Earlier Activation of Interferon and Pro-Inflammatory Response Is Beneficial to Largemouth Bass (Micropterus salmoides) against Rhabdovirus Infection

Round 1

Reviewer 1 Report

The results of this study indicate that early activation of interferon and pro-inflammatory response is beneficial against MSRV infection. Although a quite expected result, it is interesting to see this correlation documented, although the results do not document a causal relationship.

The description of the experimental design is vague and must be improved significantly. It is hard to understand immediately what the groups U1, P1 and U3 are. Apparently, the use of the term "species"(l.9, l. 65) is wrong in this context. The species name is, in all three cases, M. salmoides (note that species name shall be with small caps, l.3 - this error is repeated several places in the text, also with other species, like Litopeneaus vannamei - l.- 64). One is left wondering whether these (U1, P1, U3) are breeding groups, or just strains of different origin. Contrary to this, it is also stated that the "fish were divided into 3 groups (U1, P1 and U3 group)" (Line 96). I would interpret the term "divided" as an active measure taken by the scientists to divide into experimental groups. Although this Reviewer does not have English at his first language, it must be stated that the manuscript needs significant linguistic improvement, also regarding the correct use of scientific terms.

Certain methods are listed without any references (line 135, 138). 

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Reviewer 2 Report

The article entitled “Earlier activation of interferon and pro-inflammatory response is beneficial to largemouth bass (Micropterus Salmoides) against rhabdovirus infection” by Runzhen He et al. discusses the pro-inflammatory response against rhabdovirus infection.

The article is homogeneous and well-argued and the results are presented in a clear and straightforward way.

The conclusions are consistent with the results.

I suggest that authors re-check the manuscript, implement the introduction and create a summary table of results to improve the quality of the article and make it easier to understand the data obtained.

Author Response

Thank you for your review. We have rechecked and revised some mistakes and deficiencies in the manuscript.

Reviewer 3 Report

The manuscript “Earlier activation of interferon and pro-inflammatory response is

beneficial to largemouth bass (Micropterus Salmoides) against rhabdovirus infection” from He et al. is about host response of Micropterus salmoides against MSRV. The findings are underlined with gene expression data, enzyme data and in vivo data. The presented work has major flaws, especially in study design. The authors described well their findings but missed to include necessary data. With the presented work an important topic in aquaculture is emphasized. The presented manuscript needs further improvement before considering publication.

In general:

Comparison of RGNNV vs. MSRV was made, but the basis or explanation is missing.

In this study data on type I IFN are missing. Different types of IFN work in different direction, which was not mentioned sufficiently.

Why not using fry or compare your data with data from fry? You mentioned that mainly fry is affected.

Why doing artificial infection (i.p.) and not by immersion, which represents a natural route?

Why haven’t you tested different organs for gene expression?

Data from Gao et al were mentioned several times, however no sufficient discussion of your data and those of Gao was done. At all, your study design is similar to the one published by Gao, except you have in vivo data and used three types of M. salmoides and used fewer immune genes.

Micropterus salmoides -> no capital S for salmoides

Writing spelling

L33: Zhejiang -> capital Z

L38: leaded -> led

L64: Litopenaeus Vannamei -> vannamei

L65 ff: U1, U3, P1 are for sure no species, but strains or breedings

L68/69: non-specific immune -> non-specific immune what?, a word is missing

L81: healthy -> health

L190: fish were observed / dead fish were removed

L181: I think you mean U1 group

L238: IFN -> which one?

L243: SVC -> SVCV

M&M2.2:

• Which 10 tissues?
• Include qPCR protocol
• Why using fish fry for virus production and not Largemouth bass skin (LBS) cells? Ethics?

M&M2.3:

• untreated group missing
• group treated with crude tissue extract like M&M 2.2 missing

M&M2.4: production of standard curve is hardly understandable, incomplete?

M&M2.6: Ref 2-ddCt

Author Response

Please see the attachment.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

The present version is significantly improved compared to the first version I saw. Of special importance: the use of the terms "species" and "strains" have been straightened up, the species names have been corrected, and both are now consistent with the generally accepted use of the terms.  The experimental groups have been adequately explained. These improvements also make the general description of the experimental setup easier to follow. The non-referenced methods have been supplied with adequate references. 

Author Response

Thank you for your kind comments again.

Reviewer 3 Report

The revised manuscript “Earlier activation of interferon and pro-inflammatory response is

beneficial to largemouth bass (Micropterus Salmoides) against rhabdovirus infection” from He et al. is about host response of Micropterus salmoides against MSRV. The findings are underlined with gene expression data, enzyme data and in vivo data. The presented work has still major flaws, especially in study design and presentation and discussion of the results. The authors described well their findings but missed to include necessary data. With the presented work an important topic in aquaculture is emphasized. The presented manuscript needs further improvement before considering publication.

L248: IRF / IFN -> which one? Please be precise

M&M / results: The authors prepared a PBS-treated group; however, the data are not included. By this a clear presentation of the data was avoided, since treated fish and control cannot be compared, which is strongly advised.

M&M2.4: The authors used an unpublished qPCR. Why not using the opportunity and include it in this manuscript? Doing so everyone could reproduce your results.

Discussion:

In fact, the authors took the results of Gao et al. into account, however, this was not done as necessary. No IFN-I response was clearly measured or discussed. IFN-g and IRFs were added, but not mechanism behind all this.

Author Response

Please see the attachment

Author Response File: Author Response.pdf