veterinary services orbivirus activities at nvsl · 2019. 4. 18. · tx* 3 wtd wa* 20 wtd, mule...
TRANSCRIPT
Orbivirus activities at NVSL
Eileen N. Ostlund Associate Director
National Veterinary Services Laboratories
U.S. Department of Agriculture Animal and Plant Health Inspection Service
Veterinary Services Science, Technology and Analysis Service
October, 2015
Veterinary Services
Topics
• 2014 Calendar Year and 2015 Year-to-
date Bluetongue and Epizootic
Hemorrhagic Disease Virus observations
at NVSL
• African Horse Sickness Virus PCR
International Ring Trial
2
3
BTV Identifications 2014 (NVSL, except where noted)
BTV not typed
State No. Species
ID 2 alpaca, deer*
MO 1 cattle
WA* 7 cattle, sheep, deer
BTV-11
State No. Species
CO 1 goat
BTV-17
State No. Species
NE 2 WTD
NJ 3* WTD
*2 isolates from SCWDS BTV-10
State No. Species
NE 1 bighorn sheep
ID (WI)† 1 cattle
†WI cattle in quarantine recently
shipped from ID
BTV-18
State No. Species
FL 1* WTD
*Isolate from SCWDS, first report of
BTV-18 in the U.S.
*testing done at WADDL
4
BTV Identifications 2015 (NVSL, except where noted)
BTV not typed
State No. Species
ID* 15 deer, sheep,
cattle, bighorn
NV* 1 cattle
OR* 1 sheep
TX* 3 WTD
WA* 20 WTD, mule deer,
elk, bighorn
BTV-13
State No. Species
OK 1 sheep
NV* 1 cattle
BTV-10
State No. Species
AZ 1 bighorn sheep
CA 5 sheep
FL 1 WTD *BTV identified by WADDL, typing NVSL
BTV activity has also been reported
in TX, but information regarding
numbers and types is not available.
*BTV identified by WADDL, typing NVSL
BTV-17
State No. Species
CA 3 sheep/ *mule
deer
ID* 8 cattle, WTD,
sheep, yak
NV* 3 bighorn sheep
WA* 9 WTD, mule deer
*BTV identified by WADDL
5
BTV Identifications 2015 (NVSL, except where noted)
BTV-3
State No. Species
TX 1* Cattle
TX 1* White-tailed deer
BTV-19
State No. Species
FL 1 White-tailed Deer
*BTV identified at TVMDL, typing NVSL
First report of BTV-3 in TX
BTV-24 previously identified in FL in
2007
BTV-22
State No. Species
FL 1* White-tailed Deer
*BTV identified at TVMDL, typing
NVSL; BTV-22 previously identified
in FL in 2002 & 2005 BTV-6
State No. Species
FL 1* White-tailed Deer
BTV-19 previously identified in FL in
2003
*Also positive EHDV-6; BTV-6
previously identified in FL in 2006
BTV-24
State No. Species
FL 1 White-tailed Deer
NVSL Bluetongue Virus Identification
(endemic serotypes) 1992-2015
FLORIDA
MISSISSIPPI
WEST
VIRGINIA
RHODE ISLAND
WASHINGTON
OREGON
IDAHO
MONTANA
WYOMING
NORTH
DAKOTA
SOUTH
DAKOTA
NEBRASKA IOWA
MINNESOTA
WISCONSIN
ILLINOIS
INDIANA OHIO
MISSOURI KANSAS
COLORADO
UTAH
NEVADA
CALIFORNIA
ARIZONA NEW MEXICO
OKLAHOMA ARKANSAS
KENTUCKY
VIRGINIA
TEXAS
MASSACHUSETTS
GEORGIA ALABAMA
SOUTH
CAROLINA
NORTH CAROLINA TENNESSEE
PENNSYLVANIA
MARYLAND
NEW JERSEY
NEW YORK
CONNECTICUT
VERMONT
NEW HAMPSHIRE
MAINE
DELAWARE
LOUISIANA
MICHIGAN
2*
10
11
13
17
2, 10
11
10,11, 17
13
17
13
17
11, 17
11,17
10
13, 17
11, 17
10, 11,
13, 17
11, 13, 17 13, 17
11, 17
11, 13, 17
10, 11,
17
BTV isolate / PCR positive
17 BTV serotype identified
11, 17
11, 13, 17
17
11, 13,
17
11
13, 17 11
13,
17
*BTV-2 isolated and identified by UC Davis, CA
17 11
17
10†
10†
†BTV-10 WI, cattle originated in ID
13, 17
‘Exotic’ BT serotypes in U.S. (1999-2015)
3
3
3
3
3
3 3
3 3
3
22
22
14
14
19
5 1
1 6
24
12
12
9
9
BTV-1: 2004, 2010, 2013
BTV-3: 1999, 2001, 2002, 2003, 2006,
2008, 2009, 2012, 2013, 2015
BTV-5: 2003
BTV-6: 2006, 2015
BTV-9: 2008, 2012
BTV-12: 2008, 2010, 2012
BTV-14: 2003, 2009
BTV-18: 2014
BTV-19: 2003, 2015
BTV-22: 2002, 2005, 2015
BTV-24: 2007, 2015
12
12
12
12
1
3
3 3
18
19
3 22
6
24
BTV-1: 10, 13
BTV-2: 99, 03, 05, 06, 13
BTV-3: 99, 01, 02, 03, 13
BTV-5: 03
BTV-6: 06, 15
BTV-9: 08, 12
BTV-10: 02, 03, 15
BTV-11: 13
BTV-12: 10, 12
BTV-13: 99
BTV-14: 03
BTV-17: 99
BTV-18: 14
BTV-19: 03, 15
BTV-22: 02, 05, 15
BTV-24: 07, 15
1 2
3
3
5 6
9
9
11 12
12
14
19
22
22
24
Florida 1999-2015:
BT serotype identifications
(NVSL)
2
2
3
17
13 3
3 10 3
3 10
2
2
1 18
19
22
10
6
24
9
EHD Virus Identifications, 2014
EHD 2
State No. Species
FL 1 deer
TX 1 Eld’s deer
EHD 6
State No. Species
FL 1 WTD
NC 2 WTD
EHD not typed
State No. Species
ID 3* WTD
NE 1† bison
WA 3* WTD
*EHD identified by WADDL
†Suspect
• Sept 23, 2015 ProMed
reports EHD outbreak in
Illinois, with 47 suspected
cases in white-tailed deer.
