Quantitative Ethnomedicinal Status and Phytochemical Analysis of Berberis lyceum Royle

Round 1

Reviewer 1 Report

This manuscript describes a valuable dataset, however its scientific presentation should be greatly improved. Please follow basic instructions about writing scientific papers and Instructions for Authors provided by the journal. I added some more specific comments to the pdf-file.

Comments for author File: Comments.pdf

Author Response

List of amendment

Comments and Suggestions for Authors

 

Dear Editor/ reviewer,

Thank you very much for kind efforts and comments concerning our manuscript. We appreciate the hard work of reviewers as they fairly pointed out errors and mistakes in our manuscript. We have tried to revise the manuscript in line with comments of the reviewers. Corrections made have been highlighted as Blue.

Please find below the point by point responses to the reviewer’s comments and suggestions.

Reviewer 1 report:

 

Abbreviation unexplained

• Answer: The abbreviation has been explained in the revised manuscript accordingly.

In the last sentence of the abstract please bring out the main conclusions about your study questions not the aim of the study.

• Answer: Worthy reviewer the main conclusion of the study has been added in the abstract. The whole abstract has been rephrased and hopefully it will be ok now.

The Introduction should show knowledge gaps in your study topic and explain why your study is important and interesting for international readers of the scientific journal. Please read Instructions for Authors provided by Agronomy.

• Answer: The required detail have been incorporated in the revised paper accordingly. We have tried our best to cover the gaps and make it interesting for readers.

Why you want to publish the research in international scientific journal if your aim is to inform local community?

• Answer: Worthy reviewer, the paper not only covers the mentioned aspect of informing the local community it also contain knowledge about its phytochemical composition and biological activities which are important to all communities around the globe. Also to best of our knowledge this plant is endangered in many other parts of the world as well. Through this study the researchers there will get encourage and will perform such or better studies. Plants belonging to Berberis are reported in several folklore medicinal pharmacopeias and are used in traditional medicines in Asia and European countries. So, our aim is not only to inform the local community but to convey the knowledge related Berberis lyceum Ethnobotany is an integral part of indigenous/local knowledge of a particular society. Different societies or communities have their own knowledge about plants and their uses. However the statement ‘‘The aims of this ethnobotanical study is to aware the local community of District Shangla about the conservation and folk uses of this important medicinal plant Berberis lyceum’’ which confuse the readers that the aim of the study limited to the local community has been removed and the statement has been improved in the revised manuscript which now provide a broader meaning that our main reason to document the folkloric uses of Berberis lyceum.

The quality of the figure should be increased.

• Answer: The quality of all the figures has been improved in the revised manuscript accordingly.

‘‘The District Shangla is surrounded in the North by District Kohistan, in the East by District Batagram and the Tribal area of Kala Daka (Black mountains of District Hazara), in the West by District Swat and in the South by District Buner (32). The whole geographical map of the study area was presented in Fig.1’’ unnecessary information. Instead of those administrative details, the distribution of B. lyceum should be described.

• Answer: Worthy reviewer suggestion has been honoured and the mentioned statement has been removed and the paragraph related the distribution of lyceum has been added in the introduction section.

A and B contain the same information. Delete B. C should be in esults.

• Answer: The figure 1 is now figure 2 in the revised manuscript. Worthy reviewer the section that shows the same information has been removed in the revised figure accordingly.

Informant demographic status

A total of 100 informants of District Shangla were interviewed with aged range between 30 to ≥50 years. The people were divided into three age groups. Among these one age group was from 30 to 40 years having 21%. The second group was from 41 to 50 year having 60% while the third group from above fifty years old which were 19% (Figure 2).

Do not repeat the information that is presented in the figures or tables

• Respected reviewer the information in the text has been removed from the revised manuscript.

this is a case study not a review

• Answer: Respected reviewer the word review has been changed by a case study.

Why this column is given if it has all the same values?

• Answer: Worthy reviewer in Table 4 the Column related reference standard was deleted accordingly

Reviewer 2 Report

The article by Nausheen Nazir and co-authors "Quantitative ethnobotanical status and phytochemical analysis of Berberis lyceum Royle. In the District Shangla Khyber Pakhtunkhwa, Pakistan" is devoted to the assessment of the ethnobotanical status of the plant Berberis lyceum Royle. 

- Line 22-23

"... home-made medication used against various diseases"

I think it is better to indicate here either specific diseases, a group of diseases, or the therapeutic effects of this plant.

 

- Line 23

"Ethno botanical status"

ethnobotanical is written in one word, please correct the same for the title

 

- References to literature are not made in accordance with the requirements of the journal. It needs to be redone.

 

- Line 43 -45 World Health Organization (WHO) designates that due to poverty and lack of access to modern medicine almost 65-80% world’s populace of the developing countries mostly used medicinal plants for their primary healthcare (2).

I think it would be appropriate here to present the official report of the World Health Organization (WHO) postulating this thesis.

 

- Line 53 -55 Medicinal plants show beneficial applications because of their important constituents called the secondary metabolites that include phenolics, alkaloids, steroids, and tannins which are stored in a specific part or in the whole plant.

There is not enough link to this proposal.

