. "HSS 68118was" should be "HSS 68118 was"

23) Lines 198,199. I suggest to replace "volatile oil" with "essential oil" because the whole oil is not volatile.

24) Line 220. Please replace "Esential Oil" with "essential oil"

25) Line 229-230. Please, do not generalize. This study only informed the identification of the compound in C. fuscatum EO.

26) Line 233. Please, replace "in this paper" with "in this plant"

27) Lines 233-237. The authors conclude that validation of the ethnobotanical use of this plant has been supported with experimental chemical data collected by GC-MS and with bioactivity data published in the literature. However, the ethnobotanical use of any plant cannot be validated only with the identification of its essential oil components. Many other studies related with phytochemistry, pharmacological activities and toxicity are needed.

28) The authors should correct the references 7 (Is it a book? Please, add more information), 8-10 (Scientific name in Italic), and 11 (delete the Italic format)

Author Response

General comments:

In the manuscript entitled “Revalorization of a phytogenetic resource: the Western Mediterranean   chamomile [Chamaemelum fuscatum (Brot.) Vasc.]”, the authors propose as main objective of this study to undertake a detailed characterization of Chamaemelum fuscatum essential oils from flowers, steam and whole plant by GC-MS analysis to be correlated with the ethnobotanical information currently available in Spain. In general, the manuscript is well organized. However, this study is very preliminary, and some major issues have to be addressed before publication.

Referee is right, this is a first study on this topic and, as stated in Conclusions, our intention is to go on with the study of the effect of different factors (cultivation region, climate, extraction method, etc) on composition and bioactivity of C. fuscatum. However, as there was so little information published on this topic, we considered worthwhile to communicate the results of this preliminary study to the scientific community.

1) The article title is not clear. It is not representing the content of the article. Yes, it had a mistake in the first word. It has been changed.

2) Line 45. Figure 1 is not a major contribution to the study. Authors consider that this figure can be useful for differentiation of this species from other chamomiles. However, we let the editor make the decision on this matter.

3) Line 49, In table 1. The author should indicate the IECTB meaning (Inventario Español de los Conocimientos Tradicionales relativos a la Biodiversidad). It has been included as a footnote to this table.

4) In Table 1, First row, fourth column, please delete comma after Province. Done.

5) Line 57. The authors should add what part of the plant was used to identified sesquiterpene lactones and aliphatic esters. In addition, they should add what extract was obtained to carry out this objective. Done.

6) Line 61. The IECTB meaning should be before the table 1. Done.

7) Lines 58-68 are redundant. Lines 58-64 mean the same thing as line 66-68. Text has been rewritten according to the referee’s comment.

8) Lines 74-76 and lines 78-79 does not mean the same. In the first statement, the authors say that only 71,42% of components identified in the steam EO are present in other plant part, but in the second statement (It is right according to the table 2), the authors say that 71.42 % of components in the steam EO was identified. Ok. Corrected

9) Line 80 and 117. When you say "important" and "some", respectively. Do you refer to the most abundant compounds"? Please, clarify it. Done.

10) In Table 2. The author should add a column with Linear retention index (I) from literature. A new column has been added to Table 2 to include retention data (I^T) from literature.

11) Line 91. "...(10.39-14.97%)..." it should be (14.97-10.39%). Done.

12) Lines 92 and whole manuscript. When the authors say, "other compounds" or "Others" (in Table 2), they should indicate that long chains fatty acids are the major compounds classified in "other compounds".

Text has been rewritten to clarify this point.

13) Lines 94, 96, 99, 100. The author use "concentration" in a wrong way (check it in whole manuscript). The results should be presented as relative abundance (%). It indicates the amount of a compound about the total IDENTIFIED compounds. However, in the present study, the total identified compounds were different in each EO. Then, compare these values is not correct, but compare concentrations (e.g. mg/L) is possible to do.

Referee is right and expression of results in this new version of the manuscript has been done as relative abundance, percent data, etc. Concentrations cannot be calculated from the present study as no internal standard was added to the essential oils.

