2013 International Symposium on Ranaviruses by Anke Stohr

1. Ranavirus associatedRanavirus associatedRanavirus associatedRanavirus associated
dermatitis in lizardsdermatitis in lizards
Anke C StöhrAnke C Stöhr11 Kim O HeckersKim O Heckers22 Jutta WiechertJutta Wiechert3†3† Helge BehnckeHelge Behncke44Anke C. StöhrAnke C. Stöhr11, Kim O. Heckers, Kim O. Heckers22, Jutta Wiechert, Jutta Wiechert3†3†, Helge Behncke, Helge Behncke44,,
Karina MathesKarina Mathes55, Pascale Günther, Pascale Günther55, Inna Ball, Inna Ball11, Rachel E. Marschang, Rachel E. Marschang11
11Fachgebiet für UmweltFachgebiet für Umwelt-- und Tierhygiene, University of Hohenheim, Germanyund Tierhygiene, University of Hohenheim, Germany
22Laboklin GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, GermanyLaboklin GmbH & Co. KG, Laboratory for Clinical Diagnostics, Bad Kissingen, Germany
33Reptilium TerrarienReptilium Terrarien-- und Wüstenzoo, Landau, Germanyund Wüstenzoo, Landau, Germany
44ImportImport--Export Peter Hoch GmbH, Waldkirch, GermanyExport Peter Hoch GmbH, Waldkirch, Germany
55Clinic for Pets Reptiles Pet and Feral Birds University of Veterinary Medicine Hannover GermanyClinic for Pets Reptiles Pet and Feral Birds University of Veterinary Medicine Hannover Germany55Clinic for Pets, Reptiles, Pet and Feral Birds, University of Veterinary Medicine Hannover, GermanyClinic for Pets, Reptiles, Pet and Feral Birds, University of Veterinary Medicine Hannover, Germany

2. Ranaviruses in lizards
Lit t
Ranaviruses in lizards
• Literature:
• Leaf-tailed gecko (Uroplatus fimbriatus):
l ti ti i l iti- ulcerative-necrotizing glossitis
- granulomatous lesions on the tail
- focal necrosis in the liver (Marschang et al 2005)focal necrosis in the liver (Marschang et al., 2005)
• Iberian mountain lizard (Lacerta monticola):
- no overt disease (Alves de Matos et al., 2011)
-> Screening of lizards for the presence of ranaviruses
- 2010: n=141 -> all negative2010: n 141 > all negative
- 2011: n =190 -> 1 positive (0.53%)
- 2012: n =60 -> 6 positive (10%)

3. First case:First case:
J l l didJ l l didJapalura splendidaJapalura splendida
•• GreenGreen stripedstriped treetree dragonsdragons importedimported fromfrom
southwesternsouthwestern China via FloridaChina via Florida toto Germany inGermany in
spring 2011spring 2011spring 2011spring 2011
•• ClinicalClinical signssigns:: dehydratationdehydratation,, apathyapathy,, skinskin lesionslesions,,
centralcentral nervousnervous disordersdisorders highhigh mortalitymortalitycentralcentral nervousnervous disordersdisorders,, highhigh mortalitymortality
200
50
100
150
alive animals
0
50
15-Feb
2-M
ar
17-M
ar
1-Apr
16-Apr
01-M
ay
16-M
ay

4. First case:First case:
J l l didJ l l didJapalura splendidaJapalura splendida
•• ParasitologicalParasitological examinationexamination::•• ParasitologicalParasitological examinationexamination::
FlagellataFlagellata sppspp.,., nematodesnematodes ((HeterakisHeterakis sppspp.,., SpiruridaSpirurida sppspp.),.),
coccidiacoccidia ((CholeoeimeriaCholeoeimeria sppspp.), larval.), larval ascaridsascarids inin thethe pharynxpharynx
•• PathologicalPathological examinationexamination::
FibrinousFibrinous granulomatousgranulomatous necroticnecrotic dermatitisdermatitis,,
catarrhal purulent to granulomatous sinusitiscatarrhal purulent to granulomatous sinusitis hyperemic oedematouscatarrhal purulent to granulomatous sinusitiscatarrhal purulent to granulomatous sinusitis, hyperemic oedematous
GIT, vacuolar tubulonephrosis of the distal renal tubules, hyperemia
and necrosis in the liver, basophilic intracytoplasmatic inclusion bodies

