Championing Line Breeding and Hybridization in Aquaculture to Safeguard Intellectual Property

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The manuscript proposes biological protection of intellectual property rights through genetic improvement techniques such as strain breeding and hybridization, providing scientifically valuable and commercially feasible solutions for aquaculture. It proposes an innovative concept of strengthening traditional intellectual property protection legal means by controlling parent lines through complex genetic improvement to avoid reverse engineering and imitation, which also has enlightening significance.

Overall, there are relatively few cases of strengthening IP protection through line breeding and line hybridization of aquaculture in the manuscript, while non aquaculture cases such as crops and livestock account for a larger proportion. Suggest adding examples of line breeding and line hybridization of aquaculture strains, which will have more practical guidance significance.

Specific revisions:

Line 26: Add a full stop at the end of the sentence.

Lines 108-110: For potential disadvantages, consider proposing 1-2 concrete management recommendations.

Tables 1 & 2: Reduce plant case listings appropriately, while expanding aquaculture species examples through a dedicated table to emphasize the manuscript's aquaculture-specific insights.

Author Response

Reviewer 1

The manuscript proposes biological protection of intellectual property rights through genetic improvement techniques such as strain breeding and hybridization, providing scientifically valuable and commercially feasible solutions for aquaculture. It proposes an innovative concept of strengthening traditional intellectual property protection legal means by controlling parent lines through complex genetic improvement to avoid reverse engineering and imitation, which also has enlightening significance.

Overall, there are relatively few cases of strengthening IP protection through line breeding and line hybridization of aquaculture in the manuscript, while non aquaculture cases such as crops and livestock account for a larger proportion. Suggest adding examples of line breeding and line hybridization of aquaculture strains, which will have more practical guidance significance.

# Thank you for your positive feedback on our manuscript. It is indeed true that, unlike crops and livestock, aquaculture has relatively few examples where line breeding and hybridization are applied. This is largely because such breeding strategies have seldom been implemented in the aquaculture industry. Our group in Singapore began applying this approach to Asian seabass breeding in 2003 and has since recognized its great potential, particularly in terms of intellectual property protection. Therefore, I would like to advocate for the broader adoption of this breeding strategy in aquaculture.

 

Specific revisions:

Line 26: Add a full stop at the end of the sentence.

# Done as suggested

Lines 108-110: For potential disadvantages, consider proposing 1-2 concrete management recommendations.

# Good suggestion. I added “To mitigate this, breeding programs should incorporate careful pedigree tracking and periodic introduction of genetic diversity to maintain vigor and overall fitness.” (see lines 111-113).

Tables 1 & 2: Reduce plant case listings appropriately, while expanding aquaculture species examples through a dedicated table to emphasize the manuscript's aquaculture-specific insights.

# As mentioned above, in aquaculture breeding, this breeding strategy has seldomly been implemented in the aquaculture industry. Therefore, not many cases on line and hybrid breeding can be listed in this table.

Reviewer 2 Report

Comments and Suggestions for Authors

This manuscript does a good job of outlining the advantages of line breeding/hybridization in aquaculture. However, although mentioning some major downsides of this approach to genetic improvement it does not indicate clearly how the downsides can be avoided, mitigated or delayed. I suggest adding a paragraph or two to fill this gap.

The major problem that needs addressing is merely mentioned in the paper under review. For example "... inbreeding depression in line breeding risks reducing vigor if not managed properly." [line 109]. The very last and most important line of the paper under review concludes, "Despite risks like inbreeding depression, careful management ensures ... a sustainable future]." Nowhere is "careful management" for this purpose described or even outlined.

Suggestion: On lines 51 - 58 the authors mention that clear legal frameworks and international agreements are needed for sharing genetic resources. I recommend a new paragraph or two to outline how that might be accomplished in aquaculture -- if the industry were ever to grow up. 

There are examples from poultry, swine and cattle breeding where genetic diversity is shared among enterprises that commercially compete with each other. It is understood to be essential to the growth and long-term adaptability of the industry. Genomics of course comes into play at many places in the practical implementation of these genetic sharing strategies.

