The Antioxidant and Antimicrobial Activities of Two Sun-Dried Fig Varieties (Ficus carica L.) Produced in Eastern Morocco and the Investigation of Pomological, Colorimetric, and Phytochemical Characteristics for Improved Valorization

Round 1

Reviewer 1 Report

Manuscript entitled: „Antioxidant and antimicrobial activities of two sun-dried fig varieties (Ficus carica L.) produced in Eastern Morocco, inves-tigation of pomological, colorimetric, and phytochemical characteristics for improved valorization.”.

General suggestion:

Article in that form is not relevant for publication in International Journal of Plant Biology. The article requires a lot of changes, therefore it is not recommended for publication. 

Authors used standard analytical methods and obtained results are interesting but described and discussed not sufficiently.

What is new is not emphasized in the work? Is the place of cultivation? Is the way of drying? Whether these are new varieties of figs or well-known ones? This is missing in the introduction.

In part of Material and methods please add the information:

How many samples of the raw material were taken to analysis? The Authors took samples in in one year – 2022 from different places. Please explain whether the places of sampling has an influence on the composition of the figs. How environmental factors affect the composition of figs? There is no information on the characteristics of the cultivation method.

In the part of Results and discussion:

There is no information why total polyphenols and total flavonoids were determined using three different solvents? It is not explained in the introduction or in the methodology?

The obtained results of vitamin C content in dried figs are very low – 1,03-1,51 (%). It cannot be said that figs are the source of this vitamin.

The discussion should be elaborated in a better way. The results presented in tables are more described by the Authors, than discussed.

The conclusion is that figs have a high energy value, where are the results?

References:

There are numerous errors in citing the literature. This needs improvement.

Author Response

Review Report round 1 : responses

Dear Editors, Dear Reviewers,

Thank you for giving us the opportunity to improve our manuscript with the revised version and thank you for your useful comments.

We really thank the Reviewers for their thorough review. We hope this revision will satisfy reviewers’ queries, and that our work will be considered for publication in International Journal of Plant Biology (IJPB). Below we have made efforts to either address or respond to each (paraphrased) requested change and communicated weakness. Major changes are highlighted in yellow text in the revision. All typos/minor concerns have been fixed in the manuscript and are otherwise not addressed in this response.

With kind regards

Dr. Mohamed Addi, and the co-Authors

 

Review Report (Reviewer 1)

General suggestion:

Article in that form is not relevant for publication in International Journal of Plant Biology. The article requires a lot of changes, therefore it is not recommended for publication. 

Authors used standard analytical methods and obtained results are interesting but described and discussed not sufficiently.

What is new is not emphasized in the work? Is the place of cultivation? Is the way of drying? Whether these are new varieties of figs or well-known ones? This is missing in the introduction.

 

Response: We sincerely appreciate your thorough evaluation of our manuscript and your valuable feedback. We have carefully considered your comments and suggestions and have made significant improvements to address the concerns raised.

To enhance the suitability of our manuscript for publication in the International Journal of Plant Biology (IJPB), we have taken several measures to enhance its scientific rigor and clarity.

Firstly, we have placed a stronger emphasis on delineating the myriad features and characteristics of the two distinct varieties of sun-dried figs found within the study area. We have employed a comprehensive set of standard analytical methods known for their ability to yield substantial and intriguing results. Our aim is to provide a meticulous description of these fig varieties, thereby facilitating a rigorous comparison of their unique features against established standards and prior research findings that we thoroughly examined.

One key aspect we have addressed is the need to highlight what is novel and distinctive about our work. In our revised manuscript, we have introduced the novelty of our study, particularly emphasizing the lack of prior investigations in the Eastern region of Morocco concerning the characteristics, features, and qualities of the Chetoui "CH" and Ghoudane "GH" sun-dried fig varieties. We consider it essential to shed light on these aspects, as they have not been previously explored in scientific literature.

