Effects of Fructooligosaccharides (FOS) on the Immune Response of the Shrimp Penaeus vannamei and on the Reduction in Vibrio spp. and Pseudomonas spp. in Cultures of Post-Larvae
, Leonardo Davier MartÃn RÃos 1, Maikelis RodrÃguez González 1, Marbelys González Salotén 1, Yeidel López ZaldÃvar 1 and Amilcar Arenal 3,*Round 1
Reviewer 1 Report
see attached
Comments for author File: 
Comments.pdf
It is essential to comply with the editorial standards of journals
Author Response
Reviewer 1
The paper ¨Effects of fructooligosaccharides (FOS) on the immune response and reduction of Vibrios and Pseudomonas in cultured postlarvae of the shrimp Penaeus vannamei¨, presents a series of inconsistencies and misinterpretations that do not allow the conclusions proposed by the authors to be reached in an appropriate way. In addition, it presents a series of basic editing errors that should be checked before sending the work to any publisher (i.e. scientific names in italics, larger font sizes for no reason (L89, L119), type and quality of graphics.
Thank you very much to the editor for their time and the comments.
The work needs an extensive revision of the English, and a restructuring and edition to be able to explain in a better way the ideas.
We review thoroughly considering reviewer comments and two native English revisions.
In the Results and discussion section, there are results that are not described in an appropriate way (a difference is noted in the type of writing between the different sections, it seems that they were written independently and were only pasted), in addition there are results that they are not even discussed (what is the answer to this?).
The work needs extensive revision of the English, and edition to be able to explain in a better way the ideas.
A series of essential information gaps are presented in the development of the experiment (i.e. how the FOS was added to the food).
We included!
Vibrios reduction was through a homogenization of the larvae and later their culture (where apparently the same organisms are not taken per tank, L181), this modifies the bacterial load of the samples. In addition, an experiment designed to verify that the reduction in vibrios is due to the direct action of adding FOS to the diet is needed (it is very risky to conclude that).
We agree is not the same organism, we are evaluating the ability of FOS to stimulate the larvae in the reduction of Vibrio load. There are some articles that discuss this possible mechanism of action. We improved our discussion and suggest further research for the mechanism.
What was the proximal composition of the diets, how to know if the modification of the results was not due to changes in the diet formulation (i.e. protein and lipids), rather than the direct effect of FOS itself.
The diet was the same in all tanks, we included.
Although, in the instructions for prebiotics it is suggested to add it to the water (an argument that lacks scientific validity due to its way of acting), do the authors also add it directly to the water, what is the basis for this?
We discussed in the MS.
Each one of the graphs has a different format, the authors call the treatments in a different way depending on the graph, even within the same figure (Fig 1), and it can even be seen that it was a screenshot since the text even appears. cursor
We made the corrections, including the recommendations of the other authors!
The figure captions are not written in an appropriate way, since it does not have the necessary information. Once again change the writing style depending on the results
L103-104: the samples are filtered, and subsequently washed (they do not explain if they refer to water or organisms)
Done
Standardization of the units, throughout the text the units are mentioned in different ways, sometimes there are spaces, different symbols, etc. (once again, the editing varies depending on the section of the result described
Done!
Reviewer 2 Report
Although the study is generally interesting and the results worth publishing, I feel that the manuscript needs to be improved throughout, e.g. with respect to the language and grammar quality and due to the generally too many mistakes, e.g. with respect to the Figure and Figure captions. Here are some selected comments that might be helpful:
-I do not know if applicable also to bacteria nomenclature, but for animals, genus and species authorities should be given upon first mentioning, including the title of a paper.
-Abstract: “Penaeus” and “P. vannamei” must be written in italics and the genus and species authorities should be added.
-Abstract: “Vibrios” and “Pseudomonas” also must be written in italics in all instances.
-Abstract: It must be “Student's t-test” instead of “t-Student’s test”.
-Keywords: keywords not already included in the title of the manuscript should be used.
-1.: “Penaeus vannamei” must be written in italics.
-Methods: significance levels need to be explained. In Figure 1, a significance level of p≤0.001 but it should be p<0.001 and why are four asterisks used? Usually, this level of significance is marked with three asterisks.
-2.1: There seems to be some variation in font type and size (see “P. vannamei”).
-2.6: There seems to be some variation in font type and size (see “x g”).