 

- Ref. 15, does not match the text quoted in the introduction.

 

- Line 81 – 84 In District Shangla among most of the plant species B. lyceum is endangered species. The local communities use it for burning to cook food due to which plant endangered. The aims of this ethno botanical study is to aware the local community of District Shangla about the conservation and folk uses of this important medicinal plant Berberis lycium.

The formulated hypothesis is not very clear, on the one hand, the introduction postulates that the population of the region actively uses this plant as a therapeutic agent, and on the other hand, the population does not know anything about this, and additional information is needed.

In addition, the plant is on the verge of extinction due to burning, but informing the population about the use of this plant as a medicine is unlikely to have a positive effect on the restoration of the population of this species.

 

- Line 95 Shahpur, Besham, Chakesar, Aloch, Olander and Damorai (31)

The linear order of presentation of links is broken.

 

- Line 97 Figure 6 Geographical location of the District Shangla (study area) (Adopted from Tabani, et al.,

The numbering of figures is violated, the resolution of the map is very low, it is impossible to read the geographical names indicated on the map.

 

-Very low quality of the graphs presented in the article.

- different fonts are used

- charts are surrounded by frames

- Figure A and B of Figure 1 actually display the same information, but expressed in different graphic forms

 

- Figure C, Figure 1

On what basis were such diagnoses made?

 

- Line 214 Table S1 (Supplementary file)

Doesn't require decryption

 

- On what basis the respondents were divided into age groups (30-40 years old, 41-50 years old, over 51 years old).

 

- Figure 2 obviously duplicates the data from table S1. By and large, 2 paragraphs (from 214 lines and from 222 lines) tell about the same thing.

 

- The abbreviation UV is misleading, in some cases it stands for «use value», and in some, more traditional, ultraviolet.

 

- I don't really understand the need for Figure 4.

In any case, most of the abscissa positions are unreadable, you can use the ordinate break.

 

- Clause 3.5.

It seems to me redundant information in the context of this study.

 

- Line 318 3.9. Future impact of the study

I think the publication of the material in the journal Agronomy is unlikely to increase the awareness of the local population about the medicinal properties of the plant.

 

- Line 319 This review will provide a sense of awareness among the local people

This is not a review.

 

The fragment with the questionnaire looks inappropriate in the context of this study, I am sure that without it the article will not lose arguments in confirming the hypothesis formulated.

 

There is no harmony and transparency in the design of the experiment. A very large set of heterogeneous data is presented, which, from the point of view of semantic content, could be divided into three categories. If supplement, qualitatively describe and discuss the obtained experimental materials, it would be possible to write 3 separate articles.

1. Biochemical analysis and antioxidant status of Berberis lyceum Royle. Taking into account the fact that the authors claim that different parts of the plant are used for medicinal needs, it is possible to carry out (if this was not done earlier) a differential analysis of plant organs. You can analyze seasonal changes in the concentration of biologically active compounds. In addition, as I understand it, there were several collection points for the plant, obviously these are places with a unique microclimate, and the researchers have the opportunity to assess how they affect the metabolic processes of plants.
2. Sociological research. Population survey. Perhaps it is worth expanding the sample of respondents (the number of people, normalizing groups by age, gender, describing their state of health in accordance with diagnostic tests).
3. Phytobiocenotic research. Researchers attempted to describe the floristic profile of the area where Berberis lyceum Royle grows. However, this section seems unnecessary in this article. If we describe in detail the terrain, the dominant species of the studied ecotopes, the ecological links between them, and assess the conditions in which this species is most productive, then such a data set (with proper description) can become the basis for a separate publication.

Author Response

List of amendment

Comments and Suggestions for Authors

 

Dear Editor/ reviewer,

Thank you very much for kind efforts and comments concerning our manuscript. We appreciate the hard work of reviewers as they fairly pointed out errors and mistakes in our manuscript. We have tried to revise the manuscript in line with comments of the reviewers. Corrections made have been highlighted as Blue.

Please find below the point by point responses to the reviewer’s comments and suggestions.

Reviewer 2 reports:

The article by Nausheen Nazir and co-authors "Quantitative ethnobotanical status and phytochemical analysis of Berberis lyceum Royle. In the District Shangla Khyber Pakhtunkhwa, Pakistan" is devoted to the assessment of the ethnobotanical status of the plant Berberis lyceum Royle. 

- Line 22-23

"... home-made medication used against various diseases"

I think it is better to indicate here either specific diseases, a group of diseases, or the therapeutic effects of this plant. 

• Answer: Worthy reviewer your suggestion was honoured and the statement has been changed accordingly in the revised manuscript.

- Line 23

"Ethno botanical status"

ethnobotanical is written in one word, please correct the same for the title

• Answer: Worthy reviewer the ethno botanical word has been corrected as one word ‘‘ethnobotanical’’ in the abstract as well as in the title.

- References to literature are not made in accordance with the requirements of the journal. It needs to be redone.

• Answer: The references have been rechecked and corrected accordingly.

- Line 43 -45 World Health Organization (WHO) designates that due to poverty and lack of access to modern medicine almost 65-80% world’s populace of the developing countries mostly used medicinal plants for their primary healthcare (2).