Furthermore, we have provided a more comprehensive introduction that clearly explains the significance of our study area, particularly the prominence of Chetoui and Ghoudane fig varieties in the eastern region of Morocco. The annual Aghbal festival held in Ahfir, dedicated to celebrating these fig varieties, exemplifies their cultural and economic significance in the region. This contextual information underscores the relevance and importance of our research objectives.

In terms of research methodology, we have expanded our explanation of the rationale behind selecting three research sites within Ahfir. This clarification enhances the transparency of our study design and strengthens the scientific foundation of our work.

Lastly, our findings have enabled us to draw conclusions regarding the suitability of sun drying as a processing method for these two distinct fig varieties. Moreover, we have identified alternative techniques that can be adopted to enhance the overall value of this fruit. These findings contribute to the practical applicability and relevance of our research within the field of plant biology.

We sincerely hope that these revisions address your concerns and that you will reconsider your recommendation regarding the publication of our manuscript in the IJPB. Your guidance and expertise have been invaluable, and we are committed to producing a high-quality contribution to the field.

In part of Material and methods please add the information:

How many samples of the raw material were taken to analysis? The Authors took samples in in one year – 2022 from different places. Please explain whether the places of sampling have an influence on the composition of the figs. How environmental factors affect the composition of figs? There is no information on the characteristics of the cultivation method.

 

Response: In the Materials and Methods section of our study, we incorporated additional information regarding the sampling process, the influence of sampling locations on fig composition, the impact of environmental factors, and the cultivation method characteristics.

Sampling Procedure: We employed a sampling strategy that involved the collection of three kilograms of sample material for each of the two types of dried figs under investigation at each of the three designated study stations. In instances where the study required the examination of a total of 30 dried figs, we selected ten pieces randomly from each of the three aforementioned samples belonging to the two types of figs being studied at each of the three study areas. For experiments conducted in triplicate, each was based on a sample derived from the grinding of one of the three kilograms of the specific fig variety under scrutiny in all examined areas.

Influence of Sampling Locations: Given that there were no prior studies available for the two dried fig varieties, and taking into account the annual renewal of leaves during photosynthesis by fig trees, we chose to conduct a comprehensive study across three distinct study stations. This approach aimed to assess the uniformity of dried figs through simultaneous sun exposure techniques. In the event of noticeable differences, we intended to investigate potential contributing factors, such as genetic variations, soil conditions, agricultural practices, and environmental variables.

For example, should fig trees experience leaf damage due to environmental factors like excessive heat, drought, or pest infestations, it can significantly impact the efficiency of photosynthesis, a crucial process in fig fruit development.

Cultivation Method Characteristics: The identification of fig trees was conducted in collaboration with local farmers who possess a deep knowledge of these trees and specifically chose them for their wide distribution and commercial significance in the region. Traditional orchards owned by local farmers were selected as sampling sites at three distinct locations, focusing on mature trees. This identification process was facilitated with the assistance of the Gharmawne cooperative, organizers of the Aghbal fig festival, and the provincial directorates of agriculture.

We believe that these additional details provide a comprehensive understanding of our sampling methodology, the significance of sampling locations, and the cultivation method characteristics employed in our study. These factors collectively contribute to the robustness and credibility of our research findings.

 

In the part of Results and discussion:

There is no information why total polyphenols and total flavonoids were determined using three different solvents? It is not explained in the introduction or in the methodology?

The obtained results of vitamin C content in dried figs are very low – 1, 03-1, 51 (%). It cannot be said that figs are the source of this vitamin.

The discussion should be elaborated in a better way. The results presented in tables are more described by the Authors, than discussed.

The conclusion is that figs have a high energy value, where are the results?

Response: In the Results and Discussion section, we appreciate your feedback and have made the necessary clarifications and elaborations as follows:

Use of Multiple Solvents for Polyphenols and Flavonoids Extraction: We utilized three distinct solvents for the extraction of polyphenols and flavonoids from the two types of dried figs to accurately determine their availability and to statistically compare the effectiveness of extraction methods. We acknowledge the importance of explaining this choice, and we appreciate your observation.

