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GREAT LAKES FISH HEALTH COMMITTEE
2014 Summer Meeting Winnipeg, Manitoba
August 6-7, 2014
Minutes (with attachments)
Submitted By:
Christina Haska Great Lakes Fishery Commission
The data, results, and discussion herein are considered provisional; permission to cite the contents of this report must be requested from the authors or their
agency.
GREAT LAKES FISHERY COMMISSION 2100 Commonwealth Blvd, Suite 100
Ann Arbor, Michigan 48105 Great Lakes Fish Health Committee
Table of Contents
List of Attendees ........................................................................................................................................... 3 Meeting Agenda ............................................................................................................................................ 4 Minutes ...........................................................................................................................................................
Welcome and introductions ............................................................................................................ 5 Approval of meeting minutes .......................................................................................................... 5 CLC update ....................................................................................................................................... 5 Model Program/Risk Assessment update ........................................................................................ 5 Lake Sturgeon nucleo-cytoplasmic large DNA virus in Manitoba .................................................... 5 Manitoba Provincial Fish Health Overview ......................................................................................6 Michigan State University research update .................................................................................... 7Fish disease cases ............................................................................................................................7 Agency updates ............................................................................................................................ 7,9 Research priorities ........................................................................................................................... 8 Technical advisors ............................................................................................................................ 8 Continuation of VHS list of susceptible species ............................................................................... 8 Thiamine injection effect to EMS in Skamania steelhead................................................................9 Update on furunculosis outbreak in Pequest, NJ ............................................................................ 9 Future meetings .............................................................................................................................11 Tour of Freshwater Institute Aquatic Animal Health Lab ..............................................................11
Appendices ...................................................................................................................................................... 1: GLFHC responses to CLC concerns about the Risk Assessment ................................................. 12 2. Nucleo-cytoplasmic large DNA virus in Manitoba Lake Sturgeon .............................................133: Manitoba: Fish health overview ................................................................................................22 4: Van Hornesville State Fish Hatchery ..........................................................................................30 5: Ontario fish disease case ...........................................................................................................36 6: Research Priorities 2014 ............................................................................................................39 7: Proposed Technical Advisors ..................................................................................................... 41 8: Thiamine injection of Skamania steelhead broodstock .............................................................42
2
List of Attendees
John Dettmers Great Lakes Fishery Commission
Christina Haska Great Lakes Fishery Commission
Sunita Khatkar Fisheries and Oceans Canada
Dave Meuninck Indiana Department of Natural Resources
Andy Noyes New York State Department of Environmental Conservation
Paula Phelps Minnesota Department of Natural Resources
Ken Phillips U.S. Fish and Wildlife Service- Wisconsin
Ling Shen Minnesota Department of Natural Resources
Gary Whelan Michigan Department of Natural Resources
Coja Yamashita Pennsylvania Fish and Boat Commission
Other attendees included:
Jeff Long Manitoba Fisheries Branch Sharon Clouthier Fisheries and Oceans Canada Martha Wahlagamood Michigan Department of Natural Resources
3
Great Lakes Fish Health Committee Meeting August 6-7, 2014
Winnipeg, Manitoba Draft Agenda
Wednesday, August 6
1:00 pm – 1:10 pm Welcome & Introductions (Shen)
1:10 pm – 1:20 pm Approval of Meeting Minutes (Shen)
1:20 pm – 1:30 pm CLC update (Dettmers)
1:30 pm – 2:00 pm Model Program/Risk Assessment Update (Shen/Dettmers)
2:00 pm- 2:45 pm Lake Sturgeon nucleo-cytoplasmic large DNA virus in Manitoba (Dr. Clouthier)
2:45 pm – 3:05 pm Manitoba Provincial Fish Health Overview (Jaff Long/Laureen Janusz)
3:05 pm – 3:20 pm Break
3:20 pm – 4:15 pm MI State University research updates (Dr. Faisal/Dr. Loch)
4:15 pm – 4:45 pm Fish disease cases (Andy Noyes, Sunita Khatkar)
4:45 pm – 5:30 pm Agency Updates— (All)
Thursday, August 7
8:30 am – 9:00 am Research Priorities (Dettmers)
9:00 am – 10:00 am Technical advisors (Shen and Subcommittee)
10:00 am – 10:30 am Update on A.s. situation in Pequest, NJ (Dr. Lovy)
10:30 am – 10:45 am Break
10:45 am – 11:05 am Thiamine injection effect to EMS in Skamania steelhead (Dave Meuninck)
11:05 am – 11:50 am Agency Updates— (All)
11:50 am – 12:00 pm Future meetings (Shen)
12:00 am – 1:00 pm Lunch
1:00 pm – 3:30 pm Tour Freshwater Institute Aquatic Animal Health Lab
4
1. Welcome & introductions (Shen)
2. Approval of meeting minutes (Shen)
Minutes were approved pending changes.
3. CLC update (Dettmers)
Ling gave a presentation to the CLC in April about recommended changes to the risk assessment (see update below).
The lake committees are assisting with efforts related to increasing connectivity in watersheds (e.g., removing or modifying dams and barriers). This may affect fish health down the road. Outside the Great Lakes basin, Manitoba is removing small-head dams in agro-systems, and they are curious about lessons learned from the states (e.g., how to address pathogens, AIS, hydrology, etc). In the U.S., there’s a recognition that this issues need to be incorporated upfront to make better decisions, but people are just starting to get involved. There’s a lot of ecosystem benefits to having connectivity in Michigan’s perspective, realizing they are on the edge of opinion.
4. Model Program/Risk Assessment update (Shen/Dettmers)
Ling presented the risk assessment to the CLC, whose members had a number of questions. As a result, the writing subcommittee had a conference call with three members of the CLC to address their concerns. See Appendix 1 for more information. The new risk assessment was sent to the committee for final approval.
One potential change was to increase the degree of risk if a fish is infected with more than one pathogen. There were multiple ways to address this, including doing a risk assessment for each pathogen and using the one with highest risk. Instead, it was decided to take a precautionary approach and increase the risk to the next level if more than one pathogen is present.
This risk assessment is a living document and may be changed in the future after agencies use it. The committee reached consensus that it is ready to go online. Ken will check on the progress of automating the forms.
5. Lake Sturgeon nucleo-cytoplasmic large DNA virus in Manitoba (Clouthier)
See Appendix 2 for the presentation.
Discussion points: • It is expected that some eggs have the virus on the surface, and most sturgeon
management hatchery plans disinfect the eggs’ surfaces to minimize risk of transmission. It is unknown if iodophore affects the virus.
5
• The virus is doubly encapsulated. Hendrick looked at cross species and there were someevidence the virus was retained, but there was no evidence of transmission betweenother salmonids.
• Without a massive die-off, it is hard to determine what the virus is doing. There areseasonal variations in virus load, and it is most likely endemic in Manitoba. Infectionscan be acute or chronic, and these events can be reduced by decreasing the viral load inthe water. The fish’s barbells can be affected, which prevent the fish from finding food,causing starvation and death.
• The expectation is the virus is widespread across the provinces. Future testing willdetermine that, but unfortunately only her lab is surveying the virus. Various stockingprograms and sturgeon projects have helped with data collection.
o USFWS in WI has a stocking program and has some histology information. Theycould collaborate on sample gathering and transfer.
• It is recommended that testing for the virus be required when moving fish betweenstates. The outbreaks occur in the hatcheries.
• Sometimes coinciding with this virus are sturgeon herpesvirus and iridoviruses.
6. Manitoba provincial fish health overview (Long/Janusz)
See Appendix 3 for the presentation.
Discussion points: • There is limited interaction with other provinces, more so with AIS managers than
hatcheries, but there are no clear connections with other fish health people. Manitoba is at the bottom of a huge drainage basin, so things show up that are in other provinces, and the province is most similar to Saskatchewan.
• Alberta, Manitoba, and Saskatchewan do not have a very good fish health program. CFIAis coming into the picture, and it does not want DFO testing anything other than what is sent to them. The focus is on a limited number of pathogens and agriculture. As a result, Manitoba cannot go to them for guidance. With the ambiguous mandate for Manitoba, they end up doing things under the radar.
o In the U.S., the GLFHC exists, and the Northeastern Fish Health Committee isfollowing its footsteps. The committee recommendation is to network.
• The general consensus was the province is doing what it can. It’s hard to justify testingfor pathogens that have never shown up in the province BUT collecting that data is not a waste of time and provides a good data set. Never underestimate the power of private aquaculture being able to affect your work. There’s always alternative vectors and routes for pathogens that are not being monitored. Improving biosecurity was recommended.
• There’s a debate about whether or not disinfecting for VHS helps or hinders fish. TheUSFWS has found disinfecting walleye eggs for VHS can help improve overall health because it gets rid of other pathogens. In Manitoba, there are competing opinions over this, but the two experts have very different water supplies.
6
7. Michigan State University research update (Faisal/Loch)
Mohamed updated the committee about work being done in his lab at MSU.
8. Fish disease cases (Noyes/Khatkar)
See Appendix 4 for Andy’s presentation.
Discussion points:
• The water supply was 48°F year-round. Water hardens around 1400 and is supplied bytwo springs.
• Mortality at its worst was 1% for about 5 days pre-treatment. There was a problem withmedicated feed arrival, which extended mortality. After psycrophalum jumped in, therewas some bacterial coldwater disease signs.
See Appendix 5 for Sunita’s presentation.
Discussion points:
• CFIA says they have to grow the fungus to meet the demands of their and OIE protocols,but OIE wants issues reported but not investigated. They only care about certainpathogens.
9. Agency updates (All)
New York State DEC: In 2012, the Rome hatchery had furunculosis, and the hatchery lost about 800k fish. The hatchery classification was downgraded, and a two-year abatement plan included disinfection and depopulation of susceptible lots. The last inspection takes place next month. The hatchery does not plan to get fish outside of the state of NY again. CLT was approved for warmwater fish, walleye, and salmonids. Andy is working with Jim Balker to address targeted studies to move along the approval for esocids. There are promising data, but tests need to be repeated.
Pennsylvania FBC: CTv was not found; however, three hatcheries had large die-offs of Lake Trout finglerlings due to IPN. A vaccination program began for furunculosis, and they don’t see larger die-offs like previously. There were no problems with BKD this year except for one hatchery that always has whirling disease. Eden City hatchery was Class A, but new broodstock were brought in for production of northern pike and furunculosis was found. The progegny were negative. The hatchery is changing policies so broodfish are not in same facility as production fish. Finally, in 2010 a picture was posted on Facebook of what looked to be a Bighead Carp at a pay lake in southwestern PA. In May 2014, another was posted. The lake was surveyed, and a diploid Grass Carp was found as well as some endangered species. The lake’s owner received fish from a Lake Erie commercial fisherman and didn’t know if the
7
fish were tested prior to stocking. The FBC sampled bluegill, carp, and largemouth bass for VHS and all were negative. Now, the lake owner has to seine out his lake (less than ½ acre in size) and keep the fish he wants, rotenone the lake, and then re-stock. Pennsylvania was not prepared for this type of situation.
