Chemical Compounds, Bioactivities, and Applications of Chlorella vulgaris in Food, Feed and Medicine

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The revised text uses accessible language and addresses a fascinating topic; however, there is a large amount of literature covering this same subject. Since the 1990s, Chlorella vulgaris has been proposed as a potential food source or dietary supplement. Nevertheless, its production presents multiple challenges. Although numerous experimental studies have been conducted in the laboratory, none have taken place in field conditions. Additionally, the maximum photosynthetic efficiency of Chlorella is only achieved under shaded conditions with artificial lighting, as direct exposure to sunlight reduces its yield to levels only slightly above those of traditional crops.

While its medicinal use has been suggested, we still lack evidence to support its efficacy in humans. Claims about its health benefits, without solid scientific backing, could enable unethical vendors to market dietary supplements.

These points deserve to be explored in greater depth in a review of this kind.

Author Response

Reviewer 1

The revised text uses accessible language and addresses a fascinating topic; however, there is a large amount of literature covering this same subject. Since the 1990s, Chlorella vulgaris has been proposed as a potential food source or dietary supplement. Nevertheless, its production presents multiple challenges. Although numerous experimental studies have been conducted in the laboratory, none have taken place in field conditions. Additionally, the maximum photosynthetic efficiency of Chlorella is only achieved under shaded conditions with artificial lighting, as direct exposure to sunlight reduces its yield to levels only slightly above those of traditional crops.

While its medicinal use has been suggested, we still lack evidence to support its efficacy in humans. Claims about its health benefits, without solid scientific backing, could enable unethical vendors to market dietary supplements.

These points deserve to be explored in greater depth in a review of this kind.

 

Reply: Thank you for your thoughtful and detailed feedback. We appreciate your recognition of our efforts to present the topic in accessible language and for highlighting the extensive literature on Chlorella vulgaris as a food source and supplement.

We agree that Chlorella vulgaris production faces significant challenges, especially regarding photosynthetic efficiency under direct sunlight and the absence of large-scale field trials. While this review primarily focuses on the bioactive compounds and health potential of Chlorella vulgaris, we acknowledge the relevance of these production constraints. Addressing them fully would require additional studies and more specialized discussion, which is beyond the current scope. Nonetheless, we emphasize that overcoming these production challenges is essential for advancing Chlorella’s practical application as a sustainable supplement.

Your point regarding the current limitations in clinical evidence for Chlorella’s medicinal efficacy is highly pertinent. Although promising experimental results exist, mainly in controlled settings, we recognize the importance of avoiding overstated health claims in the absence of robust evidence from large-scale human trials. Misrepresenting Chlorella’s benefits could indeed lead to ethical concerns with supplement marketing. In line with your suggestions, we have expanded our discussion to include a more comprehensive overview of available human trials and meta-analyses, focusing on evidence-based benefits.

We have integrated the following studies in our review, which provide insight into the effects of Chlorella vulgaris on various health outcomes. The listed studies, while encouraging, highlight the need for continued clinical trials to build stronger evidence:

    Panahi et al. (2012): Investigated Chlorella vulgaris supplementation in non-alcoholic fatty liver disease patients, noting beneficial effects on liver health (Hepato-gastroenterology, 59(119), 2099–2103, doi: 10.5754/hge10860).

    Panahi et al. (2012): Studied Chlorella vulgaris as adjunct therapy for dyslipidemia, observing improvements in lipid profile (Nutr. Diet. 69(1), 13-19, doi: 10.1111/j.1747-0080.2011.01569.x).

    Panahi et al. (2012): Examined antioxidant effects in patients with obstructive pulmonary diseases (Sci Pharm, 80, 719–730).

    Panahi et al. (2013): Analyzed Chlorella vulgaris’s role in oxidative stress modulation among smokers (Clin Lab, 59, 579–587).

    Panahi et al. (2015): Evaluated Chlorella supplementation in major depressive disorder, noting mood improvements (Complementary Therapies in Medicine, 23(4), 598–602, doi: 10.1016/j.ctim.2015.06.010).

    Sherafati et al. (2022): Meta-analysis on Chlorella’s impact on lipid profile, emphasizing the need for dosage standardization (Complementary Therapies in Medicine, 66, 102822, doi: 10.1016/j.ctim.2022.102822).

