Fatty Acid Profile, Mineral Composition, and Health Implications of Consuming Dried Sago Grubs (Rhynchophorus ferrugineus)

Round 1

Reviewer 1 Report

The article needs some revision to be suitable for publication

-The abstract should be informative and include the main findings.

-Language should be revised. 

-Figures should be represented in higher resolution

 

- The introduction should be enriched with recent references (2016-2022)

Author Response

Response to Reviewer 1 Comments

 

We thank the Reviewer for their comments about our manuscript.

 

1. The abstract should be informative and include the main findings.

We believe our abstract is succinctly informative, in providing key findings that capture the gist of the study. The abstract is written in a way to stand alone in bibliometrics indexing websites, to encourage readers to explore the whole manuscript.

Abstract: A comprehensive evaluation of fatty acid and mineral composition of Rhynchophorus ferrugineus (sago grub powder, SGP) pre-treated to meet New Zealand biosecurity and import requirements was investigated. Palmitic acid (42.5% FA), oleic acid (39.0% FA) and linoleic acid (1.02% FA), were the most abundant saturated fatty acid, monounsaturated fatty acid, and polyunsaturated fatty acid, respectively. Lipid nutritional indices of SGP fats were ω6/ω3 (2.17), hypocholesterolemic/hypercholesterolemic acid ratio (0.88), atherogenicity index (1.01), thrombogenicity index (1.65), and health-promoting index (0.99). The mineral analysis profile of the SGP found 11 essential, 29 non-essential minerals, and 4 heavy metals. The most abundant minerals were potassium (1657 mg/kg DW), magnesium (805.3 mg/kg DW), iron (23 mg/kg DW) and manganese (8.8 mg/kg DW). Heavy metals such as arsenic (0.17 mg/kg DW), cadmium (0.04 mg/kg DW), lead (0.56 mg/kg DW), and vanadium (0.01 mg/kg DW) were detectable in SGP, but below acceptable toxicity limits. These findings indicate that SGP pre-treated for export is safe for consumption and contain appreciable nutrients. However, the elevated saturated fatty acid and the low polyunsaturated fatty acid content of SGP should be a nutritional consideration.

 

2. Language should be revised.

The manuscript has been reviewed by a fluent English speaker.

3. Figures should be represented in higher resolution 

There is only one figure in the manuscript, and was provided at 300dpi, which was uploaded separately.

4. The introduction should be enriched with recent references (2016-2022)-

We aimed to include all papers relevant to the species (Rhynchophorus ferrugineus), and to the best of our knowledge consider that our literature review is appropriately inclusive. Six out of the eight references cited in the Introduction are recent.

1. Chaijan, M.; Panpipat, W. Techno-biofunctional aspect of seasoning powder from farm-raised sago palm weevil (Rhynchophorus ferrugineus) larvae. Journal of Insects as Food and Feed 2021, 7, 187-195.
2. Hanboonsong, Y.; Jamjanya, T.; Durst, P.B. Six-legged livestock: edible insect farming, collection and marketing in Thailand. RAP publication 2013, 3.
3. Chaijan, M.; Chumthong, K.; Kongchoosi, N.; Chinarak, K.; Panya, A.; Phonsatta, N.; Cheong, L.; Panpipat, W. Characterisation of pH-shift-produced protein isolates from sago palm weevil (Rhynchophorus ferrugineus) larvae. Journal of Insects as Food and Feed 2022, 8, 313-324.
4. Chinarak, K.; Chaijan, M.; Panpipat, W. Farm-raised sago palm weevil (Rhynchophorus ferrugineus) larvae: Potential and challenges for promising source of nutrients. Journal of Food Composition and Analysis 2020, 92, 103542, doi:https://doi.org/10.1016/j.jfca.2020.103542.
5. Leatemia, J.; Patty, J.; Masauna, E.; Noya, S.; Hasinu, J. Utilization of sago grub (Rhynchophorus ferrugineus Olivier)(Coleoptera: Curculionidae) as an alternative source of protein. In Proceedings of the IOP Conference Series: Earth and Environmental Science, 2021; p. 012028.
6. Nirmala, I.R.; Pramono, M.S. Sago worms as a nutritious traditional and alternative food for rural children in Southeast Sulawesi, Indonesia. Asia Pacific Journal of Clinical Nutrition 2017, 26.
7. Yhoung-Aree, J. Edible insects in Thailand: nutritional values and health concerns. Edible forest insects 2010, 201-216.
8. Bang, A.; Courchamp, F. Industrial rearing of edible insects could be a major source of new biological invasions. Ecology Letters 2021, 24, 393-397.