14) In Figure 2. The authors should number the chemical structures according to the Table 1 and delete "acid" in the second row. Additionally, they should draw the aliphatic esters with the organic acid part on the same side, in order to make a better structural comparison.

Authors consider that grouping of compounds by chemical class rather than by elution order is more useful for the reader to draw conclusions on the chemical composition of C. fuscatum essential oils. The rest of changes suggested by the referee have been done.

15) In Table 3. The data in the second column is redundant. Moreover, Why the authors chose only these compounds to report its biological activity? Considering that these compounds are not the most abundant in the Chamomile EOs. Second column of Table 3 has been removed. Text and legend of Table 3 have been changed to clarify that C. fuscatum compounds included in this table are only those for which bibliographic references endorsing their pharmacological activity have been found.

16) Lines 124-126 is not clear. Please clarify it. Text has been rewritten.

17) Lines 134-140. This statement was not based on properly reference. Additionally, what is the mechanism of this physiological role? Is it related with lipophilicity of the compounds? Please, improve this assumption. New information has been included in this paragraph

18) Lines 142-143. The authors say that information related with essential oils of related species to C. fuscatum is scarse. However, more information has been reported in literature related with it. Text has been rewritten to clarify this point.

19) Lines 143-153. The cited studies were done using different extraction methodologies. For example, in [10] the plant material was extracted with n-hexane. In this study the EOs were obtained by hydrodistillation. A non-polar solvent like n-hexane is capable to extract non-polar compounds, leaving the more polar compounds. Non-polar compound and more polar compounds can be extracted by hydrodistillation. So, this comparison is unsuitable. Text has been changed to provide details on the extraction methods employed in each previous reference and to clarify the compounds previously reported in the literature.

20) Lines 154-155. This statement has not any relation with the aim of this study. It has relation, because it is useful to demonstrate C. fuscatum can be used as a plain chamomile infusion.

21) Lines 156-160. The hydrodistillation involve heated the plant material and water until 100 °C approximately (during 8h in this case). So, if hydrolysis of esters occurs probably you results show it. We agree with referee. Text in the new version of the manuscript takes into account this fact.  

22) Lines 194. "HSS 68118was" should be "HSS 68118 was". Done.

23) Lines 198,199. I suggest to replace "volatile oil" with "essential oil" because the whole oil is not volatile. Done.

24) Line 220. Please replace "Esential Oil" with "essential oil". Done.

25) Line 229-230. Please, do not generalize. This study only informed the identification of the compound in C. fuscatum EO. We have rewritten the text to take into account the referee’s suggestion.

26) Line 233. Please, replace "in this paper" with "in this plant". Done.

27) Lines 233-237. The authors conclude that validation of the ethnobotanical use of this plant has been supported with experimental chemical data collected by GC-MS and with bioactivity data published in the literature. However, the ethnobotanical use of any plant cannot be validated only with the identification of its essential oil components. Many other studies related with phytochemistry, pharmacological activities and toxicity are needed.

We agree with the referee and the experimental evaluation of C. fuscatum bioactivity is required to fully support the conclusions here presented. It is our intention to address this evaluation in our future research on this topic. However, the results here presented are a preliminary contribution to correlate chemical composition of C. fuscatum essential oils and their bioactivity/ethnobotanical use. In this new version of the paper, abstract, main text and conclusion sections have been rewritten to clarify this.

28) The authors should correct the references 7 (Is it a book? Please, add more information) Thesis 8-10 (Scientific name in Italic), and 11 (delete the Italic format). Done.

Reviewer 4 Report

Comments on the manuscript

 

Manuscript ID:           molecules-436784

Recommendation:    Reject and resubmit

The authors of manuscript “Revalorization of a phytogenetic resource: the Western Mediterranean chamomile [Chamaemelum fuscatum (Brot.) Vasc.]” had the aim to give insight into chemical and ethnobotanical characterization C. fuscatum essential oil. The plant has been used traditionally over the centuries and was known for its wide variety of bioactivities, what might be real base for its application in the pharmacological, cosmetic and food industries.