    Kwak et al. (2012): Reported immunostimulatory benefits, enhancing natural killer cell activity in short-term supplementation (Nutrition Journal, 11, 53, doi: 10.1186/1475-2891-11-53).

While these studies offer promising insights, we acknowledge the need for further randomized controlled trials to substantiate Chlorella vulgaris's efficacy in human health. We have highlighted these limitations in the manuscript to promote transparency in the current understanding of Chlorella’s health benefits.

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript provides a comprehensive review of Chlorella vulgaris, covering its chemical composition, bioactivities, and applications across multiple sectors, including food, feed, and medicine. The review is well-researched and presents a broad range of findings, particularly highlighting the functional food and therapeutic potential of this microalga. However, there are several areas that require further refinement to improve the manuscript's scientific rigor and practical relevance.

1. This article discusses a variety of potential health benefits of Chlorella vulgaris, including antioxidant, anti-inflammatory and anti-cancer effects. However, most of these benefits are based on in vitro or animal experiments, and there is a lack of evidence from clinical studies or human trials. It is recommended to add more clinical data based on human studies or clarify that these benefits still need clinical validation to enhance the scientific rigor of health benefit conclusions.

 

2. The article mentioned many bioactive functions (such as antioxidation, immune regulation, etc.), but the discussion of the molecular mechanism behind these biological activities was relatively rough, especially in anti-cancer, antiviral and other applications.

 

3. Although the article mentioned future research directions, the discussion was relatively general, such as "optimizing culture methods" and "improving bioavailability", without further detailing specific research steps. Specific technical challenges or research questions are suggested to be clearly proposed in future research directions, such as how to improve the extraction efficiency of key components in Chlorella vulgaris or how to improve their bioavailability in humans.

 

 

4. Although the article covers the application of Chlorella vulgaris in many fields, the analysis of the benefits and limitations of each field is relatively shallow, and there is a lack of comparison of its actual performance and applicability in different fields. It is suggested to analyze the advantages and challenges of different application fields through more detailed comparison, especially the practical benefits and technical bottlenecks of Chlorella vulgaris in the food and pharmaceutical fields.

Author Response

Reviewer 2

The manuscript provides a comprehensive review of Chlorella vulgaris, covering its chemical composition, bioactivities, and applications across multiple sectors, including food, feed, and medicine. The review is well-researched and presents a broad range of findings, particularly highlighting the functional food and therapeutic potential of this microalga. However, there are several areas that require further refinement to improve the manuscript's scientific rigor and practical relevance.

 

1. This article discusses a variety of potential health benefits of Chlorella vulgaris, including antioxidant, anti-inflammatory and anti-cancer effects. However, most of these benefits are based on in vitro or animal experiments, and there is a lack of evidence from clinical studies or human trials. It is recommended to add more clinical data based on human studies or clarify that these benefits still need clinical validation to enhance the scientific rigor of health benefit conclusions.

 

Reply: Thank you for your thoughtful and detailed feedback.

This point regarding the current limitations in clinical evidence for Chlorella’s medicinal efficacy is highly pertinent. Although promising experimental results exist, mainly in controlled settings, we recognize the importance of avoiding overstated health claims in the absence of robust evidence from large-scale human trials. Misrepresenting Chlorella’s benefits could indeed lead to ethical concerns with supplement marketing. In line with your suggestions, we have expanded our discussion to include a more comprehensive overview of available human trials and meta-analyses, focusing on evidence-based benefits.

We have integrated the following studies in our review, which provide insight into the effects of Chlorella vulgaris on various health outcomes. The listed studies, while encouraging, highlight the need for continued clinical trials to build stronger evidence:

    Panahi et al. (2012): Investigated Chlorella vulgaris supplementation in non-alcoholic fatty liver disease patients, noting beneficial effects on liver health (Hepato-gastroenterology, 59(119), 2099–2103, doi: 10.5754/hge10860).

    Panahi et al. (2012): Studied Chlorella vulgaris as adjunct therapy for dyslipidemia, observing improvements in lipid profile (Nutr. Diet. 69(1), 13-19, doi: 10.1111/j.1747-0080.2011.01569.x).

    Panahi et al. (2012): Examined antioxidant effects in patients with obstructive pulmonary diseases (Sci Pharm, 80, 719–730).

