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Risk Assessment of Transgenic Fluorescent Ornamental Fishes to the United States Using FISK v2 Jeffrey E. Hill Larry L. Lawson Scott Hardin Pictures from Glofish.com What are Glofish® Ornamental fishes that express a fluorescent protein Protein originally isolated from jellyfish (Aequorea victoria) and IndoPacific sea anemone (Discosoma sp) Microinjection into embryos at 1- or 2-cell stage Colors highly expressed in skeletal muscle Zebra Danio -Most studied of the three -Tropical tolerances -Native to India, Bangladesh, and Nepal -Small bodied -Diet of small invertebrates -Large scale commercial production Black Tetra -Native to Rio Guapore, Brazil and Bolivia and Rio Paraguay, Argentina -Small bodied -Diet of small invertebrates -Large scale commercial production Tiger Barb -Native to Borneo and Sumatra -Small bodied -Diet of small invertebrates -Large scale commercial production So Who Cares? Balancing Environmental Safety and Economic Value Critics Supporters • • • • • • • • Media Advocacy groups Regulatory process Scientists Marketing Sales Producers Scientists ??? • “We meddle with this at our peril.” • “I don’t think we know enough to say they are safe” • “There is no way of predicting any species fitness in the absence of environmental settings, because species do not behave as predicted by models” • “We think the feds should step in whether or not [GloFish] pose a threat.” • “It shows the big holes in our regulatory system…” Current Regulatory Status • Multiple risk evaluations completed – Food and Drug Administration (FDA) 2003 – California Department of Fish and Game (2003) – Florida Department of Food and Agriculture Services (2004) • Concluded that fluorescent red zebra danios presented little to no risk to the United States • FDA reported no increased risk to environment or public health Fish Invasiveness Scoring Kit v2 • FISK: 49 questions in total (each with guidance notes) – 13 Qs Biogeography/History • Domestication/cultivation (3) • Climate and distribution (5) • Invasive elsewhere (5) – 36 Qs Biology and ecology • • • • • Undesirable traits (12) Feeding guild (4) Reproduction (7) Dispersal (8) Persistence (5) FISK Scoring and Product • Question scoring – Each response assigned a numerical value – Some questions are weighted • Total score – Sum of response values – Range (-11 to 54) depending on responses – determines risk category Why Use FISK • Recently updated and evaluated for use in peninsular Florida • Identifies potentially invasive species in a timely and cost-effective manner • First use of FISK on a transgenic animal • Transparent, peer reviewed, internationally vetted, published • Two Assessors with 1 expert reviewer Zebra Danio Score Black Tetra Tiger Barb -0.5 (-1/0) -3.5 (-6/-1) -2.5 (-3/-2) Risk Category Low Low Low Biogeography/Historical 0/-1 0/2 2/1 1. Domestication/cultivation 0/0 0/0 0/0 2. Climate and distribution 2/1 2/2 1/0 3. Invasive elsewhere -2/-2 -2/0 1/1 Biology/Ecology -1/1 -6/-3 -5/-3 4. Undesirable traits 1/2 0/2 1/2 5. Feeding guild 0/0 0/0 0/0 6. Reproduction 2/2 0/0 0/0 7. Dispersal mechanisms -1/-1 -1/-1 -1/-1 8. Tolerance attributes -3/-2 -5/-4 -5/-4 Zebra Danio 100 80 60 40 20 0 VU MU 100 VC MC VC Black Tetra 80 % Responses MC 60 40 20 0 VU MU 100 Tiger Barb 80 60 40 20 0 VU MU MC VC Conclusions • Low risk of invasiveness for the United States • No evident risk of gene transmission or hybridization with native species • Limited ability to even establish in U.S. • Risk from the transgenic varieties are not different than the wild-types. Conclusions Cont. • In most cases, transgene reduced fitness – Reduced fecundity (Gong et al. 2003; Jiang et al. 2011 – Reduced thermal tolerances (Cortemeglia and Beitinger 2005) – Increased predation vulnerability (Hill et al. 2011) • FISK not picking up on reduced fitness attributes Issues • Climate matching different for the 3 fishes • Invasion History • Assessor variability – Interpretations of questions and data quality – Overall, agreement and certainty was high Benefits • Results indicate little need for further investment into risk mitigation • Identified few data gaps, important uncertainty areas • Low-cost timely information to managers • Expert reviewer identified issues