Phytochemical Investigation of Myrcianthes cisplatensis: Structural Characterization of New p-Coumaroyl Alkylphloroglucinols and Antimicrobial Evaluation against Staphylococcus aureus
, Elisabetta Buommino 2,*, Leslie Landrum 3, Rosita Russo 1, Francesca Lembo 2, Antonio Fiorentino 1,† and Brigida D’Abrosca 1,*,†Round 1
Reviewer 1 Report
1、The author mentioned that the antibacterial potential of the methanolic extract of M. cisplatensis leaves against two S. aureus ATCC strains (29213 and 43300) were investigated in Figure 1. Why the inhibitory effects were best at 64 μg/ml rather than at 128 μg/ml.
2、Single factor analysis of variance should be done for relevant data in Figure 1 and Figure 7.
3. The leaves were collected at flowering stage, what is the growth state of the leaves, juvenile or mature, top or bottom, or the whole plant.
4. Whether the content is different in different growth region and different mature stages？
Author Response
1、The author mentioned that the antibacterial potential of the methanolic extract of M. cisplatensis leaves against two S. aureus ATCC strains (29213 and 43300) were investigated in Figure 1. Why the inhibitory effects were best at 64 μg/ml rather than at 128 μg/ml.
Ans Considering the standard deviations within the group the results can be considered comparable.
2、Single factor analysis of variance should be done for relevant data in Figure 1 and Figure 7.
Ans Single factor analysis of variance has been performed, the results added in figures 1 and 7. In materials and method section the paragraph -3.6.3 Statistical analysis –has been added.
- The leaves were collected at flowering stage, what is the growth state of the leaves, juvenile or mature, top or bottom, or the whole plant.
Ans Most of the leaves were full grown and mature. The leaves stay on the plant for over a year so it always has mature leaves. The leaves of the whole plant were collected.
- Whether the content is different in different growth region and different mature stages？
Ans In this manuscript we report the phytochemical investigation of leaves collected in Arizona in May 2022. In general, the cultivated plant in Arizona grows in a climate similar to Uruguay, but drier. Arizona is hot in Summer (30-40 C common temperatures) and cool in winter, with freezing temperatures rare. M. cisplatensis grows in Arizona well here but needs irrigation. The plant was growing along a stream in Uruguay.
Reviewer 2 Report
1. Please provide the name of the botanist who identified the plant material
2. Most of the methods are without proper citations. Please provide the references
3. "Each tested compound was dissolved in methanol" what is the final concentration of the methanol in the mixture?
4. Final concentration of the methanol will affect the overall antimicrobial assay? How does the author control this?
5. "MIC was defined as the lowest concentration of drug that caused a total inhibition of microbial growth". How the author determined the MIC concentration. Not clear and please explain this.
6. If the possible author should conduct statistical analysis for MIC findings.
7. Please suggest at least one suggestion for future study in the conclusion sections.
Author Response
- Please provide the name of the botanist who identified the plant material
Ans The plant have been identified by prof. Leslie Landrum, co-author of this paper. Their name has been added in 3.1 section -plant material.
- Most of the methods are without proper citations. Please provide the references
Ans Methods should be described or proper citations heve been added
- "Each tested compound was dissolved in methanol" what is the final concentration of the methanol in the mixture?
Ans Each tested compound was dissolved in methanol (1mg/200µl) to give a stock solution and diluted in Mueller-Hinton (MH).
- Final concentration of the methanol will affect the overall antimicrobial assay? How does the author control this?
Ans Each experiment included negative control wells containing bacteria in Mueller–Hinton broth plus the amount of vehicle (methanol) used to dilute each compound. We added a sentence in the manuscript, results section, paragraph 2.1, lines 86-87
- "MIC was defined as the lowest concentration of drug that caused a total inhibition of microbial growth". How the author determined the MIC concentration. Not clear and please explain this.
Ans The compounds were added to the bacterial suspension in each well yielding a final cell concentration of 1 × 106 CFU/ml and a final compound concentration ranging from 128 to 16µg/ml. The procedure was reported as previously published. Anyway, we added a sentence in material and methods section to better specify the assay. Paragraph 3.6.2
- If the possible author should conduct statistical analysis for MIC findings.
Ans Statistical analysis has been added in the figures 1 and 7, and in test. Furtheremore in materials and method section the paragraph -3.6.3 Statistical analysis –has been added.
- Please suggest at least one suggestion for future study in the conclusion sections.
Ans In conclusion section, we have included one suggestion for future plan.
Reviewer 3 Report
The manuscript is interesting and well drafted, however, there are some suggestions that need to be incorporated before publication.
Comments for author File: 
Comments.pdf
Author Response
- Family name should be in italic font
Ans. Done. Myrtaceae has been italicize in manuscript
- Line 20 also in Brazil and Paraguay as per the following link
https://powo.science.kew.org/taxon/urn:lsid:ipni.org:names:30076995-2
Ans. Thanks for your suggestion. The sentence about plant distribution has been added in abstract as well as in introduction section.
- Line 27 How the authors have analyzed this MIC value?
Ans The procedure was reported as previously published. Anyway, we added a sentence in material and methods section to better specify the assay. Paragraph 3.6.2
- Line 40
Ans Correction has been made
- Lines 85-86Rewrite . complex statement lines 84-85
Ans The sentence has been rephrased
- Line 87 what about another control oxacillin
Ans Oxacillin was not able to inhibit the growth of the MRSA (data not showed). we added this sentence in paragraph 2.1
- Line 245 Add Statistical analysis part in methodology
Ans Statistical analysis has been added in the figures 1 and 7, and in test. Furtheremore in materials and method section the paragraph -3.6.3 Statistical analysis –has been added.
- Line 274 Have u measured the particle size?
Ans For our extraction method, a precise measure of particle size in not necessary. The pulverization is functional to increase the exchange surface between the plant material and the solvent
- Lines 368-368 Elaborate the methodology
Ans The procedure was reported as previously published- ref- 14. Anyway, we added a sentence in material and methods section to better specify the assay. Paragraph 3.6.2
10-. Line 375 Draw significant conclusion of your research, not generalized statements here
Ans Thanks for your suggestion. We insert a specific conclusion for this manuscript
- Line 379 Repetition of the intro line
Ans The sentence has been deleted
Round 2
Reviewer 3 Report
All the suggestions have been incorporated.
