In Vitro Antioxidant, Antimicrobial, Anticoccidial, and Anti-Inflammatory Study of Essential Oils of Oregano, Thyme, and Sage from Epirus, Greece
, Virginia Marugán-Hernández 2, Ioannis Skoufos 3, Ilias Giannenas 1,*, Eleftherios Bonos 3, Kensilandia Aguiar-Martins 3, Diamanto Lazari 4, Theodora Papagrigoriou 4, Konstantina Fotou 5, Katerina Grigoriadou 6, Damer P. Blake 3 and Athina Tzora 5
Round 1
Reviewer 1 Report
The manuscript focus on evaluation of the in vitro antioxidant, antimicrobial, anti-inflammatory and antiparasitic activity of the essential oils obtained from Greek oregano, thyme and sage plants. The experiments were designed reasonably, and the main finds and results were expressed clearly. There are some points should be considered or revised before its publication.
1. Line 270-273: what's the initial concentration of bacterium used during evaluation of antimicrobial activity?
2. Line 291: EO solutions? it's not a proper expression. In addition, EOs are hydrophobic, while the surface of bacterial cells are hydrophilic, there is a conflict/contradiction of improve the antimicrobial activity of EOs.
3. Line 289 CFUml-1
4. Table 4, what do C=5%....C=100% mean?
5. Table 5 or Table 6, please choose only one.
6.Figure 6, this picture is blurry
7. Too much References, please delete some of them.
8. Writing issues: for example, in Line 40, the first usage of LOX needs full spelling.
Author Response
Comments and Suggestions for Authors
Dear Editor
We would like to thank you and the reviewers for the help to improve the presentation of our work. All comments are accepted and all changes are made in the text in green color to be easily checked.
Further you may find some replies.
Reviewer 1:
- Line 270-273: what's the initial concentration of bacterium used during evaluation of antimicrobial activity?
Bacterial suspensions of each bacterial ATCC strain in saline solution of 0.5 McFarland (density of approximately 1.5×108 CFU ml-1) turbidity were streaked onto Mueller-Hinton agar (MHA) with a sterile swab.
- Line 291: EO solutions? it's not a proper expression. In addition, EOs are hydrophobic, while the surface of bacterial cells are hydrophilic, there is a conflict/contradiction of improve the antimicrobial activity of EOs.
Bacterial suspension was diluted 1/10 in saline solution and finally 100 µl was added to each well of each column over the EOs which were mixed in the previous stage (or before??) in MHB containing 5% DMSO.
- Line 289 CFUml-1 (corrected)
- Table 4, what do C=5%....C=100% mean?
Pure EΟs were diluted to 50, 20 and 5% concentration in DMSO 5% (v/v) (line 266) for Disc Diffusion assay and then each sterile filter disc was impregnated with these EOs concentrations (15 µl/disc) (line 270).
So…
C=100% means that pure EO (100% concentration, no diluted with DMSO) was impregnated the sterile filter disc
C=50% means that 50 volumes of each EO was diluted in 50 volumes of DMSO
C=20% means that 20 volumes of each EO was diluted in 80 volumes of DMSO and
C=5% means that 5 volumes of each EO was diluted in 95 volumes of DMSO
- Table 5 or Table 6, please choose only one.
We choose Table 6.
Reviewer 2:
"Fig.5 and Fig.6 both need replacement with clearly levelled one.
Nothing visible in Fig.6 you replace with clear photograph"
We will try to improve
Reviewer 3:
"In M &M please mention what is the source of microbial strains and is it deposited in the culture collection."
All the reference bacterial strains that were used for the experiment were purchased from American Type Culture Collection (ATCC) (line 263)
6.Figure 6, this picture is blurry
This figure could not be improved.
- Too much References, please delete some of them.
Done
- Writing issues: for example, in Line 40, the first usage of LOX needs full spelling.
Done
We appreciate the authors for their nice and exhaustive work. Below you can find suggestions to improve the manuscript:
- In material method section 2.1 it is not clearly mentioned about age of the plant from which plant materials taken for extraction of oil. Age of the plant and season play moprtant role for essential oil conten. Please includethe these data in the section.
It has been inserted in the text
- GC chromatograms are missing in the the manuscript. You add in the manuscript or give in supplementaty file.