    Panahi et al. (2013): Analyzed Chlorella vulgaris’s role in oxidative stress modulation among smokers (Clin Lab, 59, 579–587).

    Panahi et al. (2015): Evaluated Chlorella supplementation in major depressive disorder, noting mood improvements (Complementary Therapies in Medicine, 23(4), 598–602, doi: 10.1016/j.ctim.2015.06.010).

    Sherafati et al. (2022): Meta-analysis on Chlorella’s impact on lipid profile, emphasizing the need for dosage standardization (Complementary Therapies in Medicine, 66, 102822, doi: 10.1016/j.ctim.2022.102822).

    Kwak et al. (2012): Reported immunostimulatory benefits, enhancing natural killer cell activity in short-term supplementation (Nutrition Journal, 11, 53, doi: 10.1186/1475-2891-11-53).

While these studies offer promising insights, we acknowledge the need for further randomized controlled trials to substantiate Chlorella vulgaris's efficacy in human health. We have highlighted these limitations in the manuscript to promote transparency in the current understanding of Chlorella’s health benefits.

 

2. The article mentioned many bioactive functions (such as antioxidation, immune regulation, etc.), but the discussion of the molecular mechanism behind these biological activities was relatively rough, especially in anti-cancer, antiviral and other applications.

 

Reply: Thank you for highlighting the need for a deeper exploration of the molecular mechanisms underlying the bioactive functions of Chlorella vulgaris, particularly in anti-cancer, antiviral, and related applications.

In response, we have revised the discussion to provide a more detailed analysis of these mechanisms, focusing on key pathways involved in antioxidation, immune modulation, and specific anti-cancer and antiviral activities. For example, we now elaborate on Chlorella’s influence on signalling pathways like NF-κB and MAPK, its role in enhancing immune responses via cytokine modulation, and its effects on apoptosis and cell cycle regulation in cancer cells. This added depth aligns with your suggestion and strengthens our review of Chlorella vulgaris's bioactive potential.

 

3. Although the article mentioned future research directions, the discussion was relatively general, such as "optimizing culture methods" and "improving bioavailability", without further detailing specific research steps. Specific technical challenges or research questions are suggested to be clearly proposed in future research directions, such as how to improve the extraction efficiency of key components in Chlorella vulgaris or how to improve their bioavailability in humans.

 

Reply: Thank you for your valuable feedback. In response, we have refined our discussion on future research directions to include specific research steps and address critical technical challenges. For instance, we now suggest targeted approaches to optimize Chlorella vulgaris cultivation, such as adjusting light spectra and nutrient formulations to enhance compound yields. We also discuss advanced extraction techniques like enzymatic and ultrasound-assisted methods to improve the extraction efficiency of key bioactive compounds. Furthermore, we propose exploring nano-encapsulation and formulation technologies to enhance bioavailability in humans. These additions provide a clearer roadmap for future research, aligning with your recommendations.

 

4. Although the article covers the application of Chlorella vulgaris in many fields, the analysis of the benefits and limitations of each field is relatively shallow, and there is a lack of comparison of its actual performance and applicability in different fields.

It is suggested to analyze the advantages and challenges of different application fields through more detailed comparison, especially the practical benefits and technical bottlenecks of Chlorella vulgaris in the food and pharmaceutical fields.

 

Thank you for your insightful feedback and valuable suggestions. We have made substantial revisions to enhance the depth of analysis across application fields for Chlorella vulgaris and address your concerns as follows:

1. Comparative Analysis of Application Fields: The revised manuscript now includes a detailed comparison of the practical benefits and limitations of Chlorella vulgaris across sectors, focusing on specific advantages and technical bottlenecks in the food and pharmaceutical industries. This comparison highlights its nutritional and functional contributions alongside key production and formulation challenges.
2. Potential Adverse Effects: We have expanded the discussion on potential negative side effects, particularly in food and pharmaceutical applications, addressing issues such as gastrointestinal discomfort and other health considerations. This balanced analysis aims to present a more comprehensive view of Chlorella's practical applicability.
3. Human Trials: We have incorporated additional information on existing human trials, noting the current limitations in this research area. Given the relatively small number of human studies, we highlight the insights derived from animal trials that help guide and inform our understanding of Chlorella’s effects in human health contexts, especially concerning its bioactive compounds.