 

Author Response File: Author Response.docx

Reviewer 2 Report

In my opinion, the manuscript entitled ,,Fatty acid profile, mineral composition, and health implications of consuming dried sago grubs (Rhynchophorus ferrugineus) is a good one. The introduction is quite good and the results are discussed and compared with the current state of the art. I have some comments, as follows:

 

1. Materials and methods should be better explained and detailed. For instance, which was the protein conversion factor, considering that for insect the factor is different (see literature such as Boulos et al., https://doi.org/10.3389/fnut.2020.00089)?

2. Please better described the method used for fatty acid analysis. For instance, which was the injection temperature and the temperature of the ion source and the GC-MS interface? How fatty acids were expressed?

3. Which were the detection limits of minerals? Which were the working conditions such as air:acetylene ratio, nebulizer uptake rate? Please better explained the method.

4. Are the SGP larvae safe from the microbiological point of view? Even they were oven dried and bolied, did authors made some microbiological analysis such as Enterobacteriaceae or even Escherichia coli?

Comments for author File: Comments.docx

Author Response

Response to Reviewer 2 Comments

In my opinion, the manuscript entitled ,,Fatty acid profile, mineral composition, and health implications of consuming dried sago grubs (Rhynchophorus ferrugineus) is a good one. The introduction is quite good and the results are discussed and compared with the current state of the art.

 

We thank the Reviewer for their positive comments about our manuscript.

 

I have some comments, as follows:

1. Materials and methods should be better explained and detailed.

For instance, which was the protein conversion factor, considering that for insect the factor is different (see literature such as Boulos et al., https://doi.org/10.3389/fnut.2020.00089)?

 

The Materials and Methods section has been reviewed. We are aware of the study by Boulos et al. and in fact we used a conversion factor of 5.6 which was reported by Chaijan et al., 2022; Chinarak et al., 2021. This has been added to the manuscript.

 

Chaijan, M., Chumthong, K., Kongchoosi, N., Chinarak, K., Panya, A., Phonsatta, N., Cheong, L., Panpipat, W., 2022. Characterisation of pH-shift-produced protein isolates from sago palm weevil (Rhynchophorus ferrugineus) larvae. Journal of Insects as Food and Feed 8, 313-324.

 

Chinarak, K., Panpipat, W., Summpunn, P., Panya, A., Phonsatta, N., Cheong, L.-Z., Chaijan, M., 2021. Insights into the effects of dietary supplements on the nutritional composition and growth performance of sago palm weevil (Rhynchophorus ferrugineus) larvae. Food Chemistry 363, 130279.

2. Please better described the method used for fatty acid analysis. For instance, which was the injection temperature and the temperature of the ion source and the GC-MS interface? How fatty acids were expressed?

Done. As per the Reviewer’s comments, more details have been added to the manuscript.

3. Which were the detection limits of minerals?

The experimental % recovery, detection limits and limit of quantification for SGP are reported in Table 2 and Table S1, respectively.

 

4. Which were the working conditions such as air: acetylene ratio, nebulizer uptake rate? Please better explained the method.

 

The method used in this study is ICP-MS, a type of mass-spectrometry that uses inductively coupled plasma to ionise, more details have been added to the manuscript.

 

5. Are the SGP larvae safe from the microbiological point of view? Even they were oven dried and boiled, did authors made some microbiological analysis such as Enterobacteriaceae or even Escherichia coli?

We understand the Reviewer’s comment. We did not carry out any microbiological analyses since the sago grub product has been treated using stringent conditions prescribed by New Zealand Biosecurity Regulatory Authorities (https://www.mpi.govt.nz/biosecurity/) that deemed the product safe to be consumed without any concern for microbial safety.

 

Author Response File: Author Response.docx

Reviewer 3 Report

The MS “Fatty acid profile, mineral composition, and health implications of consuming dried sago grubs (Rhynchophorus ferrugineus)” by Ruchita Rao Kavle, Ellenna Tamsin Maree Pritchard, Alan Carne, Alaa El-Din Ahmed Bekhit, Dominic Agyei.

  

The authors of this manuscript investigated the FA and the mineral composition and content in pre-treated sago grub larvae. The authors measured some indicators (FA ratios) that are used to assess the quality of a product for human health and compared the content of elements with the minimum and maximum values allowed in human food. However, despite the fact that many FA indicators showed the low value of this product for human health, the authors focused on its benefits, which is unacceptable and indicates the authors' biased attitude towards their results. This manuscript needs to be substantially revised, showing the true quality of these insects, and not trying to veil it with beautiful phrases like "These findings indicate that SGP pre-treated for export are safe for consumption and contain appreciable nutrients that are essential for health".