Although the subject of manuscript is worth thorough investigation, as essential oil had not be investigated since eighties in the past century, as the plant has been traditionally used and showed the numerous beneficial effects on human health, the chosen methodology presented in the paper could not enable the achievement of the proposed goal. The paper has several shortcomings that made the work inadequate for the publishing in the present form.

       [1]       Extensive editing of English language and style are required. Please, before submitting, the authors are asked to check with the native English language speaker for polishing the text

 

       [2]       Abstract is unclear, and poorly written, needing the complete rewriting. Some of the abstract parts that required revision and rewriting:

1.      The statement: “For this reason, it is proposed in this paper to get insight into the scientific validation of the traditional knowledge of this plant with the aim of taking advantage of its anti-inflammatory, gastroprotective and antinociceptive activities, among others..” does not correspond to the investigation conducted. Namely, authors did not validate the traditionally recognized activities, but rather had been involved into chemical composition investigation of three different parts of the plant known for their application in human and veterinary medicine.

2.      Authors throughout the text use “essence” for essential oil, and it should be corrected, taking into account the meaning of the word “essence” and what “essential oil” represent.

3.      The statement:  “In addition to other aliphatic esters previously reported, several compounds of this group such as isobutyl methacrylate and isoamyl butyrate were identified for the first time in this paper”, it is unclear if the mentioned compounds were for the first time identified in plant material overall, or for the first time in the plant investigated.

4.      Afterwards, authors introduced benzyl methacrylate as compound not identified earlier, but in Abstract it has not been mentioned.

 

       [3]       Introduction should give more information regarding the earlier performed research

1.      In section Introduction, it should be specifically pointed out the differences into up-to-now performed investigation and the one presented in the manuscript - namely, the volatile compounds have not been investigated earlier, except in order to investigate the antioxidative properties “Comparative antioxidant evaluation of the essential oils from chamomile's commercial samples” presented at the congress, Planta Med 2007; 73 - P_066, DOI: 10.1055/s-2007-986848 (not pure C. fuscatum essential oil), and the use of NMR techniques to identify the methacrylate esters present in this interesting plant, and of course, in the work presented in the cited literature No. 10.

2.      Authors should stress the novelty in their approach to investigation of the chemical composition of essential oil, clearly pointing out the aim.

3.      When first mentioned, the Family name of the investigated species should be given

 

       [4]       Results and Discussion

1.      The main concern is for presented quantitative composition of investigated essential oils. For quantification of the detected compounds present in essential oils MS could not be used - the combination of GC/MS and GC/FID is required. As there is no available literature about C. fuscatum essential oil chemical composition comprehensive investigation, it is a reason more to use GC/FID for quantitative calculations.

2.      Please, throughout the text use flower, or flowerheads, not both.

3.      “Essence” is not essential oil

4.    The classification of the identified compounds should be corrected - authors choose to represent monoterpenoids, sesquiterpenoids… in the table 2; monoterepnes, sesquiterpenes … represent hydrocarbons, without oxygen in molecules.

5.      As authors mentioned identification of benzyl methacrylate, please give the literature if that compound was identified in nature, at all.

6.      Lines 96-97 - as authors stressed the new findings in their investigation regarding the presence of isobutyl methacrylate and isoamyl butyrate, why was only the presence of isobutyl methacrylate stressed, taking into account that the other mentioned compound was detected in relatively high percentage (3.6 and 1.64% in F and WP, respectively).

7.      Authors are asked to make reasonable the selection of just several compounds from the list of the identified compounds present in the investigated essential oils in Figure 2. Please, correct the “monoterpenes”, “sesquiterpenes”….