I think it would be appropriate here to present the official report of the World Health Organization (WHO) postulating this thesis.

• Answer: Respected reviewer the reference related the above mentioned statement has been replaced by the official report of the World Health Organization (WHO) in the revised manuscript.

- Line 53 -55 Medicinal plants show beneficial applications because of their important constituents called the secondary metabolites that include phenolics, alkaloids, steroids, and tannins which are stored in a specific part or in the whole plant.

There is not enough link to this proposal.

• Answer: Respected reviewer the mentioned paragraph has been expanded in the revised manuscript which I think provide better link to this proposal. Also the revised paper has totally been rephrased and hopefully it will be ok now.

- Ref. 15, does not match the text quoted in the introduction.

• Answer: Worthy reviewer the reference 15 has been replaced by relevant one.

- Line 81 – 84 In District Shangla among most of the plant species B. lyceum is endangered species. The local communities use it for burning to cook food due to which plant endangered. The aims of this ethno botanical study is to aware the local community of District Shangla about the conservation and folk uses of this important medicinal plant Berberis lycium.

The formulated hypothesis is not very clear, on the one hand, the introduction postulates that the population of the region actively uses this plant as a therapeutic agent, and on the other hand, the population does not know anything about this, and additional information is needed.

• Answer: Worthy reviewer the paper has been revised very carefully through expert and such type of discrepancies has been removed. Hopefully it will be ok now.
• Respected reviewer, some of the people of District Shangla are uneducated and they did not know about the conservation of this important medicinal plant. So to cover both educated and uneducated groups a new term less educated has been used in the revised manuscript. The whole manuscript has been revised to remove such types of contradictions. 

In addition, the plant is on the verge of extinction due to burning, but informing the population about the use of this plant as a medicine is unlikely to have a positive effect on the restoration of the population of this species.

• Answer: Worthy reviewer, it is common practice throughout the world that valuable woody plant like olive, pinus etc and fruit plant are not used for burning to cook food. They are used when the tree dies. It is because they are valuable. If the community is informed about its value they will definitely stop its use for burning to cook food. Instead they will try to preserve it to get maximum benefits. As I have clarified that some of the people of District Shangla are uneducated and they exploit this important medicinal specie by using it for burning purpose too in spite of its uses for healing purposes. So the aim of this current investigational study was to inform the local communities of District Shangla about the high therapeutic value of B. lyceum.

- Line 95 Shahpur, Besham, Chakesar, Aloch, Olander and Damorai (31)

The linear order of presentation of links is broken.

• Answer: Worthy reviewer, sorry for the mistake the details about sub tehsils have been moved to first sentence of paragraph with corrected alphabetical order as Besham, Shahpur, Aloch, Chakesar, Damorai and Olander in the revised manuscript.

- Line 97 Figure 6 Geographical location of the District Shangla (study area) (Adopted from Tabani, et al.,

The numbering of figures is violated, the resolution of the map is very low, it is impossible to read the geographical names indicated on the map.

• Answer: Worthy reviewer the figures number has been changed in the revised manuscript. The resolution of the map which is now figure 1 has been increased.

-Very low quality of the graphs presented in the article.

• Answer: The quality of all the graphs has been improved.

- different fonts are used

• Answer: The font sizes has been corrected. However in some tables it has been retained as the table then look somewhat awkward.

- charts are surrounded by frames

• Answer: The frames surrounded the charts has been removed.

- Figure A and B of Figure 1 actually display the same information, but expressed in different graphic forms

• Answer: Section B of figure 1 has been removed. Figure 1 has now become figure 2 in the revised manuscript.

- Figure C, Figure 1

On what basis were such diagnoses made?

• Answer: Figure 1 has been revised which now becomes figure 2 in the revised manuscript. The analysis has been made from the data collected from the local population of District Shangla on the basis of folk uses of lyceum they have used it for curing of various diseases.

- Line 214 Table S1 (Supplementary file)

Doesn't require decryption

• Answer: The worthy reviewer suggestion has been honoured and the description related supplementary file has been removed from Table S1.

- On what basis the respondents were divided into age groups (30-40 years old, 41-50 years old, over 51 years old).

• Answer: In designing ethnobotanical data collection age is primarily the main factor that explains the differences between the respondents and medicinal plant knowledge. Respondents were categories in different age groups to know about their knowledge related medicinal plants usage in their traditional health care. As elder people have great knowledge related the use of medicinal plants because elder age group people recognized more medicinal plants than all other groups.  Elder participants were able to describe the specifics of the remedy and were more likely to list medicine as a use or reason for conservation. Also to decide whether the folkloric has been transferred from top age group to lower groups or not.

- Figure 2 obviously duplicates the data from table S1. By and large, 2 paragraphs (from 214 lines and from 222 lines) tell about the same thing.

• Answer: Respected reviewer the paragraph (from 222 lines) that was added mistakenly as duplicates with the paragraph (from 214 line) has been removed from the revised manuscript. Respected editor as Figure 2 and Table S1 has same data but the table show all parameters like Frequency of citation  (FC), The total number of use mentioned by each Informant (∑Ui),  the total number of informant (N), use value, and % use value in detail, while figure 2 show only the percent use value. However to reduce the confusion to the readers we have remove the figure and insert the table in the manuscript instead presenting in the supplementary file. 

