the wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3
DESCRIPTION
2013 International Symposium on Ranaviruses by Maria ForzanTRANSCRIPT
![Page 1: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/1.jpg)
Rana sylvatica (Lithobates sylvaticus) as a model to study
FV3
María J. Forzán1,2, Kathleen M. Jones2
1Canadian Cooperative Wildlife Health Centre2Atlantic Veterinary College
2nd Ranavirus SymposiumTennessee, July 2013
![Page 2: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/2.jpg)
Laboratory vs. field research
• Laboratory vs. field research
Source: Yale University Historical Library
![Page 3: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/3.jpg)
Species selection
• Native species • Widespread• Sympatric• Suitable for captive breeding
![Page 4: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/4.jpg)
• Native species • Widespread• Sympatric• Suitable for captive breeding
Species selection
the wood frog
![Page 5: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/5.jpg)
Wood frog
• Fam. Ranidae (Rana or Lithobates)• 9 species Canada
• Terrestrial• Spread across NA• Broadly sympatric
• Naturally host of ranavirus• Life span >3-5 y
![Page 6: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/6.jpg)
Wood frogs in the lab
• Housing/feeding of pre- & post-metamorphic frogs• breeding in captivity?
• Dosing of infective virus (FV3)• Collection and evaluation of blood samples• cellular and humoral responses
• Collection of tissues• pathogenesis• PCR• virus isolation• histopathology• immunohistochemistry
![Page 7: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/7.jpg)
Eggs for tadpoles and metamorphs
![Page 8: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/8.jpg)
Eggs for tadpoles and metamorphs
![Page 9: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/9.jpg)
Eggs for tadpoles and metamorphs
![Page 10: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/10.jpg)
Froglets from tadpoles
![Page 11: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/11.jpg)
Froglets from tadpoles
![Page 12: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/12.jpg)
Froglets from tadpoles
![Page 13: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/13.jpg)
Housing tadpoles-metamorphs
![Page 14: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/14.jpg)
Adults
![Page 15: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/15.jpg)
Adults
@AVC
![Page 16: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/16.jpg)
![Page 17: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/17.jpg)
![Page 18: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/18.jpg)
Housing
• Temperature 17-23°C
• Adults fed
• CK, MW, EW
• Froglets fed
• Dm, Dh, CK, MW
• Tadpoles fed
• FF, Blet
Extremely hard!• 75% adults (44/59, 6mo)• 72% froglets (81/112, 8mo)
Date (mm-dd)
Cage Frogs Food left Fed Comments
Alive Dead
C01 4x 1CK Ø 8MW Not hungry
C02
C04
C05
C06
C07
C08
C09
C10
C03
![Page 19: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/19.jpg)
Oral dosing 107.7 pfu / ml
![Page 20: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/20.jpg)
Oral dosing
0.25ml / adult0.05ml / 1-y-old
![Page 21: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/21.jpg)
LD50
1-year-old frogs (3.5 cm SV)• raised from tadpoles• oral infection – 0.05ml• euthanized when terminal (*)
Dose (pfu/frog)
105.4
104.4
103.4
102.4
101.4
100.4
Control
![Page 22: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/22.jpg)
LD50
1-year-old frogs (3.5 cm SV)• raised from tadpoles• oral infection – 0.05ml• euthanized when terminal (*)
Dose (pfu/frog)
%(*)
105.4 100
104.4 100
103.4 80
102.4 20
101.4 0
100.4 0
Control 0
103.5 pfu/frogor
104.8 pfu/ml
![Page 23: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/23.jpg)
Pathogenesis
• Adults (wild-caught)• 5.3 cm SV
• Killed at 0.25, 0.5, 1, 2, 4, 9 and 14 days post-infection• Blood, tissues and abdominal fluid
collected
![Page 24: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/24.jpg)
Blood samples
![Page 25: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/25.jpg)
Blood cells: N, Eo, B, Lym, Th, Ery
![Page 26: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/26.jpg)
End day
![Page 27: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/27.jpg)
Shedding – fecal swab (PCR)
![Page 28: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/28.jpg)
Immunity – Abs, IF-α/β
![Page 29: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/29.jpg)
Pathogenesis – PCR, H&E , IHC
![Page 30: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/30.jpg)
Pathogenesis – PCR, H&E , IHC
![Page 31: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/31.jpg)
Pathogenesis – PCR, H&E , IHC
![Page 32: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/32.jpg)
Spleens: Ctrl(-), 9 dpi, 14 dpi
![Page 33: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/33.jpg)
Done / In progress
Housing and feeding of pre- and post-metamorphic frogs
Dosing of infective virus (FV3)
Collection and evaluation of blood samples
• cellular and humoral responses
• Pathogenesis
• PCR, virus isolation, histopathology, immunohistochemistry
![Page 34: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/34.jpg)
Acknowledgments
Unaccredited images from:Snailtail.com, Silkwormshop.com, Flukefarms.com, Animal Planet , CALA, Acadia University and Env. Canada websites
![Page 35: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/35.jpg)
![Page 36: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/36.jpg)
Rana sylvatica (Lithobates sylvaticus) as a model to study FV3
• Native to North America• Fam. Ranidae (Rana or Lithobates)
• Widespread (Canada, Eastern USA)• 9 species Canada • broadly sympatric
• Suitable for captive breeding (possibly)• Naturally host of ranavirus• Life span >3-5 y
![Page 37: The wood frog as a model to study the pathogenesis and host-pathogen interactions of frog virus 3](https://reader038.vdocuments.site/reader038/viewer/2022103018/558c73bad8b42a72098b475d/html5/thumbnails/37.jpg)
Wood frogs in the laboratory• Housing/feeding of pre- and post-metamorphic frogs• recommendations• breeding in captivity (?)
• Dose of infective virus (FV3)• LD50• oral dosing
• Blood samples• cellular & humoral responses
• Tissues• pathogenesis• PCR, virus isolation• histopathology, immunohistochemistry