Vitamin C Content in Dried Figs: The results of our analysis indicate a comparaison between the  varieties. In fact, the vitamin C content in dried figs is relatively low, with values ranging from 1.13 to 1.14 mg/100g for CH dried figs and from 0.99 to 1.00 mg/100g for GD-dried figs. Therefore, it is not accurate to consider dried figs as a significant source of this vitamin. Additionally, we have noted that the percentage values (1.03-1.51%) mentioned in the previous version relate to the lipid content in the two dried fig varieties, with approximately 1.50-1.51% for CH and 1.03-1.04% for GH dried figs.

Discussion Elaboration: In response to your feedback, we have enhanced the elaboration of the discussion. We recognize that the results presented in the tables should be discussed in greater detail. Our intention is to provide a comprehensive discussion that not only describes the results but also interprets their significance in the context of our study objectives. We have strived to offer a more comprehensive analysis of the findings and their implications, emphasizing comparisons with previous research and established benchmarks.

Concluding Remarks on Energy Value: While our previous version did not explicitly mention the energy value, we appreciate your observation. The energy value of the dried figs is indeed an important aspect of our study. By presenting the nutritional value and benefits provided by the components under investigation, we aim to provide a more holistic understanding of the overall nutritional profile and energy content of the figs.

We hope that these clarifications and enhancements contribute to a more robust and informative Results and Discussion section in our manuscript. Thank you for your valuable feedback, which has helped us improve the quality and depth of our analysis.

 

References:

There are numerous errors in citing the literature. This needs improvement.         

Response: Thank you for your diligent review of our references. We acknowledge that there were errors in citing the literature, and we apologize for any inconvenience this may have caused. The citations were initially added using a specialized program, but we are committed to rectifying these issues.

We have taken your feedback seriously and have made the necessary revisions to ensure the accuracy and correctness of our references. We understand the importance of proper citation in maintaining the integrity of our research.

We genuinely appreciate your valuable input, and we hope that these improvements will positively influence your decision regarding the publication of our article. Your thorough evaluation has been instrumental in enhancing the quality of our manuscript, and we are committed to upholding the highest standards of academic rigor.

Thank you once again for your time and consideration.

 

Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript is extensive and well-written with a lot of results adequately presented. In conclusion, some minor mistakes should be addressed:

Check the English grammar, fonts, and punctuation extensively. Names of bacteria should be in italics.

Add at least the country of origin for each mentioned instrument (e.g. KERN PCB 2000-1).

Chapter 2.3. is very difficult to read and follow, I would suggest starting in a new paragraph when a description of a new method is mentioned.

Table 3 and Table 5 have the same title. One should be changed.

Lines 482-488 - Please write the Units in which IC50 values are expressed

Tables 3, 4, 5, 7, and 8 - The decimal point needs to be uniform. In English, it is a dot, not a notch.

Please check the main comments.

Author Response

Review Report round 2 : responses

Dear Editors, Dear Reviewers,

Thank you for giving us the opportunity to improve our manuscript with the revised version and thank you for your useful comments.

We really thank the Reviewers for their thorough review. We hope this revision will satisfy reviewers’ queries, and that our work will be considered for publication in International Journal of Plant Biology (IJPB). Below we have made efforts to either address or respond to each (paraphrased) requested change and communicated weakness. Major changes are highlighted in yellow text in the revision. All typos/minor concerns have been fixed in the manuscript and are otherwise not addressed in this response.

With kind regards

Dr. Mohamed Addi, and the co-Authors

 

Review Report (Reviewer 2)

- The manuscript is extensive and well-written with a lot of results adequately presented. In conclusion, some minor mistakes should be addressed:

Response: We deeply appreciate your thorough review of our manuscript and your constructive comments. We are pleased to inform you that we have diligently addressed each of your concerns, as outlined below:

- Check the English grammar, fonts, and punctuation extensively. Names of bacteria should be in italics.