According to Instructions for authors (Applied Sciences), the manuscript have to include a Result section and a Discussion section, but there is no separation between these two sections in the manuscript. I believe that the Editor should evaluate whether this is acceptable in the journal.

  

Special comments.

 

Abstract

This section needs a major revision. The values of the indicators do not need to be given here. Authors should write what follows from all the measurements they have made. Indicate by what indicators these larvae are harmful to human health, and by what indicators they are useful.

 

Introduction

The introduction contains only general information about these larvae, including their anatomical structure, but no more important information about the value of the larvae. It is necessary to add a description that the larvae are considered as a source of protein (primarily), their value as a food product.

 

Methods

the type of detector of the chromatograph (Agilent 6890 N, CA, USA) should be written.

 

Results

1.     table 1. arachidonic acid is 20:4n-6, but arachidic acid is 20:0.

It is not clear why the common names of the FAs in the table are given. This makes the table bulky and inconvenient. Column 2 should be deleted, and the abbreviated names should be written in the note to the table below it. The letter C in the names of FAs is not needed, but cis should be written in the beginning of the name of FAs. Authors should reduce the use of common names in the text of the manuscript.

2.     The abbreviated name is written at the first mention, the authors should check this.

3.     As a rule, data from tables is not duplicated in the text of the article.

People are deficient in omega-3 PUFAs, not omega-6 and omega-9. The studied larvae contained an insignificant amount of omega-3 PUFAs, moreover, only 18:3n-3. The authors do not comment on this. Referring to the WHO, the authors do not indicate, for example, that myristic and palmitic acids increase risk of CVD (the studied sago grub had 42% of palmitic acid!) and “Diets should provide an adequate intake of PUFAs, i.e. in the range 6-10% of daily energy intake. There should also be an optimal balance

between intake of n-6 PUFAs and n-3 PUFAs, i.e. 5-8% and 1-2% of daily energy intake, respectively. The ratio is not just <10 but 2.5-8.

4.     The abbreviations UFA, MUFA and PUFA are not introduced, but are used by the authors.

5.     Authors should check the literature citations, as I found an error: the authors cite source 26, but the WHO citation is 25.

6.     Line 163. Authors should check the value 0.01, maybe it should be 0.1

7.     The authors focus only on the benefits, but do not write about the harm when they analyze PUFA:SFA, IT, IA, HA, and HPI, but almost all of them indicated low quality of such food.

8.     Line 232-236. The authors evaluate the product according to all criteria, but when evaluating Ca:P they write that the sago grubs could be better according to this criterion, but the processing could lead to such a result. This is the wrong approach, if the authors evaluate the product, then they need to write about the product, and not about how it could be.

9.     Line 331-333. I don't understand why this sentence is needed?

 

Conclusion

This section needs a major revision. Authors should write what follows from all the measurements they have made, but not just list the results. Moreover, data that are presented as beneficial are not so on closer examination.

 

Author Response

Response to Reviewer 3 Comments

The MS “Fatty acid profile, mineral composition, and health implications of consuming dried sago grubs (Rhynchophorus ferrugineus)” by Ruchita Rao Kavle, Ellenna Tamsin Maree Pritchard, Alan Carne, Alaa El-Din Ahmed Bekhit, Dominic Agyei.

The authors of this manuscript investigated the FA and the mineral composition and content in pre-treated sago grub larvae. The authors measured some indicators (FA ratios) that are used to assess the quality of a product for human health and compared the content of elements with the minimum and maximum values allowed in human food.

We thank the Reviewer for their comments.

However, despite the fact that many FA indicators showed the low value of this product for human health, the authors focused on its benefits, which is unacceptable and indicates the authors' biased attitude towards their results.

1. This manuscript needs to be substantially revised, showing the true quality of these insects, and not trying to veil it with beautiful phrases like "These findings indicate that SGP pre-treated for export are safe for consumption and contain appreciable nutrients that are essential for health

We understand the Reviewer’s comment. Part of the reason for the study was that there is a lack of information in the literature in relation to the lipid nutritional indices. Hence, the results obtained in the present study were compared to other information in the literature on other food products.

