Effect of FASN, SCD, and GH Genes on Carcass Fatness and Fatty Acid Composition of Intramuscular Lipids in F1 Holstein × Beef Breeds
Round 1
Reviewer 1 Report (Previous Reviewer 3)
The manuscript is well written. There are valuable results and information evaluating meat quality depending on genotype.
Author Response
We thank Reviewer for the comments.
The reviewer has no suggestions for improving the manuscript, so we have no specific response.
Author's
Author Response File: 
Author Response.pdf
Reviewer 2 Report (Previous Reviewer 1)
The reviewer reviewed the improved version of the manuscript. The reviewer recognizes that the manuscript has been significantly improved.
However, the reviewer feels the need of additional minor modifications for the manuscript. The reviewer requests the authors to add superscripts(a,b,c...) to the values in each table to indicate which values are significantly different from the others among breeds or genotypes based on statistical multiple comparisons.
The reviewer has found several grammatical errors in the manuscript (e.g., L.267- "In Table 4 is present effect of SCD genotypes and ...", "EUROP conformation classification was significant effected...". ). English re-proofreading is requied.
Totally, the reviewer's decision is "Minor revision".
Author Response
We thank the reviewer for his useful observations and suggestions.
There is a corresponding response to each suggestion (in the text below).
Point 1: However, the reviewer feels the need for additional minor modifications to the manuscript. The reviewer requests the authors to add superscripts (a,b,c...) to the values in each table to indicate which values are significantly different from the others among breeds or genotypes based on statistical multiple comparisons.
Response 1: The authors sincerely thank the reviewer for the useful suggestion. The suggestion is fully accepted. We indicate values that are significantly different from the others among breeds or genotypes based on statistical multiple comparisons. We have highlighted the differences in the rows (p<0.05) with small letters (a, b, c), and for consistency, we kept p‑values (Table 2 - 5).
Point 2: Several grammatical errors in the manuscript (e.g., L.267- "In Table 4 is present effect of SCD genotypes and ...", "EUROP conformation classification was significant effected...".). English re-proofreading is required.
Response 2: We thank the reviewer for the useful suggestion. We have reread the manuscript and corrected some grammatical errors that we noticed. If the editors decide, we will accept additional proofreading of the manuscript text.
Author's
Author Response File: Author Response.pdf
Reviewer 3 Report (New Reviewer)
For me, major revisions are needed.
Comments for author File: Comments.pdf
Author Response
We thank the reviewer for his useful observations and suggestions.
There is a corresponding response to each suggestion (in the text below).
Point 1: The abstract should contain essential information from Introduction, Material and Methods, Results and Discussion/Conclusions. Please add an introductory sentence into the Abstract. Moreover, in the abstract should not include acronimus without an explanation, such as NIT or MLD...
Response 1: The authors sincerely thank the reviewer for the useful suggestion. We add an introductory additional sentence to the Abstract (lines 10-11). We have corrected the acronyms (NIT, MLD) (lines 16-17).
Point 2: The chosen keywords are included in the title as well. Please consider it acceptable to use different keywords.
Response 2: We thank the reviewer for the useful suggestion. We propose new keywords (lines 26-27).
Point 3: As the manuscript describe the importance of crossbreed the introduction is lacking of the importance of milk specializide or dual porpouse breed. I suggest to add more information regarding that and I suggest to include this reference: 10.1080/1828051X.2022.2032850
Response 3 We thank the reviewer for the suggestion. We add new information in the text of the manuscript (line 51-56) and cite four additional papers (line 545-554; reference [9], [10], [11], [12]).
Point 4: These meat fatty acid modifications are not only determined by the genetics of the animals but also the diet play a kye role. Please add this imformation and cite these papers: 10.3390/antiox11050827 and 10.3389/fvets.2021.662079, where thanks the diet they were able to change the fatty acide profile of the meat.
Response 4: We thank the reviewer for the suggestion. We add some new information (lines 56-62) and cite three additional papers (lines 555-563, reference [13], [14], [15]).
Point 5: Also in the discussion I suggest to discuss the importance of the cross breed in the cattle breeding, moreover, I suggest to include some practical implication of the results on genetic improvment of the cattle breed.
Response 5: We thank the reviewer for the useful suggestion. We have included some practical implications of the results for genetic improvement of the cattle breed (lines 448-455).
Author Response File: Author Response.pdf
Round 2
Reviewer 3 Report (New Reviewer)
Dear authors, the paper improved a lot with the revisions
This manuscript is a resubmission of an earlier submission. The following is a list of the peer review reports and author responses from that submission.
Round 1
Reviewer 1 Report
Pleasae see the attached file.