Discussion: Should there be a new list to reference since APHIS has rescinded theirs? Now it’s up to each state to create their own list, not the GLFHC. New York assumes all fish are susceptible to VHS, which takes care of testing requirements. CFIA has list of susceptible species on its website for Canadians to reference.
Action Item: Create fact sheets for major pathogens and post them on the GLFHC website. Refer to the Blue Book to reduce confusion. Pathogen descriptions should be written for the Model Program pathogens that are not included.
10. Research Priorities (Dettmers)
See Appendix 6 for the revised Research Priorities document.
11. Technical Advisors (Shen and Subcommittee)
A list of potential technical advisors was created and approved by the committee (Appendix 7). Ling will contact the scientists to see if they would be interested in serving the committee. This list will be revisited on a regular basis and may be revised in the future.
12. Continuation of VHS list of susceptible species
Minnesota changed its VHS rules:
1. Rearing ponds are being tested every two years instead of every year.2. Fish are undergoing PCR testing.3. The state is being divided into two districts: the Lake Superior watershed
(regulations are staying the same) and rest of state (where VHS has not beenfound).
4. Sampling size has increased to 60 fish (from 30 right now).
The state is continuing active surveillance and analysis of fish kills.
Conversation regarding a potential place to house the original APHIS order
OIE still has a version of the list, and that could be referenced. The USFWS does not have regulatory authority and would not want to house the list, as they refer to state regulations. The Service is revising Title 50, which deals with the importation of salmonids. Pathogens make the fish potentially injurious.
8
The AFS Fish Health section also does not have regulatory authority, and its Blue Book only makes recommendations for families of fish, not specific species.
For now, it would be best to check with APHIS and see if they plan to update the list when a new species is found to be susceptible and what that process would be. Will they upkeep it or get rid of it eventually? Action Item: Ling will contact the agency and ask.
13. Thiamine injection effect to EMS in Skamania steelhead (Meuninck)
See Appendix 8 for the presentation.
Discussion points:
• The process of injecting 100 wild broodstock fish takes about 45 min.• The fish also get an OTC injection to prevent furunculosis. The thiamine injection goes in
the dorsal sinus.• Inject every year? Just coming off the truck. Spawning females are sacrificed. Fresh
broodstock is captured every year.• There are tradeoffs between treating the fish or doing flow-through treatment of the
eggs, such as expense costs and survival of the fish, likely.• Fish get injected when coming into the facility and are held for a few months prior to
spawning.
14. Update on furunculosis outbreak in Pequest, NJ (Lovy)
Discussion points:
• Andy offered to send Rome strain brook trout that are resistant to furunculosis. It wouldneed to be worked out among the management folks.
o A spread test of Rome-strain fish with A. sal showed the infection is gone after 3weeks.
o Coja has experience with the Rome strain, and they are resistant in PA.• Jan did cultures of the fish that were not responding to treatment. Likely the outbreaks
were occurring because some fish ate the medicated feed and other did not.
14. Agency updates, cont. (All)
U.S. Fish and Wildlife Service: Smallmouth Bass were infected with Yersinia ruckerii at Genoa this spring. The Iron River hatchery will be Class A if the inspection this month is clean. Eric Griese has made some Asian carp cell lines in conjunction with the USGS lab to screen potential chemical and viral controls. Construction should start soon at the Jordan River hatchery to
9
cover raceways and update the early-life stage rearing building. There will be room for coregonid culture as well – not sure what species yet.
Indiana DNR: There were no BKD detections in any coldwater units, but hatcheries still get occasional A. sal positive detections without any clinical signs of disease, which then disappear in the following inspection. EMS was found in Coho in the winter runs from Michigan this year. Treatment did not seem to help initially but the fish did recover. A local Department of Corrections has a walleye culture facility which grew Channel Catfish for children’s fishing events. It began culturing Walleye but ran into problems and lost all of them. Analysis is being done at Purdue now. There was a large die-off of Gizzard Shad this winter in northern Indiana. There was concern about whether or not Muskie spawning would happen on time, but the ice melted and they had a good egg take. In southern Indiana, there is evidence that Gizzard Shad are causing reproductive failure in Largemouth Bass. A comparison study between a lake with and without Shad showed different thiamine levels in Bass between the reservoirs. Eagles around the area also eat Shad and are sometimes found to be lethargic. After receiving thiamine injections for a vet, their energy returns.
Fisheries and Oceans Canada: All hatcheries in Manitoba and Ontario are clean according to CFIA testing methods. A new process requires accreditation for labs, which is being pursued now. On the research side, SVCv is being looked for and an assay is being developed. CFIA is taking over domestic issues so DFO has to rescind their orders. There is concern this will affect the states – if anyone imports from Canada and has issues, let Sunita know. It’s unclear what the issues will be as this transition occurs. It might be helpful to invite CFIA to the winter meeting and give an update. They could not be a formal member but could be invited regularly to ensure open communication. Action Item: Sunita will find a contact person. Might be helpful to find someone at APHIS to invite regularly as well.
Michigan DNR: There were limited mortalities in the wild this year except in shallow lakes. Oden hatchery had an IPN outbreak in broodstock and caused mortality in brown trout. It was initially detected in production fish but it disappeared, which is raising some concerns. The hatchery is taking an aggressive management approach: biosecurity, testing, culling out positives, and reducing stressors on fish to try to reduce the load. It’s unclear why this is an issue this year. There was a trace of IPN in Marquette, which has the pathogen in the supply water, but it disappeared. A UV system needs to be installed for production fish (already have one for broodstock). Private aquaculture wants to be a bigger industry in Michigan. The strategic plan is vague and unspecific but wants an expansion from $5-10 M/year to a billion dollar agency by 2025. The plan is to culture 80% of fish in cages in the Great Lakes. This has created lots of work for the state with permitting and disease control. The DNR is unenthusiastic, as are constituent/commercial/angling groups.
10
Minnesota DNR: A few fish kills involving Crappies and other sunfish were due to Columnaris. The Lanesboro hatchery had Rainbow Trout yearling mortalities from Aeromonas hydrophila. The fish were treated with Oxytech medicated feed twice before moving them to an outdoor pond where they would have more space. No more mortalities were observed. A few weeks ago, Crystal Springs hatchery sent pictures of Lake Trout with clinical signs of disease. The fish are kept in covered raceways with panels for daylight to come through. Hatchery personnel do not think it’s sunburn, but it may be frostbite. The issues seems to be strictly in the dermis and does not penetrate the body cavity. Histology has not been done yet.
15. Future meetings (Shen)
The Winter 2015 meeting will be in West Lafayette, IN from February 3-4, 2015.
The Summer 2015 meeting will be in Syracuse, NY from July 28-29, 2015 (including an Oneida Lake field trip).
The Winter 2016 will be in Lansing, MI.
16. Tour of the Freshwater Institute Aquatic Animal Health Lab
17. Adjourn
11
GLFHC Writing Subcommittee Responses to CLC Concerns Regarding the Risk Assessment
May 2014
Questions from S. LaPan’s email (5/12/2014)
Form RA-2, question 5: Given that this criterion addresses pathogens not currently in the Great Lakes,
and a necessary score above 733 is required to recommend against stocking, is the maximum, weighted
value of this criterion (100) adequate?
The value for Absence was increased to 30 points.
Form RA-2, questions 5, 6 and 7: Recognizing our ignorance of fish pathology, we know there are
distinctions between these criteria, but would greatly benefit from a discussion (conference call) regarding
their significance/weighting. Further to the previous question, the introduction of a new pathogen into the
Great Lakes would receive a combined score of only 450 for criteria 4 through 8 and 12. Would we not
need to recommend against stocking based on these 6 criteria alone?
The scoring was changed for questions 5, 6, 7, and 12. The writing subcommittee is considering
changing the wording of #12 as well. This can be discussed during the call.
Form RA-2, questions 6 and 7: Suggest revised wording: “Will introduction of these fish likely
increase….”
Done.
Form RA-2, question 10. Several of the criteria are auto-correlated and therefore appear
unnecessary. For instance, if the “wild” receiving water is being considered for stocking, we can assume
that there is fish stocking and likely recreational fishing in the receiving water. Also, are there any wild
receiving waters that don’t have the potential for predators?
This question could potentially be eliminated.
Form RA-2, question 17. Under “Comprehensive/Continual….”/ “No other Model Program pathogen(s)
detected,” why would these selections generate a score other than “0”?
In general, the value of ‘1’ was used to show there is still a degree of risk. Several GLFHC
committee members felt this distinction was important and disapproved initially of having zero
values; however, the writing subcommittee decided to follow your suggestion and made this
change.
Form RA-3. Once this form is signed by the GLFHC, who receives copies of the form?
This is mentioned in the introduction of the document: “The summary report will be provided to
all member agencies, the appropriate lake committee(s), and the CLC.”
Comments raised during the CLC meeting
If a fish is infected with a pathogen, does that make it more susceptible to other pathogens?
This is very likely and will depend on the number of other active infections along with the
virulence of the pathogens involved.
How does the risk assessment take into account if a fish is infected with more than one pathogen?
The RA was not intended to take into account more than one pathogen because most cases are
focused on single pathogens. Multiple pathogen cases would likely not get to this decision
stage. If the CLC thinks it would be beneficial to have an additional risk assessment that would
analyze multiple pathogens, this could be developed in the future.