These revisions aim to provide a more nuanced, comparative perspective on Chlorella vulgaris's role across various applications, aligning with your recommendations.

Reviewer 3 Report

Comments and Suggestions for Authors

Comments and suggestions for Authors in the attached file

Comments for author File: Comments.pdf

Comments on the Quality of English Language

The quality of the English language in this manuscript is generally acceptable, but some areas require improvement. Some sentences are overly complex, and the phrasing can be convoluted at times, which affects readability. Simplifying the sentence structure and improving the flow would enhance clarity. Additionally, there are minor grammatical errors that should be addressed to ensure the language is smooth and professional throughout the manuscript. Overall, while the content is understandable, revisions are needed to polish the language.

Author Response

Reviewer 3

Overall, this manuscript demonstrates scientific rigour and effectively aims to highlight both the benefits and potential risks associated with C. vulgaris consumption. However, several aspects could be improved. Some sections are overly complex or convoluted, which impacts readability. Simplifying the language and presenting the key points more clearly and concisely would enhance the manuscript's clarity. As a result, major revisions may be necessary to refine these areas and strengthen the manuscript before it is ready for publication.

 

1. Comment 1. Introduction

 

The introduction is well-written and clearly and comprehensively covers the key points regarding the importance and potential of Chlorella vulgaris. However, a few small suggestions exist to improve the flow and correct minor language imperfections.

 

• Improving transitions:

Lines 45-48: "Notably, C. vulgaris has shown particularly noteworthy is C. vulgaris's high protein content, which has positioned it as a viable alternative protein source in food products..."

This sentence starts with "notably" and then immediately continues with "particularly noteworthy," creating redundancy and breaking the flow.

 

Reply: Thank you, we made the necessary changes.

 

Lines 68-73: "Additionally, C. vulgaris has demonstrated potential in improving lipid metabolism and immune function and detoxifying heavy metals from the body [21,22]. However, the impact of cultivation methods, environmental conditions, and processing techniques on the stability and potency of these bioactive compounds remains an ongoing subject of investigation."

The shift from "C. vulgaris' benefits" to "cultivation methods" is abrupt, without a clear connection between the concepts.

 

Reply: Thank you, we made the necessary corrections.

 

Lines 76-82: "This manuscript aims to provide a comprehensive assessment of the chemical composition, bioactivities and diverse applications of C. vulgaris across the food, nutrition medicine, and animal feed sectors. By synthesizing current research from reference databases, such as Google Scholar, PubMed, Scopus, and Web of Science, this manuscript highlights the health-promoting effects of C. vulgaris’s bioactive compounds..."

The transition from the manuscript’s goal to the research synthesis is not perfectly smooth.

 

Reply: Thank you, we revised the sentences.

 

• 2. Unclear concept:

Lines 51-54: "Chlorella vulgaris contains 43% to 61% protein by dry weight, depending on growth conditions [9,10], with approximately 20% of these proteins associated with the cell wall, 50% located inside the cells, and the remaining 30% continuously migrating in and out of the cell [11]."

The idea of proteins "migrating in and out of the cell" is not very clear and could confuse the reader.

 

Reply: Thank you for your valuable feedback. To clarify the concept of protein movement within Chlorella vulgaris, we have revised the text to explain that the proteins are dynamic and participate in cellular functions. For further details on this topic, the reader may refer to the original sources cited.

 

• 3. Making the concept more direct:

Lines 61-62: "Research into C. vulgaris has also explored its antioxidant and anti-inflammatory activities."

The sentence is somewhat generic and could be made more direct.

 

Reply: Thank you. We have made the sentence more direct

 

2. Comment 2. Section 2. Chemical Composition of Chlorella vulgaris

The section is well-written and informative, but the content can be improved for clarity and flow.

 

• Repetition and Redundancy:

Lines 111-113 "The digestibility of C. vulgaris protein is also an important factor. Studies indicate that the protein digestibility of C. vulgaris is comparable to that of other plant proteins, with digestibility rates reported between 80% to 90% [30]."

The phrase “digestibility of C. vulgaris protein” is repeated twice within a short span, which affects the flow.

 

Reply: Thank you for your comments, we have improved the sentence.