8.      Afterwards, the biological activity of other compounds was presented Table 3 - please, explain what were the criteria for choosing the mentioned compounds…

9.      Although at the beginning of the manuscript, authors gave the evidence that all investigated parts of the plant were used in traditional medicine, in Discussion section, in lines 126-127 authors stated that only flowers/flowerheads were medicinally used - please, give the sustainable explanation.

10.   Lines 132-140 should be rewritten - English should be polished.

11.   Aliphatic esters constituted, according to the results presented, major part of flower and aerial part essential oils - please give the literature data about their pharmacological potential, as they were not isolated and tested for specific biological activities. For the statements, lines 156 -162, the literature should be provided.

12.   Please, precise the butyrates detected in Roman chamomile (lines 152-154), and other Anthemis species, as they were considered as potential chemotaxonomy markers. Literature data, if any, might be helpful confirming the statement.

13.   Give the full Latin name for Chamaemelum nobile

14. Taking into account that Roman and German chamomile have their monographs in numerous Pharmacopoeias, the comparison with them might be valuable (their chemical profile, not only essential oils composition, but non-volatile fractions, like flavonoids and sesquiterpene lactones)

15.   The bibliography data regarding the pharmacological application are very interesting, but the obtained results could not be used to evaluate the mentioned activities, as the experiments were not conducted. Such approach is based on theoretical compilation, what is not acceptable in research article. Such approach might be used in reviews.

 

       [5]       Conclusion should point out the main results obtained in the experiments, stressing the importance of the experimentally obtained results.

 

Overall, the impression is that authors had an interesting staring material, but somehow did not succeed into achieving the scientific soundness.

Author Response

Recommendation: Reject and resubmit.

The authors of manuscript “Revalorization of a phytogenetic resource: the Western Mediterranean chamomile [Chamaemelum fuscatum (Brot.) Vasc.]” had the aim to give insight into chemical and ethnobotanical characterization C. fuscatum essential oil. The plant has been used traditionally over the centuries and was known for its wide variety of bioactivities, what might be real base for its application in the pharmacological, cosmetic and food industries.

Although the subject of manuscript is worth thorough investigation, as essential oil had not be investigated since eighties in the past century, as the plant has been traditionally used and showed the numerous beneficial effects on human health, the chosen methodology presented in the paper could not enable the achievement of the proposed goal. The paper has several shortcomings that made the work inadequate for the publishing in the present form.

       [1]       Extensive editing of English language and style are required. Please, before submitting, the authors are asked to check with the native English language speaker for polishing the text. English has been revised by MPDI Services.

        [2]       Abstract is unclear, and poorly written, needing the complete rewriting. Some of the abstract parts that required revision and rewriting:

1.       The statement: “For this reason, it is proposed in this paper to get insight into the scientific validation of the traditional knowledge of this plant with the aim of taking advantage of its anti-inflammatory, gastroprotective and antinociceptive activities, among others..” does not correspond to the investigation conducted. Namely, authors did not validate the traditionally recognized activities, but rather had been involved into chemical composition investigation of three different parts of the plant known for their application in human and veterinary medicine. We agree with the referee and the experimental evaluation of C. fuscatum bioactivity is required to fully support the conclusions here presented. It is our intention to address this evaluation in our future research on this topic. However, the results here presented are a preliminary contribution in which experimental data on chemical composition of C. fuscatum essential oils has been correlated with bioactivity data previously reported in the literature. In this new version of the paper, abstract, main text and conclusion sections have been rewritten to clarify this.

2.      Authors throughout the text use “essence” for essential oil, and it should be corrected, taking into account the meaning of the word “essence” and what “essential oil” represent. This change has been done through the text.

3.      The statement:  “In addition to other aliphatic esters previously reported, several compounds of this group such as isobutyl methacrylate and isoamyl butyrate were identified for the first time in this paper”, it is unclear if the mentioned compounds were for the first time identified in plant material overall, or for the first time in the plant investigated. This sentence has been changed to clarify that these compounds were detected for the first time in the plant investigated.

