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