Type not reported.
• Increased EHD 2 activity in
IA and OK in 2015; has not
been observed to date in NE
or SD.
10
EHD 2
State No. Species
IA 7 WTD, cattle
IL 1 WTD
KS 1 WTD
OK 3 elk, WTD
EHD not typed
State No. Species
KY 1* cattle
*EHD identified by WADDL
EHD Virus Identifications, 2015
EHD 6
State No. Species
FL 1 WTD
NVSL Epizootic hemorrhagic disease virus (EHDV)
identification 1992-2015
FLORIDA
MISSISSIPPI
WEST
VIRGINIA
RHODE ISLAND
WASHINGTON
OREGON
IDAHO
MONTANA
WYOMING
NORTH
DAKOTA
SOUTH
DAKOTA
NEBRASKA IOWA
MINNESOTA
WISCONSIN
ILLINOIS
INDIANA OHIO
MISSOURI KANSAS
COLORADO
UTAH
NEVADA
CALIFORNIA
ARIZONA NEW MEXICO
OKLAHOMA ARKANSAS
KENTUCKY
VIRGINIA
TEXAS
MASSACHUSETTS
GEORGIA ALABAMA
SOUTH
CAROLINA
NORTH CAROLINA TENNESSEE
PENNSYLVANIA
MARYLAND
NEW JERSEY
NEW YORK
CONNECTICUT
VERMONT
NEW HAMPSHIRE
MAINE
DELAWARE
LOUISIANA
MICHIGAN
WASHINGTON, DC
EHDV 2
EHDV 1 and 2
EHDV 6
EHDV type unknown
NVSL EHD/BT Identifications (by county), 2013
(U.S. cases)
2 6
FLORIDA
MISSISSIPPI
WEST
VIRGINIA
RHODE ISLAND
WASHINGTON
OREGON
IDAHO
MONTANA
WYOMING
NORTH
DAKOTA
SOUTH
DAKOTA
NEBRASKA IOWA
MINNESOTA
WISCONSIN
ILLINOIS
INDIANA OHIO
MISSOURI KANSAS
COLORADO
UTAH
NEVADA
CALIFORNIA
ARIZONA NEW MEXICO
OKLAHOMA ARKANSAS
KENTUCKY
VIRGINIA
TEXAS
MASSACHUSETTS
GEORGIA ALABAMA
SOUTH
CAROLINA
NORTH CAROLINA TENNESSEE
PENNSYLVANIA
MARYLAND
NEW JERSEY
NEW YORK
CONNECTICUT
VERMONT
NEW HAMPSHIRE
MAINE
DELAWARE
LOUISIANA
MICHIGAN
BT
EHD
EHD
2
10
BT
2
2
2
2
17
BT
2
17
2
2
17 2 2
2
2 2
17
17 17
2 2 2
2
2
2
1
11
11
13
13
2 17
11
2
2 2
2
2 17
13
2
11
1
6
2
2
2
2
2 2
2
2
17
13
BT
2
11
2
11
2
1
13
1 2
2 2
2 2
17 13
2
11
13
11
EHD
2
3
11
3
BT
2
EHD/BT Identifications (by county), 2014 (U.S. cases, identified by, or reported to the NVSL)
17
6
FLORIDA
MISSISSIPPI
WEST
VIRGINIA
RHODE ISLAND
WASHINGTON
OREGON
IDAHO
MONTANA
WYOMING
NORTH
DAKOTA
SOUTH
DAKOTA
NEBRASKA IOWA
MINNESOTA
WISCONSIN
ILLINOIS
INDIANA OHIO
MISSOURI KANSAS
COLORADO
UTAH
NEVADA
CALIFORNIA
ARIZONA NEW MEXICO
OKLAHOMA ARKANSAS
KENTUCKY
VIRGINIA
TEXAS
MASSACHUSETTS
GEORGIA ALABAMA
SOUTH
CAROLINA
NORTH CAROLINA TENNESSEE
PENNSYLVANIA
MARYLAND
NEW JERSEY
NEW YORK
CONNECTICUT
VERMONT
NEW HAMPSHIRE
MAINE
DELAWARE
LOUISIANA
MICHIGAN
BT
EHD
BT
BT
11
17
6
2 6
2
17
10
EHD
10
18
BT
BT
BT
EHD
EHD
EHD/BT Identifications (by county), 2015 (U.S. cases, identified by, or reported to the NVSL)
6
FLORIDA
MISSISSIPPI
WEST
VIRGINIA
RHODE ISLAND
WASHINGTON
OREGON
IDAHO
MONTANA
WYOMING
NORTH
DAKOTA
SOUTH
DAKOTA
NEBRASKA IOWA
MINNESOTA
WISCONSIN
ILLINOIS
INDIANA OHIO
MISSOURI KANSAS
COLORADO
UTAH
NEVADA
CALIFORNIA
ARIZONA NEW MEXICO
OKLAHOMA ARKANSAS
KENTUCKY
VIRGINIA
TEXAS
MASSACHUSETTS
GEORGIA ALABAMA
SOUTH
CAROLINA
NORTH CAROLINA TENNESSEE
PENNSYLVANIA
MARYLAND
NEW JERSEY
NEW YORK
CONNECTICUT
VERMONT
NEW HAMPSHIRE
MAINE
DELAWARE
LOUISIANA
MICHIGAN
BT
EHD
2 2
2
2
BT
10
2
2
19
3
2 17
17
17
17
17
17
17
17
17
17
BT
BT 17
13
10 22
2
2
2
BT
BT
EHD
10
10
6
24
6
13
10
17
African Horse Sickness
International Ring Trial
AHSV detection by PCR
15
African Horse Sickness
