Transcript Addressing sustainability for aquaculture though the

Continuing the journey along the path of
aquaculture sustainability with the
Integrated Multi-Trophic Aquaculture (IMTA)
project in the Bay of Fundy - 2008.
S.M.C. Robinson, B. Glebe, B. Forward, M.
Liutkus, T.R. Lander, G.K. Reid, J.D. Martin
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Continue from last year at Scotia Pride
Brief IMTA familiarisation
Issues
Fish health experiments
Planktonic food
Ecological implications
Future directions
Integrated Multi-trophic
Aquaculture
Organic Extractive
Fed
Inorganic Extractive
IMTA in the Bay of Fundy
IMTA site fly-through
Species Interactions
Aquatic Organism Health
• Will IMTA sites act as reservoirs for
disease? ISAV?
– Norway, Newfoundland
• Are all diseases/pathogens the same?
• Have we already done the experiment
inadvertently?
• What are the risks?
– Skar and Mortensen (2007)
• What about parasites?
ISAV Observations (2003)
• Cross transmission of disease (NB, Norway,
Scotland)
Effect of ISA on Invertebrates
• Experiment – October
2003
As Reservoirs
– Shawn Robinson, Brian Glebe, Mike Beattie, RPC
Mussels
– RNA extracted (Trizol Reagent (Gibco BRL))
– 2 μg RNA (standardised) added to Ready to Go RT-PCR
Control
Treatment
Nylund
FA-3 and RA-3 primers
(3 replicates, 12°C, with
24 h static,
6 d flow-through)
(3 replicates, 12°C, 24 h static, 6 d flow-through)
– T0: none detected (controls and treatment)
0 hrs
0 hrs
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T1:
none detected (controls and treatment)
N=5 PCR, 10IFAT)
(n=5 PCR, 10 IFAT)
– T7: none detected (controls and treatment)
24 hrs
• Corroborating
results
(n=10 PCR, 20 IFAT)
24 hrs
(n=10 PCR, 20 IFAT)
– S. Ellis and L. Gustafson (USDA)
7 days
7 days
– Cem
Giray et al. (Micro Technologies Inc.)
(n=10
PCR, 20 IFAT)
(n=10 PCR, 20 IFAT)
Experiment 2 Protocols (2008)
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4 viral treatments:
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2 bacterial treatments:
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Infectious Pancreatic Necrosis Virus (IPNV),
Infectious Salmon Anemia Virus (ISAV, two strains)
Nodavirus.
Aeromonas salmonicida (strain FFA 21) atypical furunculosis
Renibacterium salmoninarum (strain FFA 198) bacterial kidney disease (BKD)
Virology: 1 ml concentrated sample of seawater from each tank was collected at 0, 4,
8 and 24 hour time points, filter sterilized and delivered to RPC for analysis.
Bacteriology: 2 ml water sample was taken at 0, 4, 8 and 24 hrs post inoculation and
mixed with 2 ml of Phosphate Buffered Saline (PBS) containing 40% glycerol. The
resulting 20% glycerol sample was then frozen in liquid nitrogen and stored at -80°C
until further analysis.
Water samples were serially diluted and plated on appropriate indicator cell line (ASK
II for ISAV, CHSE for IPN, and SSN for Nodavirus) in order to determine the presence
of active virus.
Growth of atypical Aeromonas Salmonicida strain FFA21 was at 20°C for 40 hrs in 1L
of Tryptone Soy Broth (TSB) supplemented with 2% NaCl with shaking at 90 rpm.
Growth of Renibacterium Salmoninarum strain FFA198 was on SKDM-2 plates
overlayed with dialysis membrane at 14°C for 4 weeks.
Viral Trials – 2008
Live virus – after 24h
IPNV
ISAV-4
ISAV-2
Nodavirus
60%
40%
Viral Reduction
20%
0%
-20%
-40%
-60%
Control
-80%
Treatment
-100%
Bacterial Trials
Residual Bacteria in Mussels
R. salmoninarum
10
7.50E+04
7.50E+08
Culture volume (L)
T0 water titre (#/ml)
# bacteria
A. salmonicida
10
1.25E+04
1.25E+08
Mean mussel wt (g)
# mussels
Total wt (g)
# Bacteria/g
# bacteria
2.02
50
101
3.00E+02
3.03E+04
1.33
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66.5
1.40E+05
9.31E+06
% bacteria retained
0.02%
1.24%
Conclusions
• Mussels can deal with certain microscopic
particles/organisms
• Some microbes are not affected
• Some pathogenic viruses are different
than pathogenic bacteria
• What mechanism is working on microbes?
• What is their ultimate fate?
IMTA Particle Fields
Bacteria on Glass Slides
in the Water Column
Re-Packaging Machine
Future Investigations
• What role does IMTA play in the interactions of
organisms at micro to macro-scales?
• What is the fate of pathogens that have been
bound by macro-organisms? (Pietrak, Bricknell)
• What is impact of microbial activity on the scale
of organic loads delivered by farms?
• Is there a succession of microbes on the farm
and what properties do they have that could be
valuable to IMTA? Can we adapt to this?
• The potential role of CIMTAN
Thank you!
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Atlantic Silver/Heritage Salmon/Cooke Aquaculture Inc.
Acadian Seaplants Ltd.
AquaNet
AIF – Atlantic Innovation Fund
ACRDP – Aquaculture Collaborative Research Development
Program
SABS – St. Andrews Biological Station
CIAS – Centre for Integrated Aquaculture Science
UNB – University of New Brunswick
CFIA – Canadian Food Inspection Agency
EC – Environment Canada
• ISEAS – International Sustainable Ecological Aquaculture Society