However, based on the Reviewer’s comment, the comparison was mostly limited to other insects, to allow for a comparable assessment. That said, in terms of the possibility of edible insects as an alternative food source, a comparison with other foods would be of interest to readers.

We have modified statements in the Abstract and Conclusion sections accordingly. 

2. According to Instructions for authors (Applied Sciences), the manuscript have to include a Result section and a Discussion section, but there is no separation between these two sections in the manuscript. I believe that the Editor should evaluate whether this is acceptable in the journal.

To minimise repetition we considered that the best way to report our analyses in the present study was to merge the Results and Discussion. The following links are to recently published articles in Applied Sciences: https://www.mdpi.com/2076-3417/12/24/12695; https://www.mdpi.com/2076-3417/12/19/10043, in which the Results and Discussion are combined.

 

Special comments.

Abstract

1. This section needs a major revision. The values of the indicators do not need to be given here.

We are of the opinion that the values should be retained so that the abstract provides a succinct summary of the key results from the study for the benefit of the reader.

2. Authors should write what follows from all the measurements they have made.

These findings indicate that SGP pre-treated for export contain appreciable nutrients at levels that could be considered safe at moderate for consumption. Lines 23-24 have been included to communicate this in the abstract.

3. Indicate by what indicators these larvae are harmful to human health, and by what indicators they are useful.

We have included a statement in the Abstract to the effect that SGP pre-treated for export are safe for consumption and contain appreciable nutrients. However, the elevated saturated fatty acid content and the low polyunsaturated fatty acid content of SGP should be a nutritional consideration. 

Introduction

1. The introduction contains only general information about these larvae, including their anatomical structure, but no more important information about the value of the larvae. It is necessary to add a description that the larvae are considered as a source of protein (primarily), their value as a food product.

We thank the Reviewer for their comments. We have re-cited in the Introduction and indicated the protein values of sago grubs (Line 35-38).

 

Methods

1. the type of detector of the chromatograph (Agilent 6890 N, CA, USA) should be written.

The wording has been changed to the following:

According to Kavle, et al. [9] the FAMEs were analysed using a gas chromatography device outfitted with a flame ionisation detector (Agilent 6890 N, CA, USA) and an autosampler (Agilent 7683, China) (2021). In order to conduct the analysis, 1 μL of the FAME samples were injected into a GC-FID device. The ionisation detector port was set at 250°C, and the injector detector ports were configured with a 10:1 inlet split ratio. On a BPX-70 silica column (Phenomenex, Torrance, California, United States) (length: 50 m; inner diameter: 0.32 mm; film thickness: 0.25 m), the FAME was successfully separated. The temperature program for the column oven was as follows: 40°C for three minutes, then 10°C/min up to 225°C at a rate of 3°C/min, and finally 10°C/min to a final temperature of 250°C. By comparing retention times with recognised FAME standards (FAMQ-005) from NuCheck Prep, Elysian, Minnesota, and Sigma, St. Louis, Missouri, it was possible to identify the FAME chromatogram peaks. The percent peak area data from the GC data processing software were used to determine the percentage composition of each detected FAME in the sample. Using an internal standard and theoretical relative FID response corrections, the FAME values from the GC-FID were converted to free fatty acid (FFA). The values were expressed as % fatty acid and g fatty acid / 100 g dried SGP insect material.

 

Results

1. table 1. arachidonic acid is 20:4n-6, but arachidic acid is 20:0.

We thank the Reviewer for their comment. This correction has been made.

2. It is not clear why the common names of the FAs in the table are given. This makes the table bulky and inconvenient. Column 2 should be deleted, and the abbreviated names should be written in the note to the table below it.

We thank the Reviewer for their comment. We consider that it is appropriate to retain the general names in Table 1 and main text for the benefit of a generalist reader.

3. The letter C in the names of FAs is not needed, but cis should be written in the beginning of the name of FAs. Authors should reduce the use of common names in the text of the manuscript.

We have reconsidered the Reviewer’s comment and have reorganized Table 1 and removed the ‘C’. However, we are of the opinion that it is useful for readers to retain the common name and details of the fatty acids.

4. The abbreviated name is written at the first mention, the authors should check this.

We have checked the manuscript to make sure that the full name occurs at first use and then introduction of the abbreviation.

5. As a rule, data from tables is not duplicated in the text of the article.

We understand the Reviewer’s comment, but we consider that reiteration of the values in the main text allows for efficient communication of information on reading the manuscript.