• African Horse Sickness (AHS) is an acute or subacute Culicoides-borne, viral disease of horses and other equids that is often fatal. Clinical signs and lesions are compatible with impaired respiratory and circulatory function.
• There are 9 serotypes of AHSV
• AHSV is not present in the Americas
• If introduced, could be spread by native Culicoides spp.
• Current import requirements for equids from affected areas = 60 day quarantine
– Impetus for decreasing quarantine with AHSV PCR tests
Host susceptibility • All solipeds are susceptible to infection.
Susceptibility to disease varies by species. Horses > Mules > Donkeys > Zebras
75-90% 50% 5-10% no illness
• Dogs can become infected and ill by eating infected horse meat.
Clinical Disease
• The incubation period is usually 5-14 days
• AHSV replicates in endothelium, especially heart and lung
• Fever up to 41C (106F), congestion of conjunctivae
• Pulmonary, cardiac, mixed forms of disease
• Differentia Diagnosis includes EIA, EVA, Piroplasmosis, etc.
2015 AHS PCR Ring Trial
• OIE AHS ad hoc group requested opinion on adoption of an RT-PCR as a prescribed test for international trade
• Ring Trial administered by AHS OIE Reference Laboratories in the UK and Spain
• Variety of methods being used by AHS testing labs – compare results on provided sample set.
• Participants from 4 OIE AHS reference laboratories: South Africa, UK, Spain (2), plus EU reference center for equine infectious diseases (other than AHS), Deltammune in South Africa and NVSL = 7 laboratories
19
2015 AHS PCR Ring Trial • Panel 1 (not tested by NVSL)
• Panel 2 = 45 samples (40 positive)
– Inactivated AHSV-spiked horse blood samples
– Panel encompassed all AHSV serotypes
• Reference strains and recent outbreak isolates
– Serial dilutions of some AHSV serotypes
– One equine encephalosis virus spiked sample
– AHSV-negative horse blood samples
• Each laboratory used their own internally validated
and accredited test
– Laboratories could also provide results for additional
methods
– 10 different PCR methods reported 20
2015 AHS PCR Ring Trial • NVSL tested by 3 methods
– Fernández-Pinero (FP) [2009] conventional RT-PCR
• FP conventional missed several positive samples
– Fernández-Pinero (FP) [2009] real time RT-PCR
• Correct results on nearly all samples (a few highly diluted samples were missed by most labs).
– Guthrie [2013] real time RT-PCR
• Correct results on nearly all samples (a few highly diluted samples were missed by most labs).
• The Guthrie method was slightly more sensitive.
• Current NVSL method
• NVSL performed serotyping PCRs on positive samples
– All AHSV typing correct
21
2015 AHS PCR Ring Trial
• Aguero 2008 method is used by many
EU laboratories
• Dossier presented to OIE Biological Standards
Commission in September 2015
– Accepted Aguero 2008 method as validated test with
understanding that other PCR tests may yield equivalent results
– Will be incorporated in OIE manual pending May 2016 OIE
meeting
• Guthrie 2013 method undergoing separate validation
testing
22
Questions?