Appendix 1
12
NU
CLE
O-C
YTO
PLA
SMIC
LA
RG
E D
NA
VIR
US
IN
MA
NIT
OB
A L
AK
E ST
UR
GEO
N
Shar
on
Clo
uth
ier*
, DFO
Fre
shw
ater
Inst
itu
te
0.4
CIV
IIV
3FV3
ISK
NV
LDV
MV
DP
AV
ASF
V
PO
V
AC
MV
NV
WSI
VC
roV
MV
Ch
ileM
VC
ou
rA
PM
V-1
AP
MV
-2
OLP
V-2
PP
VC
EV
HA
V
ATC
VP
BC
V
MP
VO
TV
FSVES
V
Irid
ovi
rid
ae
Ph
yco
dn
avi
rid
ae
Un
clas
sifi
edA
sco
viri
da
e
Asf
arv
irid
ae
Mim
ivir
ida
e
Un
cla
ssif
ied
m
imiv
iru
s-lik
e
Ove
rvie
w o
f p
rese
nta
tio
n
Vir
us
dis
cove
ry
Cu
rren
t in
vest
igat
ion
s
Bir
thd
ay R
apid
s ca
se
In v
ivo
ch
alle
nge
stu
dy
Dia
gno
stic
tes
t d
evel
op
men
t Dis
ease
man
agem
ent
stra
tegi
es
0.4
CIV
IIV
3FV3
ISK
NV
LDV
MV
DP
AV
ASF
V
PO
V
AC
MV
NV
WSI
VC
roV
MV
Ch
ileM
VC
ou
rA
PM
V-1
AP
MV
-2
OLP
V-2
PP
VC
EV
HA
V
ATC
VP
BC
V
MP
VO
TV
FSVES
V
Irid
ovi
rid
ae
Ph
yco
dn
avi
rid
ae
Un
clas
sifi
edA
sco
viri
da
e
Asf
arv
irid
ae
Mim
ivir
ida
e
Un
cla
ssif
ied
m
imiv
iru
s-lik
e
Na
ma
ovi
rus
(NV
)
Na
ma
ovi
rus
(NV
)&
u
ncl
assi
fie
d, I
rid
ovi
rid
ae
(WSI
V, M
RSI
V, B
CW
SV, S
NSV
)
NC
LDV
s•d
ie o
ffs
•co
nse
rva
tio
n s
tock
ing
MA
NIT
OB
A
Hu
dso
n B
ay
Win
nipe
g R
iver
Lake
stu
rgeo
n (A
cip
ense
rfu
lves
cen
s) in
Ma
nit
ob
a, C
an
ad
a
5 p
op
ula
tio
ns
con
sid
ered
fo
r SA
RA
list
ing
•W
. Hu
dso
n B
ay –
DU
1•
Sask
atch
ewan
R –
DU
2•
Nel
son
R –
DU
3•
Red
& A
ssin
ibo
ine
R t
o L
kW
inn
ipeg
–D
U4
•W
inn
ipeg
R t
o E
ngl
ish
R –
DU
5
Co
nse
rvat
ion
sto
ckin
g, 1
992
-20
14
•St
akeh
old
ers:
•Fi
rst
Nat
ion
s•
Pro
vin
cial
go
vern
men
t•
Man
ito
ba
Hyd
ro•
Un
ive
rsit
y o
f M
anit
ob
a•
Fed
eral
go
vern
men
t
Riv
er•G
amet
es c
olle
cted
fro
m w
ild b
roo
dst
ock
str
eam
sid
e
Hat
cher
y
•Fe
rtili
zati
on
of
eggs
•La
rval
rea
rin
g
Riv
er
•R
elea
se o
f ju
ven
iles
in s
um
mer
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all o
f h
atch
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r
•R
elea
se o
f ju
ven
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in s
pri
ng
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g ye
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Pro
cess
DU
1
DU
3
DU
2 DU
4
DU
5
MA
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A
Hu
dso
n B
ay
Appendix 2
13
Vir
us-
ass
oci
ate
d o
utb
rea
ks in
lake
stu
rgeo
n
0102030405060708090100
14
710
1316
1922
2528
3134
3740
4346
49
Tim
e (
d)
Cumulativemortality (%)
0
10
20
30
40
50
60
70
80
90
10
0
6/19/09
7/17/09
8/14/09
11-09-09
9-10-09
6-11-09
4-12-09
1-01-10
1/29/10
2/26/10
3/26/10
4/23/10
5/21/10
Dat
e
Cumulativemortality (%)
Gra
nd
Rap
ids
Hat
che
ry
Un
iver
sity
of M
anit
ob
a
Nel
son
Riv
er 2
009
Win
nip
eg R
ive
r 2
009
Mo
rtal
ity
99
.6%
9½
mo
nth
s
99%
62%88
%
64%
Win
nip
eg R
ive
r 2
0083
½ m
on
ths
Mo
rtal
ity
75
%
10
day
s
Wild
bro
od
sto
ck
Na
ma
ovi
rus
dia
gn
ost
ic t
ests
his
tolo
gy,
ele
ctro
n m
icro
sco
py,
PC
R
SKIN
GIL
L
NC
Dia
gno
stic
tes
tin
g
•H
isto
logy
•ep
ith
elio
tro
ph
ic(o
ral,
nas
al, b
arb
els,
ski
n, g
ill, f
in)
•p
ath
ogn
om
on
ic c
han
ges
•en
larg
ed e
pit
hel
ial c
ells
•cy
top
lasm
ic in
clu
sio
n b
od
ies
•El
ectr
on
mic
rosc
op
y•
ico
sah
edra
l-sh
aped
•24
2 n
m d
iam
ete
r•
Co
nven
tio
nal
PC
R•
Maj
or
Cap
sid
Pro
tein
•21
9 b
pam
plic
on
GIL
L
Mo
lecu
lar
test
m
eth
od
a
Dia
gno
stic
re
sult
s2
00
8 Y
C2
00
9 Y
CU
MA
AH
FG
ran
d R
apid
s H
atch
ery
UM
AA
HF
Win
nip
eg
Riv
er
Ne
lso
n R
ive
rW
inn
ipe
g R
ive
r W
inn
ipe
g R
ive
rN
V c
PC
R7
/83
/33
5/3
71
4/1
5W
SIV
qP
CR
0/8
0/3
0/3
50
/12
MR
SIV
qP
CR
0/8
0/3
0/3
50
/12
0.4
CIV
IIV
3FV3
ISK
NV
LDV
MV
DP
AV
ASF
V
PO
V
AC
MV
NV
WSI
VC
roV
MV
Ch
ileM
VC
ou
rA
PM
V-1
AP
MV
-2
OLP
V-2
PP
VC
EV
HA
V
ATC
VP
BC
V
MP
V
OTV
FSVES
V
Irid
ovi
rid
ae
Ph
yco
dn
avi
rid
ae
Un
clas
sifi
ed
Asc
ovi
rid
ae
Asf
arv
irid
ae
Mim
ivir
ida
e
Un
cla
ssif
ied
m
imiv
iru
s-lik
e
Ph
ylo
gen
etic
an
alys
is
•M
ajo
r ca
psi
d p
rote
in•
5 sN
CLD
Vs
& 2
5 o
ther
NC
LDV
s•
Co
mm
on
evo
luti
on
ary
pas
t•
Mim
ivir
ida
e•
sNC
LDV
ssu
b-s
pec
ies
leve
l
NV
ta
xon
om
y
Cu
rre
nt
nam
eG
eo
grap
hic
ran
geR
ive
r d
rain
age
Firs
t re
po
rt
WSI
V
Cal
iforn
iaSa
cram
ento
Riv
er1
99
0O
rego
nC
olu
mb
ia R
iver
19
94
Idah
o –
No
rth
Koo
ten
ai R
iver
Idah
o -
Sou
thSn
ake
Riv
er
BC
WSV
Bri
tish
Co
lum
bia
Fras
er R
iver
20
03
MR
SIV
No
rth
& S
ou
thD
ako
taM
isso
uri
Riv
er2
01
0
SNSV
New
Bru
nsw
ick
St J
oh
n R
iver
20
14
NV
Man
ito
ba
Nel
son
Riv
er2
01
3W
inn
ipeg
Riv
er
Val
ley
Cit
y N
FH
Gav
ins
Po
int
NFH
Gar
riso
n D
am N
FH
Mile
s C
ity
SFH
htt
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/mri
.usd
.ed
u/e
du
cati
on
_net
wo
rk/
htt
p:/
/ww
w.n
wd
-mr.
usa
ce.a
rmy.
mil/
rcc/
MR
SIV
Stu
rgeo
n N
CLD
Vs –
geo
gra
ph
ic r
an
ge
SNSV
htt
p:/
/ww
w.n
icke
rso
nla
ke.c
o/
BC
WSV
WSI
V
Fras
er R
Sacra
men
to R
Up
per
Co
lum
bia
WS
Rec
ove
ry P
lan
20
02
Appendix 2
14
Stu
rgeo
n N
CLD
Vs –
Bio
log
ica
l ch
ara
cter
isti
cs
Ch
arac
teri
stic
Stu
rge
on
NC
LDV
NV
WSI
VM
RSI
VB
CW
SV
Ho
stla
ke s
turg
eon
wh
ite
stu
rgeo
np
allid
& s
ho
veln
ose
wh
ite
stu
rgeo
n
Shap
eh
exag
on
al
Size
(si
de
to s
ide,
nm
)2
42
26
22
54
26
1
Gen
om
ed
sDN
A
Tiss
ue
tro
pis
mIn
tegu
men
t &
gill
Cel
l tro
pis
mep
ith
elia
lce
lls (
Mal
pig
hia
n&
mu
cosa
l cel
ls)
Cel
l cu
ltu
ren
oSo
met
imes
no
N/A
Cel
l typ
esn
on
e id
enti
fied
1w
hit
e st
urg
eon
sple
en,
gon
ad1
no
ne
iden
tifi
ed1
N/A
Tem
per
atu
re (
°C)
15
15
21
5N
/A
1.
LS g
ill o
rgo
nad
pri
mar
yce
ll lin
es &
2 W
S ce
ll lin
es;
8 W
S ce
ll lin
es e
stab
lish
ed;
PS/
SS p
rim
ary
cell
lines
2.