 

• Simplifying Complex Sentences:

Lines 167-169: "The insoluble fibres in C. vulgaris promote bowel regularity by increasing stool bulk and facilitating movement through the digestive tract, while also providing a substrate for beneficial gut microbiota [7]."

While the sentence is informative, the use of multiple clauses makes it slightly cumbersome to read.

 

Reply: Thank you for your comments, we have revised the sentence.

 

• Clarifying Ambiguous Concepts:

Lines 119-121: "In contrast, while animal proteins provide high-quality protein, they may also come with higher levels of saturated fats and cholesterol, which can be a concern for some consumers [3]."

The phrase “which can be a concern for some consumers” is vague. It's unclear why this is a concern.

 

Reply: Thank you for your comments. We have revised the sentence to clarify why saturated fats and cholesterol are a concern, explicitly linking them to an increased risk of cardiovascular diseases and other health issues.

 

• Refining Transition Between Sections:

Lines 122-123 "Due to the rigid cellulose cell wall of C. vulgaris, pre-treatment processes are often necessary to enhance protein digestibility and bioavailability. Mechanical methods such as bead milling, sonication, and high-pressure homogenization have proven effective..."

The transition from the benefits of protein to discussing mechanical processes is abrupt and could be more clearly defined.

 

Reply: Thank you for your suggestion. We have revised the sentence and added a linking phrase to connect the two concepts more smoothly

 

• Improving Flow and Conciseness:

Lines 163-166 "These fibres are primarily composed of insoluble cellulose and hemicellulose, which contribute to the structural integrity of the cell walls. The presence of these fibres is essential not only for maintaining cellular structure but also for their beneficial role in human digestion and gut health [3]."

The first two sentences can be combined to improve flow and reduce repetition.

 

Reply: Thank you for your comments. We have combined the sentences as suggested.

 

• Enhancing Scientific Precision:

Lines 146-147: “The lipid profile of C. vulgaris has garnered attention due to its potential health benefits, including anti-inflammatory and cardiovascular protective properties [31].”

“Potential health benefits” is vague and could be more specific.

 

Reply: Thank you. We have improved the sentence and made it more specific.

 

• In subsection 2.4, while you mention the antioxidant and detoxifying properties of chlorophyll, it could be useful to provide more detail on how these mechanisms work or cite specific studies.

The discussion of lutein and zeaxanthin to AMD (age-related macular degeneration) is well-structured, but it could benefit from some quantitative data on how much consumption of these pigments influences the risk of AMD, if available.

 

Reply: Thank you for your comment. We have improved the subsection according to the given feedback.

 

• The sentence in 2.6 (Line 111): "The digestibility of C. vulgaris protein is also an important factor..." is repeated from earlier sections. It might be unnecessary to repeat this concept unless adding new insights. Consider reducing redundancy to streamline the content.

 

Reply: Thank you for your insightful feedback. We believe that the information regarding the digestibility of C. vulgaris protein is relevant and contributes to the overall context of the text. However, we appreciate your point about redundancy. In general, we have aimed to improve reading fluency and clarity throughout the manuscript based on the reviewers' feedback, and we will consider your suggestion for further refinement.

 

• Lines 221-223 "The nucleotides present in Chlorella vulgaris can support gut health by promoting the growth of beneficial gut microbiota..." would benefit from specifying which bacteria or types of microbiota are supported by the presence of nucleotides, if known.

 

Reply: Thank you for your valuable feedback. We appreciate your suggestion to specify which bacteria or types of microbiota are supported by the presence of nucleotides in Chlorella vulgaris. However, to our knowledge, specific information regarding the exact bacterial species or types of microbiota influenced by nucleotides in C. vulgaris has not been well-documented in the literature.

 

• Lines 193-194 "Chlorella vulgaris is also a notable source of minerals such as iron, calcium, magnesium, and zinc, all of which contribute to its health-promoting properties” . The transition between the discussion of vitamins and minerals could be made smoother.

 

Reply: Thank you for your suggestion. We have improved the transition.

 

• Sections 2.5 and 2.7 are both brief and discuss related topics—essential nutrients and bioactive compounds. Combining them into a single section could improve the flow of the text and offer a more comprehensive discussion of the health benefits of C. vulgaris. For instance, both vitamins/minerals and secondary metabolites (such as polyphenols

and sterols) contribute to the antioxidant, anti-inflammatory, and immune-supporting effects of C. vulgaris .