-2.7.6: “Pseudomonas” must be written in italics.
-3.9: “Pseudomonas” and “Vibrios” must be written in italics in all instances
-4.: “P. vannamei”, “Vibrios” and “Pseudomonas” must be written in italics.
-Caption to Figure 1: This needs to be revised as there are missing commas, missing explanation of abbreviations, mistakes in capitalization of letters, significance level, asterisks, missing elements, and general wording.
-Caption to Figure 2: This also needs to be revised, e.g. with respect to the parentheses and general wording.
-Caption to Figure 3: It must be 0.001 instead of 0,001. And see also previous comments regarding significance levels and asterisks.
-Caption to Figures 4-6: as mentioned previously, all Figure captions need to be rewritten.
-Figures: The Figures are apparently screenshots with the formatting characters included, which is unacceptable for publication.
-Write “References” instead of “references”
Please see above.
Author Response
Although the study is generally interesting and the results worth publishing, I feel that the manuscript needs to be improved throughout, e.g. with respect to the language and grammar quality and due to the generally too many mistakes, e.g. with respect to the Figure and Figure captions. Here are some selected comments that might be helpful:
Thank you very much to the editor for their time and the comments.
-I do not know if applicable also to bacteria nomenclature, but for animals, genus and species authorities should be given upon first mentioning, including the title of a paper.
Done
-Abstract: “Penaeus” and “P. vannamei” must be written in italics and the genus and species authorities should be added.
Done!
-Abstract: “Vibrios” and “Pseudomonas” also must be written in italics in all instances.
Done!
-Abstract: It must be “Student's t-test” instead of “t-Student’s test”.
Done!
-Keywords: keywords not already included in the title of the manuscript should be used.
Done!
-1.: “Penaeus vannamei” must be written in italics.
Done!
-Methods: significance levels need to be explained. In Figure 1, a significance level of p≤0.001 but it should be p<0.001 and why are four asterisks used? Usually, this level of significance is marked with three asterisks.
Done!
-2.1: There seems to be some variation in font type and size (see “P. vannamei”).
Done!
-2.6: There seems to be some variation in font type and size (see “x g”).
Done!
-2.7.6: “Pseudomonas” must be written in italics.
Done!
-3.9: “Pseudomonas” and “Vibrios” must be written in italics in all instances
Done!
-4.: “P. vannamei”, “Vibrios” and “Pseudomonas” must be written in italics.
Done!
-Caption to Figure 1: This needs to be revised as there are missing commas, missing explanation of abbreviations, mistakes in capitalization of letters, significance level, asterisks, missing elements, and general wording.
Done!
-Caption to Figure 2: This also needs to be revised, e.g. with respect to the parentheses and general wording.
Done!
-Caption to Figure 3: It must be 0.001 instead of 0,001. And see also previous comments regarding significance levels and asterisks.
Done!
-Caption to Figures 4-6: as mentioned previously, all Figure captions need to be rewritten.
Done!
-Figures: The Figures are apparently screenshots with the formatting characters included, which is unacceptable for publication.
Done!
-Write “References” instead of “references”
Done!
Reviewer 3 Report
The authors claim the addition of FOS during Penaeus farming exhibits positive effects on its growth and resistance to bacterial pathogens, the conclusion is basically making sense based on results from the growth index, enzymatic activity assay, and microbiota analysis. See the following detailed comments about this manuscript:
1. Authors claim that "Changes in the microbiota could be observed, with a reduction of Vibrios and Pseudomonas (Control 1.1E03±0.3E02; FOS 2.4E03±0.5 E02).", the exact mean of authors may be that addition of FOS can decrease microbiota, but data supplied here is showing a higher microbiota in FOS group, please confirm this, this issue also appears in "3.9. Bacteriological analysis" lines 2-3.
2. It looks like there are only 6 Figures in this manuscript with one Figure missing. Please correct the Figure number as well as the legend.
3. Authors claim there is no stimulation of superoxide dismutase and amylase activities in "3.3. Superoxide dismutase specific enzymatic activity, and 3.8. Amylase specific enzyme activity", can you show these data? it may be important for others' future work.
Author Response
The authors claim the addition of FOS during Penaeus farming exhibits positive effects on its growth and resistance to bacterial pathogens, the conclusion is basically making sense based on results from the growth index, enzymatic activity assay, and microbiota analysis. See the following detailed comments about this manuscript:
Thank you very much to the editor for their time and the comments.