- The abbreviation UV is misleading, in some cases it stands for «use value», and in some, more traditional, ultraviolet.

• Answer: The worthy reviewer suggestion has been honoured and to remove the confusion related the abbreviation UV has been replaced by the word ‘‘use value’’.

- I don't really understand the need for Figure 4.

In any case, most of the abscissa positions are unreadable, you can use the ordinate break.

• Answer: Worthy reviewer figure 4 has been removed and the table S2 from supplementary file has been inserted in the manuscript file instead.

- Clause 3.5.

It seems to me redundant information in the context of this study.

• Answer: Worthy reviewer suggestion has been honoured and the mentioned clause 3.5 has been removed from the revised manuscript. The whole manuscript has been revised to remove redundant data.

- Line 318 3.9. Future impact of the study

I think the publication of the material in the journal Agronomy is unlikely to increase the awareness of the local population about the medicinal properties of the plant.

• Answer:Worthy reviewer, the paper not only covers the mentioned aspect of informing the local community it also contain knowledge about its phytochemical composition and biological activities which are important to all communities around the globe. Also to best of our knowledge this plant is endangered in many other parts of the world as well. Through this study the researchers there will get encourage and will perform such or better studies. However worthy reviewer suggestion has been honoured and this section has been removed from the revised manuscript.
• As agronomy is science and technology of producing and using plants in agriculture for food, fuel, fibre, and land restoration. As it is a hilly area if plant production is restored the land restoration will automatically be achieved. Worthy reviewer as the world has become a global village similarly different science field have overlap their boundaries. Hope the worthy reviewer, better knows it as we are expert in biochemistry and botany. As request if worthy reviewer knows its better reason. We will be grateful if he guide us in this issue.

- Line 319 This review will provide a sense of awareness among the local people

This is not a review.

• Answer: Thank you worthy reviewer for valuable comment, the word review has been removed by a case study.

The fragment with the questionnaire looks inappropriate in the context of this study, I am sure that without it the article will not lose arguments in confirming the hypothesis formulated.

There is no harmony and transparency in the design of the experiment. A very large set of heterogeneous data is presented, which, from the point of view of semantic content, could be divided into three categories. If supplement, qualitatively describe and discuss the obtained experimental materials, it would be possible to write 3 separate articles.

1. Biochemical analysis and antioxidant status of Berberis lyceum Royle. Taking into account the fact that the authors claim that different parts of the plant are used for medicinal needs, it is possible to carry out (if this was not done earlier) a differential analysis of plant organs. You can analyze seasonal changes in the concentration of biologically active compounds. In addition, as I understand it, there were several collection points for the plant, obviously these are places with a unique microclimate, and the researchers have the opportunity to assess how they affect the metabolic processes of plants.
2. Sociological research. Population survey. Perhaps it is worth expanding the sample of respondents (the number of people, normalizing groups by age, gender, describing their state of health in accordance with diagnostic tests).
3. Phytobiocenotic research. Researchers attempted to describe the floristic profile of the area where Berberis lyceum Royle grows. However, this section seems unnecessary in this article. If we describe in detail the terrain, the dominant species of the studied ecotopes, the ecological links between them, and assess the conditions in which this species is most productive, then such a data set (with proper description) can become the basis for a separate publication.

• Answer: Thank you so much for giving such kind suggestion related the manuscript data. The way the worthy reviewer given the suggestion clearly indicates his expertise in the field. Worthy reviewer, being living in a third world countries we do not have the facilities to complete and fully evaluate one study area. Therefore, we attempted a new approach. In the previous version the link between the studied aspect was poor which now has been revised by expert and connection between the aspect have been made that hopefully will be satisfactory to the worthy reviewer.
• The aim in introduction has been revised as: “The aim of present study is therefore to revive the medicinal importance of the selected plant which is an endangered species. The collected ethnopharmacological data have been correlated with its phytochemical composition and antioxidant potentials. This ethnobotanical study in form of a case report will help researchers to recognize the latent and patent potentials of lyceum. The report will open a new way for further studies on its chemical profile and biological activities.” In other part also link has been established.
• We are very thankful for the valuable suggestion of worthy reviewer. Hope the worthy reviewer will have understand our problems as well.

 

Reviewer 3 Report

• Considering the structure of the article, it seems to be a research article. However, the authors name this paper a review on multiple occasions:
• L 249 in the present review study
• L 319 this review
• L 312 this review study
• L 115 This ethanobotanical review
• L 109 Fig. 1(B) presents the distribution of 100 people into 3 groups, the third group consisting of people above 51 years of age (22%, 62% and 16%). L122-123 "ages ranging in between 30 to 50 years". L196-197 states the distribution as 21%, 60% și 19%. In table 1 Supplementary material we can see the number of participants was 123. The authors are required to clarify these aspects
• L 150-152 Please present more details concerning the extraction process (preliminary treatment, S-L ratio, agitation etc)

• L 163 What is gm? Please use the correct unit. To the best of our knowledge, quantification through the external standard method is performed via the preparation of a calibration curve which will be the source of an equation. The method used here is questionable.
• L 176 Equation 4 – perhaps the authors are reffering to blank sample absorbtion, and not standard (ascorbic acid). Also, the preparation of negative control samples was not described.