4.      Afterwards, authors introduced benzyl methacrylate as compound not identified earlier, but in Abstract it has not been mentioned.

According to referees, authors have decided to reinforce the interest of the other compounds that are more abundant, so this has changed.

 

       [3]       Introduction should give more information regarding the earlier performed research.  Done.

. 1.      In section Introduction, it should be specifically pointed out the differences into up-to-now performed investigation and the one presented in the manuscript - namely, the volatile compounds have not been investigated earlier, except in order to investigate the antioxidative properties “Comparative antioxidant evaluation of the essential oils from chamomile's commercial samples” presented at the congress, Planta Med 2007; 73 - P_066, DOI: 10.1055/s-2007-986848 (not pure C. fuscatum essential oil), and the use of NMR techniques to identify the methacrylate esters present in this interesting plant, and of course, in the work presented in the cited literature No. 10.  Ok. Done

2.      Authors should stress the novelty in their approach to investigation of the chemical composition of essential oil, clearly pointing out the aim

3.      When first mentioned, the Family name of the investigated species should be given. Done.

 

       [4]       Results and Discussion

1.      The main concern is for presented quantitative composition of investigated essential oils. For quantification of the detected compounds present in essential oils MS could not be used - the combination of GC/MS and GC/FID is required. As there is no available literature about C. fuscatum essential oil chemical composition comprehensive investigation, it is a reason more to use GC/FID for quantitative calculations. We will use GC/FID in our next projects on this plant for offer better quantitative calculations.

2.      Please, throughout the text use flower, or flowerheads, not both. Done.

3.      “Essence” is not essential oil. Changed.

4.    The classification of the identified compounds should be corrected - authors choose to represent monoterpenoids, sesquiterpenoids… in the table 2; monoterepnes, sesquiterpenes … represent hydrocarbons, without oxygen in molecules. This has been changed throughout the text.

 

5.      As authors mentioned identification of benzyl methacrylate, please give the literature if that compound was identified in nature, at all. This has been changed.

 

6.      Lines 96-97 - as authors stressed the new findings in their investigation regarding the presence of isobutyl methacrylate and isoamyl butyrate, why was only the presence of isobutyl methacrylate stressed, taking into account that the other mentioned compound was detected in relatively high percentage (3.6 and 1.64% in F and WP, respectively). This compound has also been highlighted in the new version of the paper.

 

7.      Authors are asked to make reasonable the selection of just several compounds from the list of the identified compounds present in the investigated essential oils in Figure 2. Please, correct the “monoterpenes”, “sesquiterpenes”…. Nomenclature has been corrected. Text has been changed to clarify which compounds were selected to be included in Figure 2.

 8.      Afterwards, the biological activity of other compounds was presented Table 3 - please, explain what were the criteria for choosing the mentioned compounds.  

Text and legend of Table 3 have been changed to clarify that C. fuscatum compounds included in this table are only those for which bibliographic references endorsing their pharmacological activity have been found.

9.      Although at the beginning of the manuscript, authors gave the evidence that all investigated parts of the plant were used in traditional medicine, in Discussion section, in lines 126-127 authors stated that only flowers/flowerheads were medicinally used - please, give the sustainable explanation. Flowerheads are the most used and preferred part according to IECTB.  This has been corrected in the text.

10.   Lines 132-140 should be rewritten - English should be polished. English has been revised by MPDI Service.

11.   Aliphatic esters constituted, according to the results presented, major part of flower and aerial part essential oils - please give the literature data about their pharmacological potential, as they were not isolated and tested for specific biological activities. For the statements, lines 156 -162, the literature should be provided. No reference has been found on bioactivity of these compounds. Authors will address this evaluation in future work on this topic. Text has ben changed to clarify this.

12.   Please, precise the butyrates detected in Roman chamomile (lines 152-154), and other Anthemis species, as they were considered as potential chemotaxonomy markers. Literature data, if any, might be helpful confirming the statement. In the present paper, comparison of non-volatile fraction of all these species has not been done as this was not the objective of the paper..

