Streptococcus thermophilus and Lactobacillus bulgaricus Attributes as Influenced by Carao (Cassia grandis) Fruit Parts
, Allan Avila 1, Dany Avila 1, David Picha 3, Roberto Cedillos 2, Shirin Kazemzadeh 4, Leyla K. Pournaki 5, Ajitesh Yaday 6, Jhunior Marcia 1 and Aryana Kayanush 2,*
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsGeneral comments:
The manuscript highly resembles an already published manuscript, entitled “Probiotic Characteristics of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus as Influenced by Carao (Cassia grandis)” by Paz et al. yet the manuscript does not cite the already published article. https://www.mdpi.com/2311-5637/8/10/499 For example, the “plant material” section in both papers even mention the exact dates!!! The reviewer is highly concerned about significant overlaps between the manuscript and Paz et al. 2022.
Streptococus thermophilus and Lactobacillus bulgaricus are used to make yogurt. The term “probiotic(s)” and “starter culture(s)” are used interchangeably in the manuscript. Does Chr. Hansen sell Streptococus thermophilus ST-M5 and Lactobacillus bulgaricus LB-12 as starter cultures with probiotic properties?
The manuscript unnecessarily concentrates on “Materials and Methods.” Much of this section can be shorted.
Many of the figures are confusing because the authors mention “values with different letters are significantly different” while there are no letters anywhere. Also, it’s very difficult to differentiate error bars.
The “Results and Discussion” section is not well articulated.
Specific comments
Page 1
Line 3: consider deleting “different” from the title of the article for a better read.
Line 4: “castro” should start with a capital C instead of lower-case c.
Line 16-17: “gas-trointestinal” should be spelled “gastrointestinal”.
Line 24: “succinic” should read “succinic acid”.
Line 28: “Fructose” is written with capital F. Be consistent when you write sugars. Use lower case letters.
Line 32-34: confusing sentence. Rewrite. Odd word choices, such as “the crust” and “hurt”.
Page 2
Line 53-55: this information is repeated. See line 45-47 on the same page.
Line 56: consider “can provide” instead of “provide”. Not all LAB have probiotic properties.
Line 59-60: “immuneglobulin” is misspelled.
Line 64: consider deleting “items”.
Line 66-69: line 66-67 mentions dairy fermentation. The following sentence that starts with “This category…” mentions many dairy products that are NOT fermented!
Line 77-78: reference is missing.
Line 90: “diverse dairy items” is misleading. I assume the authors have meant yogurt and select cheeses.
Page 3
Line 109: provide a timeline instead of “until it underwent lyophilization.”
Line 116: what do you mean by “initial cultures”?
Line 118: how did you decide on “a concentration of 2%”? Is this your work? If not, include information. If not, include references.
Page 4
Line 145: “G” is not how the relative centrifugal force is written.
Line 154: what is the name of the “resultant blue compound”?
Line 164: The plants tissues extracts” does not read well.
Line 172: “The plants tissues extracts” does not read well.
Page 5
Line 193: consider “filtrate” instead of “filtrated”
Line 193: consider “transferred” instead of “transfer”
Line 198" Multiple (3?) spectrophotometers (different brands, manufacturers, models, etc.) are being mentioned throughout “Materials and Methods.” (line 198, line 306) Why is that?
Line 209: “For plants tissues extracts preparation” does not read well. Also, is this your method? If not, you will need a reference.
Line 209: consider “was” instead of “were.”
Line 213: need to include unit after “0.45”. I assume this is micrometer.
Line 281: Beckman needs to be written with capital B.
Line 224: need company information for the column.
Line 229-230: be consistent in how you write these organic acids. Either use capital letters or use lowercase letters (preferred).
Page 6
Line 235: delete “was used” here.
Line 237” add “was used after “214 nm.”
Line 256: the target organisms are called “starter cultures” here. They are called “probiotics” or “probiotic cultures” or “cultures” at times. Consider consistency.
Line 257” consider “their ability.”
