Abstract
As the world increasingly relies on aquaculture operations to meet rising seafood demands, reliable biocontainment measures for farmed fish stocks are desired to minimize ecological impacts arising from interactions of cultured fish with wild populations. One possible biocontainment strategy is to induce a dietary dependence on a vitamin, such as thiamine (vitamin B1), required for survival. Fish expressing thiaminase (an enzyme that degrades thiamine) within a confined aquaculture facility could receive supplemental thiamine to allow survival and normal growth, whereas escapees lacking this dietary rescue would die from thiamine deficiency. To test the concept and efficacy of such a dietary dependency system (for potential future use in larger aquaculture species), we expressed thiaminase in zebrafish as a test model. We drove the expression of thiaminase under the strong ubiquitous and constitutive control of the CMV promoter which resulted in non-viable fish, indicating that the thiaminase sequence kills fish. However, the CMV promoter is too strong to allow conditional survival since the lethality could not be rescued by exogenous thiamine provided as a supplement to typical food. In addition, microinjection of 0.5 pg of thiaminase mRNA in zebrafish embryos at the one-cell stage resulted in 50% larval mortality at 5 days post-fertilization (dpf), which was partially rescued by thiamine supplementation. Evaluating the efficacy of biocontainment strategies helps assess which methods can reliably prevent ecological impacts arising from breaches in physical containment systems that release engineered organisms to nature, and consequently provides critical information for use in regulatory risk assessment processes.
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Abbreviations
- Dpf :
-
days post-fertilization
- dw/dw :
-
dry weight/dry weight
- ef1a :
-
elongation factor-1 alpha
- EMS :
-
early mortality syndrome
- Ere :
-
estrogen response element
- GM :
-
genetically modified
- Hpf :
-
hours post-fertilization
- MO :
-
morpholino
- SIT :
-
self-limiting approach
- SP :
-
signal peptide
- vtg1 :
-
vitellogenin 1
- WT :
-
wild type
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Acknowledgements
We thank Evan Byers, Fei Fei Shao, Marine Arsouze, and Hugo Chevalon for assistance with microinjecting the thiaminase mRNA and DNA constructs and data collection. Discussions with Jim Powell regarding the thiaminase system as a possible containment measure are appreciated. This work was supported by funding from the Canadian Department of Fisheries and Oceans and the Natural Sciences and Engineering Research Council of Canada (NSERC).
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All experiments were performed according to the guidelines of the Canadian Council on Animal Care and were approved by the University of Ottawa Animal Care Committee.
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The authors declare that they have no conflict of interest.
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Noble, S., Saxena, V., Ekker, M. et al. Expression of Thiaminase in Zebrafish (Danio rerio) is Lethal and Has Implications for Use as a Biocontainment Strategy in Aquaculture and Invasive Species. Mar Biotechnol 19, 563–569 (2017). https://doi.org/10.1007/s10126-017-9776-2
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DOI: https://doi.org/10.1007/s10126-017-9776-2