People are deficient in omega-3 PUFAs, not omega-6 and omega-9. The studied larvae contained an insignificant amount of omega-3 PUFAs, moreover, only 18:3n-3. The authors do not comment on this. Referring to the WHO, the authors do not indicate, for example, that myristic and palmitic acids increase risk of CVD (the studied sago grub had 42% of palmitic acid!) and “Diets should provide an adequate intake of PUFAs, i.e. in the range 6-10% of daily energy intake. There should also be an optimal balance between intake of n-6 PUFAs and n-3 PUFAs, i.e. 5-8% and 1-2% of daily energy intake, respectively. The ratio is not just <10 but 2.5-8.

We thank the Reviewer for this comment and agree that it is omega-3 PUFA that most people are deficient in. The sago larvae SGP indeed contained a relatively small amount of omega-3 PUFA, but, being an insect (rather than a marine) material, we did not expect SGP to contain a substantial amount of omega-3 PUFA. It is also correct that SGP was found to have relatively high levels of palmitic and myristic acids. The role of these fatty acids in the aetiology of CVD is implicated in the index of thrombogenicity (IT). The Reviewer will observe that the formula for IT takes into consideration the levels of myristic and palmitic acids. We discussed in the manuscript the IT of SGP by saying that:

The index of thrombogenicity (IT) measures the risk of clot formation in blood vessels based on the net contribution of prothrombic FAs and anti-prothrombic FAs [12]. In the present study, the IT value for SGP (1.65) was comparable to that of the medium size Huhu grub (1.63) [11]. However, these values are higher than Acheta domestica (1.25) [34], locusts (1.51) [35], R. ferrugineus (1.35) [17], and plant oil (soybean oil, 0.4) [36]. The lower the IT value, the better the nutritional quality and reduced risk of developing coronary heart diseases, although no specific values are recommended for IA or IT [26].

Again, we agree with the Reviewer’s comment about omega-6 and omega-3 and PUFA:SFA ratios, and have discussed the importance of omega-6 and omega-3 and PUFA:SFA ratios in Sections 3.2.1 and 3.2.2.

6. The abbreviations UFA, MUFA and PUFA are not introduced, but are used by the authors.

Done. Line 106-111.

7.  Authors should check the literature citations, as I found an error: the authors cite source 26, but the WHO citation is 25.

This has been fixed.

 

8. Line 163. Authors should check the value 0.01, maybe it should be 0.1

We have double checked the value. The below link indicates that 0.01 is correct.

https://www.sciencedirect.com/science/article/pii/S0308814622013164

 

9. The authors focus only on the benefits, but do not write about the harm when they analyze PUFA: SFA, IT, IA, HA, and HPI, but almost all of them indicated low quality of such food.

We understand the Reviewer’s comment. Part of the reason for the study reported in the manuscript is that there is a lack of information in the literature in relation to the lipid nutritional indices. Hence, the results obtained in the present study were compared to information in the literature on other food products.

However, based on the Reviewer’s comment, the comparison was mostly limited to other insects, to allow for a more directly comparable assessment. That said, in terms of the possibility of edible insects as an alternative food source, a comparison with other foods would be of interest to readers.

Hence, we have modified statements in the abstract and conclusion accordingly. 

10. Line 232-236. The authors evaluate the product according to all criteria, but when evaluating Ca:P they write that the sago grubs could be better according to this criterion, but the processing could lead to such a result. This is the wrong approach, if the authors evaluate the product, then they need to write about the product, and not about how it could be.

We have reconsidered that section of the text and have modified the text accordingly.

 

11. Line 331-333. I don't understand why this sentence is needed.

We have reviewed Lines 331-333 in the original submitted manuscript and we don’t understand the Reviewer’s comment.

 

Conclusion

1. This section needs a major revision. Authors should write what follows from all the measurements they have made, but not just list the results. Moreover, data that are presented as beneficial are not so on closer examination.

We have reconsidered the text in the Conclusion section and have re-drafted accordingly.

 

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

 Accept in present form

Author Response

We thank the Reviewer for their time to review our manuscript and constructive comments.

Reviewer 2 Report

In my opinion, the authors highly improved the quality of the paper, and I believe that now it can be published in the present form.

Author Response

We thank the Reviewer for their time to review our manuscript and constructive comments.

Reviewer 3 Report

1.       I still recommend the Authors to improve the Introduction

2.       Line 111 – ω is described incorrectly. It’s the position of the first double bond from methyl end of the molecule. Moreover, in this MS additionally to omega-3/6 there are omega-9 and omega-7.