 

Author Response

Reviewer 2

This manuscript does a good job of outlining the advantages of line breeding/hybridization in aquaculture. However, although mentioning some major downsides of this approach to genetic improvement it does not indicate clearly how the downsides can be avoided, mitigated or delayed. I suggest adding a paragraph or two to fill this gap.

 

The major problem that needs addressing is merely mentioned in the paper under review. For example "... inbreeding depression in line breeding risks reducing vigor if not managed properly." [line 109]. The very last and most important line of the paper under review concludes, "Despite risks like inbreeding depression, careful management ensures ... a sustainable future]." Nowhere is "careful management" for this purpose described or even outlined.

# I added “To mitigate this, breeding programs should incorporate careful pedigree tracking and periodic introduction of genetic diversity to maintain vigor and overall fitness.” (see lines 111-113).

Suggestion: On lines 51 - 58 the authors mention that clear legal frameworks and international agreements are needed for sharing genetic resources. I recommend a new paragraph or two to outline how that might be accomplished in aquaculture -- if the industry were ever to grow up.

There are examples from poultry, swine and cattle breeding where genetic diversity is shared among enterprises that commercially compete with each other. It is understood to be essential to the growth and long-term adaptability of the industry. Genomics of course comes into play at many places in the practical implementation of these genetic sharing strategies.

#I added the following paragraph “Clear legal frameworks and international agreements are essential for the fair and transparent sharing of genetic resources in aquaculture line breeding and hybridization. As aquaculture continues to expand globally, the movement and use of valuable genetic materials—such as broodstock, gametes, and improved lines—have become increasingly common across borders. Without clear legal mechanisms, there is a risk of biopiracy, unfair benefit distribution, and disputes over intellectual property rights. International frameworks like the Nagoya Protocol provide guidelines for access and benefit-sharing of genetic resources, but their application to aquatic species, especially hybrids and selectively bred lines, remains inconsistent and underdeveloped. A harmonized legal approach is crucial to ensure that both provider and user countries can participate in and benefit from genetic advancements. This promotes trust, encourages investment in breeding programs, and supports conservation efforts by valuing genetic diversity. Ultimately, strong legal and policy structures foster sustainable and equitable innovation in aquaculture breeding.” (see lines 127-139).

Reviewer 3 Report

Comments and Suggestions for Authors

Technical Comments

1. [Page 1, Line 14] – "Traditional intellectual property (IP) protections, such as patents, are difficult to enforce..."

Comment: Consider specifying why aquaculture poses unique challenges compared to other sectors in enforcing IP—perhaps due to open-water rearing systems or unregulated broodstock exchanges.

2. [Page 2, Line 41] – "...enhanced growth, or improved feed efficiency."

Comment: The sentence discussing CRISPR-Cas9 ends abruptly. Consider adding an example or reference where such traits were successfully edited in aquaculture species.

3. [Page 3, Line 72] – "increased omega-3 content.[19]."

Comment: The punctuation is inconsistent—use either “[19]” or “[19].” without a period after the bracket.

4. [Page 4, Line 87] – (Gjedrem, 2005).

Comment: This citation is not included in the reference list in its current form (see Line 164). Ensure in-text citations match the reference formatting style of the journal (e.g., number-based or author-year system).

5. [Page 4, Lines 90–91] – "$1.2 million" cost for shrimp breeding in Thailand.

Comment: Consider including a source reference here to support this figure, ensuring consistency with the rest of the manuscript's citation style.

6. [Page 5, Line 98] – "was replicated illegally..."

Comment: The use of "illegally" implies legal judgment. Consider rephrasing to "was reproduced without authorization" unless a legal case is cited.

7. [Page 5, Lines 106–107] – "four-line systems...offer greater trait diversity"

Comment: This statement needs clarification or citation. While four-way crosses can combine more traits, there is a trade-off in complexity—elaborate or reference this claim.

8. [Page 6, Line 125] – "...in combination with novel technologies including MS, GS and GE..."

Comment: These abbreviations are clear but not defined again in the conclusion. For clarity, consider restating their full forms briefly.

9. [Page 6, Lines 135–136] – "Author Contributions..."

Comment: Since this is a single-author paper, this section may be redundant. You might consider removing or combining with the acknowledgments, unless required by journal formatting.