5. First case:First case:
J l l didJ l l didJapalura splendidaJapalura splendida
•• Virological examination (5 animals):Virological examination (5 animals):
–– Detection ofDetection of ranavirus (RV), adenovirus (AdV), invertebrateranavirus (RV), adenovirus (AdV), invertebrate
i id ii id i (IIV) i PCRi PCRiridovirusiridovirus (IIV) via PCRvia PCR
–– Isolation of RV and IIV on IgHIsolation of RV and IIV on IgH--2 cells2 cells
RV AdV IIV Virus isolationRV AdV IIV Virus isolation
Animal 1 - intestine skin, lung -
Animal 2 - intestine
skin, lung, liver/kidney,
i t ti
IIV (skin, lung, liver/kidney,
i t ti )
Animal 2 intestine
intestine intestine)
Animal 3 lung, intestine -
skin, lung, liver/kidney,
intestine
RV + IIV (skin, lung,
liver/kidney, intestine)
Animal 4 skin lung liver/kidneyAnimal 4 - - skin, lung, liver/kidney -
Animal 5 - intestine skin, lung, intestine
IIV (lung, liver/kidney,
intestine)

6. Second case:Second case:
D i iliD i iliDopasia gracilisDopasia gracilis
•• 2012: Asian glass lizards illegally imported into2012: Asian glass lizards illegally imported into
Germany from Asia and confiscatedGermany from Asia and confiscated
•• Clinical signs:Clinical signs:•• Clinical signs:Clinical signs:
–– Increasing mortalityIncreasing mortality
–– Skin lesionsSkin lesions
•• Detection of RV via PCR (skin + mixed organs) and virusDetection of RV via PCR (skin + mixed organs) and virus
isolation (skin, IgHisolation (skin, IgH--2)2)
Reptilia

7. Third case:Third case:
A li li iA li li iAnolis carolinensisAnolis carolinensis
•• Green anoles from a population in Miami repeatedlyGreen anoles from a population in Miami repeatedly•• Green anoles from a population in Miami repeatedlyGreen anoles from a population in Miami repeatedly
imported into Germanyimported into Germany
•• winter/spring 2012:winter/spring 2012:•• winter/spring 2012:winter/spring 2012:
–– Increasing mortalityIncreasing mortality
–– Moderate parasite loadModerate parasite loadModerate parasite loadModerate parasite load
–– Catarrhal enteritis, skin lesionsCatarrhal enteritis, skin lesions

8. Third case:Third case:
A li li iA li li iAnolis carolinensisAnolis carolinensis
•• Virological examination (1 animal):Virological examination (1 animal):
RV AdV IIV Virus isolation
ki (RV)skin + - + + (RV)
liver + - - + (RV)
S llSmall
intestine
+ + - + (RV)

9. Fourth case:Fourth case:
I iI iIguana iguanaIguana iguana
•• Adult male and female green iguanas from different originsAdult male and female green iguanas from different origins
collected to a single group in a private zoo in Germanycollected to a single group in a private zoo in Germany
O 5 i di id l i l t dl d l dO 5 i di id l i l t dl d l d•• Over 5 years, individual animals repeatedly developedOver 5 years, individual animals repeatedly developed
hyperkeratotic skin lesionshyperkeratotic skin lesions interrupted by asymptomaticinterrupted by asymptomatic
periodsperiodspp
• Further examination: bacterial infection, dermatomycosis
• No treatment effective
• May 2012: last iguana developed disease
-> skin biopsy for virological testing
RV iti (PCR)-> RV positive (PCR)

