Effect of the Potential Probiotic Vibrio proteolyticus DCF12.2 on the Immune System of Solea senegalensis and Protection against Photobacterium damselae subsp. piscicida and Vibrio harveyi
Round 1
Reviewer 1 Report
The Ms. need minor corrections before final publication.
1. Line no. the scientific name must be in italics.
2. Line 21-23 rewrite the statement for better understanding.
3. line 101 correct 109 to 108
4. figure 2 letter indicated significant differences between groups were not not mentioned in all the groups.
5.fig 5 quality is poor, need to replace with high quality fig.
6. Authors need to improve probable mechanism to improve the immune response in discussion.
Satisfactory
Author Response
Response to reviewer 1 comments:
The Ms. need minor corrections before final publication.
- Line no. the scientific name must be in italics.
Response: We thank the Reviewer for its efforts in revising the manuscript. We have revised the manuscript adding italics where necessary (mainly abstract section).
- Line 21-23 rewrite the statement for better understanding.
Response: We thank the Reviewer its suggestion. We have revised the statement accordingly.
- line 101 correct 109 to 108
Response: Thank you. We have corrected the error
- figure 2 letter indicated significant differences between groups were not not mentioned in all the groups.
Response: Letters have only been added where there were significant differences (p < 0.05) between treatments.
- figure 5 quality is poor, need to replace with high quality fig.
Response: The figure has been increased in definition from 600 to 1200 dpi (attached document Figure 5 v2).
- Authors need to improve probable mechanism to improve the immune response in discussion.
Response: We have revised the manuscript according to its suggestions.
Reviewer 2 Report
The article by Medina et al. is an interesting study on probiotic activity on immune system. I think that a chart on probiotic effects may be useful to better understand the range of its biological activity. Why did the authors not think about performing immunohistochemical analysis to evaluate the immune cells response to the probiotic?
I would suggest the authors to further check Materials and Methods section and add to the conclusion a critical note on the eventual limitation of this study.
However, I suggest the publication of the article after minor revisions.
English grammar and spell check is suggested.
Author Response
Response to reviewer 2 comments:
The article by Medina et al. is an interesting study on probiotic activity on immune system. I think that a chart on probiotic effects may be useful to better understand the range of its biological activity. Why did the authors not think about performing immunohistochemical analysis to evaluate the immune cells response to the probiotic?
Response: Thanks for the suggestion. Our intention is to continue working with this strain, so in future research we will consider the possibility of carrying out immunohistochemical studies.
I would suggest the authors to further check Materials and Methods section and add to the conclusion a critical note on the eventual limitation of this study.
Response: We thank the Reviewer for its efforts in revising the manuscript. We have revised the manuscript according to its suggestions.
However, I suggest the publication of the article after minor revisions.
Comments on the Quality of English Language
English grammar and spell check is suggested.
Response: Thanks for the suggestion. We have revised the manuscript to improve the English language.
Reviewer 3 Report
Dear authors:
In this section: "2.2. Experimental diets and feeding trial" I recommend being clearer and justifying why you used the given doses. Why the dose in immersion was 1x107 instead of 1x109 as IP and oral administration? Similarly, explain why in oral administration your schedule consisted of 5 days of feeding and 2 days of rest.
In this section: "2.4. Detection of antibodies in serum, intestinal mucus, and skin" Please specify the second antibody used: origin, type, and any information that you consider relevant for this purpose.
What was the water's pH? Please discuss little bit because an administration route was the bath immersion.
Probably authors would like to propose some genes to analyze immune response in future studies with DCF12.2 strain in S. sole as potential biomarkers of immune response.
Author Response
Response to reviewer 3 comments:
In this section: "2.2. Experimental diets and feeding trial" I recommend being clearer and justifying why you used the given doses. Why the dose in immersion was 1x107 instead of 1x109 as IP and oral administration? Similarly, explain why in oral administration your schedule consisted of 5 days of feeding and 2 days of rest.
Response:
The concentration used intraperitoneally was the same as that used in Medina et al, 2020, cited in the manuscript.
In the bath treatment, according to other researchers, probiotic administration in water ranges between 105 and 108 cfu mL-1 (Jahangiri, L.; Esteban, M.Á. Administration of Probiotics in the Water in Finfish Aquaculture Systems: A Review. Fishes 2018, 3, 33. https://doi.org/10.3390/fishes3030033).
Similarly, the concentration of probiotics administered in the diet is consistent with that used by other authors: Sultana S, Saifuddin M, Kaizer M (2020) Potential probiotic and health fostering effect of host gut-derived Enterococcus faecalis on freshwater prawn, Macrobrachium rosenbergii. Aquacul Fish https://doi.org/10.1016/j.aaf.2020.10.004.
These references have been added to the manuscript.
On the other hand, the feed supply system was conditioned by technical reasons. The fish were fed for 5 days with the treatment (Monday to Friday) supervised by the researchers. On weekends the feeding was carried out by the technical staff, who were not allowed to implement the research procedures.
In this section: "2.4. Detection of antibodies in serum, intestinal mucus, and skin" Please specify the second antibody used: origin, type, and any information that you consider relevant for this purpose.
Response: Thanks for the suggestion. The antibody detection methodology has been enhanced, giving details of the materials used.
What was the water's pH? Please discuss little bit because an administration route was the bath immersion.
Response: The pH of the water did not change during the maintenance of the fish or with the inclusion of strain DCF12.2, remaining in a range between 7.8 and 7. This pH range has been included in Material and Methods.
Probably authors would like to propose some genes to analyze immune response in future studies with DCF12.2 strain in S. sole as potential biomarkers of immune response.
Response: Thank you for the suggestion. We are currently continuing to work with this strain in fish, detecting expression in other genes related to metabolism, stress and immune response.