Line 264: why did you pick “8 hours” as incubation time?
Line 269: why did you use a single pH (pH 2.0 as the only pH adjustment? Would it not make sense to work with a range of pH values?
Line 279: the target organisms are called “starter cultures” here. They are called “probiotics” or “probiotic cultures” at times. Consider consistency.
Line 279: the genera and species names need to be italicized.
Page 7
Line 281: include details of the anaerobic incubation.
Line 287: the target organisms are called “starter cultures” here. They are called “probiotics” or “probiotic cultures” or “cultures” at times. Consider consistency.
Line 295: the genera and species names need to be italicized.
Line 297: the genera and species names need to be italicized.
Line 304: consider “to be used” instead of “to use”.
Line 314: there is an issue with how the chemical was spelled and where the company name was included.
Line 323: What is “Streptococcus thermophilus agar”? There is no such medium! Did you mean M17?
Page 8
Line 332: what do you mean by “getting different dilution”?
Line 334: consider “were” instead of “was.”
Line 337: what is “Lactobacillus bulgaricus agar”? There is no such medium! Did you mean Lactobacillus MRS Agar?
Table 1: “different letters” in the table need to be in superscript.
Table 1 results: What is the significance of your findings?
Table 2 result: What is the significance of your findings?
Page 9
Line 371: use lower case “s” in “Seed.”
Table 3 results: What is the significance of your findings?
Line 383-387: Are you trying to compare your results to others’ here?
Line 389: “the cell membrane is remarkably powerless to bile salts” is an odd expression.
Line 393: “a substantial decrease in ST counts”?
Page 10:
Line 403-404: what results?
Figure 1: while the manuscript mentions “values with different letters are significantly different”, there are no letters anywhere. Also, the reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Line 408-409: what is the basis of the statement?
Page 11
Figure 2: while the manuscript mentions “values with different letters are significantly different”, there are no letters anywhere. Also, the reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Page 12:
Figure 4: while the manuscript mentions “values with different letters are significantly different”, there are no letters anywhere. Also, the reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Figure 5: The reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Page 13
Figure 6: The reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Page 14
Figure 7: The reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Page 15:
Figure 9: The reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Figure 10: The reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Page 16
Line 534: the genus and species names need to be in italics.
Line 534-543: confusing paragraph that makes statements about proteolytic microorganisms in the large intestine and then proteolysis in fermented milk.
Line 545-547: Not following the statement. Could you have not determined properties of these organisms in the absence of carao components?
Line 547-550: what is the importance/significance (if any)?
Comments on the Quality of English LanguageThe manuscript needs work as indicated in specific comments made above.
Author Response
Reviewer 1
The manuscript highly resembles an already published manuscript, entitled “Probiotic Characteristics of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus as Influenced by Carao (Cassia grandis)” by Paz et al. yet the manuscript does not cite the already published article. https://www.mdpi.com/2311-5637/8/10/499 For example, the “plant material” section in both papers even mention the exact dates!!! The reviewer is highly concerned about significant overlaps between the manuscript and Paz et al. 2022.
Answer: The research was done in the same time by 2 different visiting scholars from Honduras. This manuscript was delayed because of writing and editing improvements by authors. The difference is that Paz et al. worked with different parts of the plant material. Paz et al. 2022. Is cited in this manuscript.
Streptococus thermophilus and Lactobacillus bulgaricus are used to make yogurt. The term “probiotic(s)” and “starter culture(s)” are used interchangeably in the manuscript. Does Chr. Hansen sell Streptococus thermophilus ST-M5 and Lactobacillus bulgaricus LB-12 as starter cultures with probiotic properties?
Answer: The term “probiotic(s)” was changed to “culture(s)” throughout the manuscript.
The manuscript unnecessarily concentrates on “Materials and Methods.” Much of this section can be shorted.
Answer: details were provided to clarified the meth dodgy approach to the editor and reviewers.
