Effect of Heat Starvation Stress on Physiological Immunity and Metabolism of Mizuhopecten yessoensis
Round 1
Reviewer 1 Report
This is an interesting study about the regulation of the metabolism pathway in M. yessoensis under food deprivation.
Although the research topic is more interesting, there are few suggestions to improve the content of the manuscript as follows:
1. Abstract needs modification with these sentences to improve the scientific writing, (a) In quest of healthy aquaculture system, much attention was attracted on the environmental 13 factors. (b) KEGG analysis 21 showed that DEGs in striated adductor muscle enriched in pathways involved in metabolism.
2. Similarly, in the methods sections, authors may read carefully and make corrections with improvement in English writing. For instance,
In Line 145, “After that, the software bowtie2 145 (v2.2.5) was applied to align the clean reads to the coding gene set and the expression level …..
3. Other minor Changes are, in discussion section, line 351-352 ..
“Analysis of data in striated adductor muscle showed no significant up-or down- regulation appeared in expression of genes encoding key enzymes in citrate cycle and glycogen biosynthesis/ degradation” needs to be modified.
Author Response
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Author Response File: 
Author Response.pdf
Reviewer 2 Report
In this study, the authors compared the starved and control scallops in the changes in meat yield, number of haemocytes, and several biological markers, including activities of acid phosphatase and lysozyme, and plasma protein concentration (BAC), total antioxidant capacity of plasma (T-AOC), and cellular malondialdehyde (MDA) concentration. They also performed mRNA-seq analyses to compare the gene expression levels between starved and control scallops. The present study contains some novel findings and useful information for scallops aquaculture industries, but the reviewer found several major and minor concerns which should be addressed before publication.
Major concern
For figures 1 through 7, please indicate the data of initial stage (day 0). The data of day 0 is quite important to understand the effects of starvation.
Please indicate the data of body weight (total weight) of scallops on days 0, 15, 30 and 45 for control and starved groups. The data on body weight gain/loss during the experiments are also quite important to understand the effects of starvation.
The authors say in the abstract (lines 25 to 26) that “The down-regulation of PGD and tktA genes on pentose phosphate pathway suggested that the carbohydrate decomposition and utilization was decreased in starved scallops.”, and also inDiscussion (lines 522 to 525) that “In this study, the down-regulated PGD and ktkA gene in pentose phosphate pathway indicated the decrease of ribose-5P and NADPH, pointing to the negative effect on synthesis of nucleotide and amino acid biosynthesis“, but there is no description on this result in the Result section.
The authors showed significant difference in Figure 1 through 7, but the methods for statistical analyses are not described in Materials and Methods section.
Minor concerns and suggestions
Line 101; Please show the size of cement pools, and volume of water.
Lines 167-173
What is "fertilizer fullness", “fullness” and “fertility”. These words should be defined somewhere in the manuscript.
Lines 171 to 172;
The fertility of the control group was significantly decreased under starvation stress for 30 d and 45 d (P<0.05).
The reviewer does not understand this description, because control group was not starved.
p.7, Table 1
The reviewer suggests this Table can be shown as a supplemental material.
Lines 253 to 256
The reviewer suggests the authors to provide the list of 27 up-regulated and 41 down-regulated genes of haemolymph and 426 up-regulated and 255 down-regulated genes of adductor muscles as a supplemental Table (as an excel file if possible). It must be quite informative for the readers.
p.9, Figure 9
Please indicate the top figure is for haemolymph and bottom is for striated adductor muscle.
Line 286; Figure 4 may be Figure 10.
Figure 13;
Please show the log2FoldChange values of hesokinase. pfkA, fda, gpmA and pyk, even their changes are not significant.
Lines 486 to488
The authors say that “On the other side, the gene encoding fbp which catalyzes the reaction of fructose-1,6P2 towards fructose-6P only in gluconeogenesis showed significant down-regulation, consistent with the trend of other DEGs on the pathway.”
The reviewer suggests the authors to show the log2FoldChange values of glucose-6-phosphatase (G6PC), and phosphoenolpyruvate carboxykinase (pckA) to support their assumption on the down-regulation of gluconeogenesis.
Author Response
Please see the attachment.
Author Response File: Author Response.pdf
Round 2
Reviewer 2 Report
Line 188
What is "fertilizer fullness"? The reviewer searched PubMed and google scholar for "fertilizer fullness", but no result was obtained. Please check again.
Line 190
What are fatworthiness and “fat fill”? Are these words different from “Meat yield”? If they are different, please explain the difference. If no difference, please use “Meat yield” instead of these word.
The reviewer suggested in the first round of review as follows,
9. Lines 486 to488，The authors say that “On the other side, the gene encoding fbp which catalyzes the reaction of fructose-1,6P2 towards fructose-6P only in gluconeogenesis showed significant down-regulation, consistent with the trend of other DEGs on the pathway.”The reviewer suggests the authors to show the log2 Fold Change values of glucose-6-phosphatase (G6PC), and phosphoenolpyruvate carboxykinase (pckA) to support their assumption on the down-regulation of gluconeogenesis.
And the authors responded as follows,
Response: Accept the expert's modification opinions, and it has been added in L104-105. “Figure 14. Log2 fold change of expression of DEGs involved in pentose phosphate pathway, and related results”(L375).
But the reviewer did not find the log2 Fold Change values of G6PC, and pckA in the revised manuscript. Please check it again, but if the authors think the log2 Fold Change values of G6PC, and pckA do not add any support to their assumption, it is OK.
Author Response
请参阅附件。
Author Response File: Author Response.pdf