Comments for author File: Comments.pdf
Author Response
The Review Report (Reviewer 1)
Comments to the article “agriculture-2134102”
This manuscript deals with the effect of polymorphisms in FASN, SCD and GH genes on carcass fatness, intramuscular fat content and its fatty acid composition in longissimus dorsi muscles (LMD) of Holstein crossbred bulls and heifers. The polymorphisms have already been reported to be involved in fatty acid compositions for other beef breeds (Japanese Black, Korean, Fleckvieh etc.), and the author stated that there are several significant relationships between the polymorphisms and fatty acid compositions in the cattle studied.
The reviewer finds serious problems in the manuscript and will request the editorial board to reject the manuscript to be published.
Reviewer: First, the authors analyzed the data from the eighty cattle all together, without taking the effect of the breed into account. There maybe significant differences among the breeds in carcass traits and fatty acid compositions even though here would be no differences in FASN, SCD and GH gene allele frequencies among the breeds, although the authors did not mention the effect of the breeds at all. The heads for each breed (twenty) is not enough to elucidate the effect of the polymorphisms. The use of single breed and the increase in head (e.g. eighty for one breed) would be better for such studies to exclude the effect of breed.
Author's: We would like to sincerely thank the reviewer for useful suggestions. Corrected according to reviewer suggestion.
The authors are aware that it is better to perform each study on a larger number of cattle. Based on the evaluation of the required sample size, we decided to use 80 animals. Scientific and practical interest (interest of breeders) was the incentive to include four beef breeds in the crossbreeding program with Holstein cattle to obtain an answer to the effects of genotype on carcass fatness and fatty acid content of meat (they influence carcass and meat quality and market value). Sex was determined as one of the factors, as we collected male and female calves. Conventional fattening technology is used in beef fattening.
The authors thank the reviewer for pointing out the need to refine the model. We included genotype and sex as variables and age as a covariate in the model.
The refined model is described in the manuscript (lines 200 to 204). Before the refined model, all values (LS mean±SE and p-value) in Tables 3 to 5 were recalculated (all LS Mean±SE and p-values in Tables 3 to 5 were corrected). The observed corrected effects of the influence of the FASN, SCD, and GH genes on carcass fatness and fatty acid composition are discussed in the “Results” and “Discussion” text. Application of the improved refined model provided new insights into the relationship between SCD gene polymorphism and fatty acid composition of intramuscular lipids.
Consistent with the reviewer's recommendation, we added a table of observed values for carcass fatness and fatty acid composition by genotype and sex. We have added Table 2 (lines 244 to 250) and the text that follows the indicated Table 2 (lines 222 to 242).
Because our discussion focuses on the "Effect of FASN, SCD, and GH genes on carcass fatness and fatty acid composition of intramuscular lipids," we have added a commentary on the influence of sex on carcass fatness and fatty acid composition (lines 368 to 375). The authors have added two additional references (43 and 44; lines 492 to 495).
The reviewer notes that "Heads for each breed (twenty) are not enough to clarify the effect of polymorphisms," but we are not able to increase the number of animals in the research at the moment. We believe that 80 individuals with the added calculation model (genotype and sex as variables, age as a covariate) is sufficient for a reliable evaluation of the effects studied (relationships of FASN, SCD and GH genes on carcass fatness and fatty acid composition of intramuscular lipids).
We will consider the reviewer recommendation when we are able to extend the research.
Reviewer: Second, the authors did not take sex into account in the data analysis, either. Supposed that the allele frequencies for the three genes do not differ between bulls and heifers in a single breed, there may be significant differences in carcass traits and fatty acid compositions between both sexes. The author slaughtered the bulls at 19 -20 months of age, while the heifers were slaughtered at 14 to 15 months of age. The younger age at slaughter for the heifers than the bulls may also result in notable difference in body weight at slaughter between both sexes. The age will be involved in the difference in fatty acid composition. Generally, older and heavier cattle tend to exhibit higher unsaturated fatty acid content in LMD. The effect of sex should be included in the model formula.
Author's: We would like to sincerely thank the reviewer for suggestions.
Corrected according to reviewer suggestion.
The authors apologize for not immediately providing accurate information on the slaughter ages of bulls and heifers (our big omission). We analyzed the actual slaughter ages of bulls and heifers and found the following: Young bulls were slaughtered at an average age of 489.8±12.9 days, and heifers at 468.0±8.2 days (lines 98 to 99).
The effect of age on carcass fatness and fatty acid composition of intramuscular lipids was tested and was not significant. When calculating the influence of sex on carcass fatness and fatty acid composition of intramuscular lipids, the influence of age was included as a covariate. We included genotype and sex as variables and age as a covariate in the refined model.
Reviewer: Totally, the authors should re-analyze the data using more appropriate model formula, including breed and sex as fixed effects. If there are wide dispersions in final body weight within bulls or heifers, it should also be included as a random effect in the formula.
Author's: We would like to thank the reviewer again for pointing out a gap in the analysis of the effects of the FASN, SCD, and GH genes on carcass fatness and fatty acid composition of intramuscular lipids. We corrected the model formula by including breed and sex as fixed effects and age as covariates (lines 200 to 204) and recalculated all values in Tables 3 to 5 (including Table 2).