Many of the figures are confusing because the authors mention “values with different letters are significantly different” while there are no letters anywhere. Also, it’s very difficult to differentiate error bars.
Answer: values with different letters are significantly different’ statement was deleted.
The “Results and Discussion” section is not well articulated.
Specific comments
Page 1
Line 3: consider deleting “different” from the title of the article for a better read.
Answer: recommendation was followed.
Line 4: “castro” should start with a capital C instead of lower-case c.
Answer: recommendation was followed.
Line 16-17: “gas-trointestinal” should be spelled “gastrointestinal”.
Answer: recommendation was followed.
Line 24: “succinic” should read “succinic acid”.
Answer: recommendation was followed.
Line 28: “Fructose” is written with capital F. Be consistent when you write sugars. Use lower case letters.
Answer: recommendation was followed.
Line 32-34: confusing sentence. Rewrite. Odd word choices, such as “the crust” and “hurt”.
Answer: recommendation was followed. Sentence was rewrite.
Page 2
Line 53-55: this information is repeated. See line 45-47 on the same page.
Answer: recommendation was followed. Lines 45-47 were deleted.
Line 56: consider “can provide” instead of “provide”. Not all LAB have probiotic properties.
Answer: recommendation was followed.
Line 59-60: “immuneglobulin” is misspelled.
Answer: recommendation was followed.
Line 64: consider deleting “items”.
Answer: recommendation was followed. word was changed.
Line 66-69: line 66-67 mentions dairy fermentation. The following sentence that starts with “This category…” mentions many dairy products that are NOT fermented!
Answer: recommendation was followed. Sentence was rewrite.
Line 77-78: reference is missing.
Answer: recommendation was followed. Reference was placed.
Line 90: “diverse dairy items” is misleading. I assume the authors have meant yogurt and select cheeses.
Answer: recommendation was followed. word was changed.
Page 3
Line 109: provide a timeline instead of “until it underwent lyophilization.”
Answer: recommendation was followed. Sentence was rewrite.
Line 116: what do you mean by “initial cultures”?
Answer: The word initial was deleted.
Line 118: how did you decide on “a concentration of 2%”? Is this your work? If not, include information. If not, include references.
Answer: recommendation was followed. Reference was placed.
Page 4
Line 145: “G” is not how the relative centrifugal force is written.
Answer: recommendation was followed. Units were changed.
Line 154: what is the name of the “resultant blue compound”?
Answer: It has no name. word was changed
Line 164: The plants tissues extracts” does not read well.
Answer: recommendation was followed. word was changed.
Line 172: “The plants tissues extracts” does not read well.
Answer: recommendation was followed. word was changed.
Page 5
Line 193: consider “filtrate” instead of “filtrated”
Answer: recommendation was followed. word was changed.
Line 193: consider “transferred” instead of “transfer”
Answer: recommendation was followed. word was changed.
Line 198" Multiple (3?) spectrophotometers (different brands, manufacturers, models, etc.) are being mentioned throughout “Materials and Methods.” (line 198, line 306) Why is that?
Answer: Analysis were done in different places to validate results.
Line 209: “For plants tissues extracts preparation” does not read well. Also, is this your method? If not, you will need a reference.
Answer: recommendation was followed. Reference was placed. Word was changed.
Line 209: consider “was” instead of “were.”
Answer: recommendation was followed. word was changed.
Line 213: need to include unit after “0.45”. I assume this is micrometer.
Answer: recommendation was followed. Units were added.
Line 281: Beckman needs to be written with capital B.
Answer: recommendation was followed. word was changed. Line 218 not 281.
Line 224: need company information for the column.
Answer: recommendation was followed. Company information was placed.
Line 229-230: be consistent in how you write these organic acids. Either use capital letters or use lowercase letters (preferred).
Answer: recommendation was followed.
Page 6
Line 235: delete “was used” here.
Answer: recommendation was followed.
Line 237” add “was used after “214 nm.”
Answer: recommendation was followed.
