Taro in West Africa: Status, Challenges, and Opportunities

Round 1

Reviewer 1 Report

The review is a valuable contribution for research on taro on Africa. It covers a good range of literature on taro in Africa, or relevant to taro in Africa.

Relevant literature has been cited to support most statements, but in some cases a cited source does not seem likely to be a primary source, or the most significant source. For example, to highlight the ornamental uses or taro, it might be good to also cite Cho (2004) on breeding ornamental taros.

Cho, J. (2004). Breeding Hawaiian Taros for the Future. Third Taro Symposium. 21-23 May 2003, Tanoa International Hotel, Nadi, Fiji Islands. L. Guarino, M. Taylor and T. Osborn. Suva, Secretariat of the Pacific Community: 192-196.

Also, it seems that too much text (multiple sentences in some cases) have been copied and pasted from cited sources without such text being identified as direct quotation. One consequence of this is that some of the content of cited sources has not been fully analysed and related to the situation in Africa. A review is not just a compilation of text from cited sources, but an interpretation of the cited sources, with  direct quotations made (and marked as quotations) when needed.

Botanical details should presented with correct use of botanical terms. Photo legends for plants should describe location, habitat, photographer, date, specific identification of the plant (e.g. cultivar name, or "wild"). Figures 3 & 4 appear to be the same plant or patch of plants, and are best combined as parts of one figure. The term "inflorescence" should be used, as the photo shows an entire inflorescence, with spathe and spadix. The flowers are not visible as they are enclosed in the lower (green) spathal tube.

Figures 5-6 are also poorly labelled (source not identified, etc.) and are obviously distorted as a result of the angle of view (Fig. 5) and digital compression (both).

There is also a structural problem with "taro breeding' reviewed in two different sections (8.3 and 10) leading to some dupication of content.

The annotated text attached has many other points that the author can consider in order to make this paper more focused, useful and easier to read.

To revise your text, I highly recommend making your own notes on a printed full copy, and reading each sentence out loud to yourself or another person, rather working within the limits of a computer screen.

Comments for author File: Comments.pdf

Author Response

The authors would like to thank the reviewer for all the suggestions and comments on the manuscript which provided the opportunity to revise the manuscript by incorporating the changes and improving the quality of the manuscript.

1. The review is a valuable contribution for research on taro on Africa. It covers a good range of literature on taro in Africa, or relevant to taro in Africa.

Relevant literature has been cited to support most statements, but in some cases a cited source does not seem likely to be a primary source, or the most significant source. For example, to highlight the ornamental uses or taro, it might be good to also cite Cho (2004) on breeding ornamental taros.

Cho, J. (2004). Breeding Hawaiian Taros for the Future. Third Taro Symposium. 21-23 May 2003, Tanoa International Hotel, Nadi, Fiji Islands. L. Guarino, M. Taylor and T. Osborn. Suva, Secretariat of the Pacific Community: 192-196.

Response: We appreciate the observation of the reviewer and made necessary changes for references using most relevant ones in the text and also made necessary changes in the Reference section. We have also included the reference of Cho et al (2004) to reflect the breeding efforts in Hawaii.

2. Also, it seems that too much text (multiple sentences in some cases) have been copied and pasted from cited sources without such text being identified as direct quotation. One consequence of this is that some of the content of cited sources has not been fully analysed and related to the situation in Africa. A review is not just a compilation of text from cited sources, but an interpretation of the cited sources, with  direct quotations made (and marked as quotations) when needed.

Response: We fully agree with this observation of the reviewer. We have now gone through the manuscript critically and tried to analyze each and every section to ensure that the information taken from the references are well organized and valuable information is presented in our own language.  

3. Botanical details should presented with correct use of botanical terms. Photo legends for plants should describe location, habitat, photographer, date, specific identification of the plant (e.g. cultivar name, or "wild"). Figures 3 & 4 appear to be the same plant or patch of plants, and are best combined as parts of one figure. The term "inflorescence" should be used, as the photo shows an entire inflorescence, with spathe and spadix. The flowers are not visible as they are enclosed in the lower (green) spathal tube.

Figures 5-6 are also poorly labelled (source not identified, etc.) and are obviously distorted as a result of the angle of view (Fig. 5) and digital compression (both).

