A Preliminary Study of the Effects of Gaseous Ozone on the Microbiological and Chemical Characteristics of Whole-Plant Corn Silage

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The paper is a preliminary study on the effects of gaseous ozone (O3) on the microbial and chemical properties of whole-plant corn silage. The overall scale of the study is quite large and has strong practical guidance. The study shows that gaseous ozone is a promising additive for improving the microbial quality of corn silage, providing an effective alternative to traditional chemical preservatives. However, I have some questions that need to be addressed by the authors:

1. The environment in which the authors applied the treatment needs to be described in detail, preferably with photos of the setup, to facilitate a better evaluation of this technique. For instance, in what kind of environment was the experiment conducted? Was it sealed? Was the silage compacted before ozone treatment?

2. Will there be any oxygen leakage during the ozone treatment, posing a hazard to the operators? How is the uniformity of ozone treatment ensured? What is the cost of ozone treatment? Can a cost analysis be provided?

3. The paper contains relatively few data points. Is this a short communication?

4. The data should include standard deviations (SD) and should undergo t-tests or one-way ANOVA for statistical analysis.

5. How does ozone treatment affect the aerobic stability of silage? It would be best to evaluate this or reflect it in subsequent experiments.

Author Response

Dear reviewer,

We thank the reviewer for the important observations made regarding our manuscript.

Below are our point-by-point responses.

1. The environment in which the authors applied the treatment needs to be described in detail, preferably with photos of the setup, to facilitate a better evaluation of this technique. For instance, in what kind of environment was the experiment conducted? Was it sealed? Was the silage compacted before ozone treatment?

The experiment was carried out on a commercial farm (l. 76 – 78). Due to the shortage of personnel (l. 94 – 117), during the study we were not able to take photos of the setup. However, we included more details regarding the reviewer’s questions in the Materials and Methods section of the manuscript to elucidate our procedures.

1. Will there be any oxygen leakage during the ozone treatment, posing a hazard to the operators? How is the uniformity of ozone treatment ensured? What is the cost of ozone treatment? Can a cost analysis be provided?

More details regarding the reviewer’s questions in the Materials and Methods section of the manuscript, including the procedure for including the ozone in the silage bag (l. 101 – 105).  Experimental procedures could be improved in future studies to provide better diffusion capacity of the gas inside silage bale. However, as stated in our manuscript, this was a preliminary study with limited funding, so other studies are required for better understanding of this mechanism.

In these experimental conditions, the cost of ozone treatment and analysis cost was difficult to be made.

1. The paper contains relatively few data points. Is this a short communication?

This paper is not a short communication, however if the reviewers and the editor-in-chief choose to publish only as a short communication, we would consider.

1. The data should include standard deviations (SD) and should undergo t-tests or one-way ANOVA for statistical analysis.

A new analysis was made following the reviewer’s suggestion, adapting for non-parametrical data (Figure 1; l. 224 – 226).

1. How does ozone treatment affect the aerobic stability of silage? It would be best to evaluate this or reflect it in subsequent experiments.

We agree with the reviewer that it is necessary to evaluate the aerobic stability of the silage. Our idea with this study was to begin with the limited funding we had available, but also to encourage other studies based on our procedures.

Best regards,

Reviewer 2 Report

Comments and Suggestions for Authors

The study called A preliminary study on the effects of gaseous ozone on the microbiological and chemical characteristics of whole-plant corn silage is a very well conducted and documented one, with a number of 56 recent bibliographic references. However, I have some curiosities:

 

What motivated you to choose this ozone concentration range, given the novelty of the study?

How did you choose proper bags for ensiling?

Please note in the manuscript the potential risks of using ozone in animal feed.

Good luck!

Author Response

Dear reviewer,

We thank the reviewer for the important observations made regarding our manuscript.

Below are our point-by-point responses.

 

What motivated you to choose this ozone concentration range, given the novelty of the study?

We adapted due to the calibration of the machine (machine generated 10g/hours). We added this information in the Materials and Methods section (l. 106)

How did you choose proper bags for ensiling?

All procedures and equipment were determined and used according to our limited budget.

Please note in the manuscript the potential risks of using ozone in animal feed.

Typically, ozone toxicity in cattle is associated with respiratory issues due to inhalation. The ingestion of ozone is less likely as ozone is a gas and does not persist in feed or water. However, as mentioned in our manuscript (l. 274 – 275), it is important for future studies to evaluate possible impacts on animal health.

 

Best regards,