Line 256: the target organisms are called “starter cultures” here. They are called “probiotics” or “probiotic cultures” or “cultures” at times. Consider consistency.
Answer: recommendation was followed.
Line 257” consider “their ability.”
Answer: recommendation was followed.
Line 264: why did you pick “8 hours” as incubation time?
Answer: Reference was placed.
After the meal is eaten, it takes about six to eight hours for food to pass through your stomach and small intestine where bile salts are mostly present.
Line 269: why did you use a single pH (pH 2.0 as the only pH adjustment? Would it not make sense to work with a range of pH values?
Answer: Reference was placed (standardized method).
The normal volume of the stomach fluid is 20 to 100 mL and the pH is acidic (1.5 to 3.5).
Line 279: the target organisms are called “starter cultures” here. They are called “probiotics” or “probiotic cultures” at times. Consider consistency.
Answer: recommendation was followed.
Line 279: the genera and species names need to be italicized.
Answer: recommendation was followed.
Page 7
Line 281: include details of the anaerobic incubation.
Answer: time and temperature were provided with no CO2.
Line 287: the target organisms are called “starter cultures” here. They are called “probiotics” or “probiotic cultures” or “cultures” at times. Consider consistency.
Answer: recommendation was followed.
Line 295: the genera and species names need to be italicized.
Answer: recommendation was followed.
Line 297: the genera and species names need to be italicized.
Answer: recommendation was followed.
Line 304: consider “to be used” instead of “to use”.
Answer: recommendation was followed.
Line 314: there is an issue with how the chemical was spelled and where the company name was Answer: the chemical was spelled and company information provided.
Line 323: What is “Streptococcus thermophilus agar”? There is no such medium! Did you mean M17?
Answer: recommendation was followed. Wording was changed.
Page 8
Line 332: what do you mean by “getting different dilution”?
Answer: Wording was deleted.
Line 334: consider “were” instead of “was.”
Line 337: what is “Lactobacillus bulgaricus agar”? There is no such medium! Did you mean Lactobacillus MRS Agar?
Answer: recommendation was followed. Wording was deleted.
Table 1: “different letters” in the table need to be in superscript.
Answer: recommendation was followed.
Table 1 results: What is the significance of your findings?
Answer: Pulp of the carao has the highest moisture (12.55± 0.44%), ash (6.45±0.15%), lipids (0.66±0.07%) and proteins (16.56±0.21%). Crust of the carao has the lowest moisture (10.23±0.38%) and protein (2.41±0.45%). Seed of the carao has the lowest lipids (0.15±0.09%) and ash (1.97±0.34%).
Table 2 result: What is the significance of your findings?
Answer: Seed of carao has the highest TPC (766.87±11.56 μg GAE/mL), TCC (7.43±0.31 mg Q/Ml) and antioxidant activity (40.76±1.87%). Crust of carao has the lowest TPC (245.55±10.48 μg GAE/mL)’ TCC (4.73±0.33 mg Q/mL) and antioxidant activity (22.88±2.84%).
Page 9
Line 371: use lower case “s” in “Seed.”
Answer: recommendation was followed.
Table 3 results: What is the significance of your findings?
Answer: Pulp had the highest amount of sucrose and fructose (9.07±0.78 and 3.76±0.06 g/100g, re-spectively) compared to seed and crust. Also, maltose was not detected in seed and crust significantly.
Line 383-387: Are you trying to compare your results to others’ here?
Answer: Yes.
Line 389: “the cell membrane is remarkably powerless to bile salts” is an odd expression.
Answer: recommendation was followed. Wording was changed.
Line 393: “a substantial decrease in ST counts”?
Answer: Wording was changed.
Page 10:
Line 403-404: what results?
Answer: Line 403-404 are describing the effects of bile salts in cell wall.
Figure 1: while the manuscript mentions “values with different letters are significantly different”, there are no letters anywhere. Also, the reviewer cannot differentiate the error bars for control, crust, pulp, seed.\
Answer: The phrase “values with different letters are significantly different” was deleted.
