PONE-D-19-26666

Toxicopathological studies on the effects of T-2 mycotoxin and their interaction in juvenile goats

PLOS ONE

Dear Dr Basalingappa,

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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5. Review Comments to the Author

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Reviewer #1: In this study entitled " Toxicopathological studies on the effects of T-2 mycotoxin and their interaction in juvenile goats", Nayakwadi et al. have delineated the mechanisms by which T-2 toxin induced toxicosis in goats. The authors demonstrated that T-2 toxin elicited significant haematological, biochemical and histological alterations in several tissues including liver, intestine, kidney, spleen, brain etc. They reported altered expression of HSPs, pro-apoptotic and pro-inflammatory cytokines upon exposure to this toxin. Finally, they concluded that the T-2 toxicosis is mediated through oxidative, apoptotic and inflammatory mechanisms in goats. This study is very important as this toxin has been considered to elicit fatal reactions among animals and humans upon consumption. The findings from this study may open a new avenue in understanding the T-2 toxicosis in goats, which may be helpful in diagnosis and prevention of diseases in goats.

The study is well designed and the manuscript is fairly written, however, few queries need to be answered.

Major Comments:

1. Did the authors observe any pathological lesions in rumen or other compartments of stomach during 15th and 30th day of toxin treatment? Any additional data related to this may strengthen the current findings.

2. Under the Materials and Methods, in the lines 169-170, DAPI is mentioned as chromogen. However, the authors have used DAB as chromogen in TUNEL kit. This needs to be corrected. Also the sentences in the lines 183-186 should be removed as the authors have performed real-time RT-PCR and therefore sequencing is not required in mRNA expression studies.

3. Under the results, in line 215, the authors mentioned that "no mortality was observed during the experimental period". Did they observe any abnormal signs in the toxin treated groups during the study? Supportive evidences may be provided in this section.

4. What is the rationale for assessing IL-1apha than IL-1 beta gene expression, since IL-1beta is more potent than IL-1alpha in stimulating IL-6 release in several tissues?

5. The discussion is too descriptive and need to shortened with straightforward discussion.

6. The conclusion needs to be rewritten as it is not clear.

Minor Comments:

1. The authors should mention the name of the authors while referring to the methodology. e.g. Under Materials and Methods, in line 95, the authors mentioned "produced by fermentation of maize and wheat mixture as per the method described by [32]". Similarly, in line 139, the authors mentioned "MLN and brain tissue homogenates for all the animals on 15th and 30th day according to [33]". The authors name should be mentioned in those places.

2. In Figure legend 15: Correct the spelling of " sattellitosis".

3. Cross check the image magnification (both bright field and EM) in all the figures.

4. In the Table 1, under Control, with respect to Route of administration, it has been mentioned "Oral (Mixed with feed)". Did the control feed was mixed with any vehicle?

5. The Table 5 legend reads "T-2 Toxin induced histological alterations (score card) in different organs of young kids on 15th and 30th days fed with 10ppm and 20ppm dose". Remove the word "young kids" and change to "goats".

6. Under "Table 9: Effect of T-2 toxin on expression of heat shock protein (HSP) genes in different tissues in 10ppm and 20ppm groups on 30th day", the P value of some of the HSP mRNA expression levels are greater than P<0.05, which suggest no significant difference between the 10 and 20 ppm toxin treated goats and control. However, the "Expression pattern" suggests "up regulation". So check all the parameters.

7. The authors should strictly follow the references as per the journal format.

8. The manuscript should be checked for grammatical corrections and uniformity to match with the journal format.

Reviewer #2: This study throws light on the effect of T-2 mycotoxin contamination in juvenile goats and holds promise in goat husbandry considering their importance in economy and international trade. The authors have extensively studied the impact of T-2 mycotoxin in goats with respect to their growth and pathophysiology. Following are the queries that the authors need to address before it goes for publication.

1. The authors have stated that such studies have been carried out in other animals like sheep, pigs and poultry. In that case, what is the novelty of this study other than the organism used?

2. It has been mentioned that the toxin was mixed with the feed at specific concentrations. Is there any way the authors could assess the amount of toxin ingested by each animal?

3. The authors could give a possible explanation for the elevated levels of HSP molecules in the brain of control animals as opposed to those treated with the toxin.

4. The figure legends are obscure and very cryptic. It should be more descriptive for the readers to follow the results.

5. There are a few typographical errors that need to be corrected.

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Reviewer #1: No

Reviewer #2: No

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Toxicopathological studies on the effects of T-2 mycotoxin and their interaction in juvenile goats

PONE-D-19-26666R1

Dear Dr. Basalingappa,

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With kind regards,

Natarajan Aravindan

Academic Editor

PLOS ONE

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