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Review
Peer-Review Record

Revolutionizing Veterinary Vaccines: Overcoming Cold-Chain Barriers Through Thermostable and Novel Delivery Technologies

Appl. Microbiol. 2025, 5(3), 83; https://doi.org/10.3390/applmicrobiol5030083
by Rabin Raut 1,*, Roshik Shrestha 2,3, Ayush Adhikari 4, Arjmand Fatima 1 and Muhammad Naeem 1,*
Reviewer 1: Anonymous
Appl. Microbiol. 2025, 5(3), 83; https://doi.org/10.3390/applmicrobiol5030083
Submission received: 23 June 2025 / Revised: 19 July 2025 / Accepted: 8 August 2025 / Published: 19 August 2025

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

Raut et al, provide a comprehensive overview of the challenges faced in veterinary vaccinology and how these challenges are being tackled through advances in vaccine thermostability and delivery technologies. The authors provide detailed examples of how these advances are helping to overcome issues surrounding cold-chain and vaccine delivery whilst also providing a synopsis of the impact on One Health. Finally, the authors provide a forward look towards the potential of AI in the field of veterinary vaccinology, highlighting its previous successes and how AI could be used to improve antigen selection, particularly in LMICs where missteps in vaccine selection can have wide-ranging implications. Overall, this is an excellent review which would be of interest to the field. I have made a couple of minor suggestions that may improve the manuscript prior to publication.

Comments

It would be of interest to the readership to potentially offer some of the drawbacks to the use of inorganic nanoparticles in vaccinology. Perhaps commenting on whether these inorganic metals are known to be excreted following vaccination, or whether these may build up within the animal and have known or unknown side-effects on immune competency?

The authors highlight the potential of different technologies used to stabilise vaccines, however I believe this review could be further improved by providing some examples of antigen stabilisation as well as drying technologies. For example, picornaviruses have been shown to become more thermostable through the introduction of specific mutations, such as FMDV (https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1003255) and with regard to human vaccines, poliovirus (https://pmc.ncbi.nlm.nih.gov/articles/PMC5245777/ and https://www.nature.com/articles/s41467-025-56118-z). I believe that the adding examples such as these to this review will provide a fuller picture for the readership.

Author Response

It would be of interest to the readership to potentially offer some of the drawbacks to the use of inorganic nanoparticles in vaccinology. Perhaps commenting on whether these inorganic metals are known to be excreted following vaccination, or whether these may build up within the animal and have known or unknown side-effects on immune competency?

The authors highlight the potential of different technologies used to stabilise vaccines, however I believe this review could be further improved by providing some examples of antigen stabilisation as well as drying technologies. For example, picornaviruses have been shown to become more thermostable through the introduction of specific mutations, such as FMDV (https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1003255) and with regard to human vaccines, poliovirus (https://pmc.ncbi.nlm.nih.gov/articles/PMC5245777/ and https://www.nature.com/articles/s41467-025-56118-z). I believe that adding examples such as these to this review will provide a fuller picture for the readership.

Thank you so much for your valuable time and constructive feedback. We sincerely appreciate your insightful comments, which have helped us improve the clarity and quality of our manuscript. Below, we provide point-by-point responses to each of your suggestions and explain the corresponding revisions made in the manuscript.

Author Response File: Author Response.docx

Reviewer 2 Report

Comments and Suggestions for Authors

Lanes 44-46, explain whether these statements are covered in the field of veterinary medicine. For example, if you're talking about hospitalization, what species are you talking about?


Lanes 92-94 mention that failure in the cold chain causes losses of 50%, assuming the product is discarded, a situation that surely occurs in many public health institutions.
In veterinary medicine, does this failure in logistics result in the disposal of vaccines or their continued use, regardless of whether the cold chain has been compromised? It would be appropriate to discuss this.


Lanes 102-103 argue that the use of injectable vaccines is one of the problems, and also mention that trained personnel are required. In many countries, this work is performed by a veterinarian, who has sufficient training. Other people who lack the training should not be responsible for administering vaccines or medications to animals, according to the legislation in many territories. Occupational risk is discussed, and an example is presented, indicating a recurrence rate of needlestick accidents in the region of 80%. This seems like an extreme case; a success story could be presented as a counterpoint.


Lane 123-125 should clarify and separate when discussing vaccines for viruses and those for bacteria or other microorganisms.


Section 3 does not specifically show innovations. This section should be improved or its title modified.


In Section 4, you should complete the options for organic or other nanoparticles; two options described are very limited.


Section 4.2 should provide a robust explanation of how a microneedle vaccine works, indicate what is being stimulated, and also describe this system and its ease of application in the region where it poses challenges with access to technologies and vaccine conservation.


Section 4.3, as in the previous section, should provide a more robust rationale for how immunity is induced at the mucosal level and provide examples of the adjuvants studied. Lanes 266-268, 290-292, please indicate if it's an attenuated virus or what origin? What type of adjuvants? Reference 80 is weak.


Section 7 should provide more examples using the names of the main AIs used in vaccinology and how this would be applied in the complex scenario of developing countries, as the authors originally mentioned. I mean, if veterinarians suffer 80% of needle accidents when administering a vaccine, what would be the point of using AI?

Author Response

Thank you very much for your valuable time and constructive feedback. We sincerely appreciate your insightful comments, which have significantly improved the quality of our work. Please find our detailed point-by-point responses in the attached Word file.

Author Response File: Author Response.docx

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

The attention to the comments is appreciated; the manuscript has improved in a positive way.

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