Effects of γ-Irradiated Date (Phoenix dactylifera) Fruit on Growth, Immunological and Antioxidant Parameters of Goldfish (Carassius auratus)
Round 1
Reviewer 1 Report
The manuscript would be interesting for the readers provided the authors made the suggested modifications and corrections attached in the file.
Comments for author File: 
Comments.pdf
Author Response
Reviewer 1
Follow the word limit mentioned in the guideline.
Answer: Some parts were removed.
Generally it is practiced to keep the fish starved before the measurement! Was it followed??
Answer: it was applied (line 107)
Mention the concentration.
Answer: it was applied (line 124).
Mention the CFU concentration (line 143)
Answer: it was applied (line 140).
Rectify it
Answer: it was applied (line 149-150).
Mention the Kit IDs (superoxide dismutase and glutathione peroxidase)
Answer: it was applied (line 155).
Compare the result statistically
Answer: it was applied (line 188).
followed by a Tukey post hoc test. Mention the N value in each table.
Answer: it was applied in all tables.
It is not clearly explained why IPFE5 has better result than IPFE10 though IPFE10 has higher concentration of date extract!
Answer: a paragraph related was added to line 316-326.
Author Response File: Author Response.pdf
Reviewer 2 Report
The Authors of this article examined the effects of γ-irradiated date fruit extract on growth, mucosal immunity and liver antioxidant parameters of goldfish. The experiment was well designed, the selected parameters were appropriate and their evaluation was correct. The results are very interesting and promising. Based on the novelty of this research, the proper experimental design and promising results, this article is really worth for publication. However, there are some really minor errors that should be corrected before publication, therefore this manuscript can only be accepted after a minor revision.
Lines 152-156: It is not written that this method is for measuring lipid peroxidation product. This is mentioned only in the Results section.
Table 2: Significant differences are not marked in this table. Or if there are no significant differences between the extracts, it is not mentioned in the text.
After these modifications are done, I highly recommend this article for publication.
Author Response
Reviewer 2
Lines 152-156: It is not written that this method is for measuring lipid peroxidation product. This is mentioned only in the Results section.
Answer: it was applied (line 148).
Table 2: Significant differences are not marked in this table. Or if there are no significant differences between the extracts, it is not mentioned in the text.
Answer: it was applied (line 188).
Author Response File: Author Response.pdf
Reviewer 3 Report
Within this study, the effect of date fruits as dietary additives for ornamental pet goldfish on growth and immune parameters in mucosa and liver was investigated. The major weakness of the study is that no information on the actual feed intake was provided. The differences between the positive and negative control indicate that there could already be impacts of the date fruits without gamma-irradiation. IPFE analysis would also need to be done of the control feed (CTR) to verify the potential of this feed. In general, the material/methods section has several errors. Several improvements are needed here. The same applies to the results - no sensible headings, lots of abbreviations, no description of these and generally a superficial description.
Details:
Authors: L5: There is a missing letter in the name. “steban, MA” means Maria Ángeles Esteban.
Abstract
The abstract is a bit too long (200 words are recommended) The results part could be slightly shortened.
L29: I think the term “titrated” is used in a wrong context.
L31: The key word “goldfish (Carassius auratus)” is already included in the title and should be removed from the list.
Introduction
L43-45: It is difficult to follow the sentence and the dietary additives are the most important part. I recommend making two sentences and putting the dietary additives at the beginning.
L49-50: All plants contains water, protein, fat and so on. Be more specific.
Line 56 ff – there is really nobody else on the whole world doing these kind of studies? I can’t believe this.
L59: Where is the contrast (on one hand)?
Material and Methods
L71: How much date seed is used?
L73: I think the correct term is Moulinex. Give details of designation and manufacturer for all instruments used, as in L77. See also L83 ff.
L83: Did you mean a meat grinder with a 2 mm perforated disc?
L86: The table 1 is splitted into two parts. Correct this or repeat the heading on the next page.
L97: Correct to Folin-Ciocalteu method.
L102: Means this the initial weight and length after 10 days of acclimatisation? The values are missing in text or tables.
L107: The fish were fed ad libitum. How much feed per day and fish or how much feed per gram of fish was used?
L113: “ …length and weight of each fish were estimated” or measured in mm and gram?
L115: The formula is wrong but I hope your calculations are correct.
L117: What do you mean with “mucus obtention”?
L159/L164: How was the absorption measured? In which machine?
Results
In general, the headings of the tables should include and explain all abbreviations used in the table so that the table is understood as such.
Furthermore, the description of the results in the text is very weak and could be improved.
In Table 2, the values for the CTR feed are missing. This is important to understand the benefits of the IPFE feed and forms the basis for the following tables.
L191: To understand the differences between CTR and IPFE0, it would be important to know how much feed the fish actually uptake. This is because feed intake plays an important role in growth parameters.
The table 4 is splitted into two parts. Correct this or repeat the heading on the next page.
Discussion
The discussion is extensive and includes the topics: antioxidant activity, growth performance in fish, microbiota/gut microbiota, mucosa surfaces of fish in context with immune parameters, and date fruit as a feed additive due to the antioxidant effect. The statements on the authors' own data remain on the surface and could be deepened.
L223/L293: On one hand … on the other hand. Contrast!