- Fig.5 and Fig.6 both need replacement with clearly levelled one. Nothing visible in Fig.6 you replace with clear photograph.
This article deals with the evaluation of focuses on the evaluation of the in vitro antioxidant, antimicrobial, and anti-inflammatory. This article deals with the evaluation of focuses on the evaluation of the in vitro antioxidant, antimicrobial, anti-inflammatory, and antiparasitic activity of the essential oils obtained from Greek oregano, thyme, and sage plants. In this context, the authors identified the bioactive compounds in these species and proposed their further use based on their composition. Thus, the main objective of this research is obvious and well-elaborated.
The results in tables and figures are well presented, so I have no objection to their presentation. The results are adequately discussed and compared with other works. The conclusion confirms the obtained results and the references inserted in the main document show a good connection between this investigation and other works.
The material and methods, as well as the results and discussion part, are understandable. The article is good from a grammatical and structural point of view, and from my perspective is acceptable for publication in this journal.
Minor errors:
- In the section Plant material:
A voucher specimen must be deposited in a recognized herbarium.
IPEN number of the plants is mentioned. International Plant Exchange Network accession number (IPEN) indicates that the plant with the specific accession number is maintained in an official Botanic Garden and voucher specimen with the same number is deposited in the herbarium of this Botanic Garden. In our cases both living plants and herbarium specimens are maintain and deposit at the collection of Balkan Botanic Garden of Kroussia (41°05'44.3"N 23°06'33.7"E) belonging in the Institute of Plant Breeding and Genetic Resources, Hellenic Agricultural Organization-DIMITRA, in Greece.
Add a photo of the plant if possible
Map of area o collocation of collection area.
This has been inserted.
Figure 1: a) Origanum vulgare ssp.hirtum, b) Salvia fruticosa, c) Thymus vulgaris, d) Balkan Botanic Garden of Kroussia
Reviewer 2 Report
I appreciate to authors for their nice exhaustive work. I have suggestions to improve the manuscript:
1. In material method section 2.1 it is not clearly mentioned about age of the plant from which plant materials taken for extraction of oil. Age of the plant and season play moprtant role for essential oil conten. Please includethe these data in the section.
2. GC chromatograms are missing in the the manuscript. You add in the manuscript or give in supplementaty file.
3. Fig.5 and Fig.6 both need replacement with clearly levelled one. Nothing visible in Fig.6 you replace with clear photograph.
Author Response
Comments and Suggestions for Authors
Dear Editor
We would like to thank you and the reviewers for the help to improve the presentation of our work. All comments are accepted and all changes are made in the text in green color to be easily checked.
Further you may find some replies.
Reviewer 1:
- Line 270-273: what's the initial concentration of bacterium used during evaluation of antimicrobial activity?
Bacterial suspensions of each bacterial ATCC strain in saline solution of 0.5 McFarland (density of approximately 1.5×108 CFU ml-1) turbidity were streaked onto Mueller-Hinton agar (MHA) with a sterile swab.
- Line 291: EO solutions? it's not a proper expression. In addition, EOs are hydrophobic, while the surface of bacterial cells are hydrophilic, there is a conflict/contradiction of improve the antimicrobial activity of EOs.
Bacterial suspension was diluted 1/10 in saline solution and finally 100 µl was added to each well of each column over the EOs which were mixed in the previous stage (or before??) in MHB containing 5% DMSO.
- Line 289 CFUml-1 (corrected)
- Table 4, what do C=5%....C=100% mean?
Pure EΟs were diluted to 50, 20 and 5% concentration in DMSO 5% (v/v) (line 266) for Disc Diffusion assay and then each sterile filter disc was impregnated with these EOs concentrations (15 µl/disc) (line 270).
So…
C=100% means that pure EO (100% concentration, no diluted with DMSO) was impregnated the sterile filter disc
C=50% means that 50 volumes of each EO was diluted in 50 volumes of DMSO
C=20% means that 20 volumes of each EO was diluted in 80 volumes of DMSO and
C=5% means that 5 volumes of each EO was diluted in 95 volumes of DMSO
- Table 5 or Table 6, please choose only one.
We choose Table 6.
Reviewer 2:
"Fig.5 and Fig.6 both need replacement with clearly levelled one.
Nothing visible in Fig.6 you replace with clear photograph"
We will try to improve
Reviewer 3:
"In M &M please mention what is the source of microbial strains and is it deposited in the culture collection."