Response: We thank the reviewer for this critical observation, and accordingly we have deleted Figure 1 and provided all necessary information on the remaining figures. We also merged both Figure 5 and 6 into one figure. We also tried to improve the quality of the figures so that they don’t look distorted and compressed. 

4. There is also a structural problem with "taro breeding' reviewed in two different sections (8.3 and 10) leading to some dupication of content.

The annotated text attached has many other points that the author can consider in order to make this paper more focused, useful and easier to read.

Response: We fully agree with this observation of the reviewer, and merged the two sections (8.3 and 10). We also revised the section to ensure that there is no duplication of information. We also tried to add more information so that the manuscript covers useful information on taro situation in Africa.

5. To revise your text, I highly recommend making your own notes on a printed full copy, and reading each sentence out loud to yourself or another person, rather working within the limits of a computer screen.

Response: We appreciate this valuable suggestion of the reviewer, each of us read the manuscript critically and made necessary changes.

Relevant Comments/Suggestions from the text: Below we have tried to respond to most of the comments/suggestions on the manuscript, while we have addressed to all the observations of the reviewer in the revised version of the manuscript.

Line 29-30: 3, 4 seem weak secondary sources wrt to global production

Response: We have now changed these two references using direct sources.

Line 59: 22 is out of date source and not primarily, on topic cf  Ahmed, I., P. J. Lockhart, E. M. G. Agoo, K. W. Naing, D. V. Nguyen, D. K. Medhi and P. J. Matthews (2020). "Evolutionary origins of taro (Colocasia esculenta) in Southeast Asia." Ecology and Evolution 10: 13530–13543.

Response: We agree with the suggestion of the reviewer, and changed the reference accordingly.

Line 66: “Lebot” is not proper citation;  cite Ivancic, A. and V. Lebot (2000). The genetics and breeding of taro. Monpellier, CIRAD. for description of insect pollination and natural outcrossing by taro. “Hybridisation” refers to crossing between very ditinct parents or species; not all breeding produces hybrids. Check use of terms.

Response: We thank the reviewer for this suggestion, and changed the reference in the text as well as reference section.

Line 78: 22 may have written these sentences, but other papers by same author are better sources; avoid direct copy and paste of text, or use quotation marks

Response: We appreciate the suggestion of the reviewer and included other recent sources by the same author.

Line 143: Fig. 1 is OK but Grimaldi, I. M. (2016) can be cited for more complete map of distribution in Africa: (2016). Taro across the oceans, journeys of one of our oldest crops. News from the Past, Progress in African Archaeobotany. Proceedings of the 7th International Workshop on African Archaeobotany in Vienna, 2 – 5 July 2012. U. Thanheiser. Groningen, Barkhuis: 67-81.

Response: We appreciate the observation of the reviewer, and we have included the description of Grimaldi (2016) in the text, however, we feel that the current Figure 1 represents the most recent taro production areas in Africa from 2016-2020. Therefore, we didn’t remove this figure and retained it in the text. We hope this will be agreed by the reviewer.

Line 232 – 245: This entire paragraph is lacking cited sources; Is this all copied from 42?  These are Pacific diseases; please relate to Africa…e.g is there a threat of introduction?

Response: We appreciate the observation of the reviewer, we have reviewed and revised the entire paragraph. We have also added different references and added information on diseases from Africa. We also deleted the diseases from the Pacific which may not have relevance to Africa. We also added information on the necessity for pre-emptive breeding and preparedness in the region in relation to climate change and introduction of planting materials.

Line 254-258: Incorrect description of inforescence. The infl. is composed of an outer spathe and inner spadix borne on a peduncle. The inflorescence as a whole is protogynous because the female flowers (on lower spadix) mature before the male flowers (upper spadix) .average age at maturity (define maturity in new sentence preceding: e.g. for production purposes, the crop is considere mature when the fmother crom is fully expanded..).

Response: We fully agree with your suggestions and we have made necessary changes in the text.

Line 279-296: Cite relevant sources more frequently in relation to specific areas of information, not just at the end of a paragraph that covers multiple topics. 63, 66 may be secondary sources in relation to ornamental taros

Response: We appreciate the suggestion of the reviewer and we have made necessary changes in the text.

Line 333-359: Summarise more briefly, then note the connections made with Africa (reports of introduction of resistant breeding stocks to Africa from the Pacific).