 

Reply: Thank you for your insightful suggestion regarding the potential to combine Sections 2.5 and 2.7. While we appreciate the rationale behind this idea, we believe that keeping these sections separate allows for a more focused discussion of the essential nutrients and bioactive compounds of Chlorella vulgaris while maintaining clarity for readers.

 

3. Comment: Section 3. Bioactivity of Chlorella vulgaris Compounds

The content in this section on the bioactivity of Chlorella vulgaris is well-organized and provides a comprehensive overview of its health-promoting properties. However, there are some areas where the text can be improved for clarity, depth, and flow.

 

• 3.1 Antioxidant Activity:

The explanation of the difference between non-enzyme and enzyme-promoted antioxidants is a bit abrupt. You could expand on this distinction at the beginning of the section to clarify their roles in more detail.

For instance, the discussion on superoxide dismutase (SOD) and catalase (CAT) could be further enriched by mentioning how these enzymes specifically mitigate oxidative stress at a cellular level, perhaps with references to studies measuring the increase in SOD/CAT activity after supplementation with C. vulgaris.

There is a repeated mention of the antioxidant roles of lutein and beta-carotene. This information could be consolidated in the non-enzyme-promoted antioxidants subsection and omitted from other parts.

 

Reply: Thank you for your suggestion. We have explained the difference and made a smoother transition in the text as suggested. We also added studies as recommended.

 

• 3.2 Anti-inflammatory Effects:

You could expand on the specific pathways in which C. vulgaris acts on COX-2 inhibition. Perhaps include more detail on how extracts of C. vulgaris have been studied in vitro or in vivo to determine its impact on these enzymes and cytokines.

There’s an opportunity to introduce examples of clinical or preclinical trials that demonstrate these anti-inflammatory effects, which would add more concrete evidence to support these claims.

Consider explaining the relationship between inflammation and chronic diseases like arthritis, and cardiovascular diseases, and how C. vulgaris contributes to managing these conditions through its anti-inflammatory properties.

 

Reply: Thank you. In response to your suggestions, we have expanded our discussion on the specific pathways through which Chlorella vulgaris exerts its COX-2 inhibition. We included more detail on how extracts of C. vulgaris have been studied, emphasizing their impact on various inflammatory cytokines and enzymes.

Regarding the suggestion explaining the relationship between inflammation and chronic diseases like arthritis, and cardiovascular diseases, we developed this information in subsection: 4.3. Applications in medicine

 

• 3.3 Immunomodulatory Activity:

Provide specific data from animal studies or clinical trials showing how C. vulgaris influences immune response. This would help substantiate claims about its immunomodulatory effects.

 

Reply: Thank you for your valuable feedback regarding Section 3.3 on Immunomodulatory Activity. In response to your suggestion, we have added specific data from both animal studies and clinical trials to substantiate our claims about the immunomodulatory effects of Chlorella vulgaris.

 

The text mentions that C. vulgaris improves the balance between pro-inflammatory and anti-inflammatory cytokines but could delve deeper into the mechanism by which this regulation occurs.

There is some overlap with the anti-inflammatory section regarding IL-10. Consider consolidating or distinguishing this information more clearly to avoid repetition.

You could include graphical representations of how C. vulgaris impacts various biological pathways (such as antioxidant defence, inflammation regulation, or immune modulation). It would add clarity and visual interest.

 

Reply: Thank you for the suggestion. It has been revised according to the recommendation provided.

 

• Subsections 3.4-3.9

While the document references various studies, most of the findings seem to be based on in vitro or animal models. It acknowledges that further studies are needed to confirm C. vulgaris's clinical efficacy, particularly regarding antiviral, anticancer, and heavy metal detoxification effects. More emphasis on human clinical trials would strengthen the claims and provide a clearer understanding of its effectiveness in real-world scenarios.

 

Reply: Thank you for your feedback. We have addressed your comment, which aligns with a suggestion from the other Reviewers, by clarifying that many health benefits of Chlorella vulgaris are based on in vitro or animal studies and require clinical validation. We also added references to available clinical data where applicable to strengthen the scientific rigor of our conclusions. Additionally, we acknowledge that further investigation in humans through randomized trials is necessary.