Response: We have conducted an extensive review to ensure the correctness of English grammar, fonts, and punctuation throughout the manuscript. Additionally, we have applied italics to the names of bacteria as per your suggestion.

- Add at least the country of origin for each mentioned instrument (e.g. KERN PCB 2000-1).

Response: To provide more context, we have included the country of origin for each mentioned instrument, such as the "KERN PCB 2000-1."

- Chapter  2.3. is very difficult to read and follow, I would suggest starting in a new paragraph when a description of a new method is mentioned.

Response: Your suggestion to begin a new paragraph when introducing a new method in Chapter 2.3 has been implemented to enhance readability and clarity.

- Table 3 and Table 5 have the same title. One should be changed.

Response: We have addressed the issue with Table titles. One of the tables has been changed to ensure they are distinct from each other.

- Lines 482-488 - Please write the Units in which IC50 values are expressed

Response: We have included the units in which IC50 values are expressed in lines 482-488.

- Tables 3, 4, 5, 7, and 8 - The decimal point needs to be uniform. In English, it is a dot, not a notch.

Response: The decimal point format in Tables 3, 4, 5, 7, and 8 has been standardized to a dot, aligning with the English convention.

We would like to express our sincere gratitude for your valuable input and suggestions, which have undeniably enriched the scientific quality of our revised manuscript. We hope that these improvements meet your expectations and contribute positively to the manuscript's overall quality.

Thank you for your time and diligence in reviewing our work.

 

 

Author Response File: Author Response.docx

Reviewer 3 Report

The manuscript is well written and the experiments conducted were extensive. 

 

1)     Experimental section 2.3, procedures for the determination for metals lack clarity, it will be difficult for others to follow. The extraction of target compounds with water and organic solvents lacks clarity , it was not clear how the samples were prepared and finally used. For all procedures, it was not clear how if the samples were ground, sieved and others.  In short, too many procedures were mentioned in this section and it can be hard for others to follow. The authors are required to re organize the section. 

2)     For the determination of metals in the samples , it was unclear again what are the quality control measures in place. How are the samples digested, what are the equipment used and are certified reference materials (CRM) used. 

3)     The weakness for the current manuscript was that no target compounds were quantified, chemical fingerprint were not obtained. Identification of target compounds were not done. 

4)     For DPPH, it was noted that significant changes were not observed. There are differences but significant drops were not seen. 

5)     For the bioassay, microbiological determination seems to be a challenge without the knowledge of the residual solvent such as ethanol , acetone and others present. This is due to the fact that organic solvents inhibit the growth of bacteria. 

NA 

Author Response

Review Report round 3 : responses

Dear Editors, Dear Reviewers,

Thank you for giving us the opportunity to improve our manuscript with the revised version and thank you for your useful comments.

We really thank the Reviewers for their thorough review. We hope this revision will satisfy reviewers’ queries, and that our work will be considered for publication in International Journal of Plant Biology (IJPB). Below we have made efforts to either address or respond to each (paraphrased) requested change and communicated weakness. Major changes are highlighted in yellow text in the revision. All typos/minor concerns have been fixed in the manuscript and are otherwise not addressed in this response.

With kind regards

Dr. Addi, and the co-Authors

 

Review Report (Reviewer 3)

 

The manuscript is well written and the experiments conducted were extensive.

We greatly appreciate your thorough review and insightful comments on our manuscript. Your feedback has been invaluable in improving the quality and clarity of our work. Here are our responses to your points:

1)     Experimental section 2.3, procedures for the determination for metals lack clarity, it will be difficult for others to follow. The extraction of target compounds with water and organic solvents lacks clarity, it was not clear how the samples were prepared and finally used. For all procedures, it was not clear how if the samples were ground, sieved and others.  In short, too many procedures were mentioned in this section and it can be hard for others to follow. The authors are required to re organize the section.