Vir
us
rep
licat
ion
occ
urs
at
10
, 15
an
d 2
0°C
bu
t n
ot
at 5
or
25
°C;
op
tim
um
is 1
5°C
Gen
om
e si
ze (
kb)
10
02
50
50
0
Irid
ovi
rid
ae
Asc
ovi
rid
ae
Asf
arv
irid
ae
Po
xvir
ida
e
Ph
yco
dn
avi
rid
ae
Mim
ivir
ida
e
75
01
00
01
25
0
Ma
rsei
llevi
rus
Cro
V
Mim
ivir
use
s
Meg
avi
ruse
s
NV
Ove
rvie
w o
f p
rese
nta
tio
n
Vir
us
dis
cove
ry
Cu
rre
nt
inve
stig
atio
ns
Bir
thd
ay R
apid
s ca
se
In v
ivo
ch
alle
nge
stu
dy
Dia
gno
stic
tes
t d
evel
op
men
t Dis
ease
man
agem
ent
stra
tegi
es
Scre
enin
g la
ke s
turg
eon
bro
od
sto
ck, g
am
etes
& p
rog
eny
for
NV
Win
nip
eg R
iver
dra
ina
ge
con
serv
ati
on
sto
ckin
g p
rog
ram Sl
ave
Falls
Stu
rgeo
n F
alls
Poin
te d
u B
ois
Fal
ls
Win
nip
eg R
iver
(M
an
ito
ba
)
Sam
ple
so
urc
e
Win
nip
eg
Riv
er
Dra
inag
e
Ye
ar a
nd
Ori
gin
of
Bro
od
sto
ck
Stu
rge
on
Fal
lsSl
ave
Fal
lsP
oin
te d
u B
ois
Po
inte
du
Bo
isP
oin
te d
u B
ois
Po
inte
du
Bo
isP
oin
te d
u B
ois
20
05
to
20
07
20
08
20
09
20
10
20
11
20
12
20
13
PC
RH
isto
PC
RH
isto
PC
RH
isto
PC
RH
isto
PC
RH
isto
PC
RH
isto
PC
RH
isto
Mal
e b
roo
ksto
ckfi
n c
lip0
/50
/50
/30
/30
/90
/91
/70
/7
Fem
ale
bro
od
sto
ckfi
n c
lip0
/60
/60
/30
/30
/90
/90
/50
/5
Milt
0/5
0/4
0/1
2
Eggs
0/6
0/8
0/5
Larv
ae0
/24
0/2
40
/40
0/4
00
/60
0/1
00
/30
0/3
0
Juve
nile
s fi
n c
lip3
/91
30
/13
03
2/3
25
/50
/50
/10
Tota
l nu
mb
er o
f sa
mp
les
3/9
13
0/1
30
32
/32
5/4
00
/40
0/5
70
/46
0/1
00
0/2
81
/59
0/4
2
NV
dia
gno
stic
res
ult
s: h
isto
logy
, qP
CR
Sam
ple
co
llect
ion
sit
es
Scre
enin
g la
ke s
turg
eon
bro
od
sto
ck, g
am
etes
& p
rog
eny
for
NV
Nel
son
Riv
er d
rain
ag
e co
nse
rva
tio
n s
tock
ing
pro
gra
m
Bu
rntw
oo
dR
iverBir
thd
ay R
apid
s
Lan
din
g R
iver
Nel
son
Riv
er (
Ma
nit
ob
a )
Sam
ple
so
urc
e
Ne
lso
n R
ive
r D
rain
age
Ye
ar a
nd
Ori
gin
of
Bro
od
sto
ck
Lan
din
g R
ive
r n
ear
th
e N
els
on
Riv
er
Bu
rntw
oo
d R
ive
rB
irth
day
Rap
ids
20
10
20
11
20
12
20
13
20
14
20
13
20
14
PC
RH
isto
PC
RH
isto
PC
RH
isto
PC
RH
isto
PC
RH
isto
PC
RH
isto
PC
RH
isto
Mal
e b
roo
ksto
ck0
/20
/20
/20
/20
/20
/21
/2
Fem
ale
bro
od
sto
ck0
/20
/22
/20
/21
/20
/21
/1
Milt
0/2
0/1
0/2
0/2
Eggs
0/2
0/2
0/2
0/1
Larv
ae0
/20
0/5
0/6
01
/59
Juve
nile
s0
/94
0/7
90
/60
0/3
0/6
00
/12
00
/60
Tota
l nu
mb
er o
f sa
mp
les
0/9
40
/79
0/8
40
/12
2/6
80
/40
/12
30
/60
1/6
80
/43
/62
Sam
ple
co
llect
ion
sit
es
NV
dia
gno
stic
res
ult
s: h
isto
logy
, qP
CR
Appendix 2
15
Ove
rvie
w o
f p
rese
nta
tio
n
Vir
us
dis
cove
ry
Cu
rren
t in
vest
igat
ion
s
Bir
thd
ay R
apid
s ca
se
In v
ivo
ch
alle
nge
stu
dy
Dia
gno
stic
tes
t d
evel
op
men
t Dis
ease
man
agem
ent
stra
tegi
es
A c
ase
stu
dy –
20
14
Bir
thd
ay
Ra
pid
s la
ke s
turg
eon
Nel
son
Riv
er c
on
serv
ati
on
sto
ckin
g p
rog
ram
06.1
2.2
014
06.2
1.2
014
06.2
3.2
014
Bro
od
sto
cksa
mp
ling
(n=3
)
Pect
ora
l fin
clip
G
amet
esEg
g fe
rtili
zati
on
Hat
cher
y
Sam
plin
g p
roge
ny f
or
NV
tes
tin
g
Wh
ole
larv
ae:
29
F1
xM1
W
ho
lela
rvae
: 3
0 F
1xM
2
Hat
ch
F1M
2F1
xM2
Bro
od
sto
ckFa
mily
F1M
1F1
xM1
fin
++
+
+-
-
NV
qP
CR
ga
met
es-
-
fin
NV
qP
CR
ga
met
es-
-
09
dp
f1
1 d
pf
07.m
id.2
014
Rel
ease
~ 3
0 d
pf
Dis
ease
eco
log
y
Envi
ron
men
t•
Wa
ter
leve
ls,
flo
w r
ate
& t
emp
era
ture
•St
ock
ing
den
sity
Ho
st•
susc
epti
bili
ty•
lifec
ycle
•im
mu
ne
sta
tus/
fitn
ess
•st
ock
sta
tus
Pa
tho
gen
•lif
ecyc
le•
tra
nsm
issi
on
•re
serv
oir
s•
viru
len
ce
Fact
ors
to
co
nsi
der
for
dis
ease
ma
na
gem
ent
or
con
tro
l
Ris
kFa
cto
rs20
14YC
Bir
thd
ay R
apid
s
Incr
easi
ng
ho
st p
op
ula
tio
ns
un
kno
wn
Ch
ange
sin
lan
d/w
ater
use
√
Fish
mig
rati
on
/mo
vem
ent
√
Rap
id,
larg
e sc
ale
envi
ron
men
tal
chan
ges
√
Rap
id,
lon
g d
ista
nce
tran
spo
rt√
Trad
e in
wild
an
imal
s an
d m
eat
X
Path
oge
n e
volu
tio
nu
nkn
ow
n
Dri
vers
of
dis
eas
e e
mer
gen
ce
Ho
st &
En
viro
nm
ent –
Bir
thd
ay
Ra
pid
s la
ke s
turg
eon
po
pu
lati
on
Nel
son
Riv
er (
DU
3)
CO
SEW
IC &
SA
RA
Hu
dso
n B
ay
Lake
Win
nip
egMU
1
MU
2
MU
3B
irth
day
Rap
ids
LS
MU
4
MU
5
MU
6
Co
nse
rva
tio
n s
tatu
s o
f st
ock
*
MU
Po
pu
lati
on
tr
ajec
tory
Imp
ort
ance
to
D
U r
eco
very
Rec
ove
ry
po
ten
tial
1In
crea
sin
gLo
wLo
w
2St
able
Hig
hM
od
erat
e
3U
nkn
ow
nH
igh
Mo
der
ate
4U
nkn
ow
nLo
wLo
w
5U
nkn
ow
nLo
wLo
w
6u
nkn
ow
nH
igh
Hig
h
Sto
ck a
sses
smen
t 2
01
0*
*Fro
mC
leat
or,
H.,
K.A
.M
arti
n,
T.C
.P
ratt
an
dD
.M
acd
on
ald
.2
01
0.
Info
rma
tio
nre
leva
nt
toa
reco
very
po
ten
tial
asse
ssm
en
to
fLa
keSt
urg
eon
:N
elso
nR
iver
po
pu
lati
on
s(D
U3
).D
FOC
an.S
ci.A
dvi
s.Se
c.R
es,D
oc.
20
10
/08
2.v
i+3
3p
.
Un
cert
ain
ty o
f an
nu
al s
tock
sta
tus
&
bio
mas
s fo
r st
ock
pro
du
ctiv
ity
dec
lines
Hyd
roel
ectr
ic G
S (n
=5)
Appendix 2
16
Pa
tho
gen
-tr
an
smis
sio
n
Ro
ute
s o
f tr
an
smis
sio
n
•H
ori
zon
tal –
infe
cted
to
naï
ve in
div
idu
als
•D
irec
t co
nta
ct w
ith
infe
cted
ind
ivid
ual
s•
Exp
osu
re t
o w
ater
co
nta
inin
g vi
rus
•V
erti
cal –
adu
lt t
o o
ffsp
rin
g vi
a th
e eg
g•
Egg
surf
ace
•In
tra-
ovu
m
Fish
den
sity
Gen
etic
susc
eptib
ility
of h
ost
Viru
s loa
d
Stre
ss
Det
erm
inan
ts
On
set
&
seve
rity
of
dis
ease
Dis
eas
e
Vir
us
Tran
smis
sio
n
DIS
EASE
MA
NA
GEM
ENT
STR
ATE
GIE
S
VIR
US
TRA
NSM
ISSI
ON
PA
TTE
RN
S
Pa
tho
gen
-ep
ith
elio
tro
pis
m&
ho
rizo
nta
l tra
nsm
issi
on
of
NV
4 days
Livi
ng
cells
Dea
d c
ells
Epit
hel
ial
cell
Epit
hel
ial c
ells
Epid
erm
is
Bas
emen
t m
emb
ran
e
Der
mis
Epit
hel
ial c
ells
Skei
n c
ell
Skei
n c
ell
Hyp
od
erm
is
Ch
rom
ato
ph
ore
Slo
ug
hin
g o
f d
ead
cel
ls
Ellio
t D
G F
un
ctio
nal
Mo
rph
olo
gy o
f th
e In
tegu
men
tary
Sys
tem
in F
ish
es
Epit
he
lial
cell
= m
alp
igh
ian
cell,
ep
ider
mal
cel
l, fi
lam
ent-
con
tain
ing
cell,
fila
men
tou
s ce
ll,
po
lygo
nal
cel
l, p
oly
hed
ral c
ell,
kera
tocy
te,
kera
tin
ocy
te,
pri
nci
pal
cel
l, an
d c
om
mo
n c
ell.
Metabolically active
Pa
tho
gen
-V
erti
cal t
ran
smis
sio
n o
f N
V?
20
10
20
11
20
12
20
13
Win
nip
eg R
ive
r
Bro
od
sto
ck-
--
+
Gam
ete
sn
/a-
--
Larv
ae/f
ry/y
ear
lings
+-
--
20
11
20
12
20
13
20
13
20
14
Ne
lso
n R
ive
r
Bro
od
sto
ck-
+n
/a+
+
Gam
ete
sn
/a-
--
-
Larv
ae/f
ry/y
ear
lings
--
--
+
Po
inte
du
Bo
is
Lan
din
g R
iver
Bu
rntw
oo
dR
ive
rB
irth
day
Rap
ids
Noevidence
Vertical or horizontal from unknown source
Pa
tho
gen
-re
serv
oir
s
Res
ervo
ir=
one
or
mo
reep
idem
iolo
gic
ally
con
nec
ted
po
pu
lati
on
so
ren
viro
nm
ents
inw
hic
hth
ep
ath
oge
nca
nbe
per
ma
nent
lym
ain
tain
eda
nd
fro
mw
hic
hin
fect
ion
istr
an
smit
ted
toth
ed
efin
edta
rget
po
pu
lati
on
•2
ho
st p
op
•M
ain
tain
pat
ho
gen
by
tran
smis
sio
n•
Res
ervo
ir i
s m
ain
ten
ance
co
mm
un
ity
of
two
dif
fere
nt
spec
ies
•Tr
ansm
issi
on
to
tar
get
occ
urr
ing
thro
ugh
on
ly o
ne
po
pu
lati
on
•2
ho
st p
op
•M
ain
ten
ance
in
on
e h
ost
po
p o
nly
•Tr
ansm
issi
on
to
tar
get
occ
urr
ing
thro
ugh
eit
her
po
pu
lati
on
NV
res
ervo
ir -
un
kno
wn
Re
serv
oir
sce
nar
ios
OIE
(2
01
0)
Trai
nin
g m
anu
al o
n W
ildlif
e D
isea
ses
and
Su
rvei
llan
ce
Am
oeb
a
Aq
uat
ic s
pec
ies
LS
Am
oeb
a
Aq
uat
ic s
pec
ies
LS
Appendix 2
17
Ove
rvie
w o
f p
rese
nta
tio
n
Vir
us
dis
cove
ry
Cu
rren
t in
vest
igat
ion
s
Bir
thd
ay R
apid
s ca
se
In v
ivo
ch
alle
nge
stu
dy
Dia
gno
stic
tes
t d
evel
op
men
t Dis
ease
man
agem
ent
stra
tegi
es
In v
ivo
ch
alle
ng
e st
ud
yO
bje
ctiv
es•
Esta
blis
h c
ausa
l rel
atio
nsh
ip b
etw
een
NV
an
d d
isea
se•
Esta
blis
h if
ho
rizo
nta
l vir
us
tran
smis
sio
n o
ccu
rs•
Esta
blis
h t
issu
e tr
op
ism
, vir
us
load
an
d in
fect
ion
dyn
amic
s
Co
ntr
ol t
anks
•M
ock
imm
ersi
on
fish
(n
=63)
•U
ntr
eate
d fi
sh (
n=6
2)
•1
tan
k: w
eekl
y sa
mp
ling;
dia
gno
stic
tes
tin
g•
1 ta
nk:
no
sam
plin
g; c
um
ula
tive
per
cen
t m
ort
alit
y
Vir
us
exp
ose
d t
anks
•10
0% im
mer
sio
n c
hal
len
ge (n
=126
)•
Co
hab
itat
ion
(vir
us
imm
ersi
on
ch
alle
nge
d fi
sh (
n=6
3)
& n
aïve
fis
h (
n=6
2))
•1
set
of
tan
ks: w
eekl
y sa
mp
ling;
dia
gno
stic
tes
tin
g•
1 se
t o
f ta
nks
: no
sam
plin
g; c
um
ula
tive
per
cen
t m
ort
alit
y
T3Im
m/n
aive
n =
63
/62
T6Im
m/n
aive
n =
63
/62
T2Im
m/n
aive
63/6
2
T1Im
mer
sio
n12
6
Co
ntr
ol t
anks
No
sam
plin
gW
eekl
y sa
mp
ling
Vir
us
chal
len
ge t
anks
No
sam
plin
gW
eekl
y sa
mp
ling
T5Im
m/n
aive
63/6
2
T4Im
mer
sio
n12
6
Envi
ron
me
nta
l co
nd
itio
ns
•70
L f
low
th
rou
gh a
qu
aria
•W
ater
tem
per
atu
re 1
5°C
•Fl
ow
rat
e 0
.4 L
/min
•St
ock
ing
den
sity
15.