Author's reply to the Review Report (Reviewer 1)
We thank the reviewer for useful observations and suggestions.
There is a corresponding response to each suggestion (in the text above).
Author Response File: Author Response.pdf
Reviewer 2 Report
Modelling of the beef quality begins at the stage of rearing animals for slaughter, and the final quality of meat is influenced by many factors, which can generally be divided into intra-vital and post-slaughter factors. Among the pre-slaughter factors, genetic and non-genetic conditions play an important role, such as: breed, sex, age and rearing system, nutrition and pre-slaughter turnover. Among other factors, the quality characteristics of the meat are also greatly affected by the natural differences between the muscles that are related to their physiological function and structure. The most important post-slaughter factors include the handling of carcasses and meat after slaughter, including chilling, carcass cutting, packaging, storage and maturation of meat. Despite the slight variation in the quality of meat between breeds of cattle, it is possible to use individual breeds or their hybrids to produce beef with the desired characteristics. This also applies to the fatty acid profile. Animal fat (including beef fat) is perceived very one-sidedly as a source of unhealthy fatty acids and cholesterol. However, beef is a rich source of animal protein, and some of its valuable physicochemical properties do not deteriorate even in the conditions of 30-day freezing storage. The variability of the chemical composition of beef depends, among others, on the type of muscle, animal breed or sex. According to many authors, the share of fatty acids in beef may also be determined by environmental and physiological factors, as well as the breed of the animal.I believe that the manuscript "Effect of FASN, SCD and GH genes on carcass fatness and fatty acid composition of intramuscular lipids in F1 Holstein x beef breeds" has both scientific and industrial importance. The knowledge contained in it will be successfully used both in the agricultural sector during the production of livestock and among food technologists. It should also attract the attention of dieticians who need to broaden their knowledge of food ingredients of animal origin. I accept the manuscript "Effect of FASN, SCD and GH genes on carcass fatness and fatty acid composition of intramuscular lipids in F1 Holstein x beef breeds" and recommend it for publication in Agriculture (ISSN 2077-0472).
Author Response
Author's reply to the Review Report (Reviewer 2)
We thank Reviewer 2 for his useful comments.
The reviewer has no suggestions for improving the manuscript, so we have no specific response.
Reviewer 3 Report
The manuscript is well written. There are valuable results and information evaluating meat quality depending on genotype.
Some points need to be clarified:
Line 66: What means „USFA”? It is the first time in the text so you should translate the abbreviation.
Lines 70-73. It is not clear. I think you should share this sentence on two sentences. For example: Since farmers and especially consumers are interested in producing meat with favorable nutritional and sensory characteristics, some of the solutions to improve meat quality are needed/looking. Some of these could be the selection of specific breeds for crossbreeding, genotyping, and selection for specific candidate genes in breeding animals, and the use of sexed semen to control the sex of calves.
Line 80: more often instead of „Piedmontese” is used „Piemontese”. I propose that short names should be next to the name of breed
Line 218: „No significant effect on carcass fatness was observed among the FASN genotypes.” Add (Table 2) because it is data from this table already.
Line 412: „Oprzadek” not „Oprazadek”
Line 433: There is no this publication in the text
Author Response
The Review Report and Author's Reply to the Review Report (Reviewer 3)
Comments and Suggestions for Authors
The manuscript is well written. There are valuable results and information evaluating meat quality depending on genotype.
Some points need to be clarified:
Reviewer: Line 66: What means „USFA”? It is the first time in the text so you should translate the abbreviation.
Author's: Corrected: The USFA abbreviation has been clarified (line 68).
Reviewer: Lines 70-73. It is not clear. I think you should share this sentence on two sentences. For example: Since farmers and especially consumers are interested in producing meat with favorable nutritional and sensory characteristics, some of the solutions to improve meat quality are needed/looking. Some of these could be the selection of specific breeds for crossbreeding, genotyping, and selection for specific candidate genes in breeding animals, and the use of sexed semen to control the sex of calves.
Author's: Corrected according to reviewer suggestion (line 72 to 76).
Reviewer: Line 80: more often instead of „Piedmontese” is used „Piemontese”. I propose that short names should be next to the name of breed
Author's: Corrected according to reviewer suggestion (line 83).
Reviewer: Line 218: „No significant effect on carcass fatness was observed among the FASN genotypes.” Add (Table 2) because it is data from this table already.
Author's: Corrected, according to reviewer suggestion (line 252).
Reviewer: Line 412: „Oprzadek” not „Oprazadek”
Author's: Corrected (line 451).
Reviewer: Line 433: There is no this publication in the text
Author's: Reference Oh et al. [32] are cited in text of manuscript, in line 305, 331 and 346.
Author's reply to the Review Report (Reviewer 3)
We thank the reviewer for his useful observations and suggestions.
There is a corresponding response to each suggestion (in the text above).
Author Response File: Author Response.pdf
