A Review of Veterinary Drug Residue Detection: Recent Advancements, Challenges, and Future Directions
Round 1
Reviewer 1 Report
Follow the attached document for improvements.
Comments for author File: 
Comments.pdf
Careful proof reading is required.
Author Response
REVIEWER1# ' COMMENTS TO AUTHOR:
General comments: The titled manuscript, "A Review of Veterinary Drug Residue Detection: Recent Advancements, Challenges, and Future Direction," provides an interesting and informative study. However, there are some areas where the manuscript could be improved:
Reply: Done. Thanks for your good suggestions. Please refer to the revised manuscript for detailed changes made.
Specific comments：
1． Title: Ok.
Reply: Thanks！
Abstract:
- The abstract mentions that few studies have reviewed the advancements in veterinary drug residue detection, but it does not indicate the significance or the knowledge gap that this study aims to address. Including a sentence highlighting the importance of filling this research gap would strengthen the rationale for conducting this study.
Reply: Done. Thanks for your good suggestions. Please refer to the revised manuscript for detailed changes made.
- The abstract concludes by stating that the review provides new ideas and strategies for the rapid development of the animal husbandry industry and for protecting consumers' physical health and food safety. However, it does not mention any specific recommendations or findings from the review. Adding a sentence summarizing the key findings or recommendations would enhance the abstract's impact.
Reply: Thanks for your good suggestions. Please refer to the revised manuscript for detailed changes made.
This review provides more technical methods for detecting veterinary drug residues, and proposes ideas and strategies for rapid development of animal husbandry industry, protecting consumers' physical health and food safety, and accelerating the improvement of technical route of veterinary drug residue detection.
- Finally, it is essential to proofread the abstract for grammatical errors (Line 21- were recommend?) and improve the overall flow of the text to ensure clarity and readability.
Reply: Done. Line 21- “were recommend” is revised to “were recommended”.
Introduction
- The introduction should begin with a strong and attention-grabbing opening sentence to engage readers and emphasize the importance of the topic. It would be beneficial to provide a brief context or statistic related to the global increase in the demand for animal-derived food.
In sentences 30-33, the significance of veterinary drugs for animal well-being and production is mentioned. However, it would be helpful to elaborate on the potential consequences of improper use or excessive residues of veterinary drugs in animal- derived foods.
Reply: Thanks for your good suggestions. Please refer to the revised manuscript for detailed changes made.
- Sentence 35 introduces the definition of veterinary drug residues, but it could be rephrased to provide a clearer explanation. Consider simplifying the sentence structure and providing examples to enhance understanding.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- While the potential risks and consequences of veterinary drug residues are briefly mentioned in sentences 38-41, it would be beneficial to provide more specific examples or references to support these claims. This will help readers understand the gravity of the issue.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- In sentences 44-53, the importance of monitoring and controlling veterinary drug residues is highlighted. However, the text could be further improved by explaining why effective detection methods are crucial for achieving regulatory objectives and ensuring food safety. Emphasize the need for accurate, sensitive, and efficient testing techniques.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- The mention of specific detection methods and studies (e.g., Zhao et al., Stolker et al., Kaufmann et al.) in sentences 55-62 is valuable. However, it would be more effective to summarize the key findings or advancements of these studies, rather than merely listing the authors' names and techniques used. This will provide readers with a better understanding of the existing detection methods.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- In sentences 65-68, the rationale for the review is stated, highlighting the lack of systematic reviews on veterinary drug residue detection. However, it would be helpful to briefly explain why a comprehensive review of recent advancements, challenges, and future directions is timely and valuable. Specify the potential benefits of such a review for the scientific community, industry, and regulatory bodies.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- The structure and organization of the review are mentioned in sentences 73-76. It would be useful to provide a brief summary or overview of each section to give readers a clear expectation of the content covered in the manuscript.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
Sub-Heading:
- Sample pretreatment technology:
It would be helpful to provide a brief explanation of why sample pretreatment is necessary for the analysis of veterinary drugs and pesticide residues. Consider expanding on the importance of selecting appropriate sample pretreatment techniques based on the specific properties of the target analytes and the sample matrix.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- 3.1. Liquid-liquid extraction:
Consider including information on the factors that can affect the efficiency and reliability of liquid-liquid extraction, such as the choice of solvent systems and optimization of extraction conditions.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- 3.2. Solid-phase extraction:
Clarify the range of compounds that can be effectively extracted using solid-phase extraction and discuss any limitations related to analyte characteristics.
Request more information on the specific types of solid adsorbents employed in the mentioned studies, as well as any challenges encountered during method development and validation.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- 3.2.QuEChERS:
More details on the specific QuEChERS procedures used in the mentioned studies, including the types of sorbents and the ratio of sample to extraction solvent.
Consider discussing any potential interferences or challenges associated with QuEChERS and suggest possible mitigation strategies.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- Accelerated solvent extraction:
Provide a more comprehensive explanation of the benefits of accelerated solvent extraction, including its ability to extract a wide range of compounds from solid or semi-solid samples.