L252: In this sentence, you take about the uptake rate of feed from other authors but gives no information or relations to your own data.
Author Response
Reviewer 3
Details:
Authors: L5: There is a missing letter in the name. “steban, MA” means Maria Ángeles Esteban.
Answer: it was applied (line 5).
Abstract
The abstract is a bit too long (200 words are recommended) The results part could be slightly shortened.
Answer: Some parts were removed.
L29: I think the term “titrated” is used in a wrong context.
Answer: it was corrected (line 27).
L31: The key word “goldfish (Carassius auratus)” is already included in the title and should be removed from the list.
Answer: it was corrected (line 28).
Introduction
L43-45: It is difficult to follow the sentence and the dietary additives are the most important part. I recommend making two sentences and putting the dietary additives at the beginning.
Answer: it was corrected (line 41).
L49-50: All plants contains water, protein, fat and so on. Be more specific.
Answer: this sentence was omitted.
Line 56 – there is really nobody else on the whole world doing these kind of studies? I can’t believe this.
Answer: it was corrected (line 51).
L59: Where is the contrast (on one hand)?
Answer: it was corrected (line 53).
Material and Methods
L71: How much date seed is used?
Answer: information was added (line 68).
L73: I think the correct term is Moulinex. Give details of designation and manufacturer for all instruments used, as in L77. See also L83 ff.
Answer: these parts were corrected.
L83: Did you mean a meat grinder with a 2 mm perforated disc?
Answer: it was corrected (line 78).
L86: The table 1 is splitted into two parts. Correct this or repeat the heading on the next page.
Answer: it was corrected.
L97: Correct to Folin-Ciocalteu method.
Answer: it was corrected (line 91).
L102: Means this the initial weight and length after 10 days of acclimatisation? The values are missing in text or tables.
Answer: data were added to Table 3.
L107: The fish were fed ad libitum. How much feed per day and fish or how much feed per gram of fish was used?
Answer: feed intake per treatment group was added to Table 3.
L113: “ …length and weight of each fish were estimated” or measured in mm and gram?
Answer: it was corrected (line 109).
L115: The formula is wrong but I hope your calculations are correct.
Answer: sorry for the mistake. It was corrected (line 112).
L117: What do you mean with “mucus obtention”?
Answer: it was corrected (line 113).
L159/L164: How was the absorption measured? In which machine?
Answer: it was corrected (162-168).
Results
In general, the headings of the tables should include and explain all abbreviations used in the table so that the table is understood as such.
Answer: it was corrected.
Furthermore, the description of the results in the text is very weak and could be improved.
In Table 2, the values for the CTR feed are missing. This is important to understand the benefits of the IPFE feed and forms the basis for the following tables.
Answer: in fact, the extract, not a diet containing the extract, was analysed. Therefore, there was no extract available in the CTR group. For clarifying, some words were added to line 86.
L191: To understand the differences between CTR and IPFE0, it would be important to know how much feed the fish actually uptake. This is because feed intake plays an important role in growth parameters.
Answer: data was added to Table 3.
The table 4 is splitted into two parts. Correct this or repeat the heading on the next page.
Answer: it was corrected.
Discussion
The discussion is extensive and includes the topics: antioxidant activity, growth performance in fish, microbiota/gut microbiota, mucosa surfaces of fish in context with immune parameters, and date fruit as a feed additive due to the antioxidant effect. The statements on the authors' own data remain on the surface and could be deepened.
L223/L293: On one hand … on the other hand. Contrast!
L252: In this sentence, you take about the uptake rate of feed from other authors but gives no information or relations to your own data.
Answer: a paragraph was added to the end of the discussion part. The mentioned mistakes were also corrected.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Please modify the manuscript as suggested. Find the attachment for the suggestions.
Comments for author File: Comments.pdf
Author Response
Here it is written in % value where as in Table 2 it is written as μ / ml. Rectify accordingly. It was suggested to do a data transformation for % data which is not reflected in the revised manuscript.
Answer: this comment was applied.
In all explained results write in terms of significant (P<0.05) or not significant (P>0.05) increased or decreased. Because some results showed chaanges numerically but not statistically.
Answer: this comment was applied.
Author Response File: Author Response.pdf
Reviewer 3 Report
All points of criticism have been implemented in the second version of the manuscript. Now, the abstract is balanced and shortened. Minor errors have been corrected. Missing information in material and methods are added. The table headings are arranged below the table in two different font sizes. Please check if this meets the author's instructions in the journal Fishes. The topic of feed intake is not discussed further, but the data have been added in Table 3. For this, the advantages and disadvantages of gamma γ-irradiation were included in the discussion in order to make your own point of view clear. The formula has also been exchanged in the correct way.
There ist some little error in the manuscript.
L 257: A heading was left in the middle of the text, which should be removed.
no furtehr comments
Author Response
The table headings are arranged below the table in two different font sizes.
Answer: it was corrected.
The topic of feed intake is not discussed further, but the data have been added in Table 3.
Answer: a sentence was added to lines 267-230.
There is some little error in the manuscript.
Answer: it was tried to correct grammatical mistakes.
L 257: A heading was left in the middle of the text, which should be removed
Answer: it was corrected.
Author Response File: Author Response.pdf