Response: We have revised the entire section and made connections with taro germplasm collection and characterization in relation to Africa, as well as introductions from other regions.

Line 496-502: 1938 data (ref 124) is old; breeding by taro is very common when plants are allowed to reach sexual maturity; often they are harvested before sexual maturity; this review should gather reports of flowering and breeding in Africa, as far as possible.

Response: We have revised this section and made necessary changes as suggested by the reviewer.

Line 525: whole genome duplication in Araceae is discussed by Natalie Cusimano, Aretuza Sousa and S. S. Renner (2012). "Maximum likelihood inference implies a high, not a low, ancestral haploid chromosome number in Araceae, with a critique of the bias introduced by ‘x’." Annals of Botany 109: 681-692. Is this deep evolutionary history in Araceae relevant to the present review?

Response: We appreciate this observation of the reviewer and we critically assessed the relevance of the suggested reference in the context of the current review on taro improvement. We feel that the suggested reference is beyond the scope of the current review manuscript. However, the authors also acknowledge that it will be good to write a review on deep evolutionary history in Aracaceae but it may be good to first generate additional genome sequencing information on taro and other related species of this family.

 Line 611: AOCC is an important regional initiative; can more be said about it here or earlier in the review?

Response: We have added important additional information on AOCC and its relevance to taro improvement.

Author Response File: Author Response.docx

Reviewer 2 Report

The manuscript “Taro in West Africa: Status, Challenges and Opportunities” by Oladimeji et al. is a review of the current knowledge and understanding of this crop in the region. The topic is broadly interesting; however, the manuscript falls short in several areas. The text is very broad ranging and, as a result, the development of the individual sections seems somewhat superficial. I suspect that part of the issue is that there is limited information for taro in west Africa. Therefore the authors are often drawing on broader ideas and insights (e.g., the issue of P. colocasiae extend beyond west Africa, breeding issues are common to many predominantly asexually reproducing species). It is not that these issues are not important in taro rather that the authors need to be clearer about what are general issues and which, if any, are unique to west Africa. There is also a fair amount of repetition between sections (e.g., section 8.3 and 10 both cover breeding, although slightly different aspects) and this tends to reduce the impact of the text. Finally there are a relatively larger number of spelling and grammatical errors, that although not overwhelming do make the text more difficult to read. Having a native English speaker read the text for spelling a grammar would catch most of these.

Author Response

The authors would like to thank the reviewer for all the suggestions and comments on the manuscript which provided the opportunity to revise the manuscript by incorporating the changes and improving the quality of the manuscript.

1. The manuscript “Taro in West Africa: Status, Challenges and Opportunities” by Oladimeji et al. is a review of the current knowledge and understanding of this crop in the region. The topic is broadly interesting; however, the manuscript falls short in several areas.

Response: We appreciate the observation of the reviewer, and we agree that such a review article about taro will not only provide knowledge about the current status of this crop in Africa but also may generate awareness among different stakeholders including researchers and donors about the necessity to conserve this nutritionally important food security crop in the region. We tried to cover most of the areas in which research and development has been carried out in this crop, either in West Africa or in the region or elsewhere. This may not be the most comprehensive review article but it provides all necessary information that can initiate taro improvement program in this region.

2. The text is very broad ranging and, as a result, the development of the individual sections seems somewhat superficial. I suspect that part of the issue is that there is limited information for taro in west Africa. Therefore the authors are often drawing on broader ideas and insights (e.g., the issue of  colocasiae extend beyond west Africa, breeding issues are common to many predominantly asexually reproducing species).

Response: We fully agree with the reviewer about the scarcity of research information available for taro improvement in West Africa. This review is an attempt to provide information on the importance of conservation, characterization and breeding efforts in taro improvement in West Africa.

3. It is not that these issues are not important in taro rather that the authors need to be clearer about what are general issues and which, if any, are unique to west Africa. There is also a fair amount of repetition between sections (e.g., section 8.3 and 10 both cover breeding, although slightly different aspects) and this tends to reduce the impact of the text.

Response: We appreciate the valuable observations of the reviewer, and we have now merged the two sections and revised the entire section on Taro Breeding. We also revised the entire manuscript as per the suggestions of all the reviewers and addressed the comments accordingly.

