Comparative Analysis of Cyanotoxins in Fishponds in Nigeria and South Africa
, Munyaradzi Mujuru 3, Adeeyo Adeyemi 2 and Farai Dondofema 1
Round 1
Reviewer 1 Report
Comments and Suggestions for AuthorsDear authors
Happy day
The paper is fin but needs some improvement.
1- The title is so sounded and not comparable with the work amount, kindly change it. For example, you use only descriptive statistical analysis which did not reflect the amount of work you represent. The Principal component analysis is not clear.
2- Which version of XLstat you have used?
3- For figure 1.2, kindly give more information to be more understandable. Images are used to give quick and simple understand. Kindly improve.
4- Image quality need more improvement.
5- Kindly argument why you select Nigeria and South Africa based on more variables like the environment, location, temperature etc.
6- The images need improvement.
7- More references are needed.
8- Only one ref. in 2023 is existed, more recent references are needed
Comments for author File: 
Comments.pdf
Author Response
Reviewer 1
- The title is so sounded and not comparable with the work amount, kindly change it. For example, you use only descriptive statistical analysis which did not reflect the amount of work you represent. The Principal component analysis is not clear.
Response: The title has been revised
2- Which version of XLstat you have used?
Response: XLstat Version 2022.3.1.
3- For figure 1.2, kindly give more information to be more understandable. Images are used to give quick and simple understand. Kindly improve.
Response: This has been improved
4- Image quality need more improvement.
Response: Image has been improved
5- Kindly argument why you select Nigeria and South Africa based on more variables like the environment, location, temperature etc.
Response: The selection of Nigeria and South Africa for the comparative study on cyanotoxins in fishponds was based on several key variables, including environmental diversity, geographical location, and climatic conditions.
6- The images need improvement.
Response: Images improved
7- More references are needed.
Response: More recent references have been added
8- Only one ref. in 2023 is existed, more recent references are needed
Response: More references of 2023 has been added
Author Response File: 
Author Response.docx
Reviewer 2 Report
Comments and Suggestions for AuthorsPlease, see the attached document.
Comments for author File: Comments.pdf
Author Response
“Understanding regional occurrence of cyanotoxins in aquaculture: A comparative study between Nigeria and South Africa”
This study compares the distribution of cyanotoxins in aquaculture from different commercial fishponds in Nigeria and South Africa during different seasons of the year. Liquid chromatography-mass spectrometry in combination with the Global Natural Product Social Molecular Networking and CANOPUS was used to identify various cyanotoxins. This manuscript may be suitable for publication in the journal 'Microbiology Research', but it requires significant revision. The main issues are insufficiently described results, confusion in figure numbering, and lack of information in the figures.
Point 1: Give a transcript of the GNPS in the abstract.
The transcript of the GNPS has been provided in the Abstract.
Response: The Molecular networking approach coupled with the Global Natural Products Social Molecular Networking (GNPS) database and CANOPUS annotation enabled putative identification of cyanometabolites. The resulting molecular network unveiled discernible clusters representing related molecule families, aiding in the identification of both known cyanotoxins and unfamiliar analogues. Furthermore, the molecular network revealed that water samples from different fishponds shared specific metabolites including ethanesulfonic acid, pheophorbide A, cholic acid phenylalanine, amyl amine and phosphocholine (PC) and sulfonic acid despite variations in location, local climatic factors, and sampling sites. The fishponds in Nigeria showed the presence of multiple cyanotoxin classes in the dry, wet and summer seasons in the water. Aplysiatoxins were identified in all sampling sites in Nigeria (N1, N2, and N3). The Duthuni South Africa (P1, P2, P3) sampling sites exhibited the presence of Microginins and Microcystins. All the fishponds displayed a widespread occurrence of Anabaenopeptins, Aplysiatoxins, Microcolins, and Marabmids during the selected summer. In conclusion, the untargeted metabolome analysis, guided by GNPS, proved highly effective in identifying both toxic and non-toxic metabolites in fishponds.
Item 2: Page 2, lines 87-92 - check figure reference and figure numbering. Figure 1 is of poor quality; objects are not visible. Label the parts of the figure as A and B. Is this figure needed?
Response: Figure 1 removed
Point 3: Page 3, lines 97-99 - check the sentence structure.
Response: This study utilized a total of six fish ponds situated in the Vhembe District (comprising three fish ponds) and Calabar (comprising three fish ponds). The fishpond types incorporated in this investigation consisted of concrete, tarpaulin, and earthen ponds, as illustrated in Figure 1.
Point 4: Methodology - need to improve towards a concise description of methods without minor details; do not split section 2.1 into subsections.
Response: The methodology has been improved.
Point 5: page 5, line 163 - figure numbering is confusing.
Response: This has been rectified
Point 6: page 5, lines 171 - 173 - clarify which water samples are referred to
Response: The molecular network showed that water samples from Nigeria (N1, N2, and N3) and South Africa (P1, P2, P3) shared specific metabolites despite differences in location, local climatic factors, and sampling sites.
Point 7: page 5, lines 181-183 - no figure below the text.
Response: This has been rectified
Point 8: Page 6, Table 1 needs to be redrafted to be consistent with the results described (lines 222-224).
Response: Table 1 has been removed
Item 9: page 6. Describe the results presented in Table 2.
Response: This table has been removed. A graphical representation has been used to replace the table.
Item 10: Figures 1.4-1.6 do not show seasonality, the results presented in these figures are not described.
Response: A new Figure has been added to represent seasonality
Round 2
Reviewer 2 Report
Comments and Suggestions for AuthorsReview
for the article entitled “Understanding regional occurrence of cyanotoxins in aquaculture: A comparative study between Nigeria and South Africa”
The revised version up to now requires corrections.
Point 1: Reference numbers should be corrected to either [1-3] or [1, 3].
Point 2: The reference list should include the full title, as recommended by the MDPI and ACS Style guide. For example:
Ciebiada, M.; Kubiak, K.; Daroch, M. Modifying the Cyanobacterial Metabolism as a Key to Efficient Biopolymer Production in Photosynthetic Microorganisms. Int. J. Mol. Sci. 2020, 21, 7204. https://doi.org/10.3390/ijms21197204
Point 3: All figures should be inserted into the main text near their first citation and numbered according to their order of appearance (Figure 1, Figure 2, Figure 3, Figure 4).
The numbering of figures is unclear oon page 5, lines 147 – 168; page 6, lines 184-194; and page 6, lines 201-210.
Comments for author File: Comments.pdf
Author Response
Reviewer 2
Point 1: Reference numbers should be corrected to either [1-3] or [1, 3].
Response: The references have been corrected to [1-3] or [1, 3].
Point 2: The reference list should include the full title, as recommended by the MDPI and ACS Style guide. For example:
Ciebiada, M.; Kubiak, K.; Daroch, M. Modifying the Cyanobacterial Metabolism as a Key to Efficient Biopolymer Production in Photosynthetic Microorganisms. Int. J. Mol. Sci. 2020, 21, 7204. https://doi.org/10.3390/ijms21197204
Response: The referencing style has been changed to MDPI and ACS Style.
Point 3: All figures should be inserted into the main text near their first citation and numbered according to their order of appearance (Figure 1, Figure 2, Figure 3, Figure 4).
Response: This has been rectified.
The numbering of figures is unclear on page 5, lines 147 – 168; page 6, lines 184-194; and page 6, lines 201-210.
Response: The figures’ numbering has been corrected.