5 g
/L
Vir
us
chal
len
ge•
Sou
rce
of
viru
s: h
om
oge
nat
e o
f w
ho
le f
ish
fro
m d
isea
se o
utb
reak
•C
hal
len
ge d
ose
3.2
x10
9o
r 9
.5 lo
g10
pla
smid
co
pie
s p
er L
wat
er•
1 h
ou
r st
atic
bat
h w
ith
aer
atio
n in
20
L su
spen
sio
n o
f h
om
oge
nat
e
In v
ivo
ch
alle
ng
e st
ud
y -
Tim
elin
e
Bat
h c
hal
len
ge (
mo
ck v
svi
rus)
1
hr
stat
ic w
ith
aer
atio
nN
aïve
fis
h a
dd
ed2
4 h
raf
ter
chal
len
ge
Targ
etin
g w
eekl
ysa
mp
ling
(5 f
ish
/tan
k) f
or
12
wp
cD
aily
rem
ova
l of
dea
d o
r m
ori
bu
nd
fis
h
0
Term
inat
ion
Sam
ple
d5
fis
h/g
rp/t
ank
17
wks
Tiss
ues
•H
ead
ski
n•
Bar
bel
•G
ill•
Oro
ph
aryn
x•
Pec
tora
l fin
•P
elvi
c fi
n•
Ab
do
min
al s
kin
•K
idn
ey
AB
Dia
gn
ost
ics –
all
fish
His
tolo
gyq
PC
R
12
wks
In v
ivo
ch
alle
ng
e st
ud
y -
pre
limin
ary
NV
qP
CR
resu
lts
0
20
40
60
80
10
0
16
11
16
21
26
31
36
41
46
51
56
61
66
71
76
81
86
91
96
10
11
06
11
11
16
Cumulative % mortality
Day
s p
ost
-ch
alle
nge
Mo
rtal
ity
curv
es f
or
all t
anks
5 o
f 6
NV
po
siti
ve f
ish
(T4
)
no
t ye
t an
alyz
ed (
T4)
Tan
ks f
or
wee
kly
sam
plin
g
Tan
ks f
or
cpm
1 o
f 6
NV
po
siti
ve f
ish
(T4
)
no
t ye
t an
alyz
ed (
T4)
12 w
eeks
Appendix 2
18
Ove
rvie
w o
f p
rese
nta
tio
n
Vir
us
dis
cove
ry
Cu
rren
t in
vest
igat
ion
s
Bir
thd
ay R
apid
s ca
se
In v
ivo
ch
alle
nge
stu
dy
Dia
gno
stic
te
st d
eve
lop
me
nt Dis
ease
man
agem
ent
stra
tegi
es
Dia
gn
ost
ic a
ssa
ys -
Vir
us
iso
lati
on
tes
t m
eth
od
Am
oeb
a
X X X
?
Dia
gn
ost
ic a
ssa
ys -
Mo
lecu
lar
test
dev
elo
pm
ent
stu
rgeo
n N
CLD
V
Q2
Vir
us
Dia
gno
stic
ass
ays
cPC
RQ
1
Q2
spec
ific
ity
spec
ific
ity
spec
ific
ity
WSI
V√
X√
BC
WSV
√√
√
MR
SIV
√√
√
SNSI
V√
√√
NV
√√
√
An
alyt
ical
sen
siti
vity
(pla
smid
cop
ies)
NV
503
5
Q1
cPC
R
Ma
jor
cap
sid
pro
tein
–a
ssa
y ta
rget
are
as
An
aly
tica
l sp
ecif
icit
y a
nd
sen
siti
vity
NC
LDV
cPC
RQ
1Q
2
Nam
aovi
rus
√√
√
BC
wh
ite
stu
rgeo
n v
iru
s√
√√
Mis
sou
ri R
iver
stu
rgeo
n i
rid
ovi
rus
√√
√
Wh
ite
stu
rgeo
n i
rid
ovi
rus
√X
√
Sho
rtn
ose
stu
rgeo
n v
iru
s√
√√
Caf
eter
ia r
oen
ber
gen
sis
viru
s (C
roV
)√
tbt
tbt
Meg
avir
us
chili
ensi
s(M
VC
hile
)√
tbt
tbt
Meg
avir
us
cou
rdo
(MV
Co
ur)
√tb
ttb
t
Aca
nth
oam
oeb
ap
oly
ph
aga
mim
ivir
us
(AP
MV
-2)
√tb
ttb
t
Org
anic
lake
phy
cod
nav
iru
s2
(O
LPV
-2)
√tb
ttb
t
Ch
ryso
chro
mu
lina
erci
na
viru
s (C
EV)
√tb
ttb
t
Het
ero
sigm
aak
ash
iwo
viru
s (H
AV
)√
tbt
tbt
Ost
eoco
ccu
sta
uri
viru
s (O
TV)
√tb
ttb
t
Ch
ile i
rid
esce
nt
viru
s (C
IV)
√tb
ttb
t0
.4
CIV
IIV
3FV3
ISK
NV
LDV
MV
DP
AV
ASF
V
PO
V
AC
MV
NV
WSI
VC
roV
MV
Ch
ileM
VC
ou
rA
PM
V-1
AP
MV
-2
OLP
V-2
PP
VC
EV
HA
V
AT
CV
PB
CV
MP
VO
TV
FSVES
V
Irid
ovi
rid
ae
Ph
yco
dn
avi
rid
ae
Un
clas
sifi
edA
sco
viri
da
e
Asf
arv
irid
ae
Mim
ivir
ida
e
Un
cla
ssif
ied
m
imiv
iru
s-lik
e
An
alyt
ical
sp
eci
fici
ty -
exp
and
ed
Dia
gn
ost
ic a
ssa
ys -
Mo
lecu
lar
test
dev
elo
pm
ent
stu
rgeo
n N
CLD
V
Appendix 2
19
Dia
gn
ost
ic a
ssa
ys –
qP
CR
ass
ay
per
form
an
ce a
sses
smen
t
1234 05
48
1216
2024
2832
360
40
1X
10
11
X1
04
1X
10
7
Co
pie
s
Ct (dRn)
Fluorescence (dRn)
12
38
R2
0.9
98
Slo
pe
-3.3
92
Effi
cien
cy 9
7.2
%
Cyc
le5X
10
7 5X
10
6 5X
10
5
5X
10
4 5X
10
3
5X
10
2 5X
10
15
X1
00Copies
Q1
wit
h p
NV
•Ef
fici
ency
(90
-11
0%:
dyn
amic
ran
ge; R
2>0
.99;
slo
pe
-3.3
)
•P
reci
sio
n (
at le
ast
3 r
eplic
ates
; SD
<0.1
67 t
o p
red
ict
2 fo
ld d
iluti
on
s 9
9.7%
of
tim
e)
•Se
nsi
tivi
ty (<
100
co
pie
s)
Cri
teri
a
QP
CR
ass
ays–
Fit
for
pu
rpo
se
Fish
h
eal
th
IN
Fish
h
eal
th
OU
T
Mo
lecu
lar
Dia
gno
stic
Te
sts
Bro
od
sto
ckJu
ven
iles
Lake
stu
rge
on
in
fect
iou
s d
isea
se
man
agem
ent
pla
n
Sust
ain
able
co
nse
rvat
ion
st
ock
ing
pro
gram
sele
ctio
n &
ce
rtif
ica
tio
n
hea
lth
sta
tus
pre
ven
tin
g
tra
nsm
issi
on
epid
emio
log
y
Hat
cher
yM
inim
izin
g i
nci
den
ce o
f d
isea
se
Re
du
ce/e
limin
ate
risk
of
intr
od
uci
ng
no
n-e
nd
emic
pat
ho
gen
s to
wild
sto
cks
of
lake
stu
rge
on
Ove
rvie
w o
f p
rese
nta
tio
n
Vir
us
dis
cove
ry
Cu
rren
t in
vest
igat
ion
s
Bir
thd
ay R
apid
s ca
se
In v
ivo
ch
alle
nge
stu
dy
Dia
gno
stic
tes
t d
evel
op
men
t Dis
eas
e m
anag
em
en
t st
rate
gie
s
Gre
ate
st r
isk
fact
or
ass
oci
ate
d w
ith
wild
life
pa
tho
gen
s =
tra
nsl
oca
tio
n•
Pri
nci
ple
ris
ks:
•R
isk
that
fis
h w
ill c
arry
pat
ho
gen
s in
to d
esti
nat
ion
env
iro
nm
ent
•R
isk
that
fis
h b
ein
g m
ove
d w
ill e
nco
un
ter
pat
ho
gen
s in
des
tin
atio
n e
nvir
on
men
t
•H
ealt
h r
isk
anal
ysis
pro
cess
:•
tran
slo
cati
on
pla
n•
hea
lth
haz
ard
iden
tifi
cati
on
an
d s
elec
tio
n f
or
asse
ssm
ent
•ri
sk a
sses
smen
t (i
.e.