Response: We genuinely thank you for highlighting the need for clarity in this section. We have reorganized it to make the procedures more comprehensible. We have included details on sample preparation, including grinding and sieving, to enhance clarity and ensure that others can follow the methods easily.

2)     For the determination of metals in the samples, it was unclear again what the quality control measures in place are. How are the samples digested, what are the equipment used and are certified reference materials (CRM) used.

Response: We have provided additional information on the quality control measures in place, including the sample digestion process, equipment used, and the use of certified reference materials (CRMs) for metal determination. ‘’Two grams of wet mass were randomly selected from each sample composite and subjected to digestion in a closed PTFE vessel (CEM MDS-200 microwave, USA) using a mixture of 65% HNO₃ (3 mL) and 30% H₂O₂ (2 mL). The resulting digested samples were diluted to a volume of 25 mL in a volumetric flask and stored in polyethylene vessels. Analysis of the samples was performed within fifteen days of their preparation. For the determination of Ca and Mg, the samples were diluted with a 0.1% (w/v) solution of lanthanum chloride. Analyses of metals of interest were conducted using a Perkin Elmer atomic absorption spectrophotometer (PE-3300) equipped with hollow cathode lamps. The flame technique was implemented for the determination of Ca, Mg, K, Na, Fe, Zn, and Cu. Duplicate solutions were created for every sample and a minimum of three distinct readings were collected to mitigate any errors. The mean values were utilized to calculate the concentrations. Merck (Darmstadt, Germany) provided standard solutions of metals (1000 mg/L in 0.1 N HNO₃) which were procured for this purpose. Stock solutions were created from the standard solutions, and working standards were generated by diluting the stock solutions. The quantification of the phosphorus amount in the dried figs was determined through the spectrophotometric molybdovanadate method.’’. This information will enhance the transparency and reliability of our metal analysis.

3)     The weakness for the current manuscript was that no target compounds were quantified, chemical fingerprint were not obtained. Identification of target compounds were not done.

Response: We acknowledge the importance of quantifying target compounds and obtaining chemical fingerprints. We want to clarify that our study aimed to investigate the overall composition and properties of the dried figs, including their nutritional aspects. However, we appreciate your suggestion, and we will consider incorporating chemical fingerprinting and quantification of specific compounds in future research.

4)    For DPPH, it was noted that significant changes were not observed. There are differences but significant drops were not seen.

Response: Thank you for your observation regarding the DPPH assay. The ANOVA statistical analysis that was conducted on DPPH has produced significant results. In particular, it was observed that the IC50 value of the ascorbic acid reference substance varied significantly from that of all CH and GD dried fig extracts. Furthermore, the DPPH IC50 has demonstrated the uniformity of each type of dried fig concerning the extraction solvent used, notwithstanding the variations in the sampling area. It is noteworthy that changing the solvent utilized for extraction, while maintaining the same sampling area for a given type of dried fig, can result in substantial differences, particularly in the case of CH. Therefore, the study's findings suggest that all the extracts tested could scavenge DPPH radicals, and the efficacy of free radical scavenging varies significantly depending on the extraction solvent and fig variety.

5)     For the bioassay, microbiological determination seems to be a challenge without the knowledge of the residual solvent such as ethanol, acetone and others present. This is due to the fact that organic solvents inhibit the growth of bacteria.

Response: We appreciate your concern regarding the presence of organic solvents in the bioassay. We would like to clarify that we have implemented a process to remove organic solvents such as ethanol and acetone via rota-vapor during the extraction process. Additionally, the use of DMSO as a solvent for mixing with the remaining extract in the bioassay does not exhibit antimicrobial properties, ensuring accurate microbial growth assessments.

We are grateful for your meticulous review and the opportunity to address these concerns. Your comments have undoubtedly enhanced the rigor and clarity of our manuscript.

 

 

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

The article can be published.