Line 408-409: what is the basis of the statement?
Answer: The sentence is a hypothesis. Sentence was rephrased
Page 11
Figure 2: while the manuscript mentions “values with different letters are significantly different”, there are no letters anywhere. Also, the reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Answer: The phrase “values with different letters are significantly different” was deleted. Statistical analysis of the main effects (ingredients vs control) is described in the result section with respective P values to indicate whether the effect was insignificant or not under an alpha value of 0.05.
Page 12:
Figure 4: while the manuscript mentions “values with different letters are significantly different”, there are no letters anywhere. Also, the reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Answer: The phrase “values with different letters are significantly different” was deleted. Statistical analysis of the main effects (ingredients vs control) is described in the result section with respective P values to indicate whether the effect was insignificant or not under an alpha value of 0.05.
Figure 5: The reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Answer: Statistical analysis of the main effects (ingredients vs control) is described in the result section with respective P values to indicate whether the effect was insignificant or not under an alpha value of 0.05.
Page 13
Figure 6: The reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Answer: Statistical analysis of the main effects (ingredients vs control) is described in the result section with respective P values to indicate whether the effect was insignificant or not under an alpha value of 0.05.
Page 14
Figure 7: The reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Answer: Statistical analysis of the main effects (ingredients vs control) is described in the result section with respective P values to indicate whether the effect was insignificant or not under an alpha value of 0.05.
Page 15:
Figure 9: The reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Answer: Statistical analysis of the main effects (ingredients vs control) is described in the result section with respective P values to indicate whether the effect was insignificant or not under an alpha value of 0.05.
Figure 10: The reviewer cannot differentiate the error bars for control, crust, pulp, seed.
Answer: Statistical analysis of the main effects (ingredients vs control) is described in the result section with respective P values to indicate whether the effect was insignificant or not under an alpha value of 0.05.
Page 16
Line 534: the genus and species names need to be in italics.
Answer: recommendation was followed.
Line 534-543: confusing paragraph that makes statements about proteolytic microorganisms in the large intestine and then proteolysis in fermented milk.
Answer: Paragraph was rephased.
Line 545-547: Not following the statement. Could you have not determined properties of these organisms in the absence of carao components?
Answer: Paragraph was rephased.
Line 547-550: what is the importance/significance (if any)?
Answer: Importance was added.
Reviewer 2 Report
Comments and Suggestions for AuthorsThis article: discusses the potential impacts of carao (Cassia grandis) on the viability and performance of Streptococcus thermophilus and Lactobacillus bulgaricus under various adverse conditions mimicking the digestive process. The study explores the influence of different parts of the carao plant on probiotic survival, considering factors such as bile tolerance, acid resistance, and protease activity.
Abstract: the abstract appears well-structured and comprehensive. It provides a clear overview of the experimental setup, including the conditions tested, the cultivation process, and the subsequent analyses of plant material. The inclusion of parameters such as total phenolic content, carotenoid content, antioxidant activity, and sugar and acid profiles adds depth to the study.
The introduction: provides a comprehensive overview of probiotics, emphasizing the importance of specific strains and quantities, as well as the need for validation through laboratory tests, animal trials, and human research, following the guidelines set by FAO/WHO. The surge in demand for functional foods containing lactic acid bacteria attributes is highlighted, particularly for their potential benefits in maintaining intestinal microbial balance and reinforcing mucosal defenses.
Materials and Methods: The research design presented in the materials and methods section appears to be thorough and well-detailed.
The results: presented results, particularly the varying effects of different carao tissues on probiotic survival in different tests, contribute to a nuanced understanding of the relationship between carao and probiotics.
Comments:
The introduction provides a substantial background on probiotics, their significance, and the potential benefits of lactic acid bacteria. It touches on the guidelines from FAO/WHO, emphasizes the importance of specific strains, and discusses the surge in demand for functional foods containing these microorganisms.