pro
bab
ility
of
haz
ard
occ
urr
ing
& m
agn
itu
de
of
neg
ativ
e co
nse
qu
ence
)•
stat
emen
t o
f u
nce
rtai
nty
•as
soci
ated
haz
ard
s &
ris
k•
risk
red
uct
ion
Wild
life
dis
ease
ma
na
gem
ent
stra
teg
ies
•P
reve
nt
new
hea
lth
pro
ble
ms
fro
m a
risi
ng
•Ta
ke n
o a
ctio
n o
r re
spo
nse
•In
terv
ene
to c
on
tro
l th
e h
ealt
h is
sue
to s
om
e ex
ten
t**
•In
terv
ene
to e
rad
icat
e th
e p
ath
oge
n o
f co
nce
rn
Bef
ore
hea
lth
issu
es a
rise
Aft
er h
ealt
h is
sues
ha
ve e
mer
ged
Ris
k m
an
ag
emen
t
He
alth
ris
k as
sess
men
t•
Ris
k•
Mag
nit
ud
e o
f co
nse
qu
ence
Ris
k an
alys
is:
qu
alit
ativ
e vs
qu
anti
tati
ve
DIS
EASE
MA
NA
GEM
ENT
PLA
N
Appendix 2
20
Dis
ease
eco
log
y
Envi
ron
men
t
Ho
stP
ath
og
en
Sum
ma
ry
Kn
ow
led
ge
ga
ps
Kn
ow
led
ge/
too
ls g
ain
edP
ath
oge
n•
iden
tity
•ta
xon
om
y•
tro
pis
m•
tran
smis
sio
n•
dia
gno
stic
to
ols
•ge
no
me
seq
uen
ce in
form
atio
n
Pat
ho
gen
•p
ath
oge
nic
ity
•p
reva
len
ce in
wild
lake
stu
rgeo
n•
geo
grap
hic
ran
ge
The
end
Ack
no
wle
dge
men
ts:
Man
ito
ba
Hyd
roN
ort
h/S
ou
th C
on
sult
ants
Inc.
Elis
sa V
anW
alle
ghem
Pro
vin
ce o
f M
anit
ob
a: F
ish
erie
s B
ran
ch o
f M
anit
ob
a W
ater
Ste
war
dsh
ipU
niv
ersi
ty o
f M
anit
ob
a
Appendix 2
21
. . .
. . .
. . .
. . .
. . .
. . .
.
Gre
at L
akes
Fis
h H
ealth
Com
mitt
ee M
eetin
g
Man
itoba
: Fis
h H
ealth
Ove
rvie
w:
Jeff
Long
/ La
uree
n Ja
nusz
MB
Fis
herie
s B
ranc
hPh
: (20
4) 9
45-7
792
Ove
rvie
w
1.
Govern
ance -
MB
’s g
uid
ing d
ocum
enta
tion
2.
Why d
oes M
B test w
ild f
ish a
nd w
hat
sto
cks
do w
e t
est?
3.
Where
& h
ow
we c
olle
ct sam
ple
s f
or
testing
•C
halle
ng
es r
e: bio
-securi
ty
4.
What dis
ease t
estin
g a
re w
e d
oin
g?
5.
Results
6.
Curr
ent
Sta
tus
•Q
: are
we o
n the
rig
ht tr
ack?
Polic
y -G
uida
nce
•M
B F
isheri
es B
ranch is the
mana
gem
ent age
ncy for
the
Pro
vin
ce’s
fisheri
es r
esourc
es. W
e h
ave g
uid
ing p
olic
y for
ou
r B
ranch:
“To
en
su
re th
e o
rde
rly u
se
of o
ur
fish
erie
s re
so
urc
es.”
An
d a
mo
ng
oth
er
thin
gs, w
e a
re:
“To
pro
vid
e a
div
ers
ity o
f a
ng
ling
op
po
rtu
nitie
s”.
•T
he F
ish C
ulture
pro
gra
m h
as p
rovid
ed
sto
cked fis
hes s
ince 1
89
3.
Our
curr
ent guid
ing p
rincip
les for
the
fis
h c
ulture
pro
gra
m inclu
de
:1
.T
o e
nsu
re h
um
an
sa
fety
fro
m b
iolo
gic
al h
arm
;
2.
Co
nse
rve
th
e h
ea
lth
of a
qu
atic e
co
syste
ms
3.
To
pro
tect fish
sto
cks (so
urc
e a
nd
re
ce
ivin
g w
ate
rs)
fro
m v
ert
ica
l a
nd
ho
rizo
nta
l tra
nsm
issio
n o
f d
ise
ase
4.
To
pro
du
ce
hig
h q
ua
lity n
ative
an
d c
ultu
red
fis
he
s fo
r sto
ckin
g in
to M
Bw
ate
rbo
die
s.
Why
Doe
s M
B T
est W
ild F
ish?
?•
At th
is p
oin
t, t
here
is n
o p
rovin
cia
l (a
nd o
nly
a tra
nsitio
nal
federa
l) legis
late
d m
andate
for
fish h
ealth.
•N
otw
ithsta
ndin
g,
the M
B F
isheries B
ranch F
ish C
ulture
pro
gra
m r
elie
s o
n w
ild b
rood s
tock to p
roduce t
he
follo
win
g s
pecie
s f
or
sto
ckin
g p
urp
oses.
Perc
idae
[25-8
0M
fry
/ a
nnum
]•
Walle
ye
–S
au
ger
(new
/ r
en
ew
ed
sp
ecie
s f
or
cu
ltu
rin
g)
Salm
on
idae
[≈ 3
50,0
00 f
ingerlin
gs /
annum
]•
Lake T
rou
t–
Sp
lake
»(L
ake T
rou
t X
Bro
ok T
rou
t)
•B
rook T
rou
t (h
atc
hery
bro
od
)–
Tig
er
Tro
ut
»(B
row
n T
rou
t X
Bro
ok T
rou
t)
•B
row
n t
rou
t -
hatc
hery
bro
od
sto
ck
•R
ain
bow
tro
ut
–im
port
ed
eg
g
•A
dditio
nally
, as t
he a
gency r
esponsib
le f
or
fish m
anagem
ent, M
B h
as a
n inte
rest
in u
nders
tandin
g t
he
dis
ease p
rofile
of sto
cks t
hat
are
subje
ct
to r
ecovery
/ m
itig
ation s
tockin
g.
Acip
enserid
ae
•L
ake S
turg
eon
(re
new
ed
sp
ecie
s)
Appendix 3
24
WH
ERE
DO
WE
CO
LLEC
T O
UR
SA
MPL
ES?
1
2
MA
NIT
OB
A’S
HA
TC
HE
RIE
S
3
Ma
nito
ba
Fis
he
rie
s B
ran
ch
Fis
h C
ultu
re H
atc
he
rie
s
& s
pa
wn
ca
mp
loca
tio
ns
1.
Wh
ite
sh
ell
Ha
tch
ery
2.
Sw
an
Cre
ek S
pa
wn
Ca
mp
3.
Cle
arw
ate
r S
pa
wn
Ca
mp
•C
urr
ently
rais
es:
•T
rout
–R
ain
bow
•T
rip
loid
/ d
iplo
id
–B
row
n•
“Tig
er”
–B
rook
–Lake / s
pla
ke
•W
alle
ye
•H
as a
lso r
ais
ed L
ake s
turg
eon
Whi
tesh
ellH
atch
ery
•R
ais
es w
alle
ye f
or
Lake M
anitoba
•R
ais
es w
alle
ye f
or
“put
& take”
fisheries in
agro
-MB
•R
ais
es w
alle
ye f
or
inte
rlake
-com
merc
ial and
recre
ationally
fis
hed la
kes.
–S
easonal opera
tion
Swan
Cre
ek H
atch
ery
Appendix 3
25
CL
EA
RW
AT
ER
LA
KE
TR
OU
TS
PA
WN
CA
MP
Wal
leye
•L
ake
Ma
nito
ba
–S
wa
n C
ree
k S
pa
wn
Cam
p –
Lu
nd
ar
MB
•C
ap
acity 8
0M
eg
gs
•F
alc
on L
ake, M
B –
•C
ap
acity 1
0-2
0M
eg
gs
–C
om
bin
ed
site
with
Sa
lmo
nid
s
SWAN
CR
EEK
-W
ALLE
YE
Appendix 3
26
WH
ITES
HEL
L -W
ALLE
YE
Appendix 3
27
CLE
ARW
ATER
–LA
KE
TRO
UT
Cap
turin
g B
rood
Sto
ckPo
und
Net
sPo
und
Net
s
Appendix 3
28
Elec
trof
ishi
ngC
aptu
ring
Bro
od S
tock
Elec
tro-
fishi
ng
Col
lect
ing
Sam
ples
•O
va
ria
n F
luid
•ta
ke
n fro
m r
un
nin
g fe
ma
les A
ND
/O
R
•T
issu
e ta
ke
n fro
m g
ravid
fe
ma
les a
nd
ne
cro
psy d
on
eo
n s
ite
•T
issu
e
•T
ake
n fro
m r
un
nin
g (
usu
ally
) m
ale
s
•N
ecro
psy d
on
e o
n s
ite
.•
Inte
ntion is to take s
am
ple
s fro
m tho
se p
are
nts
tha
t pro
du
ce
egg
s tha
t go into
the
hatc
hery
.
•C
ond
itio
ns m
ake s
am
ple
colle
ction “
bio
-securi
ty”
challe
ng
ing
Appendix 3
29
WH
ITES
HEL
L -S
ALM
ON
IDS
WH
ITES
HEL
L -S
TUR
GEO
N
Appendix 3
30
Dis
ease
s of
Inte
rest
•“D
iseases o
f In
tere
st”
to M
B a
re b
ased o
n:
–D
FO
Ma
nu
al o
f C
om
plia
nce
AN
D
–h
as m
orp
he
d to
me
et C
FIA
Na
t’l A
qu
atic A
nim
al H
ea
lth
Pro
gra
m A
ND
–A
ims to
me
et fish
cu
ltu
re a
nd
sto
ck m
an
agem
ent n
ee
ds.
•S
o, curr
ently M
B is inte
reste
d in
:•
Pe
rcid
ae
–V
HS
•S
alm
on
ida
e–
IPN
–IH
N
–B
KD
–W
hirlin
g d
isease
–IS
A (
pote
ntially
–based o
n U
V h
atc
hery
tre
atm
ent
)
•A
cip
en
se
rid
ae
–“N
am
ao
virus”
•V
ario
us fa
mili
es –
La
ke
Ma
nito
ba
–K
oiherp
es V
irus
Res
ults
•V
HS
•N
o te
stin
g d
on
e to
da
te b
y M
B –
testin
g p
en
din
g fo
r P
erc
ida
esto
cks
•N
o k
no
wn
occu
rre
nce
to
da
te in
MB
•IP
N•
Ou
tbre
ak in
Whitesh
ell
in 1
97
0-8
0s (b
ut n
on
e s
ince
)
•2
00
6 &
20
12
in G
ran
d R
ap
ids –
co
nce
rn fo
r fu
ture
sa
lmo
nid
rea
rin
g
•C
lea
rwa
ter
–re
ma
ins n
eg
ative
fo
r IP
N b
ase
d o
n M
B t
estin
g / C
FIA
ma
ps M
B a
s
IPN
po
sitiv
e.