- Authors claim that "Changes in the microbiota could be observed, with a reduction of Vibrios and Pseudomonas (Control 1.1E03±0.3E02; FOS 2.4E03±0.5 E02).", the exact mean of authors may be that addition of FOS can decrease microbiota, but data supplied here is showing a higher microbiota in FOS group, please confirm this, this issue also appears in "3.9. Bacteriological analysis" lines 2-3.
We made a mistake a switch the data, we made the correction!
- It looks like there are only 6 Figures in this manuscript with one Figure missing. Please correct the Figure number as well as the legend.
We included the figure and include the legend.
- Authors claim there is no stimulation of superoxide dismutase and amylase activities in "3.3. Superoxide dismutase specific enzymatic activity, and 3.8. Amylase specific enzyme activity", can you show these data? it may be important for others' future work.
Done!
Round 2
Reviewer 1 Report
The authors have made the suggested changes
Author Response
Thank you for your reviewing
Reviewer 2 Report
Although the manuscript has been improved, there are still many small errors, e.g. with respect to using blank spaces in significance levels (i.e., in the Abstract, three spellings are found: p<0.001, p < 0.001 and p <0.001). Therefore, a careful check of the whole manuscript needs to be carried out (and should generally have been performed prior to submission!).
Although the language quality has been improved, the authors seem to have concentrated on the examples mentioned in the reviews rather than eliminating all language problems. One example can be found directly in the first sentence of the Introduction, where simple present and simple past are mixed instead of using simple present throughout. Therefore, I once again strongly recommend a careful language revision throughout the whole manuscript.
Further issues:
-P values are missing for several comparisons in the Abstract (i.e., weight, length, survival, changes in microbiota).
-As pointed out in my last review, species authorities should be added upon first mentioning of a genus- or species-level taxon.
-There are still issues with the figures, e.g., it must be “mg”, “mm” etc. instead of “m g”, “m m” etc. Furthermore, “b” and “a” above the bars is not explained in the caption to Figure 1 and should be deleted from all figures as they are redundant in diagrams with only two bars (the significance level is already described in the Figure caption). In the text, the authors use “Control” for the control group but in the Figures and Figure captions, they use the spelling “control”. Additionally, “bars T” or “T bars” (both spellings, of which only the latter makes sense, are used) is not a commonly used term. Again, there is no consistent use of blank spaces in significance levels.
Significant improvement of the language quality is required throughout the manuscript.
Author Response
Dear reviewer and editor
Thank you very much for your comments. The English manuscript was edited by John Van der Meer, former editor of Marine Biotechnology editor.
Although the manuscript has been improved, there are still many small errors, e.g. with respect to using blank spaces in significance levels (i.e., in the Abstract, three spellings are found: p<0.001, p < 0.001 and p <0.001). Therefore, a careful check of the whole manuscript needs to be carried out (and should generally have been performed prior to submission!).
We corrected all of them!
Although the language quality has been improved, the authors seem to have concentrated on the examples mentioned in the reviews rather than eliminating all language problems. One example can be found directly in the first sentence of the Introduction, where simple present and simple past are mixed instead of using simple present throughout. Therefore, I once again strongly recommend a careful language revision throughout the whole manuscript.
Done!
Further issues:
-P values are missing for several comparisons in the Abstract (i.e., weight, length, survival, changes in microbiota).
The P values were included!
-As pointed out in my last review, species authorities should be added upon first mentioning of a genus- or species-level taxon.
We reviewed them carefully and all are declared!
-There are still issues with the figures, e.g., it must be “mg”, “mm” etc. instead of “m g”, “m m” etc. Furthermore, “b” and “a” above the bars is not explained in the caption to Figure 1 and should be deleted from all figures as they are redundant in diagrams with only two bars (the significance level is already described in the Figure caption). In the text, the authors use “Control” for the control group but in the Figures and Figure captions, they use the spelling “control”. Additionally, “bars T” or “T bars” (both spellings, of which only the latter makes sense, are used) is not a commonly used term. Again, there is no consistent use of blank spaces in significance levels.
We removed the letters from the figure. We were consistent with the blank space. We make the correction for the Control group. We were consistent with the term t bars. We did not find spaces between mg and mm units.