4. Finally there are a relatively larger number of spelling and grammatical errors, that although not overwhelming do make the text more difficult to read. Having a native English speaker read the text for spelling a grammar would catch most of these.

Response: We fully agree with this suggestion and used a professional English reader to make the corrections.

Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

This version is much better than the original draft. There are still some minor structural and language issues, and unclear matters.

Line 106 It is -> Taro is.. (Note, when "It" is used at the start of a sentence, it usually refers to the subject most-recently mentioned in the previous sentence. In this case, it is not immediately obvious which subject in the previous sentence "It" refers to: Araceae, or taro?). This problem appears in other parts of the paper too.

Line 119 -> "hyper-diversity"? The crop overall has high diversity, but "hyper-diversity" is something that may only exist in wild breeding populations in certain parts of the natural range taro

(cf. Matthews, P. J. (2017). Evolution and Domestication of Clonal Crops. Routledge Handbook of Agricultural Biodiversity. D. Hunter, L. Guarino, C. Spillane and P. C. McKeown. Oxford, Routledge: 168-191).

There are at least two main evolutionary lineages in the crop, and it is likely that hybridisation between these lineages has added to the overall diversity of cultivated forms. Much remains to be learned through analysis of both the plastid and nuclear genomes, together with more systematic morphological description.

(cf. Ahmed, I., P. J. Lockhart, E. M. G. Agoo, K. W. Naing, D. V. Nguyen, D. K. Medhi and P. J. Matthews (2020). "Evolutionary origins of taro (Colocasia esculenta) in Southeast Asia." Ecology and Evolution 10: 13530–13543).

Line 175: "huli" is a Hawaiian vernacular term specific for taro and with precise meaning; "tops" is less specific, but is understandable as common English in the context. If the authors like the term huli, then it does not need a capital letter (it describes a plant part, and is not a proper name), and its origin a Hawaiian vernacular term should be mentioned).

Line 122:  ..genetic base..."is usually narrow"... "usually" is odd in this context. --> "is likely to be narrow" or "thought to be narrow" might be better, depending on what the cited sources say.

Line 132 "In this study..." is ambiguous. The present study, or the study just cited? "The study just cited" can be expressed using (ibid).

Line 310: venation is poorly described: "...with rounded basal lobes, usually with three veins. Some prominent veins may arise from the three veins, but the overall leaf venation is reticulate (net-veined)." --->

"...with rounded basal lobes and long anterior lobe. The central veins of each lobe and primary lateral veins (ribs) are raised, and tertiary venation of the lamina is reticulate (net-veined)."

Line 320: Pollination of taro and its wild relatives is primarily [not "presumably"] by insects (drosophilid flies in the genus Colocasiomyia). See:

Sultana, F., Y.-G. Hu, M. J. Toda, K. Takenaka and M. Yafuso (2006). "Phylogeny and classification of Colocasiomyia (Diptera, Drosophilidae), and its evolution of pollination mutualism with aroid plants." Systematic Entomology 31: 684-702.

Line 351 An ambiguous "it.."

Lines 394-397 Repeats general statements and refers to "germplasm collections" in farmers' fields. Delete these lines entirely? The diversity present in farmers' fields varies greatly in different regions and has not been systematically compared across Asia, Africa, Oceania Americas. We can't say that it is always "low".

Line 421: wild plants are depended on insects for pollination, but this does not make controlled plant breeding especially difficult (in fact it might be easier than for wind pollinated plants, especially in regions where the natural pollinators are absent). Hand pollination and other breeding methods are very nicely described in:

Ivancic, A. and V. Lebot (2000). The genetics and breeding of taro. Monpellier, CIRAD.

Line 462 The last part of the sentence is not logically connected to the first part: delete: "... where it has been introduced through human migrations."

Line 489 - Is the report of a "wildtype" taro in Ethiopia reliable or based on study aimed at distinguish naturalised introductions from a natural, wild-type distribution? Does the original source qualify the observation as a "possible wild-type"? Care is needed here because extension of the natural range of taro from Asia to Africa has never been reported, and is a matter that requires field research aimed at addressing the question. For example, if it was true, then there should also be natural pollinators associated with the possible wild-type population(s).