•B
KD
•S
ee
ms to
ha
ve
ou
tbre
aks in
bro
ok tro
ut –
esp
ecia
lly d
urin
g c
row
din
g o
r o
the
r str
esso
rs a
cro
ss h
atc
he
rie
s a
nd
ove
r tim
e.
•W
hirlin
g d
ise
ase
•N
o k
no
wn
occu
rre
nce
s in
MB
•K
Hv
•O
utb
rea
k in
La
ke
MB
in
20
09
–q
uie
t e
ve
r sin
ce
(e
xce
pt 1
ca
se
in L
kW
pg).
•N
am
ao
Vir
us
•E
nd
em
ic to
DU
3 o
r M
B o
r to
LK
St p
op
ula
tio
ns in
ge
ne
ral?
Sta
tus u
nce
rta
in.
Cur
rent
Sta
tus
•C
on
tinu
e w
ork
ing
with
DF
O o
n a
wild
fis
hsu
rve
illan
ce
pro
gra
m / p
roto
co
l (d
esp
ite
no
cle
ar
leg
isla
tive
ma
nd
ate
to
do
so
).
•H
ave
se
t u
p d
ise
ase
te
sting
co
ntr
act w
/ R
PC
for
ha
tch
ery
te
sting
–co
ntinu
e to
fo
llow
ou
rd
isin
fection
, te
sting
an
d s
tockin
g B
MP
s.
•U
se
ha
tch
ery
syste
m a
s b
asis
fo
r te
sting
ba
se
d o
n th
e “
ha
tch
ery
he
alth
” d
rive
r.
•A
im to
me
et o
r b
ea
t C
FIA
co
mp
art
me
nta
lisa
tion
pro
gra
m r
e:
IPN
.
Thou
ghts
/ Com
men
ts/ Q
uest
ions
•S
o, giv
en the a
mbig
uous m
andate
under
whic
h w
e o
pera
te a
nd
the c
halle
nges o
f
sa
mp
le c
olle
ctio
n:
•Is
MB
on
th
e “
rig
ht
tra
ck”?
?
–Is
the in
form
atio
n w
e c
olle
ct usefu
l to
oth
er
jurisdic
tio
ns?
–A
re w
e c
olle
ctin
g m
isle
adin
g in
form
atio
n?
Appendix 3
31
Va
n H
orn
esvil
le S
FH
Su
mm
er
‘13
An
dre
w D
. N
oye
s
Ro
me
Fie
ld S
tati
on
New
York
Sta
te D
epart
ment
of
Environm
enta
l Conse
rvation
12
D
EC
H
atch
erie
s
9 C
old
Wate
r
12 D
EC
H
atch
erie
s
3 C
oo
l W
ate
r
12 D
EC
H
atch
erie
s
My L
ab
Appendix 4
32
12
D
EC
H
atch
erie
s1
2 D
EC
H
atch
erie
s
Van
Ho
rn
esvil
le
-Rais
e 4
00K R
T400 g
pm
Rain
bo
w T
ro
ut
fro
m
Van
Ho
rn
esville
Ju
ly,
20
13
Appendix 4
33
Rain
bow
Tro
ut
from
Van H
orn
esville
Gra
nula
r Epithelial
Hyperp
lasia
Caudal
Hyperp
igm
enta
tion
Wet
mo
un
t-S
kin
Gram
Sta
in
Wh
at
we
did
….
Check
ed for
WD
-N
OP
E
Virolo
gy
-N
OP
E
Bact
—Skin
/musc
le, G
ills,
Liv
er
O
n B
HI
and H
-S -
YE
S…
.
Gro
ws o
n H
-S
Als
o o
n B
HI
-w
hit
e c
olo
nie
s
Appendix 4
34
H-S—
lon
g r
od
sB
HI—
sh
ort
ro
ds
Gro
ws @
25
C???
Fla
vo
Tria
d
F. p
sychro
philum
F. b
ranchio
philum
F. c
olu
mnare
Fla
vo
Tria
d
F. p
sychro
philum
F. b
ranchio
philum
F. c
olu
mnare
Appendix 4
35
Sent
to K
enne
bec
Riv
er
Bio
sc
ien
ce
s…
.
For
Sequenci
ng, PCR, etc
…
Fla
vo
ba
cte
riu
mor
Ch
ryse
ob
acte
riu
m◦U
nknow
n s
peci
es
Fla
vodiv
ers
ity a
lso in M
ichig
an
◦(L
och
et
al.-
JAAH
-2013)
RT
o
rig
in
ate
@ R
and
olp
h
All R
T e
gg
s m
ove
d
to
C
H o
r V
H
VH
tra
nsfe
rs in
sp
rin
g/su
mm
er
Appendix 4
36
Dise
ase
a
pp
ea
rs
eve
ryw
he
re
b
ut R
A
…th
en
at R
A
Sta
rted a
t VH
◦m
oved fro
m t
here
O
nly
infe
cte
d R
Ts
◦D
id e
ventu
ally s
ee
concurr
ent
F. psych
G
row
s u
p t
o 2
5C
…
on H
-S a
nd B
HI
Is
ola
ted f
rom
skin
,m
uscle
*and l
iver
N
ot
seen
in
20
14
Appendix 4
37
Old
Cas
e fr
om 2
010
•H
amilt
on
Har
bo
ur,
ON
•7
Bro
wn
bu
ll h
ead
wit
h le
sio
ns-
Late
Sep
20
10
–d
eep
ulc
ers
on
: Hea
d, f
ace,
bas
e o
f fi
ns,
fin
s an
d b
elly
–u
lcer
s w
ere
dee
ply
ero
ded
wit
h r
ed c
entr
e an
d w
hit
eri
m
–So
me
of
the
ulc
ers
on
bo
dy
exp
ose
d t
he
mu
scu
latu
rean
d o
ther
s h
ad g
ran
ula
ted
tex
ture
–In
so
me
fish
ulc
ers
crea
ted
ho
le t
hro
ugh
th
e ta
il fi
n o
rin
so
me
inst
ance
s fi
ns
wer
e co
mp
lete
ly e
rod
ed??
?
•2
nd
sub
mis
sio
n (
Earl
y N
ove
mb
er)
•1
6 b
row
n b
ull
hea
ds
–Th
ree
fish
had
Sim
ilar
lesi
on
s b
ut
in r
est
of
the
fish
lesi
on
s w
ere
less
sev
ere
(Po
ssib
ly d
ue
to lo
wer
wat
er t
emp
erat
ure
)
???
Appendix 5
38
Susp
ect f
or
Ed
wa
rds
iell
ata
rda
??
?
Res
ults
•V
iro
logy
–EP
C a
nd
CH
SE a
t 1
5oC
fo
r 2
1 d
ays
•N
o C
PE
•B
acte
rio
logy
: K
idn
ey a
nd
Les
ion
s
–TS
A, B
HIA
an
d M
acC
on
key
at 2
0 a
nd
25
oC
•P
seu
do
mo
na
saa
eru
gin
osa
,
•A
ero
mo
na
sh
ydro
ph
ila,
•V
ibri
o f
luvi
alis
an
d
•P
lesi
om
on
as
shig
ello
ides
.
Res
ults
(Con
td.)
•H
isto
logy
/ M
ole
cula
r te
stin
g*–
Lesi
on
s
–Sp
leen
,
–K
idn
ey
–G
ills
–St
om
ach
* N
o s
amp
les
for
mo
lecu
lar
test
ing
wer
e co
llect
ed
fro
m 1
stsu
bm
issi
on
Sum
mar
y of
resu
lts
Sam
ple
ID
Lesi
on
co
nsi
sten
t w
ith
A. i
nvad
ans
qP
CR
(D
FO t
est)
A. i
nvad
ans
OIE
PC
R 1
, 2 a
nd
3
His
tolo
gyFu
nga
l hyp
hae
co
nsi
sten
t w
ith
A
. inv
adan
s
2010
-36
-00
1Ye
s (S
ever
e)P
osi
tive
Po
siti
veP
osi
tive
2010
-36
-00
2Ye
s (S
ever
e)P
osi
tive
Po
siti
veP
osi
tive
2010
-36
-00
3*L
esio
n a
t th
e b
ase
of
fin
an
d m
idd
le o
f ta
il fi
n w
as c
hew
ed u
p
Po
siti
ve
No
t te
sted
Po
siti
ve
2010
-36
-00
4Ye
s (S
ever
e)N
egat
ive
No
t te
sted
Po
siti
ve
2010
-36-
005
Yes
(Sev
ere)
Po
siti
veP
osi
tive
Po
siti
ve
2010
-36
-00
6N
o le
sio
ns
Neg
ativ
eN
ot
test
edN
egat
ive
2010
-36
-00
7Sm
all h
aem
orr
hag
e ar
ou
nd
mo
uth
–n
ot
sure
No
sam
ple
No
sam
ple
Po
siti
ve
2010
-36
-00
8N
o le
sio
ns
Neg
ativ
eN
ot
test
edN
egat
ive
2010
-36
-00
9Sm
all l
esio
n o
n b
elly
an
d o
per
culu
m –
no
t su
re
Neg
ativ
eN
ot
test
edN
ot
test
ed
2010
-36
-01
0*L
esio
n a
t th
e b
ase
of
pec
tora
l fin
N
egat
ive
No
t te
sted
No
t te
sted
2010
-36
-01
1N
o l
esio
ns
Neg
ativ
eN
ot
test
edN
ot
test
ed
2010
-36
-01
2Tw
o s
mal
l les
ion
s o
n b
elly
–n
ot
sure
Neg
ativ
eN
ot
test
edN
ot
test
ed
2010
-36
-01
6*L
esio
n o
n f
ins
Neg
ativ
eN
ot
test
edN
ot
test
ed
Appendix 5
39
Sum
mar
y of
Res
ults
Con
td.
Sam
ple
ID
Tiss
ue
A.
inva
dan
sq
PC
RC
t va
lues
Po
siti
ve f
or
A. i
nva
dan
s
201
0-36
-00
1L-
PC
RLe
sio
n21
.07,
20.
75P
os
201
0-36
-00
1K
-PC
RK
idn
ey29
.72,
29.
54P
os
201
0-36
-00
1S-
PC
RSp
lee
n32
.58,
32.
48P
os
2010
-36-
001L
r-P
CR
Live
r33
.11,
32.
6P
os
201
0-36
-00
1G
-PC
RG
ill28
.69,
28.
85P
os
201
0-36
-00
2L-
PC
RLe
sio
n19
.47,
19.
35P
os
201
0-36
-00
2K
-PC
RK
idn
ey31
.57,
31.
14P
os
201
0-36
-00
2S-
PC
RSp
lee
n33
.2,
32.5
7P
os
201
0-36
-00
2Lr
-PC
RLi
ver
33.3
9, 3
3.67
Po
s
201
0-36
-00
2G
-PC
RG
ill31
.23,
31.