• L 198 In this subchapter we have both diseases and general symptoms, while fig. 2 is entitled "Different diseases". Responses given by only one participant could not possibly be significant and should be removed. Are these the responses of 100 or 123 participants? 0.81% is less than 1 person.

• L 283 Table 1 HPLC-UV wavelength of detection is 320 nm, while in L160 280 nm is mentioned.

• L 346 Abbreviations should not create confusion (eg. Me. –Ext Methanolic extract, Me.Ext – Methanolic extract)

• Overall, the manuscript requires a serious language revision.

In conclusion, we recommend a major revision of this manuscript.

Author Response

List of amendment

Comments and Suggestions for Authors

 

Dear Editor/ reviewer,

Thank you very much for kind efforts and comments concerning our manuscript. We appreciate the hard work of reviewers as they fairly pointed out errors and mistakes in our manuscript. We have tried to revise the manuscript in line with comments of the reviewers. Corrections made have been highlighted as Blue.

Please find below the point by point responses to the reviewer’s comments and suggestions.

Reviewer 3 reports

Comments and Suggestions for Authors

Considering the structure of the article, it seems to be a research article. However, the authors name this paper a review on multiple occasions:

• L 249 in the present review study
• L 319 this review
• L 312 this review study
• L 115 This ethanobotanical review
• Answer: Worthy reviewer the term review in the manuscript has been corrected as case report in the revised manuscript. Thank you for your worthy suggestion.
• L 109 Fig. 1(B) presents the distribution of 100 people into 3 groups, the third group consisting of people above 51 years of age (22%, 62% and 16%). L122-123 "ages ranging in between 30 to 50 years". L196-197 states the distribution as 21%, 60% și 19%. In table 1 Supplementary material we can see the number of participants was 123. The authors are required to clarify these aspects
• Answer: Worthy reviewer Figure 1 is now figure 2 in the revised manuscript. Worthy reviewer the participants are 100 in numbers, the mistakes in presenting some data that was corrected in the revised manuscript accordingly.
• L 150-152 Please present more details concerning the extraction process (preliminary treatment, S-L ratio, agitation etc)
• Answer: Worthy reviewer the extraction process has been explained in the revised manuscript accordingly.
• L 163 What is gm?Please use the correct unit. To the best of our knowledge, quantification through the external standard method is performed via the preparation of a calibration curve which will be the source of an equation. The method used here is questionable.
• Answer: Respected reviewer the abbreviation gm means gram (The sample is taken in gram) has also been given in the abbreviation section. Now it has been replaced by correct unit symbol g.
• Respected reviewer the calibration are of two types. Multipoint point calibration and single point calibration
• The given formula below explains a single point calibration. Hopefully, the worthy reviewer will have understand our point.

(3)

Where: Cχ= concentration of sample; As= peak area of standard reference; Aχ= peak area of sample (g); Cs=concentration of standard reference (0.09 µg/ml).

 

• L 176 Equation 4 – perhaps the authors are referring to blank sample absorption, and not standard (ascorbic acid). Also, the preparation of negative control samples was not described.
• Answer: Worthy reviewer yes up course it is blank sample. Standard ascorbic acid has been used to calculate IC₅₀ In the revised paper required details have been incorporated accordingly.
• L 198 In this subchapter we have both diseases and general symptoms, while fig. 2 is entitled "Different diseases". Responses given by only one participant could not possibly be significant and should be removed. Are these the responses of 100 or 123 participants? 0.81% is less than 1 person.
• Answer: Respected reviewer upon reviewers recommendation figure has been removed as both the figure and Table has same data. Upon worthy reviewer recommendation non-significant values have been removed. The respondents were 100 which is mistakenly written as 123 which has been corrected in the revised manuscript. The whole manuscript has been revised carefully and hope will be ok now.
• L 283 Table 1 HPLC-UV wavelength of detection is 320 nm, while in L160 280 nm is mentioned.
• Answer: Worthy reviewer HPLC-UV wavelength of detection has been set at 320 nm which has been corrected in the revised manuscript. However, that column only contained a single value and have therefore been deleted. In material section that has been corrected as 320 nm.
• L 346 Abbreviations should not create confusion (eg. Me. –Ext Methanolic extract, Me.Ext – Methanolic extract)
• Answer: Worthy reviewer the abbreviation has been corrected in the revised manuscript.
• Overall, the manuscript requires a serious language revision.
• Answer: Worthy reviewer the whole manuscript has been corrected through the help of expert and hopefully will be ok now.

 

Author Response File: Author Response.pdf

Reviewer 4 Report

I read the manuscript and didn't understand why it was sent to the journal of Agronomy? This manuscript does not meet the criteria of Agronomy journal at all. As the manuscript is good, I suggest that the authors try to send it to a more appropriate journal.