3.       Authors used ω (C20: 1cω9) and n (e.g., 0.5 ×Σn-6 PUFA) in the MS. I recommend to be consistent and use one style. I also recommend to check style in the text (e.g, 18:2 ω-6, C20: 1cω9, ω6: ω3, ω-6/ ω-3… are of different style).

4.       The Authors wrote “The recommended intake ratio for these minerals ranges from 1:1 to 1.5:1 (Ca:P) [47]” It means that the optimal ratio is 1-1.5. But then the Authors wrote “The Ca:P ratio in the SGP samples was 0.67, which is an optimal ratio [13], indicating that the treated SGP is a source of Ca and P.” But 0.67<1-1.5. So, 0.67 is not an optimal ratio.

5.       The Authors wrote “the maximum allowed level of lead in plant foods, such as cereals, legumes and pulses, is 0.20 mg/kg, and the maximum permitted level in meat is 0.10 mg/kg [60].” But then they wrote “The lead content in SGP (0.56 mg/kg DW) was within the EFSA. So, everyone can see that 0.56>0.1 and >0.2. I suggest to write EFSA limits that I believe do not contradict the data obtained by the Authors.

6.       I still recommend the Authors to add in the Conclusion and in the Abstract indicators for which the larvae do not correspond to a healthy food.

Best wishes

Author Response

We thank the Reviewer for their comments.

1. I still recommend the Authors to improve the Introduction

We have add more information in the Introduction.

2. Line 111 – ω is described incorrectly. It’s the position of the first double bond from methyl end of the molecule. Moreover, in this MS additionally to omega-3/6 there are omega-9 and omega-7.

This has now been changed to:  “ω = Omega which refers to the first double bond from the methyl end of the fatty acid”

3. Authors used ω (C20: 1cω9) and n (e.g., 0.5 ×Σn-6 PUFA) in the MS. I recommend to be consistent and use one style. I also recommend checking style in the text (e.g, 18:2 ω-6, C20: 1cω9, ω6: ω3, ω-6/ ω-3… are of different style).

We have reviewed all of this terminology in the manuscript and now all ω6: ω3 has been changed to ω-6/ ω-3, and all 18:2 ω-6 has been changed to 18:2 cω-6.

4. The Authors wrote “The recommended intake ratio for these minerals ranges from 1:1 to 1.5:1 (Ca:P) [47]” It means that the optimal ratio is 1-1.5. But then the Authors wrote “The Ca:P ratio in the SGP samples was 0.67, which is an optimal ratio [13], indicating that the treated SGP is a source of Ca and P.” But 0.67<1-1.5. So, 0.67 is not an optimal ratio.

We thank the Reviewer for this comment. We have consulted the literature in relation to the reported optimum Ca:P values necessary for human bone health - and have revised the discussion and calculation on this point. We have also mentioned the nutritional and health implication that SGP is not an ideal source of Ca.  Please see below:

“The relatively high P content (2950 mg/kg DW) in SGP resulted in a Ca:P ratio of 0.16:1, which is lower than the recommended intake ratio of 1.3:1 [56], indicating that the treated SGP might not be an ideal source of well-balanced Ca and P. Therefore, SGP must be consumed as part of a well-balanced diet, with other ingredients that can supply Ca and P, thus improving the Ca:P ratio to levels needed for optimum bone health.”

5. The Authors wrote “the maximum ALLOWED LEVEL of lead in plant foods, such as cereals, legumes and pulses, is 0.20 mg/kg, and the MAXIMUM PERMITTED level in meat is 0.10 mg/kg [60].” But then they wrote “The lead content in SGP (0.56 mg/kg DW) was within the EFSA. So, everyone can see that 0.56>0.1 and >0.2.

I suggest to write EFSA limits that I believe do not contradict the data obtained by the Authors.

We thank the Reviewer for this comment.  We have included the EFSA limit for lead which is 10mg/kg for feed material (as below):

“According to the EFSA, the maximum allowed level of lead in plant foods such as cereals, legumes, and pulses is 0.20 mg/kg fresh weight, FW, and the maximum permitted level in meat is 0.10 mg/kg FW [67]. The maximum allowed level for lead content in insect materials destined for use in feed is 10 mg/kg DW [68], showing that SGP was within the maximum set limit.”

https://efsa.onlinelibrary.wiley.com/doi/pdf/10.2903/j.efsa.2010.1570]

6. I still recommend the Authors to add in the Conclusion and in the Abstract indicators for which the larvae do not correspond to a healthy food.

This has been addressed in the Abstract and Conclusion.