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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