Line 787 - keeping a database "to track genetic erosion in farmers' fields" does not seem satisfactory as a primary aim for keeping a database. The primary aim of a modern, curated and interactive database could be to help farmers and local extension officers understand the value and potential of cultivars that are present, and to learn from the experiences of other people who grow the same or similar varieties in other regions. If communities can become more positively interested the crop, then they may give more attention to keeping their local varieties and comparing them with new varieties.

 Sections 10-11 on breeding. Although the discussion of breeding has been brought together into one area, there is still repetition. It might be best to combine 10 and 11 into one section.

Line 656  The flowering ability of diploid cultivars is poorly understood but some flower very easily when allowed to reach reproductive maturity. Experimental research on flowering ability is generally lacking (but see Ivancic, A. and V. Lebot (2000). The genetics and breeding of taro. Monpellier, CIRAD).

Lines 672-673. There is no explanation of why "low production" in WA (one of the largest producers of taro globally) should contribute to a narrow genetic base. Most diversity of taro has been maintained in the past (and is still maintained) in small-scale production systems. Large-scale, modern commercial production tends to focus on a few popular cultivars. If there is a relationship between production level and genetic diversity, it is more likely to be opposite to the relationship suggest here. The key issue is how to maintain or develop commercial production systems that embrace cultivar diversity.

There may be other issues that have not been noted here: I recommend employing an editor or friendly colleague for at least a basic check of sentence construction and to avoid repetition of statements.

If the authors wish to use and rephrase any of the comments above, that is OK.

Author Response

The authors would like to express their gratitude to the reviewer for all the suggestions and comments that has truly shaped the manuscript. We have made necessary changes and also employed an English editor to go through the manuscript to ensure that the repetitions are avoided.

This version is much better than the original draft. There are still some minor structural and language issues, and unclear matters.

Line 106 It is -> Taro is.. (Note, when "It" is used at the start of a sentence, it usually refers to the subject most-recently mentioned in the previous sentence. In this case, it is not immediately obvious which subject in the previous sentence "It" refers to: Araceae, or taro?). This problem appears in other parts of the paper too.

Response: We fully agree with this observation of the reviewer and made necessary changes across the manuscript.

Line 119 -> "hyper-diversity"? The crop overall has high diversity, but "hyper-diversity" is something that may only exist in wild breeding populations in certain parts of the natural range taro

(cf. Matthews, P. J. (2017). Evolution and Domestication of Clonal Crops. Routledge Handbook of Agricultural Biodiversity. D. Hunter, L. Guarino, C. Spillane and P. C. McKeown. Oxford, Routledge: 168-191).

There are at least two main evolutionary lineages in the crop, and it is likely that hybridisation between these lineages has added to the overall diversity of cultivated forms. Much remains to be learned through analysis of both the plastid and nuclear genomes, together with more systematic morphological description.

(cf. Ahmed, I., P. J. Lockhart, E. M. G. Agoo, K. W. Naing, D. V. Nguyen, D. K. Medhi and P. J. Matthews (2020). "Evolutionary origins of taro (Colocasia esculenta) in Southeast Asia." Ecology and Evolution 10: 13530–13543).

Response: We have deleted the word ‘hyper-diversity’ and made necessary changes as per the suggestion of the reviewer. We have also included the reference suggested by the reviewer. However, we felt that it was not necessary to include the reference of Matthews P.J. (2017) and we did not include this reference.

Line 175: "huli" is a Hawaiian vernacular term specific for taro and with precise meaning; "tops" is less specific, but is understandable as common English in the context. If the authors like the term huli, then it does not need a capital letter (it describes a plant part, and is not a proper name), and its origin a Hawaiian vernacular term should be mentioned).

Response: We kept the word ‘huli’ and as per the suggestion of the reviewer, we provided the reference of Hawaiian vernacular term. We thank the reviewer for this observation and suggestion.

Line 122:  ..genetic base..."is usually narrow"... "usually" is odd in this context. --> "is likely to be narrow" or "thought to be narrow" might be better, depending on what the cited sources say.

Line 132 "In this study..." is ambiguous. The present study, or the study just cited? "The study just cited" can be expressed using (ibid).

Line 310: venation is poorly described: "...with rounded basal lobes, usually with three veins. Some prominent veins may arise from the three veins, but the overall leaf venation is reticulate (net-veined)." --->

"...with rounded basal lobes and long anterior lobe. The central veins of each lobe and primary lateral veins (ribs) are raised, and tertiary venation of the lamina is reticulate (net-veined)."

