Effect of Moderate-Intensity Endurance Exercise on Inflammatory Cytokines in Leukocytes of Dogs

Round 1

Reviewer 1 Report

Dear authors,

I recommend your manuscript for publication, but has some questions.

Line 3, 13 and other in text – don’t use term “in the blood”. Concretize it – in blood cells, in serum etc.

Line 17 – don’t use abbreviation without description.

Line 94 – you are used too small groups of dogs in case. But body weights and ages of them are too various. It was decrease significance of results. How are you solved this problem?

Line 135 – How are you prepared serum from heparinized blood? According your method it was be a plasma. You are was writing it in line 138.

Line 172 – why you are used Bonferroni correction? It was necessary only if you are having a many comparisons.

 Line 188 and other – in tables show numbers of animals per groups.

Regards,

Author Response

I sincerely appreciate your consideration for reviewing our research results. All the details pointed out significantly contributed to improving our manuscript. We have revised the manuscript accordingly and provided the following list of changes.

Author Response File: Author Response.doc

Reviewer 2 Report

Authors tried to clarify the effect of moderate-intensity short-term endurance exercise originally programmed by authors on hematology and serum biochemistry parameters and the expression of immune-related cytokine genes in the healthy dogs. However, the interpretation of the obtained results seems to be difficult under the present manuscript presentation, which should be improved focusing on the following points.

It remains unclear species differences between human and dog regarding immune system affected by exercise. Can we recognize that almost similar trends in changes of cytokines between human and dog are predicted?

Author claimed that moderate exercise would lead to positive effect on immune system in the dogs. However, what is the essential points of positive effect under the health condition is not explained in the manuscript. The decrease in mRNA expression levels of indicated cytokines are included in the “positive effect”? Authors should describe the “positive effect” more clear. Or, authors should show positive or negative control compared to the obtained results.

Why the authors focused on mRNA expression levels of the cytokines, but not the protein expression levels? Protein expression level of several cytokines essentially affect the immune responses in the dogs.

Main clear findings obtained from the present study is that moderate-intensity short-term endurance exercise decreased mRNA expression levels of TNF-α, IFN-γ, and IL-1β in the blood of the dog. Authors well discussed why the results were obtained based on enough evidences. Conversely, how obtained findings lead to advanced benefits on health of the dogs remains unclear.

Author Response

I sincerely appreciate your consideration for reviewing our research results. All the details pointed out significantly contributed to improving our manuscript. We have revised the manuscript accordingly and provided the following list of changes.

Author Response File: Author Response.doc

Round 2

Reviewer 2 Report

Several unclear points could be improved in the revised manuscript. Although I wish authors to analyze the protein expression levels of several cytokines, I could see that authors are planning to conduct a new research project including protein expression levels of indicated cytokines using muscle and blood.