While the introduction references various concepts and studies, it lacks specific citations for some statements. For instance, phrases like "There is a proposal for the utilization of carao in nutritional, pharmaceutical, and medicinal contexts [10-12]" mention citations but do not provide the actual references. Including specific citations for such claims would enhance the credibility of the information presented.
To improve the section's scientific robustness, consider providing direct references for key statements, ensuring that readers can trace the information back to reputable sources. This not only strengthens the background but also helps readers delve deeper into the referenced literature if they wish to explore specific points further.
However, there are a few points to consider for improvement:
- Clarity on Defects: The request mentions identifying the defect, but the introduction does not explicitly present any obvious defects. It appears well-structured and informative. If there are specific criteria for a defect, please provide them for a more targeted evaluation.
- Citations: The introduction references various studies and guidelines (e.g., FAO/WHO), which is good. However, providing specific citations for these references would enhance the scientific credibility of the introduction.
While the materials and methods section is comprehensive, there are a few areas where clarity or additional information could enhance the understanding of the research design:
- Lyophilization Process: The lyophilization process is well-described, but it would be beneficial to include the reasons for choosing this specific method and how it contributes to the study. Additionally, details on why the temperature range of -73 to -76 °C and pressure of 0.1-0.3 Pa were chosen could provide more insight.
- Analysis Methods Repetition: Some details related to the analysis methods appear to be repeated, such as viability evaluation, lysozyme resistance assessment, and tolerance to gastric juices. Ensure that each repetition provides unique information or clarify the specific nuances being addressed in each step.
- Antioxidant Analysis Standards: While the procedure for antioxidant analysis is well-described, it would be helpful to mention the rationale behind using gallic acid as the reference standard and Trolox for antioxidant activity. Providing a brief explanation of why these standards were chosen can add context to the analysis.
- Total Carotenoids Content Calculation: In the total carotenoids content section, the equation for calculating total carotenoid content is mentioned (Equation 2), but there is no explanation or definition of the variables (A, V, E1%1cm, P). Including a brief explanation or a reference to where these values are defined could enhance clarity.
- HPLC Determination of Sugars and Organic Acids: While the HPLC methods are well-described, it might be useful to briefly explain why the specific sugars (fructose, maltose, glucose, and sucrose) and organic acids (Citric, tartaric, L-Malic, Quinic, and Succinic) were chosen for analysis. Providing context for the selection of these components would add depth to the experimental design.
- Bile Tolerance and Acid Tolerance Tests: The modification of the methods for bile tolerance and acid tolerance tests is mentioned, but the specific modifications are not detailed. Including a brief explanation of the modifications made to the original methods would help in understanding the rationale behind these changes.
Comments for author File: 
Comments.pdf
- # There are some sentences that may need revision to make them clearer and smoother. For example, in the sentence "Various tests will be conducted to evaluate the impact of different factors on the probiotic performance," you can use the word "different" only once to improve clarity.
- Some sentences need clarification or improvement for better understanding. For instance, in the sentence "It is also important to pay great attention to the dose of probiotic used," you can clarify why this is important, for example: "To ensure the desired results."
- In the sentence "The positive effect of probiotics depends on the specific strain and quantity," you can improve the wording for better flow, such as "The positive effect of probiotics depends on the bacterial strain and the quantity used."
- Ensure proper punctuation throughout the text. For example, in the benefits list, "strengthening the mucosal barrier, and assisting in restoring the disrupted intestinal microflora," use a comma between different elements instead of "and."
- Some expressions may need refinement to avoid long sentences and make the text more explicit.
Please review the language and ensure that all phrases and expressions clearly convey the presented scientific ideas.
Author Response
Reviewer 2
This article: discusses the potential impacts of carao (Cassia grandis) on the viability and performance of Streptococcus thermophilus and Lactobacillus bulgaricus under various adverse conditions mimicking the digestive process. The study explores the influence of different parts of the carao plant on probiotic survival, considering factors such as bile tolerance, acid resistance, and protease activity.