Response: We made the necessary changes as per the suggestion of the reviewer.

Line 320: Pollination of taro and its wild relatives is primarily [not "presumably"] by insects (drosophilid flies in the genus Colocasiomyia). See:

Sultana, F., Y.-G. Hu, M. J. Toda, K. Takenaka and M. Yafuso (2006). "Phylogeny and classification of Colocasiomyia (Diptera, Drosophilidae), and its evolution of pollination mutualism with aroid plants." Systematic Entomology 31: 684-702.

Response: We fully agree with the observation of the reviewer, and added the information in the text as well as the reference.

Line 351 An ambiguous "it.."

Lines 394-397 Repeats general statements and refers to "germplasm collections" in farmers' fields. Delete these lines entirely? The diversity present in farmers' fields varies greatly in different regions and has not been systematically compared across Asia, Africa, Oceania Americas. We can't say that it is always "low".

Response: We have deleted the ‘collections’ and retained the word ‘germplasm’ to keep the meaning of the statement. We have also included the suggested statement of the reviewer.

Line 421: wild plants are depended on insects for pollination, but this does not make controlled plant breeding especially difficult (in fact it might be easier than for wind pollinated plants, especially in regions where the natural pollinators are absent). Hand pollination and other breeding methods are very nicely described in:

Ivancic, A. and V. Lebot (2000). The genetics and breeding of taro. Monpellier, CIRAD.

Line 462 The last part of the sentence is not logically connected to the first part: delete: "... where it has been introduced through human migrations."

Response: We fully agree with the reviewer and made necessary changes in the text.

Line 489 - Is the report of a "wildtype" taro in Ethiopia reliable or based on study aimed at distinguish naturalised introductions from a natural, wild-type distribution? Does the original source qualify the observation as a "possible wild-type"? Care is needed here because extension of the natural range of taro from Asia to Africa has never been reported, and is a matter that requires field research aimed at addressing the question. For example, if it was true, then there should also be natural pollinators associated with the possible wild-type population(s).

Response: We thank the reviewer for this observation, we have now deleted the part on wild-type.

Line 787 - keeping a database "to track genetic erosion in farmers' fields" does not seem satisfactory as a primary aim for keeping a database. The primary aim of a modern, curated and interactive database could be to help farmers and local extension officers understand the value and potential of cultivars that are present, and to learn from the experiences of other people who grow the same or similar varieties in other regions. If communities can become more positively interested the crop, then they may give more attention to keeping their local varieties and comparing them with new varieties.

Response: We fully agree with this suggestion of the reviewer, and added the suggested statements in the manuscript.

 Sections 10-11 on breeding. Although the discussion of breeding has been brought together into one area, there is still repetition. It might be best to combine 10 and 11 into one section.

Response: We appreciate the suggestion of the reviewer and we looked at both sections critically. We feel that these two sections should be kept separate as the information provided in these two sections are not related although some information on flowering, crossing and seed set may look repeating. We therefore kept the two sections separate.

Line 656  The flowering ability of diploid cultivars is poorly understood but some flower very easily when allowed to reach reproductive maturity. Experimental research on flowering ability is generally lacking (but see Ivancic, A. and V. Lebot (2000). The genetics and breeding of taro. Monpellier, CIRAD). 

Response: We have included the information in the manuscript as per the suggestion of the manuscript.

Lines 672-673. There is no explanation of why "low production" in WA (one of the largest producers of taro globally) should contribute to a narrow genetic base. Most diversity of taro has been maintained in the past (and is still maintained) in small-scale production systems. Large-scale, modern commercial production tends to focus on a few popular cultivars. If there is a relationship between production level and genetic diversity, it is more likely to be opposite to the relationship suggest here. The key issue is how to maintain or develop commercial production systems that embrace cultivar diversity.

Response: We fully agree with the suggestion of the reviewer and made necessary changes in the text.

There may be other issues that have not been noted here: I recommend employing an editor or friendly colleague for at least a basic check of sentence construction and to avoid repetition of statements.

Response: We appreciate the suggestion of the reviewer and recruited a English editor to critically go through the manuscript.

If the authors wish to use and rephrase any of the comments above, that is OK.

Response: We re-phrased the statements suggested by the reviewer, where necessary.

Author Response File: Author Response.docx