13.   Give the full Latin name for Chamaemelum nobile. Done.

14. Taking into account that Roman and German chamomile have their monographs in numerous Pharmacopoeias, the comparison with them might be valuable (their chemical profile, not only essential oils composition, but non-volatile fractions, like flavonoids and sesquiterpene lactones). Thank you for the idea. This comparison has been included for the essential oils of the chamomiles suggested. The non-volatile fractions are not the point of this work, so maybe we will consider them in our next projects.

15.   The bibliography data regarding the pharmacological application are very interesting, but the obtained results could not be used to evaluate the mentioned activities, as the experiments were not conducted. Such approach is based on theoretical compilation, what is not acceptable in research article. Such approach might be used in reviews.

As stated in abstract, main text and conclusion sections, the results here presented are a preliminary contribution to correlate chemical composition of C. fuscatum essential oils and their bioactivity/ethnobotanical use. We agree with the referee and the experimental evaluation of C. fuscatum bioactivity is required to fully support the conclusions here presented. It is our intention to address this evaluation in our future research on this topic.

[5]       Conclusion should point out the main results obtained in the experiments, stressing the importance of the experimentally obtained results. We have opted for giving to our experimental results an added value by the study of the scientific literature and the critical review of bibliography. Science improves by this sort of proceeding in our perspective.

 

Overall, the impression is that authors had an interesting staring material, but somehow did not succeed into achieving the scientific soundness.  

We really hope that after the corrections suggested by the five referees, we have significantly changed the quality of our manuscript.

Reviewer 5 Report

Medicinal plants and their essential oils are very popular due to return to natural medicine. Therefore, it is very important to know the composition and chemical properties of commonly used essential oils. In my opinion, the results presented in the reviewed manuscript are not enough for the work to be published. Pharmacological activity is presented only on the basis of literature data. It would also be necessary to study the activity of the Western Mediterranean chamomile. Instead ar-curcumene should be α-curcumene. The sentence in lines 74-75 is repeated in line 78.

Author Response

Medicinal plants and their essential oils are very popular due to return to natural medicine. Therefore, it is very important to know the composition and chemical properties of commonly used essential oils. OK.

In my opinion, the results presented in the reviewed manuscript are not enough for the work to be published. Pharmacological activity is presented only on the basis of literature data. It would also be necessary to study the activity of the Western Mediterranean chamomile. As stated in abstract, main text and conclusion sections, the results here presented are a preliminary contribution to correlate chemical composition of C. fuscatum essential oils and their bioactivity/ethnobotanical use. We agree with the referee and the experimental evaluation of C. fuscatum bioactivity is required to fully support the conclusions here presented. It is our intention to address this evaluation in our future research on this topic.

Instead ar-curcumene should be α-curcumene. The sentence in lines 74-75 is repeated in line 78. All these changes have been done.

Round 2

Reviewer 1 Report

The authors addresed well my comments and I recommend accepting the paper. However, English language should be carefully reviewed by an English native speaker.

Reviewer 3 Report

General comments:

In the manuscript entitled “Analysis of the Essential Oils of Chamaemelum fuscatum (Brot.) Vasc. as a Contribution to Reinforce Its Ethnobotanical Use”, the authors propose as main objective of this study to undertake a detailed characterization of Chamaemelum fuscatum essential oils from flowers, steam and whole plant by GC-MS analysis to be correlated with the ethnobotanical information currently available in Spain. In general, the manuscript was improved. However, the study is very preliminary yet, and some issues have to be addressed.

1) Considering the main objective of the manuscript. I think a review article is the best option.

2) Figure 1 is not a major contribution to the study. If the authors consider that this figure can be useful for differentiation of this species from other chamomiles, I suggest send it as supplementary material.)

3) Related with "...no information has been published on the chemical profile of C. fuscatum." Lines 58-65 and 66-67 are contradictory.

4) Line 67. When you mentioned "volatile fraction". What did you mean? Because volatile fraction and essential oils are not the same.