Abstract: the abstract appears well-structured and comprehensive. It provides a clear overview of the experimental setup, including the conditions tested, the cultivation process, and the subsequent analyses of plant material. The inclusion of parameters such as total phenolic content, carotenoid content, antioxidant activity, and sugar and acid profiles adds depth to the study.
The introduction: provides a comprehensive overview of probiotics, emphasizing the importance of specific strains and quantities, as well as the need for validation through laboratory tests, animal trials, and human research, following the guidelines set by FAO/WHO. The surge in demand for functional foods containing lactic acid bacteria attributes is highlighted, particularly for their potential benefits in maintaining intestinal microbial balance and reinforcing mucosal defenses.
Materials and Methods: The research design presented in the materials and methods section appears to be thorough and well-detailed.
The results: presented results, particularly the varying effects of different carao tissues on probiotic survival in different tests, contribute to a nuanced understanding of the relationship between carao and probiotics.
Comments:
The introduction provides a substantial background on probiotics, their significance, and the potential benefits of lactic acid bacteria. It touches on the guidelines from FAO/WHO, emphasizes the importance of specific strains, and discusses the surge in demand for functional foods containing these microorganisms.
While the introduction references various concepts and studies, it lacks specific citations for some statements. For instance, phrases like "There is a proposal for the utilization of carao in nutritional, pharmaceutical, and medicinal contexts [10-12]" mention citations but do not provide the actual references. Including specific citations for such claims would enhance the credibility of the information presented.
To improve the section's scientific robustness, consider providing direct references for key statements, ensuring that readers can trace the information back to reputable sources. This not only strengthens the background but also helps readers delve deeper into the referenced literature if they wish to explore specific points further.
However, there are a few points to consider for improvement:
- Clarity on Defects:The request mentions identifying the defect, but the introduction does not explicitly present any obvious defects. It appears well-structured and informative. If there are specific criteria for a defect, please provide them for a more targeted evaluation.
- Citations:The introduction references various studies and guidelines (e.g., FAO/WHO), which is good. However, providing specific citations for these references would enhance the scientific credibility of the introduction.
Answer: Introduction was improved.
While the materials and methods section is comprehensive, there are a few areas where clarity or additional information could enhance the understanding of the research design:
- Lyophilization Process:The lyophilization process is well-described, but it would be beneficial to include the reasons for choosing this specific method and how it contributes to the study. Additionally, details on why the temperature range of -73 to -76 °C and pressure of 0.1-0.3 Pa were chosen could provide more insight.
Answer: Reference was placed.
- Analysis Methods Repetition:Some details related to the analysis methods appear to be repeated, such as viability evaluation, lysozyme resistance assessment, and tolerance to gastric juices. Ensure that each repetition provides unique information or clarify the specific nuances being addressed in each step.
Answer: terms viability evaluation, lysozyme resistance assessment, and tolerance to gastric juices are consistent thought the materials and methods section to show clarity to editors and reviewers.
- Antioxidant Analysis Standards:While the procedure for antioxidant analysis is well-described, it would be helpful to mention the rationale behind using gallic acid as the reference standard and Trolox for antioxidant activity. Providing a brief explanation of why these standards were chosen can add context to the analysis.
Answer: Gallic acid as the reference standard for total polyphenols and Trolox for antioxidant activity were used because they are used in standard methods. Reference was placed.
- Total Carotenoids Content Calculation:In the total carotenoids content section, the equation for calculating total carotenoid content is mentioned (Equation 2), but there is no explanation or definition of the variables (A, V, E1%1cm, P). Including a brief explanation or a reference to where these values are defined could enhance clarity.
Answer: variables were define as suggested. Reference was placed.
- HPLC Determination of Sugars and Organic Acids:While the HPLC methods are well-described, it might be useful to briefly explain why the specific sugars (fructose, maltose, glucose, and sucrose) and organic acids (Citric, tartaric, L-Malic, Quinic, and Succinic) were chosen for analysis. Providing context for the selection of these components would add depth to the experimental design.