Include more information on the specific organic solvents and extraction parameters employed in the mentioned studies to ensure reproducibility.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
17.Detection analytical techniques:
Consider providing a brief overview of the importance of detection analytical techniques in the analysis of veterinary drug residues and their role in ensuring food safety.
It would be helpful to mention the specific challenges associated with the detection of veterinary drug residues, such as the need for high sensitivity and selectivity due to low residue levels and complex matrices.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- 4.2. Liquid chromatography (LC):
Expand on the advantages of using liquid chromatography as a detection technique, particularly in terms of high speed, efficiency, and sensitivity.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
- 6. Biosensor analysis:
Provide a more comprehensive explanation of biosensor technology and its potential applications in the detection of veterinary drug residues. Additional information is required on the advantages and challenges of biosensors, including their low reagent consumption, high sensitivity, and potential for future development.
The authors should consider providing more details on the biosensor methods used in the mentioned studies, including the specific techniques employed and any notable findings or advancements.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
20.In subheading 5.1, it would be helpful to provide more specific information on the limitations and challenges of the commonly used sample pretreatment techniques, such as liquid-liquid extraction, solid-phase extraction, QuECHERS, and accelerated solvent extraction. This will enhance the understanding of why these techniques may not be sufficient for detecting veterinary drug residues.
Reply: Done. Please refer to the revised manuscript for detailed changes made. These existing processing technologies have their own shortcomings (Table 1). However, some new sample pretreatment technologies with faster processing speed are relatively rarely applied, especially in the residue of multiple types of veterinary drugs, such as stir bar sorption extraction (SBSE) (48,49), molecular imprinting technology (50,51), and immunoaffinity chromatography (ICA) (52,53), and so on.
21.Subheading 5.2 discusses the lack of multi-residue detection methods, particularly for different types of veterinary drugs. While the examples provided for the detection of fluoroquinolones (FQs) and sulfonamides (SAs) are informative, it would be beneficial to mention other types of veterinary drugs and their challenges in multi- residue detection. Additionally, providing more recent references in this section would enhance the relevance and currency of the information.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
So far, there is still a lack of multiple residue detection methods for cephalosporin antibiotics and macrolide antibiotics. The greatest challenge is the growing use of new varieties of veterinary drugs, and there is still a lack of multiple residue detection methods for two or three different types of veterinary drugs.
22.In subheading 5.3, the manuscript mentions the application of quantum dots technology in veterinary drug residue detection. It would be helpful to provide specific examples or studies that demonstrate the successful application of quantum dots combined with immunochromatography, as mentioned. Additionally, the manuscript could further elaborate on the drawbacks and limitations of quantum dots technology that still need to be addressed for its wider adoption.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
The quantum dot labeling technique was applied to the development of immunochromatographic test strip for rapid detection of ractopamine (67).
Extremely tiny inorganic liquid nanocrystals known as quantum dot materials require numerous deep processing steps during preparation in order to produce perfect materials. It is challenging to establish a new method for detecting veterinary drug residues since a new quantum dot material has to be created.
23.Subheading 6.1 discusses the future direction of high-throughput and high-sensitivity veterinary drug detection technology. While the manuscript briefly mentions the use of nanotechnology and time-resolved fluorescence immunoassay (TRFIA), it would be valuable to provide more details on how these technologies can specifically contribute to improving the detection abilities of veterinary drug residues. Including specific examples or advancements in these technologies would enhance the discussion.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
Through temporal resolution and wavelength resolution, the TRFIA method successfully reduces the interference of background fluorescence by using rare earth ion chelates with extended fluorescence lifetimes and substantial stock shifts as markers.
A quick time-resolved fluoroimmunoassay for narasin and salinomycin residue screening in poultry and eggs was established (70).
24.In subheading 6.2, the manuscript mentions the development of rapid pretreatment methods for sample processing. It would be beneficial to provide examples or references of recent studies that have successfully developed rapid pretreatment methods and highlight their advantages over classical sample pretreatment methods.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
Many QuEChERS extraction methods have been established and have fast, simple, inexpensive, effective, stable, and safe characteristics (25,26).
25.Subheading 6.3 emphasizes the fusion of multiple detection methods for the analysis of veterinary drug residues. While the manuscript mentions several detection technologies, such as liquid chromatography-mass spectrometry and enzyme-linked immunosorbent detection, it would be helpful to provide specific examples of studies or applications where the integration of these technologies has been successful in improving the effectiveness and quality of detecting veterinary drug residues.
Reply: Done. Please refer to the revised manuscript for detailed changes made.
Using nanotechnology and immunochromatography, ractopamine could be detected quickly with the detection limit of 3 ng/ml, and the detection could be finished within 10 min (67).
Conclusions
26.Add clear future prospects with the conclusion section
Reply: Done.