If the emphasis is on ethnobotany, then it should bring out what are the popular names of a given plant, do these names overlap with other similar species? Looking at Table 2, there are disease names in the etical category. How did you define these diseases? People don't talk about "anti-fungal", "antibiotic" and so on. This whole list of diseases gives an unreliable impression. How did you make sure that people actually used it like that, not told you folklore? There is a lack of information on how often people use this plant once in a lifetime, once every 10 years or every year. These are quite different, either used once in a lifetime or used regularly. There are very many uses within 1%. This shows that these uses are not culturally significant. Also in Figure 4 you bring out is about a dozen within 1% of the reasons. Try to somehow categorize these little mentioned reasons. For example you show a "anthropogenic" factor, but look at the list is most of the reasons after all "anthropogenic". Why didn't you classify them as "anthropogenic"?

Author Response

List of amendment

Comments and Suggestions for Authors

 

Dear Editor/ reviewer,

Thank you very much for kind efforts and comments concerning our manuscript. We appreciate the hard work of reviewers as they fairly pointed out errors and mistakes in our manuscript. We have tried to revise the manuscript in line with comments of the reviewers. Corrections made have been highlighted as Blue.

Please find below the point by point responses to the reviewer’s comments and suggestions.

Reviewer 4 reports

Comments and Suggestions for Authors

I read the manuscript and didn't understand why it was sent to the journal of Agronomy? This manuscript does not meet the criteria of Agronomy journal at all. As the manuscript is good, I suggest that the authors try to send it to a more appropriate journal.

• Answer: worthy reviewer, As agronomy is science and technology of producing and using plants in agriculture for food, fuel, fibre, and land restoration. As it is a hilly area if plant production is restored the land restoration will automatically be achieved. Worthy reviewer as the world has become a global village similarly different science field have overlap their boundaries. Hope the worthy reviewer, better knows it as we are expert in biochemistry and botany. Respected reviewer by incorporating all the reviewers’ suggestion in the revised manuscript and as some section has been removed on worthy reviewer recommendations which makes the article good and follow the criteria of Agronomy journal.
• Worthy reviewer, we thankful for your appreciation as well.

If the emphasis is on ethnobotany, then it should bring out what are the popular names of a given plant, do these names overlap with other similar species?

• Answer: Worthy reviewer, the study covers three aspects: Biochemical analysis and antioxidant status, Sociological and Phytobiocenotic. As far the ethnobotanics aspects are concerned, respected reviewer the popular names of lyceum are Indian lyceum, Indian lycium, Indian barberry, English barberry, and "Zyarh larghai" in Pashtu and "Kashmal or Kasmal" in urdu has been added in the revised manuscript.
• Among the genus Berberis some species that have many closely related groups of species, such as,  Berberis lyceum Royle, Berberis parkeriana Schnei, Berberis huegeliane Schneid, Berberis afghanica Schneid, Berberis brevissima Jafri, Berberis calliobotrys Aitch. ex Koehne. Ahrendt with overlapping of characters, especially in leaves, stem colour, flower and berry size etc. are not lacking in our several closely related species.
• Among the above mentioned species only Berberis parkeriana Schnei species have common name of English barberry.

Looking at Table 2, there are disease names in the etical category. How did you define these diseases? People don't talk about "anti-fungal", "antibiotic" and so on. This whole list of diseases gives an unreliable impression.

• Answer: Worthy reviewer as some of the people of Shangla District are educated enough and they have tested their diseases at the medical care, and after checking their prescription we have categories this table. Here we have used a semi scientific approach

How did you make sure that people actually used it like that, not told you folklore?

• Answer: Worthy reviewer, as you know the inhabitant of this area do not have modern medical facilities and taking their patients to Peshawar and Islamabad that at least take one day to reach there. In serious infection they have been taken to there but latter on they have abandoned the use of allopathic drugs due to many reason like bad taste in mouth, quite away from Peshawar and Islamabad and poverty and have started this folkloric remedy which they pointed us in interviews. Even some of them have shown us the samples.
• First they pointed out that they are using folkloric approach which then on asking to specify which one they then have pointed out. Others they just used folkloric words have not been included in the data.

There is a lack of information on how often people use this plant once in a lifetime, once every 10 years or every year. These are quite different, either used once in a lifetime or used regularly. There are very many uses within 1%. This shows that these uses are not culturally significant.

• Answer: Worthy reviewer some people used this plant once in life while some people used it many times in life time based on their health conditions. Their use is tied with the diseased condition not with routine life. This is shown by a sign ∑Ui presented in Table 1 which describe the total number of use mentioned by each Informant.

Also in Figure 4 you bring out is about a dozen within 1% of the reasons. Try to somehow categorize these little mentioned reasons. For example you show a "anthropogenic" factor, but look at the list is most of the reasons after all "anthropogenic". Why didn't you classify them as "anthropogenic"?

• Answer: Respected reviewer as figure 4 and table 3 has same data. So upon reviewer 2 recommendation we have removed the figure 4 and insert the table 2 from supplementary file to the main manuscript.
• Thank you so much for your valuable comment, the factor that is categorise as anthropogenic are reported by most of the respondents are agriculture activities so, the word anthropogenic which confuse the readers as some other factors presented in the table 2 are also anthropogenic. Therefore to reduce the confusion specific factor that is answered by the local population are given in the revised table as agriculture activities.