 

4. Comment Section 5. Potential Negative Side Effects

The section on potential negative side effects of C. vulgaris consumption brings up several important concerns, but there are some weaknesses in the discussion that should be addressed for a more comprehensive analysis.

 

Reply: Thank you for your suggestion. We have improved section 5 taking in to account the suggestions of all the authors.

 

• Lack of Human Data: One of the major weaknesses is the absence of clinical data or references to human studies. The concerns mentioned, such as heavy metal accumulation, antinutritional factors, and toxicity, are based primarily on in vitro studies or animal models. Without human data, it is difficult to assess the real-world implications for consumers. The section could be strengthened by including any existing human studies that explore these risks or by clearly stating the gap in the current research.

 

Reply: Thank you for your feedback. We have addressed your comment, which aligns with a suggestion from ither reviewers, by clarifying that many health benefits of Chlorella vulgaris are based on in vitro or animal studies and require clinical validation. We also added references to available clinical data where applicable to strengthen the scientific rigor of our conclusions.

 

• Overgeneralization of Risks: While it is important to point out potential risks like heavy metal accumulation or pesticide contamination, the section could benefit from more specificity. For example, what levels of exposure to heavy metals or pollutants pose a real risk to consumers? Are there regulations or quality controls in place to ensure C. vulgaris supplements are free from harmful contaminants? By overgeneralizing these risks without enough detail, the text creates a sense of alarm without providing actionable information on how to mitigate these risks.

 

Reply: Thank you for your suggestion. We have improved the text accordingly.

 

• Limited Discussion of Dosage and Consumption Context: Although the text mentions potential hepatotoxicity and nephrotoxicity at high doses, it lacks specificity in terms of what constitutes a "high dose" for consumers. Offering clearer guidelines or referencing safe dosage ranges found in studies would help provide a more balanced perspective. Additionally, more attention should be paid to individual consumption contexts, such as populations that may be more susceptible to adverse effects (e.g., people with preexisting liver or kidney conditions).

 

Reply: Thank you for your insightful feedback regarding the discussion of dosage and consumption context in our manuscript. We recognize the importance of providing clear guidelines on what constitutes a "high dose" of Chlorella vulgaris for consumers. In response to your suggestion, we have included specific dosage recommendations based on existing literature, highlighting safe dosage ranges that have been identified in clinical studies.

Additionally, we have expanded our discussion to address individual consumption contexts, particularly concerning populations that may be more susceptible to adverse effects, such as individuals with preexisting liver or kidney conditions.

We also emphasize that individuals should consult healthcare professionals before starting any supplementation regimen.

 

• Inadequate Discussion of Interaction with Other Substances: The mention of C. vulgaris exacerbating toxicity when combined with substances like paracetamol is important, but more detail is needed. For instance, are there other common medications or supplements that may have similar interactions? Without further exploration, this point feels underdeveloped and leaves the reader with more questions than answers.

 

Reply: Thank you for your comments. We have improved the text.

 

• Unclear on Environmental Monitoring and Cultivation Standards: The section talks about the importance of monitoring environmental conditions during the cultivation of C. vulgaris, but it does not specify what protocols or industry standards are currently in place to ensure safe cultivation. How do producers mitigate the risks posed by heavy metals or pollutants? Providing examples of best practices in cultivation or references to certifications or regulations could enhance the reader's confidence in the safety of commercially available C. vulgaris products.

 

Reply: Thank you for your comments. We have revised the text.

 

Round 2

Reviewer 1 Report

Comments and Suggestions for Authors

The topic is not addressed objectively

Author Response

Reviewer 1

The topic is not addressed objectively.

 

Reply: Thank you for your feedback regarding the objectivity of the topic presentation. We understand the importance of presenting our findings clearly and unbiasedly. In response to your comment, we have revisited and revised key sections of the manuscript to ensure that the discussion remains focused and objective.

Reviewer 3 Report

Comments and Suggestions for Authors

I suggest simply reviewing the language to check and correct any potential typos.

Comments on the Quality of English Language

I would recommend to improve the quality of English Language

Author Response

Reviewer 3

I suggest simply reviewing the language to check and correct any potential typos.

 

Reply: Thank you for your valuable feedback and suggestions. As recommended, we have carefully reviewed the language throughout the manuscript to correct any typographical errors and improve the overall quality of the English.

We appreciate your input in enhancing the clarity and readability of our work.