a preliminary survey of viruses of bats of canada
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A preliminary survey of viruses of bats of Canada. (identification of viruses that might emerge as human pathogens). Emerging infectious diseases (EID). pathogen infects host for the first time new strains of a pathogen with distinct properties increased incidence of preexisting disease - PowerPoint PPT PresentationTRANSCRIPT
A preliminary survey of viruses of bats of Canada
(identification of viruses that might emerge as human pathogens)
Emerging infectious diseases (EID)
• pathogen infects host for the first time
• new strains of a pathogen with distinct properties
• increased incidence of preexisting disease
• new or reintroduction into area
Animals and emerging human disease
• most EID events emerge from animals• most of these events (70%) involve wild
animals
nature 21 Feb 2008
Role of bats in EID
• Confirmed or suspected in:– Nipah– Hendra– SARS– Marburg– Ebola– Menangle– Tioman– Melaka– rabies (in some areas)
“Damn! what is it about bats?”*
* David Quammen and Lynn Johnson, Deadly Contact, National Geographic, October 2007
Chiroptera “hand wing”
Epomops franqueti - www.dkimages.com/
Old order
• fossil record > 50 million years
N.B. Simmons, et al.2008. nature 451:818-821
Onchonycteris finneyi
Simmons, 2005, Science 307:527
Diversity
Megachiroptera Microchiroptera
Tim Menzies
Indian flying fox1.8 m wingspan1.6 kg
Thai bumblebee bat15 cm wingspan2 gm
Hibernation and torpor
http://www.earlham.edu/~turnbal/
Bats of Canada• Little brown bat (Myotis lucifugus)• Yuma myotis (M. yumanensis)• Long-legged myotis (M. volans)• Fringed myotis (M. thysanodes)• Northern long-eared bat (Myotis septentrionalis),• Eastern small-footed myotis (M. leibii)• Keen’s myotis (M. keenii)• Long eared myotis (M. evotis)• Western small-footed myotis (M. ciliolabrum)• California myotis (M. californicus)• Big brown bat (Eptesicus fuscus)• Silver haired bat (Lasionycteris noctivagans) • Hoary bat (Lasiurus cinereus), • Red bat (Lasiurus borealis)• Western red bat (Lasiurus blossevillii)• Eastern pipstrelle (Pipistrellus subflavus)• Spotted bat (Euderma maculatum)• Townsend’s big-eared bat (Corynorhinus townsendii)• Pallid bat (Antrozous pallidus)
Bat Conservation International
Bats in Manitoba
Little brown bat (Myotis lucifugus)
Big brown bat (Eptesicus fuscus)
Northern long-eared bat (Myotis septentrionalis),
Silver haired bat (Lasionycteris noctivagans),
Hoary bat (Lasiurus cinereus),
•Red bat (Lasiurus borealis)
Hibernating
Migrating
Derek Donald
Derek Donald
www.batcon.org
www.batcon.org
www.batcon.org
www.batcon.org
Genesis of the project
How does one find bats?
• live-trapped little brown bats
• archived rabies-negative bats at the Ottawa and Lethbridge CFIA labs
• wind farms
• submission from local veterinarian
Craig Willis
remove liver, kidney, brain, spleen, lung, intestine
fix in formaldehyde (pool tissues from same species of bats)
homogenize (pooled tissues)
freeze at -80
tissue culture(primate, rodent bats, avian cells)
Purify nucleic acidQIAmp viral RNA mini kit
(will purify RNA and DNA)
cDNA synthesis
PCR
corona(Gr 1, pan)
lyssahenipah
morbilli
WNVHanta
gelsequence
observe daily for cpe
+ -
amplify
characterize
3 passages
autoclave+ -
herpes
polyomaflavi
bunya
alphaCache Valley
Tissue Cultures
• Vero (monkey)
• NIH-3T3 (mouse)
• BHK-21 (hamster)
• TbrLu1 (bat)
• Quail
• Chick embryo
Live-trapped little brown batsbat no M/F corona polyoma bat no M/F corona polyoma
1 F 12 F +2 F 13 F
3 F 14 M +4 ? 15 F
5 M 16 F
6 M 17 M
7 F 18 F
8 M 19 F
9 M 20 M
10 F + 21 F +11 F +
Archived rabies-negative
Bat Num Bat Num
Eptesicus fuscus (BBB)
55 M. californicus 2
M. lucifugus (LBB) 10 M. evotis
(WLEB)
1
M. yumanensis 6 1 +ve polyoma
Lasionycteris noctivagans (SHB)
6
M. septentrionalis (NLEB)
3 Lasiurus cinereus
(HB)
1
Wind-farmBat No
Lasiurus cinereus(Hoary)
7*
Lasionycteris noctivagans(Silver haired)
6
Lasiurus borealis(Red)
3
* Includes 1 local submission
Strategy for recovering entire bat polyomavirus genome
product from broad-specificity PCR
primers for amplifying genome
Phylogeny of bat (Myotis) polyomavirus
• ClustalX
WU
KI
Budge
Finch
Bat
Mouse Pneumotropic
Sq monkey
BovineCrow
Goose
HamsterMouse
AG monkey
SV40
SV12Baboon
JCBK
0.1
VP1
Merkel
WU
KIBudge
Finch
Mouse Pneumotropic
Sq monkey
Bovine
Crow
HamsterMouse
AG monkey
SV40
SV12
Baboon
JCBK
0.1
Bat
VP2,3
Merkel
WUKI
BudgeFinch
Mouse Pneumotropic
Sq monkey
Bovine
Crow
Hamster
Mouse
AG monkey
SV40
SV12Baboon
JC
BK
0.1
Bat
T-Antigen
Merkel
WU
KI
Budge
Finch
Mouse Pneumotropic
Sq monkey
Crow
HamsterMouse
AG monkey
SV40
SV12Baboon
JCBK
0.1
Bat
Small t-Antigen
Merkel
What do we know about bat coronaviruses?
• SARS
• Group 1 bat coronaviruses– asian– Rocky Mountain bat coronavirus– European
• Sero-conversion in bat biologists
“…bats are the natural reservoir of all currently known coronavirus lineages.”
Vijaykrishna et al. 2007. J. Virol 81:4012-4020
What do we know about polyomaviruses?
• mouse polyomavirus - 1950s
• persistent, asymptomatic infections in immuno-competent individuals - disease in immuno-suppressed or young animals
• pathogenic avian viruses
Many Thanks!
• Tim Dumonceaux• Harvey Artsob• Robin Lindsay• Mike Drebot• Marta Sabara• Antonia Dibernardo
• Jack Dubois
• Susan Nadin-Davis
• Craig Willis