 

Round 2

Reviewer 1 Report

My main (although more general) comment, has not been addressed by authors. This was: the scientific presentation of the data should be greatly improved. Please follow basic instructions about writing scientific papers and Instructions for Authors provided by the journal.
To be more clear with this request, I added several detailed comments to the manuscript. However, for all the suggestions and corrections: analogous mistakes should be corrected or commented through the manuscript, even if these are not highlighted during the review.
In several parts the language is difficult to understand, with some additional editing needed.

Comments for author File: Comments.pdf

Author Response

Reviewer 1

Dear Editor/ reviewer,

Thank you very much for kind efforts and comments concerning our manuscript. We appreciate the hard work of reviewers as they fairly pointed out errors and mistakes in our manuscript. We have tried to revise the manuscript in line with comments of the reviewers. Corrections made have been highlighted as Blue.

Please find below the point by point responses to the reviewer’s comments and suggestions.

My main (although more general) comment, has not been addressed by authors. This was: the scientific presentation of the data should be greatly improved. Please follow basic instructions about writing scientific papers and Instructions for Authors provided by the journal.

• Answer: Sorry worthy reviewer, actually your suggestions were in PDF file and that portion the paper was revised by a junior member. I (Dr. Zahoor) saw your points now. These are quite reasonable objections. I am sorry for not consulting it on time. This time I have revised the paper by myself and hopefully it will be ok now. I am extremely thankful for the valuable suggestions of the worthy reviewer. No reviewer check so thoroughly check that you have. For which I do not have words to thanks to worthy reviewer.

To be more clear with this request, I added several detailed comments to the manuscript. However, for all the suggestions and corrections: analogous mistakes should be corrected or commented through the manuscript, even if these are not highlighted during the review.
In several parts the language is difficult to understand, with some additional editing needed.

• Answer: Worthy reviewer all the comments raised in attached file have been honoured and grammatical mistakes have been corrected accordingly. All the points raised in attached PDF has been corrected in the revised paper. Hopefully the manuscript will be ok now.

Reviewer 2 Report

The authors have done a great job in order to improve the quality of the published material. However, a number of unresolved issues remain.

- Line 110 phytochemical composition

In your study, only the antioxidant potential of plants is assessed, not the phytochemical composition. It must be said that antioxidants will have a very limited therapeutic effect for the list of diseases (or conditions) that are presented in this work.

- I'm a little worried that the province map is from a 2013 survey.

Sher, H., Yousaf, S., & Khan, K. (2013). Traditional resources evaluation of district Shangla, Pakistan. African Journal of Pharmacy and Pharmacology, 7 (46), 2928-2936.

Sampling points are marked on the map. I doubt the absolute coincidence of the places of collection of plant material.

- The maps are still of very poor quality.

A quick search on the internet led me to much better quality maps.

https://www.lcakp.org.pk/districts/district-shangla /

In addition, Shangla province also has better resolution. I attach it to the review.

Ideally, find a specialist working in qgis, ArcGIS or Surfer programs in order to make a map with an inset region for your specific research.

- 2.5. Berberis lyceum Royle collection and extraction

Why only the root was used for the analysis. Judging by the data presented in the article, almost all parts of the plant are used for therapeutic needs. In addition, as far as I understand (correct me if I am wrong), the extraction of the root for analysis is more harmful to the plant than the collection of aerial vegetative and generative parts.

- Figure 2

The quality of the illustration is still very low. The title of the picture corresponds to its signature; this is duplicate information. To the same figure, you can add a pie chart dividing respondents by gender.

- Table 1 A very large number of significant decimal places in the columns Use value,% Use value. I think only one significant decimal place will be enough. In addition, these columns actually duplicate each other (in one data are presented in shares, in the other in percent). My suggestion is to leave only% Use value and round up the data.

- The same goes for table 2.

- Since the value N (for all 3 tables) of all rows is the same, you can omit this parameter from the table body by describing it in the legend.

- I would suggest splitting the DPPH and ABTS tests into separate tables.

- It is not entirely clear what the asterisks mean in Table 5

 - Abbreviations can be placed without a table

Comments for author File: Comments.png

Author Response

Reviewer 2

Comments and Suggestions for Authors

Dear Editor/ reviewer,

Thank you very much for kind efforts and comments concerning our manuscript. We appreciate the hard work of reviewers as they fairly pointed out errors and mistakes in our manuscript. We have tried to revise the manuscript in line with comments of the reviewers. Corrections made have been highlighted as Blue.

Please find below the point by point responses to the reviewer’s comments and suggestions.

The authors have done a great job in order to improve the quality of the published material. However, a number of unresolved issues remain.

• Answer: Thank you worthy reviewer.

- Line 110 phytochemical composition

In your study, only the antioxidant potential of plants is assessed, not the phytochemical composition. It must be said that antioxidants will have a very limited therapeutic effect for the list of diseases (or conditions) that are presented in this work.

• Answer: worthy reviewer, the phytochemical composition in terms of HPLC analysis is there in the paper. That is why the claim has been made. Worthy reviewer you are right that antioxidant potential is limited to have therapeutic effect in all the mentioned diseases. However, it is a case study where folkloric knowledge and scientific approach has been combined to revive the importance of this plant in local community and attract researchers to do experiments in lined with the mentioned diseases. We have only used available resources that were HPLC analysis and antioxidant potentials. Hopefully the worthy reviewer will have understand our point.