Author Response File: 
Author Response.doc
Reviewer 2 Report
In this study the authors present a review on Veterinary Drug Residue Detection: Recent Advancements, Challenges, and Future Directions. The review will be helpful for the potential readers; however, the article has some questions.
· The authors should discuss the drawbacks of current detection techniques and future perspectives.
- It would be more convenient for readers if authors add references in table 2 for each section where possible.
- Authors dint not discuss the cost of detection technique. It is important determinant of method selection. Please discuss in your comparison.
- If possible please compare the complete method on the basis of accuracy, simplicity, timing, and cost.
- Methods are discussed in a generic way. There must be discussion on common food (milk, meat) based or class of drug based.
Author Response
REVIEWER2# ' COMMENTS TO AUTHOR:
General comments: In this study the authors present a review on Veterinary Drug Residue Detection: Recent Advancements, Challenges, and Future Directions. The review will be helpful for the potential readers; however, the article has some questions. The authors should discuss the drawbacks of current detection techniques and future perspectives.
Reply: Done. Thanks for your good suggestions. Please refer to the revised manuscript for detailed changes made.
Specific comments：
27.It would be more convenient for readers if authors add references in table 2 for each section where possible.
Reply: Done. Thanks for your good suggestions. We added corresponding references in table 1 for each section where possible.
28.Authors dint not discuss the cost of detection technique. It is important determinant of method selection. Please discuss in your comparison.
Reply: Done. We added the discuss the cost of detection technique in table 1.
29.If possible please compare the complete method on the basis of accuracy, simplicity, timing, and cost.
Reply: Thanks. We agree with your suggestions. However, we focused on the current advancements, challenges, and potential future directions of veterinary drug residue detection in this article. The advantages and disadvantages of various methods in a generic way were discussed, including accuracy, simplicity, timing, and cost. A more thorough and detailed comparison of various methods will be provided in the next article.
- Methods are discussed in a generic way. There must be discussion on common food (milk, meat) based or class of drug based.
Reply: Done. We focused on the veterinary drug residue detection of common food (milk, meat) based. We have provided clear explanations in the abstract.
Author Response File: Author Response.doc
Reviewer 3 Report
This review addresses the detection of veterinary drug residues in animal food: advances in veterinary drug residue detection technology, problems and challenges in veterinary drug residue detection and proposes future directions for veterinary drug residue detection. The article is interesting and well organised, but the main problems in the article are as follows:
1、Line 234, 5.1, the title and content need to be reworded to clearly identify the bottlenecks of each traditional method, rather than summarising them in general terms as too traditional sample pretreatment techniques；
2、Line 285, 6, the order of content needs to be adjusted to reflect the content of 5, and the review of sample preparation techniques in 6.2 is too brief, covering only microwave assisted extraction methods without summarising the relevant future solutions to the shortcomings of traditional methods in 5.1.
Author Response
REVIEWER3# ' COMMENTS TO AUTHOR:
General comments: This review addresses the detection of veterinary drug residues in animal food: advances in veterinary drug residue detection technology, problems and challenges in veterinary drug residue detection and proposes future directions for veterinary drug residue detection. The article is interesting and well organised, but the main problems in the article are as follows:
Reply: Done. Thanks for your good suggestions. Please refer to the revised manuscript for detailed changes made.
Specific comments：
- Line 234, 5.1, the title and content need to be to clearly identify the bottlenecks of each traditional method, rather than summarising them in general terms as too traditional sample pretreatment techniques；
Reply: Done. We reworded the section of 5.1 more clearly, identify the bottlenecks of each traditional method.
32.Line 285, 6, the order of content needs to be adjusted to reflect the content of 5, and the review of sample preparation techniques in 6.2 is too brief, covering only microwave assisted extraction methods without summarising the relevant future solutions to the shortcomings of traditional methods in 5.1.
Reply: Done. We reworded the section of 6.2 more clearly, giving the relevant future solutions to the shortcomings of traditional methods in 5.1.
Author Response File: Author Response.doc
Round 2
Reviewer 1 Report
Thank you for accepting all the recommendations. The response given by the authors is satisfactory.
Regards
Careful proofreading is required.
Regards
Author Response
REVIEWER1# ' COMMENTS TO AUTHOR:
General comments: Thank you for accepting all the recommendations. The response given by the authors is satisfactory.
Reply: Thanks for your good suggestions.
Specific comments：
Careful proofreading is required.
Reply: Done. Thanks for your good suggestions. Please refer to the revised manuscript for detailed changes made.
Author Response File: Author Response.doc
Reviewer 2 Report
Accept in present form
Reviewer 3 Report
Personally, I think the article has been improved according to my suggestions and can now be accepted.
Author Response
REVIEWER3# ' COMMENTS TO AUTHOR:
General comments: Personally, I think the article has been improved according to my suggestions and can now be accepted.
Reply: Thanks for your good suggestions.
Author Response File: Author Response.doc
