Review Reports

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Oxidative stress is a significant factor limiting the health and development of piglets, particularly under exposure to environmental toxins such as mycotoxins. In recent years, increasing attention has been given to the use of exogenous antioxidant enzymes, including catalase (CAT), as strategies to protect against oxidative damage. Exogenous CAT may not only directly neutralize reactive oxygen species but also influence intestinal and liver function through modulation of the gut microbiota and regulation of glycerophospholipid metabolism in the liver. The present study aimed to evaluate the effects of exogenous CAT administration on oxidative stress, intestinal and liver health in piglets exposed to mycotoxins, which may provide new insights into its protective potential in nutrition and animal welfare. Before publication, the following points should be considered:

Abstract

A clear description of the experimental groups is missing. The number of groups and the factors differentiating them should be specified (e.g., control group – CON, mycotoxin-exposed group – TOX, mycotoxin-exposed group supplemented with catalase – TOX+CAT). This also applies to the Materials and Methods section.

Information regarding the duration of the experiment is lacking. Please specify the exact start and end of the study (e.g., from weaning, day 0 of the experiment, to day 28). This should also be reflected in the Materials and Methods.

Lines 17–18: The phrase “Parameter analysis was conducted on day 28” requires clarification. It should be specified that analyses included biochemical parameters and oxidative stress markers assessed in blood and tissue samples collected from piglets on day 28 of the experiment.

Keywords

4. The term “exogenous” is ambiguous. If it refers to exogenous catalase (CAT), this should be explicitly stated, e.g., “exogenous catalase (CAT)”.

Materials and Methods

5. Lines 101–102: A brief description of sample collection should be provided (e.g., blood was collected from the jugular vein, intestinal and liver tissues were preserved for biochemical and histological analyses).
6. Catalog numbers of the kits used and the model of the equipment (biochemical analyzer, spectrophotometer) should be specified.

7. Please provide details on how protein content in the intestinal wall was measured, including references to the kit or literature.

Table 1

8. Feed conversion ratio (FCR) values for the TOX group should be checked, as they may be incorrect or inconsistent with other data.

Results and Discussion

9. Lines 168–169: The description of the results should be verified against the data presented in Table 2, as inconsistencies may be present.

10. Lines 189–190: The correctness of the p-values should be confirmed. In Table 4, the values are 0.006 and 0.018, respectively – ensure the text matches the table.

11. Consider extending the statistical analysis to include a linear trend test (linear regression or orthogonal polynomial contrast) to determine whether increasing doses of exogenous catalase result in a linear response in production, health, and biochemical parameters.

Conclusion

12. The conclusion should clearly indicate which of the tested doses of exogenous catalase may be recommended for practical use in piglet nutrition.

After addressing the above points, the manuscript may be considered for publication.

Author Response

Please see the attachment. 

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The approach of using catalase to address health issues caused by mycotoxins in animal nutrition, particularly for poultry and pigs, is quite intriguing. The research is well-designed, and the variables have been studied in line with the study's objectives. However, there are some gaps in the methodology that require further detail and clarity. Additionally, the conclusions should provide more information about this interesting study.

Comments for author File: Comments.pdf

Author Response

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Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

Dear Authors,

Your scientific manuscript, which I received for review, is very interesting. You've raised a very timely and compelling topic: exposure to mycotoxins in piglets and the protective importance of exogenous catalase. I greatly appreciate the research concept and the work you put into obtaining the results and writing the manuscript. However, as a reviewer, I have a responsibility to critically analyze your manuscript and point out any weaknesses. I'm submitting my comments and suggestions for improvement in the bullet points below.

1. My first concern concerns the keywords. Most of them overlap with the words in the title. This is unnecessary and inconsistent with global standards for scientific publications. Keywords are a second-order criterion that, in addition to the title, allows people searching for papers in your field to find your article. Matching the words in the title with the keywords won't expand your audience. If they enter the word "mycotoxins" in the Web of Science or Google Scholar search engine, even though it's in both the title and keywords, how will that expand your audience reach? Therefore, you should change words like "piglet"; "exogenous"; "catalase"; "mycotoxin"; and "oxidative stress" to other words related to your research topic.
2. The sentence "Fusarium graminearum, the most common pathogenic fungi in feed, is capable of producing mycotoxins like deoxynivalenol (DON), aflatoxin B1 (AFB1), and zearalenone (ZEA) during its growth and reproduction [1]" is very incomprehensible and conveys incorrect information. First, mycotoxins are not produced in feed; they are already present there. Fusarium graminearum infects plants growing in the field, such as cereals, whose grain is used to produce feed. If a fusarium infection occurs, it produces mycotoxins, but production ceases after the grain is harvested. Rewrite this sentence.
3. The sentence "It was common that FGM in feeds exceeded the limit range of the standard for feeds due to various reasons in production, such as improper feed storage conditions (e.g., moist or high temperature) [3]. - during feed storage, aspergillus molds develop, which produce aflatoxin, not mycotoxins produced by fusarium, such as deoxynivalenone - change this sentence.
4. Testing for mycotoxin content with an ELISA test is only a qualitative, not quantitative, assessment. I believe this method cannot be used to determine whether the actual amounts of mycotoxins you're using were the same. It's important to note that, in addition to free-form mycotoxins such as DON, there are also mycotoxins modified by glycation, which cannot be detected by an ELISA test, but only by chromatography.
5. The conclusions are very laconic, neither indicating the dose nor providing any significant information about the effect on the microbiota or the activity of individual catalase doses. This needs to be written down.

Author Response

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Author Response File: Author Response.pdf

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

Dear Authors, thank you very much for your responses and the effort you put into improving the manuscript. I have two more suggestions, which I've included below.

Your response doesn't include what I wrote. Response 2: Thanks for your suggestion. Based on your comment, we have revised this sentence in the revised manuscript. Details are listed below: Fusarium graminearum, the most common pathogenic fungi infecting cereal crops, is capable of producing mycotoxins like deoxynivalenol (DON), zearalenone (ZEA), and aflatoxin B1 (AFB1) during its growth and reproduction [1]. (See the Introduction on page 2, lines 43-45) Aflatoxin B1 is produced by Aspergillus fungi, not Fusarium. Please correct this as it is a very serious methodological error.

Response 4: Thanks for your valuable comments. Li et al. [1] tested the authenticity agricultural product samples using the ELISA kit and HPLC, and found that a good correlation of the results of HPLC (Y) and ELISA kit (X) was obtained (Y = 0.9322X + 113.78, R2 = 0.9589), which further indicated that the results of the ELISA kit were reliable. Furthermore, some researches (references 2-9) also used the ELISA kit to determine the content of mycotoxins in feed. Our previous study [10] has been shown that the contents of modified DON (3-acetyl-DON and 15-acetyl-DON) in DON-contaminated rice were low, with free-form DON being dominant. Similar to our previous study, Marquis et al. [11] I found that the contents of DON-3G and 3+15-ADON were low in the Fusarium-contaminated diets, and DON was the major mycotoxin. - Thank you for this explanation, but it would be good to provide such an explanation in the introduction of the paper, along with a description that there can be modified and free forms of mycotoxins, but ELISA kit and HPLC tests, and found that a good correlation. It would also be good to use this literature: Różewicz, M. (2024). Mycotoxins in cereal grain as a result of infection of cereals by Fusarium fungi. Polish Technical Review. DOI 10.15199/180.2024.1.2

Author Response

Please see the attachment.

Author Response File: Author Response.pdf