5) Line 87. "...yields of 0.41% (F = flowers), 0.13% (S = stems) and 0.27% (WP = whole plant) for the different parts evaluated..." I suggest: ..."yields of 0.41%, 0.13% and 0.27% for flowers (F), stems (S) and whole plant (WP), respectively".

6) In Table 2. Compounds 60 and 61 are very abundant in S and, to a lesser extent, in WP. Why it was not considered in the classification?

7) Please, check the format of Figure 2 in PDF file.

8) Line 103. "Others" compounds are more abundant than monoterpenes in WP. Please, correct the statement.

9) There are many studies about bioactivities of limonene and alcanfor. Why the authors did not include it in Table 3. Additionally, Why the authors included neryl isovalerate if they did not found information?

10) Line 144. Is not suitable use "significantly differed" because the authors did not perform any statistical analysis.

11) Lines 151-152. Is it an assertion or assumption? If it is a statement, the authors should add the properly cite. How the compounds are transported in the plant?

12) Line 173. Please, replace "apolar" with "non-polar".

Reviewer 4 Report

Manuscript ID: molecules-436784

Recommendation: Reject and resubmit.

The authors of the manuscript “Analysis of the Essential Oils of Chamaemelum fuscatum (Brot.) Vasc. as a Contribution to Reinforce Its Ethnobotanical Use” have mainly included the suggested changes, significantly improved the revised paper.

Taking into account the rang of this journal, its impact factor, the presenting only results of essential oil chemical composition is not acceptable for publishing, especially as the quantifications was based only on GC/MS method. The statement mentioned in authors' response that “GC/FID” will be “used in our next projects on this plant for offer better quantitative calculations” places this investigation into category of preliminary research not acceptable for the manuscript. Any other investigation of any of mentioned biological activities was not performed for the isolated essential oils. The hypothesis that essential oil might be responsible for the known traditional uses, is just contemplation, which was not confirmed through the performed investigation, especially as in Table 1, essential oil was not mentioned, and that the applications referred to water extracts  (infusions and decoctions).

The compounds chosen to be presented in Table 3, did not correspond to the presented results in Table 2, as there are terpenoids with greater abundance from those presented in Table 3.

Also, it might be of significant importance to compare the biological activity (microbial, anti-inflammatory, antioxidant…), of different essential oils presented in this investigation, taking into account the difference they have in their chemical composition.

The comment “… the volatile fraction of this plant showed similitude with the essential oils isolated from C. nobile, so it would work well as an alternative to this plant…” could not be confirmed, and to make such a note, not only chemical, but the biological activities investigations of volatile and others fractions are required. The manuscript did not reveal the experimentally obtained proof for these statements for the subject of the investigation.

 

Table 2, in legend still remained “monoterpenes” and “sesquiterpenes” that should be changed to “monoterpenoids”, “sesquiterpenoids”…

 

To reach the standard of the journal, the investigation regarding the essential oils should be completed - in regard to their chemical composition - qualitative and quantitative analysis, and to evaluate any of the mentioned biological activities.

Reviewer 5 Report

I suggested to study the pharmacological activity in my previous review. The authors of this manuscript replied: “the results here presented are a preliminary contribution to correlate chemical composition of C. fuscatum essential oils and their bioactivity/ethnobotanical use”.

In this case the Section “2.2 Pharmacological Activity of the Main Components of C. fuscatum Essential Oils” shouldn’t be included in the results because it contains only the literature data. And don’t contain any results obtained by the authors of the publication. (According to the instructions for authors: “Results: Provide a concise and precise description of the experimental results, their interpretation as well as the experimental conclusions that can be drawn.”)

If the work is a preliminary contribution to further research, the information presented in section 2.2 should be transferred to the section “1. Introduction”.

Also discussing the pharmacological activity in the section “3. Discussion” should be transferred to the section “1. Introduction” because it relates only to literature data regarding to other plants.