10. [Page 7, Line 145] – "MSA: Marker-assisted selection"

Comment: The abbreviation used throughout the paper is "MS", not "MSA". Please revise for consistency.

11. [Page 7, References Section] – Inconsistent formatting

Comment: Some references use author names with initials before surnames, others with surnames first. Standardize all references per journal guidelines. For example, entries like:

“FAO: The state of world fisheries and aquaculture…” (Line 147) lacks authors and should follow the same format as others.

 

 

Author Response

Reviewer 3

1. [Page 1, Line 14] – "Traditional intellectual property (IP) protections, such as patents, are difficult to enforce..."

Comment: Consider specifying why aquaculture poses unique challenges compared to other sectors in enforcing IP—perhaps due to open-water rearing systems or unregulated broodstock exchanges.

# To make it clearer, I replace it with “because biological replication and unauthorized breeding of aquatic species make it difficult to track and control proprietary genetic materials.” (see lines 15-17).

2. [Page 2, Line 41] – "...enhanced growth, or improved feed efficiency."

Comment: The sentence discussing CRISPR-Cas9 ends abruptly. Consider adding an example or reference where such traits were successfully edited in aquaculture species.

#I added two references [7, 8] after “enhanced growth, or improved feed efficiency” (see line 43).

3. [Page 3, Line 72] – "increased omega-3 content.[19]."

Comment: The punctuation is inconsistent—use either “[19]” or “[19].” without a period after the bracket.

# Done as suggested.

4. [Page 4, Line 87] – (Gjedrem, 2005).

Comment: This citation is not included in the reference list in its current form (see Line 164). Ensure in-text citations match the reference formatting style of the journal (e.g., number-based or author-year system).

# The “(Gjedrem, 2005)” was changed into [10] (see line 88).

5. [Page 4, Lines 90–91] – "$1.2 million" cost for shrimp breeding in Thailand.

Comment: Consider including a source reference here to support this figure, ensuring consistency with the rest of the manuscript's citation style.

# Thanks for the suggestions, I replaced “"$1.2 million" cost for shrimp breeding in Thailand.” with the data from our own breeding program for Asian seabass “For example, a breeding program for Asian seabass has invested over a million dollars since 2004 [42]” (see lines 90-91).

6. [Page 5, Line 98] – "was replicated illegally..."

Comment: The use of "illegally" implies legal judgment. Consider rephrasing to "was reproduced without authorization" unless a legal case is cited.

#Done as suggested (see lines 98-99).’

7. [Page 5, Lines 106–107] – "four-line systems...offer greater trait diversity"

Comment: This statement needs clarification or citation. While four-way crosses can combine more traits, there is a trade-off in complexity—elaborate or reference this claim.

# I changed the sentence into “Two-line hybridization offers a balance between cost and efficiency by requiring fewer resources than four-line systems, which, although more resource-intensive, enhance genetic improvement by maintaining and crossing multiple distinct parental lines to maximize trait diversity (see Figure 1)” (see lines 106-109).

8. [Page 6, Line 125] – "...in combination with novel technologies including MS, GS and GE..."

Comment: These abbreviations are clear but not defined again in the conclusion. For clarity, consider restating their full forms briefly.

# Done as suggested (see line 142).

9. [Page 6, Lines 135–136] – "Author Contributions..."

Comment: Since this is a single-author paper, this section may be redundant. You might consider removing or combining with the acknowledgments, unless required by journal formatting.

#This is required by the journal.

10. [Page 7, Line 145] – "MSA: Marker-assisted selection"

Comment: The abbreviation used throughout the paper is "MS", not "MSA". Please revise for consistency.

#Thanks. Done as suggested,.

11. [Page 7, References Section] – Inconsistent formatting

Comment: Some references use author names with initials before surnames, others with surnames first. Standardize all references per journal guidelines. For example, entries like:

“FAO: The state of world fisheries and aquaculture…” (Line 147) lacks authors and should follow the same format as others.

# The reference format was generated using software Endnote based on original sources from Google Scholar or Web of Science, and I have done my best to minimize any formatting errors.

Regarding the citation “FAO: The State of World Fisheries and Aquaculture…” (Line 147), which lacks authors and differs from the format of other references, this publication is authored by the FAO itself, and I have cited it accordingly.’

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

Congratulations to the Authors.