All the reference bacterial strains that were used for the experiment were purchased from American Type Culture Collection (ATCC) (line 263)
6.Figure 6, this picture is blurry
This figure could not be improved.
- Too much References, please delete some of them.
Done
- Writing issues: for example, in Line 40, the first usage of LOX needs full spelling.
Done
We appreciate the authors for their nice and exhaustive work. Below you can find suggestions to improve the manuscript:
- In material method section 2.1 it is not clearly mentioned about age of the plant from which plant materials taken for extraction of oil. Age of the plant and season play moprtant role for essential oil conten. Please includethe these data in the section.
It has been inserted in the text
- GC chromatograms are missing in the the manuscript. You add in the manuscript or give in supplementaty file.
- Fig.5 and Fig.6 both need replacement with clearly levelled one. Nothing visible in Fig.6 you replace with clear photograph.
This article deals with the evaluation of focuses on the evaluation of the in vitro antioxidant, antimicrobial, and anti-inflammatory. This article deals with the evaluation of focuses on the evaluation of the in vitro antioxidant, antimicrobial, anti-inflammatory, and antiparasitic activity of the essential oils obtained from Greek oregano, thyme, and sage plants. In this context, the authors identified the bioactive compounds in these species and proposed their further use based on their composition. Thus, the main objective of this research is obvious and well-elaborated.
The results in tables and figures are well presented, so I have no objection to their presentation. The results are adequately discussed and compared with other works. The conclusion confirms the obtained results and the references inserted in the main document show a good connection between this investigation and other works.
The material and methods, as well as the results and discussion part, are understandable. The article is good from a grammatical and structural point of view, and from my perspective is acceptable for publication in this journal.
Minor errors:
- In the section Plant material:
A voucher specimen must be deposited in a recognized herbarium.
IPEN number of the plants is mentioned. International Plant Exchange Network accession number (IPEN) indicates that the plant with the specific accession number is maintained in an official Botanic Garden and voucher specimen with the same number is deposited in the herbarium of this Botanic Garden. In our cases both living plants and herbarium specimens are maintain and deposit at the collection of Balkan Botanic Garden of Kroussia (41°05'44.3"N 23°06'33.7"E) belonging in the Institute of Plant Breeding and Genetic Resources, Hellenic Agricultural Organization-DIMITRA, in Greece.
Add a photo of the plant if possible
Map of area o collocation of collection area.
This has been inserted.
Figure 1: a) Origanum vulgare ssp.hirtum, b) Salvia fruticosa, c) Thymus vulgaris, d) Balkan Botanic Garden of Kroussia
Reviewer 3 Report
This article deals with the evaluation of focuses on the evaluation of the in vitro antioxidant, antimicrobial, and anti-inflammatory. This article deals with the evaluation of focuses on the evaluation of the in vitro antioxidant, antimicrobial, anti-inflammatory, and antiparasitic activity of the essential oils obtained from Greek oregano, thyme, and sage plants. In this context, the authors identified the bioactive compounds in these species and proposed their further use based on their composition. Thus, the main objective of this research is obvious and well-elaborated.
The results in tables and figures are well presented, so I have no objection to their presentation. The results are adequately discussed and compared with other works. The conclusion confirms the obtained results and the references inserted in the main document show a good connection between this investigation and other works.
The material and methods, as well as the results and discussion part, are understandable. The article is good from a grammatical and structural point of view, and from my perspective is acceptable for publication in this journal.
Minor errors:
- In the section Plant material:
A voucher specimen must be deposited in a recognized herbarium.
Add a photo of the plant if possible
Map of area o collocation of collection area.
- In M &M please mention what is the source of microbial strains and is it deposited in the culture collection.
The title, abstract, scheme, tables, and figures of the manuscript are adequate for the content. The experimental part gives enough details about the synthesis and spectral data. amatory, and antiparasitic activity of the essential oils obtained from Greek oregano, thyme, and sage plants.
Author Response
Comments and Suggestions for Authors
Dear Editor
We would like to thank you and the reviewers for the help to improve the presentation of our work. All comments are accepted and all changes are made in the text in green color to be easily checked.
Further you may find some replies.
Reviewer 1:
- Line 270-273: what's the initial concentration of bacterium used during evaluation of antimicrobial activity?