13P
os
201
0-36
-00
3L-
PC
RLe
sio
nN
o C
t, N
o C
tN
eg
201
0-36
-00
3K
-PC
RK
idn
eyN
o C
t, N
o C
tN
eg
201
0-36
-00
3S-
PC
RSp
lee
n35
.93,
36.
79P
os
201
0-36
-00
3Lr
-PC
RLi
ver
No
Ct,
No
Ct
Neg
201
0-36
-00
3G
-PC
RG
ill35
.13,
34.
37P
os
201
0-36
-00
4L-
PC
RLe
sio
nN
o C
t, N
o C
tN
eg
2010
-36-
004K
-PC
RK
idn
eyN
o C
t, N
o C
tN
eg
201
0-36
-00
4S-
PC
RSp
lee
nN
o C
t, N
o C
tN
eg
201
0-36
-00
4Lr
-PC
RLi
ver
No
Ct,
No
Ct
Neg
201
0-36
-00
4G
-PC
RG
ill38
.17,
No
Ct
Neg
201
0-36
-00
5L-
PC
RLe
sio
n22
.82,
22.
87P
os
201
0-36
-00
5K
-PC
RK
idn
ey34
.97,
34.
92P
os
201
0-36
-00
5S-
PC
RSp
lee
nN
o C
t, N
o C
tN
eg
201
0-36
-00
5Lr
-PC
RLi
ver
39.5
6, N
o C
tP
os
201
0-36
-00
5G
-PC
RG
illN
o C
t, N
o C
tN
eg
2014
sam
ples
(Man
itoba
)
Res
ults
201
4
Bac
teri
olo
gy
Poss
ibili
ty o
f V
ibri
o f
luvi
alis
His
tolo
gy a
nd
mo
lecu
lar
Bio
logy
Res
ult
s ar
e st
ill p
end
ing
???
Appendix 5
40
FISHERY RESEARCH PRIORITIES:
GREAT LAKES FISH HEALTH COMMITTEE Great Lakes Fishery Commission
Updated August 2014
This listing was compiled based on input from discussions within the Council of Lake
Committees (for more information go to http://www.glfc.org/lakecom.php) and the Great
Lakes Fish Health Committee http://www.glfc.org/boardcomm/fhealth/fhealth.php). Order
of listing does not imply relative ranking of priorities for the Fishery Research Program
funding.
Research Priorities • What is the ecology of fish pathogens and diseases of concern in the Great Lakes
Basin? Examples include (but are not limited to) viral hemorrhagic septicemia virus
(VHSv) genotype IVb, Heterosporis sp., Epizootic Epitheliotropic Disease virus
(EEDv), Flavobacterium sp., and emerging diseases.
• What non-lethal field sampling methods and tissue/fluid samples are equivalent to
conventional lethal field sampling methods to determine fish pathogen and/or
disease status?
• Develop and validate new methods to detect emerging fish pathogens or pathogens
of concern in the Great Lakes Basin.
Additional Research Interests 1. What is the effectiveness of the GLFHC disinfection protocols in eliminating key
pathogens of interest from fish eggs? There is a need for a reliable disinfection
methodology to prevent pathogen transmission via eggs and sperm.
2. Disease Ecology and Epidemiology
(a) What is the susceptibility of Great Lakes fish species to emerging fish
pathogens in the Great Lakes?
(b) Identification of reservoirs and vectors (including ballast water) for fish
pathogens in the Great Lakes Basin
(c) What mechanisms affect the virulence and persistence of fish pathogens?
(d) What is the effect of population size on disease expression?
(e) What are the effects of multiple pathogens or combination of pathogens and
nutritional deficiency and/or contaminant exposure on disease expression?
(f) What are the projected changes on fish pathogen prevalence and intensity as
a result of climate change?
3. Nutritional Aspects of Fish Health in the Great Lakes.
(a) What is the role of lipids or other nutrients in determining and predicting
health status?
(b) What is the role of thiaminase-producing organisms in Great Lakes
ecosystems?
(c) What affect do invasive species have on nutrient stores in the Great Lakes
and what are the associated effects on fish health?
(d) What is the effect of nutrition on reproductive success?
Appendix 6
41
(e) Does protein substitution in hatchery feeding formulations or extrusion
manufacturing methods have a negative impact on survivorship, migratory
behavior and reproductive success of hatchery-reared salmonids?
4. Fish Pathogen and Disease Management.
(a) What are the effects of fish stocking and other management decisions on the
manifestation of fish disease in the Great Lakes Basin?
(b) What effects does culling brood stock for pathogen control have on the
genetics of production fish?
(c) When should fish not be moved past barriers (from a disease perspective)?
(d) Development of an emergency response plan for disease outbreaks in the
Great Lakes Basin, including (but not limited to) training of field personnel
and preplanning.
(e) What is the effectiveness of immunostimulants against key pathogens of
interest in hatcheries?
(f) What is the effect of vaccination of hatchery fish on pathogen virulence?
Appendix 6
42
PROPOSED TECHNICAL ADVISORS
Bacteriology: Diane Elliot (USGS), Thomas Loch (MSU)
Virology: James Winton (USGS), Tom Waltzek
Molecular: Nick Phelps (University of Minnesota), Sharon Clouthier (DFO)
Nutrition: Dominique Bureau (University of Guelph), Dale Honeyfield
Quantitative Fish Health Data Analysis: Dominic Travis (University of Minnesota) Travis Brenden (Michigan State University)
Epidemiology: Lori Gustafson (USDA)
Parasitology: David J. Marcogliese (St. Lawrence Centre, Environment Canada)
Appendix 7
43
Th
iam
ine
Inje
ctio
n
of
Ska
man
ia
Ste
elh
ead
B
roo
dst
ock
Bo
din
e S
tate
Fis
h H
atc
he
ry
Ind
ian
a D
ep
art
me
nt o
f N
atu
ral R
eso
urc
es
Tw
o Q
ues
tio
ns
C
an t
hia
min
e in
jecte
d a
t harv
est
from
the w
ild im
pro
ve t
he s
urv
ival
of
adult S
kam
ania
ste
elh
ead
thro
ugh s
paw
nin
g f
our
to e
ight
month
s la
ter?
H
ow
does t
reatin
g b
roodsto
ck w
ith
thia
min
e c
om
pare
to t
reatin
g s
ac-
fry w
ith r
egard
s t
o s
wim
-up s
urv
ival
thro
ugh t
he f
irst 21 d
ays o
n f
eed?
Inje
ctio
n P
roto
col –
2005
/’06
H
alf o
f all
fem
ale
bro
odsto
ck
receiv
ed t
hia
min
e in
jectio
n
N
um
bere
d jaw
tags w
ere
used t
okeep t
rack o
f w
hic
h f
ish w
ere
inje
cte
d
A
t firs
t egg t
ake, a 2
X 2
matr
ix w
illbe u
sed t
o e
sta
blis
h f
our
gro
ups o
ffe
male
s
A
ll m
ale
s w
ere
thia
min
e in
jecte
d
D
ose w
as 9
.2 m
g T
hia
-HC
l/5 lb
s
Fem
ale
Stu
dy
Gro
up
s
U
n-in
jecte
d f
em
ale
s
S
ac f
ry n
ot
treate
d
S
ac f
ry tre
ate
d
In
jecte
d F
em
ale
s
S
ac f
ry n
ot
treate
d
S
ac f
ry tre
ate
d
Appendix 8
44
Tri
al P
erio
d
21-D
ay m
ort
alit
y p
ost
firs
t fe
edin
g
Imp
licat
ion
s
B
roodsto
ck c
arr
yin
g c
apacity is
limited
T
hia
min
e h
ydro
chlo
rid
e is
expensiv
e
B
est to
get
thia
min
e into
egg p
rior
to f
ert
ilizatio
n (
Dale
Honeyfie
ld)
Appendix 8
45
Tag
nu
mbe
rof ad
ult s
teelh
ead
bro
odsto
ck, t
ype
of
trea
tmen
t, a
nd
corr
espo
ndin
g e
gg th
iam
ine
level (
nm
ol/g
of eg
g w
eig
ht)
pri
or
to s
pa
wnin
g.
Tag
#In
jecte
dT
hia
min
e L
evel
Tag
#In
jecte
dT
hia
min
e L
evel
34
2Y
es
9.1
40
5N
o6
.3
56
Yes
8.5
42
1N
o1
0.5
10
2Y
es
14
.41
3N
o8
.6
22
6Y
es
9.4
27
3N
o9
.3
60
4Y
es
15
.92
03
No
7.7
86
Yes
9.8
68
1N
o5
.6
67
8Y
es
14
.76
83
No
3.1
19
0Y
es
10
.42
31
No
9.8
21
8Y
es
20
.75
45
No
11.2
55
6Y
es
24
.13
99
No
12
.1
332
Ye
s14.4
541
No
9.9
98
Yes
20
.06
9N
o8
.8
65
8Y
es
11.3
12
6N
o5
.2
47
2Y
es
16
.56
43
No
5.3
36
0Y
es
7.9
34
7N
o5
.2
15
4Y
es
16
.77
19
No
6.4
52
Yes
15
.46
13
No
7.5
78
Ye
s28.0
601
No
5.2
12
7Y
es
16
.42
07
No
5.0
115
Yes
24
.75
5N
o1
3.1
67
0Y
es
25
.94
55
No
7.0
39
4Y
es
20
.9118
No
7.9
47
6Y
es
17
.64
71
No
12
.3
--
-5
49
No
5.7
Mea
n16
.27.
9
Appendix 8
46
R²
= 0
.0694
0
10
20
30
40
50
60
70
80
90
10
0
% Swimup Mortality
Thia
min
e
% S
wim
up M
orta
lity
vs. T
hiam
ine
Lo
t 07
RB
ST
T
In
jecte
d F
em
ale
s
9
7.1
% 2
1 D
ay S
urv
iva
l
U
n-in
jecte
d F
em
ale
s
92.2
% 2
1 D
ay S
urv
ival
Lo
t Y
ears
201
1 to
201
4
B
rood Y
ears
2010 t
o P
resent
A
ll In
jecte
d a
t 18.4
mg /
10 lbs B
W
2011
97.8
% w
ith F
low
-Thru
2012
97.8
% w
ith F
low
-Thru
97.3
% N
o F
/T
2013
97.3
% N
o F
/T
2014
95.7
% N
o F
/T
Appendix 8
47
Eg
g T
hia
min
e A
nal
ysis
B
rood L
ot
12IN
ST
T-B
R
P
rodu
ce
d 1
3R
BS
TT
Lo
t
42 F
em
ale
s
18.8
nm
ol/gm
egg w
t
8
.9 –
29
.8
S
D 5
.0
Bro
od
sto
ck S
urv
ival
B
rood L
ots
2000 –
2013
N
o In
jectio
ns
8
Ye
ars
2
0.9
% M
ort
alit
y
•3
.2 –
35.1
% R
ang
e
In
jections
6
Ye
ars
7
.9%
Mo
rta
lity
•1.9
–2
1.1
% R
an
ge
Appendix 8
48
