In Vitro Screening of Plant Materials to Reduce Ruminal Protozoal Population and Mitigate Ammonia and Methane Emissions
Round 1
Reviewer 1 Report
Thank you for your study.
The research is interesting, but it need reworking. The text need to be reordered. The experiment is not clearly presented and not statistically representative. In particular:
Introduction would be through, with references.
Material and methods described more clearly and explained in more details, divided in chapter:
1.plant materials: number, replications (it is necessary to have more plants of the same specie and the same part).
2: Rumen fluid collection
3: In vitro gas production
4. Chemical composition
When you use acronyms for digestibility we suggest to use that indicated in the article of Animals, 2020, 10, 775 (Supplementary).
Results and discussion could be improved
References do not meet the indication of the journal.
Author Response
Response to Reviewer 1
The research is interesting, but it need reworking. The text need to be reordered. The experiment is not clearly presented and not statistically representative. In particular:
Introduction would be through, with references.
Response: Thanks for your appreciation in our work and we wish that this manuscript will benefit to the reader if can possible published. We have rewritten the introduction with thorough references as your recommendation.
Material and methods described more clearly and explained in more details, divided in chapter:
1.plant materials: number, replications (it is necessary to have more plants of the same specie and the same part).
2: Rumen fluid collection
3: In vitro gas production
- Chemical composition
Response: Thanks for your beneficial suggestion. We've also reorganized all part in the subchapters according to your suggestions. Please see in manuscript.
When you use acronyms for digestibility we suggest to use that indicated in the article of Animals, 2020, 10, 775 (Supplementary).
Response: Thank you. We have agreed and already added as your suggestion. Please see in manuscript.
Results and discussion could be improved
Response: Thank you. We have agreed and improved as your suggestion. Please see in manuscript.
References do not meet the indication of the journal.
Response: Thank you. We have agreed and thoroughly modified all as your suggestion. Please see in manuscript.
Reviewer 2 Report
The article "In vitro screening of plant herbs to mitigate ruminal protozoal, ammonia and methane emission of beef cattle" submitted for review addresses the potential for using plants to reduce methane in vitro.
Reduction of methane emission in livestock production is a necessity due to the progressing climate warming. This paper fits well with this theme and discusses the possibilities of using natural plants for this purpose.
In my opinion, the manuscript could be accepted with a few changes.
My comments and remarks are in the attachment.
Comments for author File: 
Comments.pdf
Author Response
Response to Reviewer 2
The article "In vitro screening of plant herbs to mitigate ruminal protozoal, ammonia and methane emission of beef cattle" submitted for review addresses the potential for using plants to reduce methane in vitro.
Reduction of methane emission in livestock production is a necessity due to the progressing climate warming. This paper fits well with this theme and discusses the possibilities of using natural plants for this purpose.
In my opinion, the manuscript could be accepted with a few changes.
My comments and remarks are in the attachment.
Response: Thank you. We agree with the referee's point-by-point recommendation on this issue and have changed it as our response to the referee in the attachment. Please see in manuscript.
Author Response File: 
Author Response.docx
Round 2
Reviewer 1 Report
Dear Authors the article need to be reviewed better. It was not clear the last time my indications, may be.
So I ask you to review better the paper.
L16 The organic matter digestibility (AOMDvt)
L30 I suggest to change the order of phrases:
global greenhouse emissione (GHG), that have become a major concern in recent decades. Enteric fermentation and feed production.....Methane is the....Entheric methane
L93 total mixed ration (TMR)
Material and methods need to be described better and the chapter need to have the same order that you use in results.
So I advise you to describe in this order:
2.2 Chemical composition analysis
2.3 Rumen collection
2.4 In vitro gas production. Insert after L138 the text from L155 to L177, as it describe in vitro gas method
2.5 In vitro rumen fermentation. Describe the method and materials used
2.6 Bacterial and protozoal account. Describe the method
Insert also in the chapter correct the method used for ammonia, and VFA
L174 Please use always the same acronym in the text for OM digestibility (AOMDvt)
L190 How do you compare chemical composition if there is not statistical analysis
L226 Table4
L237 Table5
L255 AOMDvt
References need to be corrected. Please see the instruction for the author.
Author Response
Response to Reviewer 1
Dear Authors the article need to be reviewed better. It was not clear the last time my indications, may be.
So I ask you to review better the paper.
L16 The organic matter digestibility (AOMDvt)
Response: Thanks for your recommendation to improve the manuscript and we wish that this manuscript will benefit to the reader if can possible published. We have been changed in this point as your recommendation.
L30 I suggest to change the order of phrases:
global greenhouse emissione (GHG), that have become a major concern in recent decades. Enteric fermentation and feed production.....Methane is the....Entheric methane
Response: Thanks for your beneficial suggestion. We've also rearranged all that in a new order of phrases based on your suggestions. Please see in manuscript.
L93 total mixed ration (TMR)
Response: We've changed in this point based on your suggestions. Please see in manuscript.
Material and methods need to be described better and the chapter need to have the same order that you use in results.
So I advise you to describe in this order:
2.2 Chemical composition analysis
2.3 Rumen collection
Response: Thanks for your beneficial suggestion. We've also reorganized all part in the subchapters according to your suggestions. Please see in manuscript.
2.4 In vitro gas production. Insert after L138 the text from L155 to L177, as it describe in vitro gas method
2.5 In vitro rumen fermentation. Describe the method and materials used
2.6 Bacterial and protozoal account. Describe the method
Insert also in the chapter correct the method used for ammonia, and VFA
Response: Thanks for your beneficial suggestion. We've also reorganized all part in the subchapters according to your suggestions. Please see in manuscript.
Insert also in the chapter correct the method used for ammonia, and VFA
Response: Thank you. The author has previously decrypted this point in the section on In vitro rumen fermentation. Please see in manuscript.
L174 Please use always the same acronym in the text for OM digestibility (AOMDvt)
Response: Thank you, however we're focusing on the chemical composition in this part, as shown in Table 3. Please see in manuscript.
L190 How do you compare chemical composition if there is not statistical analysis
Response: Thank you, as a result of the mistake in the incorrect table, which we have already corrected. We presented the chemical composition based on the average of those values.
L226 Table4
L237 Table5
L255 AOMDvt
Response: Thank you, we've changed in this point based on your suggestions. Please see in manuscript.
References need to be corrected. Please see the instruction for the author.
Response: Thank you. We have agreed and thoroughly modified all as your suggestion.
Author Response File: Author Response.pdf
Round 3
Reviewer 1 Report
Thank you Authors for your work.
Make attention to use the same acronyms in the text and in the tables. In particular TVFA/VFA/SFA, verify.
Author Response
Response to Reviewer 1
Make attention to use the same acronyms in the text and in the tables. In particular TVFA/VFA/SFA, verify.
Response: Thanks for your beneficial suggestion. We agreed and made the appropriate changes based on your recommendations. Please see in manuscript.
Author Response File: Author Response.pdf