- I'm a little worried that the province map is from a 2013 survey.

Sher, H., Yousaf, S., & Khan, K. (2013). Traditional resources evaluation of district Shangla, Pakistan. African Journal of Pharmacy and Pharmacology, 7 (46), 2928-2936.

Sampling points are marked on the map. I doubt the absolute coincidence of the places of collection of plant material.

- The maps are still of very poor quality.

A quick search on the internet led me to much better quality maps.

https://www.lcakp.org.pk/districts/district-shangla /

In addition, Shangla province also has better resolution. I attach it to the review.

Ideally, find a specialist working in qgis, ArcGIS or Surfer programs in order to make a map with an inset region for your specific research.

• Answer: Thank you worthy reviewer for you suggestions and sending a proper map. Being biochemists we are not experts in this field. However, we have tried our best to incorporate the worthy reviewer suggestions. The resolution of the map has been increased. As far the previous map and reference were concerned actually the existing map replaced on suggestion of one reviewer and mistakenly the old reference was there. Now the map taken from paper has been properly cited.

- 2.5. Berberis lyceum Royle collection and extraction

Why only the root was used for the analysis. Judging by the data presented in the article, almost all parts of the plant are used for therapeutic needs. In addition, as far as I understand (correct me if I am wrong), the extraction of the root for analysis is more harmful to the plant than the collection of aerial vegetative and generative parts.

• Answer: Worthy reviewer as the roots are the most important part of Berberis species as they contain a number of alkaloids, the most prominent one is berberine. Berberis lycium and various root extracts lower the glucose level significantly and exhibited strong antioxidant and antibacterial activity (1, 2). There is no HPLC data available on B. lyceum roots profiling therefore the extraction from roots and their HPLC profiling were studied. Mostly roots of this plant are used as therapeutic agent. For example the roots extract has many applications like it is used for the treatment of urinary tract infections, enlargement of spleen, gastric and duodenal ulcer and liver disorders. The product is mixed with butter and alum to be used as an external application for the eyelids in acute conjunctivitis.

- Figure 2

The quality of the illustration is still very low. The title of the picture corresponds to its signature; this is duplicate information. To the same figure, you can add a pie chart dividing respondents by gender.

Answer: Thank you worthy reviewer for your worthy suggestion, the figure 2 has been removed as the reviewer 1 also suggest that insert the data in text instead of presenting in the figure.

- Table 1 A very large number of significant decimal places in the columns Use value, % Use value. I think only one significant decimal place will be enough. In addition, these columns actually duplicate each other (in one data are presented in shares, in the other in percent). My suggestion is to leave only% Use value and round up the data.

- The same goes for table 2.

-Since the value N (for all 3 tables) of all rows is the same, you can omit this parameter from the table body by describing it in the legend.

• Answer: Worthy reviewer, yours suggestions related Tables has been incorporated in the revised manuscript accordingly.

- I would suggest splitting the DPPH and ABTS tests into separate tables.

• Answer: Worthy reviewer suggestion has been honoured and Table related DPPH and ABTS has been separated in the revised manuscript.

- It is not entirely clear what the asterisks mean in Table 5

• Answer: Worthy reviewer the asterisks in Table 5 shows the significant values (***P < 0.001) when sample is compared to positive control (Ascorbic acid) using student t-test.

 - Abbreviations can be placed without a table

• Answer: The table is removed from abbreviation in the revised manuscript.

 

Reviewer 3 Report

OK.

Author Response

Reviewer 3

 

Comments and Suggestions for Authors

 

Dear Editor/ reviewer,

Thank you very much for kind efforts and comments concerning our manuscript. We appreciate the hard work of reviewers as they fairly pointed out errors and mistakes in our manuscript. We have tried to revise the manuscript in line with comments of the reviewers. Corrections made have been highlighted as Blue.

Please find below the point by point responses to the reviewer’s comments and suggestions.

OK.

• Answer: Thank you so much worthy reviewer for accepting our revisions.

Reviewer 4 Report

The authors have not changed the manuscript to the requirements according to of the journal of Agronomy. https://www.mdpi.com/journal/agronomy/about 

Author Response

Reviewer 4

 

Comments and Suggestions for Authors

 

Dear Editor/ reviewer,

Thank you very much for kind efforts and comments concerning our manuscript. We appreciate the hard work of reviewers as they fairly pointed out errors and mistakes in our manuscript. We have tried to revise the manuscript in line with comments of the reviewers. Corrections made have been highlighted as Blue.

Please find below the point by point responses to the reviewer’s comments and suggestions.

The authors have not changed the manuscript to the requirements according to of the journal of Agronomy. https://www.mdpi.com/journal/agronomy/about

• Worthy reviewer, you are talking about production problem which are too early and MDPI journal have a very efficient supporting staff that correct such types of mistakes. When the paper is accepted they correct it and send the galley proof for our approval. We have tried our best to follow the guidelines of the journal fully in the revised manuscript. Hopefully it will be ok now.
• At the same time the other three reviewers are in favour of publication