Answer: the specific sugars (fructose, maltose, glucose, and sucrose) and organic acids (Citric, tartaric, L-Malic, Quinic, and Succinic) were chosen because they are the most abundant in this type on plant materials. Furthermore, this compounds plant a crucial role on the growth (Sugars) and inhibition (Organic Acids) of the cultures studied.
- Bile Tolerance and Acid Tolerance Tests:The modification of the methods for bile tolerance and acid tolerance tests is mentioned, but the specific modifications are not detailed. Including a brief explanation of the modifications made to the original methods would help in understanding the rationale behind these changes.
Answer: modifications were mention. References was placed
Comments on the Quality of English Language
- # There are some sentences that may need revision to make them clearer and smoother. For example, in the sentence "Various tests will be conducted to evaluate the impact of different factors on the probiotic performance," you can use the word "different" only once to improve clarity.
Answer: Sentence was rephrased as suggested.
- Some sentences need clarification or improvement for better understanding. For instance, in the sentence "It is also important to pay great attention to the dose of probiotic used," you can clarify why this is important, for example: "To ensure the desired results."
Answer: Sentence was rephrased as suggested.
- In the sentence "The positive effect of probiotics depends on the specific strain and quantity," you can improve the wording for better flow, such as "The positive effect of probiotics depends on the bacterial strain and the quantity used."
Answer: Sentence was rephrased as suggested.
- Ensure proper punctuation throughout the text. For example, in the benefits list, "strengthening the mucosal barrier, and assisting in restoring the disrupted intestinal microflora," use a comma between different elements instead of "and."
Answer: proper punctuation was included.
- Some expressions may need refinement to avoid long sentences and make the text more explicit.
Answer: recommendation was followed.
Please review the language and ensure that all phrases and expressions clearly convey the presented scientific ideas.
Answer: recommendation was followed.
Reviewer 3 Report
Comments and Suggestions for AuthorsThe present study aimed to assess the potential impacts of carao on the viability and performance of Streptococcus thermophilus and Lactobacillus bulgaricus under various adverse conditions simulating the digestive process. While the study provides valuable insights, there are several areas that could be addressed for improvement:
The objectives and hypotheses of the study are not explicitly stated. Clearly defining these aspects would provide a better understanding of the study's goals.
few information about cara has been given in the introduciton
The results section could benefit from a clearer organization and presentation of findings. A more structured approach, such as using tables or figures, would aid in conveying the information more effectively.
Statisitical analysis are incomplete
The discussion lacks in-depth exploration of the implications of the results. Linking the findings back to the literature and discussing the broader implications for probiotics and their survival would enhance the paper's contribution
Ensuring a consistent style throughout the paper would enhance readability
Author Response
Reviewer 3
The present study aimed to assess the potential impacts of carao on the viability and performance of Streptococcus thermophilus and Lactobacillus bulgaricus under various adverse conditions simulating the digestive process. While the study provides valuable insights, there are several areas that could be addressed for improvement:
The objectives and hypotheses of the study are not explicitly stated. Clearly defining these aspects would provide a better understanding of the study's goals.
Answer: The objectives and hypotheses of the study were stated
about carao has been given in the introduction
Answer: more information about carao has been given in the introduciton
The results section could benefit from a clearer organization and presentation of findings. A more structured approach, such as using tables or figures, would aid in conveying the information more effectively.
Answer: results were presented describing statistical results. Then, the results were discussing and the implications were added.
Statisitical analysis are incomplete
Answer: Statistical analysis section was enforced.
The discussion lacks in-depth exploration of the implications of the results. Linking the findings back to the literature and discussing the broader implications for probiotics and their survival would enhance the paper's contribution
Answer: discussion was improved.
Ensuring a consistent style throughout the paper would enhance readability
Answer: a consistent style throughout the paper was applied.
Round 2
Reviewer 2 Report
Comments and Suggestions for AuthorsAccepted
Reviewer 3 Report
Comments and Suggestions for AuthorsNo further comments