Bacterial suspensions of each bacterial ATCC strain in saline solution of 0.5 McFarland (density of approximately 1.5×108 CFU ml-1) turbidity were streaked onto Mueller-Hinton agar (MHA) with a sterile swab.
- Line 291: EO solutions? it's not a proper expression. In addition, EOs are hydrophobic, while the surface of bacterial cells are hydrophilic, there is a conflict/contradiction of improve the antimicrobial activity of EOs.
Bacterial suspension was diluted 1/10 in saline solution and finally 100 µl was added to each well of each column over the EOs which were mixed in the previous stage (or before??) in MHB containing 5% DMSO.
- Line 289 CFUml-1 (corrected)
- Table 4, what do C=5%....C=100% mean?
Pure EΟs were diluted to 50, 20 and 5% concentration in DMSO 5% (v/v) (line 266) for Disc Diffusion assay and then each sterile filter disc was impregnated with these EOs concentrations (15 µl/disc) (line 270).
So…
C=100% means that pure EO (100% concentration, no diluted with DMSO) was impregnated the sterile filter disc
C=50% means that 50 volumes of each EO was diluted in 50 volumes of DMSO
C=20% means that 20 volumes of each EO was diluted in 80 volumes of DMSO and
C=5% means that 5 volumes of each EO was diluted in 95 volumes of DMSO
- Table 5 or Table 6, please choose only one.
We choose Table 6.
Reviewer 2:
"Fig.5 and Fig.6 both need replacement with clearly levelled one.
Nothing visible in Fig.6 you replace with clear photograph"
We will try to improve
Reviewer 3:
"In M &M please mention what is the source of microbial strains and is it deposited in the culture collection."
All the reference bacterial strains that were used for the experiment were purchased from American Type Culture Collection (ATCC) (line 263)
6.Figure 6, this picture is blurry
This figure could not be improved.
- Too much References, please delete some of them.
Done
- Writing issues: for example, in Line 40, the first usage of LOX needs full spelling.
Done
We appreciate the authors for their nice and exhaustive work. Below you can find suggestions to improve the manuscript:
- In material method section 2.1 it is not clearly mentioned about age of the plant from which plant materials taken for extraction of oil. Age of the plant and season play moprtant role for essential oil conten. Please includethe these data in the section.
It has been inserted in the text
- GC chromatograms are missing in the the manuscript. You add in the manuscript or give in supplementaty file.
- Fig.5 and Fig.6 both need replacement with clearly levelled one. Nothing visible in Fig.6 you replace with clear photograph.
This article deals with the evaluation of focuses on the evaluation of the in vitro antioxidant, antimicrobial, and anti-inflammatory. This article deals with the evaluation of focuses on the evaluation of the in vitro antioxidant, antimicrobial, anti-inflammatory, and antiparasitic activity of the essential oils obtained from Greek oregano, thyme, and sage plants. In this context, the authors identified the bioactive compounds in these species and proposed their further use based on their composition. Thus, the main objective of this research is obvious and well-elaborated.
The results in tables and figures are well presented, so I have no objection to their presentation. The results are adequately discussed and compared with other works. The conclusion confirms the obtained results and the references inserted in the main document show a good connection between this investigation and other works.
The material and methods, as well as the results and discussion part, are understandable. The article is good from a grammatical and structural point of view, and from my perspective is acceptable for publication in this journal.
Minor errors:
- In the section Plant material:
A voucher specimen must be deposited in a recognized herbarium.
IPEN number of the plants is mentioned. International Plant Exchange Network accession number (IPEN) indicates that the plant with the specific accession number is maintained in an official Botanic Garden and voucher specimen with the same number is deposited in the herbarium of this Botanic Garden. In our cases both living plants and herbarium specimens are maintain and deposit at the collection of Balkan Botanic Garden of Kroussia (41°05'44.3"N 23°06'33.7"E) belonging in the Institute of Plant Breeding and Genetic Resources, Hellenic Agricultural Organization-DIMITRA, in Greece.
Add a photo of the plant if possible
Map of area o collocation of collection area.
This has been inserted.
Figure 1: a) Origanum vulgare ssp.hirtum, b) Salvia fruticosa, c) Thymus vulgaris, d) Balkan Botanic Garden of Kroussia
