Polysaccharides of Atractylodes Macrocephala Koidz Alleviate LPS-Induced Bursa of Fabricius Injury in Goslings by Inhibiting EREG Expression

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript is meaningful. However, there are still some parts need to be modified further.

1. Material and Methods

1) Line 100. “The study was conducted with 10 goslings in each group, repeated five times.” Could you please explain why the experiment was repeated 5 times? Is every group composed of 10 goslings or 50 goslings? What is the sample size per group? 

2) Line 102-106, In the in vivo experiment, why were 2mg/kg LPS and 400mg/kg PAMK chosen? Some references should be provided to support these choices. 

3) Line 187: “The goslings were anesthetized and subsequently euthanized, after which the BF was 187 collected and its cells were extracted. ” How old were the goslings at the time of collection? 

2. Results

1) The figure1B,C,E,F, Figure2, Figure 5C, and Figure 6ABC are not clearly.

 

2) Avoid repeatedly describing the methods. For example, "The experiment administered treatment with LPS at a concentration of 0.1 μg/mL and PAMK at a concentration of 10 μg/mL. Gene and protein levels of EREG, c-FOS, RAS, JNK, and p-JNK within the EREG signaling pathway were detected in the BF cells of goslings from the Control, LPS, PAMK+LPS, and PAMK groups using the WB method" should be presented without redundancy. 

3）What is the sample size per group in vitro experiment? 

3. Conclusions：AP1 was not detected in the manuscript，therefore, it should not be presented in the figures.

Author Response

 

   

   

   

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

The authors investigated the effect of polysaccharides Atractylodes macrocephala Koidz (PAMK, main active component of the Chinese medicine Atractylodes macrocephala), on lipopolysaccharide (LPS)-induced Bursa of Fabricius (BF) injury in goslings. The results showed that PAMK administration at 400 mg/kg ameliorated LPS-induced BF atrophy and structural damage in goslings. PAMK treatment also mitigated the LPS-induced increase in IL-1β, MDA and iNOS levels in BF cells. Meanwhile, KEGG enrichment pathway analysis showed that DEGs were significantly enriched in the Toll-like receptor, p53, MAPK, GnRH, and ErbB signaling pathways. EREG was involved in the action of PAMK to prevent BF injury in goslings by LPS induction. These results provide a basis for application of PAMK in poultry production.

Several issues should be considered for improvement of this manuscript:

1.       Provide the stain of goslings in this study.

2.       Provide identification of the PAMK ingredients.

3.       Make clear the dosage of PAMK at 400 mg/kg body weight or feedstuff weight, and also for LPS.

4.       Provide rational of the selection of a single dosage of PAMK (400 mg/kg) and LPS (2 mg/kg BW).

5.       The writing of this manuscript need extensive editing, i.e. L200 Change 0.1 μg/mL, 1 μg/mL, 10 μg/mL and 100 μg/mL as 0.1, 1, 10 and 100 μg/mL.

6.       Provide repetition of the experiments in section 2.15 Statistical data analysis.

7.       Add scale bar in Figure 1D rather than the amplification.

8.       Add morphology of the cultured Bursa of Fabricius cells.

9.       Provide the primers used for quantitative PCR with the gene accession numbers.

10.   Provide health assessment of the goslings after treatment with PAMK and/or LPS.

11.   The paragraphs of the discussion are too long. It needs to be concise.

12.    The format of the references need to be corrected according to the journal instruction.

Author Response

 

   

Author Response File: Author Response.pdf