10. Fifth case:Fifth case:
P ittiP ittiPogona vitticepsPogona vitticeps
•• July 2011: 5 year old male bearded dragonJuly 2011: 5 year old male bearded dragon
--> sudden appearance of pustules on the skin in the region of> sudden appearance of pustules on the skin in the region of
th k d th h dth k d th h dthe neck and the headthe neck and the head
> purulent dermatitis-> purulent dermatitis
(Micrococcus spp.)
-> antimicrobal therapy
-> 

11. Fifth case:Fifth case:
P ittiP ittiPogona vitticepsPogona vitticeps
•• October 2012:October 2012:
- inflammation and necrosis in one leg
- dark skin lesions on the head,,
neck and back

12. Fifth case:Fifth case:
P ittiP ittiPogona vitticepsPogona vitticeps
• Pathohistology:
increased growth of
ti ti (5)connective tissues (5);
Iridophores (4)
irregularly distributed,
large melanocytes (1)
Horny layer (2)
Epidermis (3)
Picture: P. Zwart, 400 x Van Gieson stain, polarised light

13. Fifth case:Fifth case:
P ittiP ittiPogona vitticepsPogona vitticeps
•• Virological examination:Virological examination:
RV AdV Virus isolation
skin + - -
muscle + - -
heart + -
Oral / cloacal swab - + -
Organs (intestine,
liver, kidney, lung)
- - -

14. Characterization and
comparison
•• Partial sequencing of MCP gene (500bp)Partial sequencing of MCP gene (500bp)
Japalura
Dopasia Iguana Pogona FV3 RTV ESV/ECV Tortoises
= Anolis
Dopasia Iguana Pogona FV3 RTV ESV/ECV Tortoises
Japalura
= Anolis
98.4 96.8 98.4 100 98.6 96.8 98.4
Dopasia 98.8 96.0 97.6 98.4 99 96 97.6Dopasia 98.8 96.0 97.6 98.4 99 96 97.6
Iguana 95.8 96.6 96.8 96.8 96.2 100 96.8
Pogona 98.2 98.2 95.2 98.4 97.8 96.8 100
FV3 100 98.8 95.8 98.2 98.6 96.8 98.4
RTV 98.8 100 96.4 98.2 98.8 96.2 97.8
ECV/ESV 95 8 96 4 100 95 2 95 8 96 4 96 8ECV/ESV 95.8 96.4 100 95.2 95.8 96.4 96.8
Tortoises 98.2 98.2 95.2 100 98.2 98.2 95.2
Upper diagonal=nt, lower diagonal=aa

15. DiscussionDiscussion
•• First detection of ranaviruses in 5 lizard speciesFirst detection of ranaviruses in 5 lizard species
–– Green striped tree dragonGreen striped tree dragon (Japalura splendida)(Japalura splendida)Green striped tree dragonGreen striped tree dragon (Japalura splendida)(Japalura splendida)
–– Asian glass lizardAsian glass lizard (Dopasia gracilis)(Dopasia gracilis)
–– Green anoleGreen anole (Anolis carolinensis)(Anolis carolinensis)
–– Green iguanaGreen iguana (Iguana iguana)(Iguana iguana)–– Green iguanaGreen iguana (Iguana iguana)(Iguana iguana)
–– Bearded dragonBearded dragon (Pogona vitticeps)(Pogona vitticeps)
•• Most RV distinct from each otherMost RV distinct from each other•• Most RV distinct from each otherMost RV distinct from each other
--> RV can infect a wide range of hosts> RV can infect a wide range of hosts
--> increasing importance of RV in lizards?> increasing importance of RV in lizards?

16. DiscussionDiscussion
•• Origin: Asia, USA, GermanyOrigin: Asia, USA, Germany
--> global distribution> global distributionglobal distributionglobal distribution
•• large groups of animals vs. single animalslarge groups of animals vs. single animals
•• high to low morbidity and mortalityhigh to low morbidity and mortalityg y yg y y
•• Coinfections with otherCoinfections with other
viruses/parasites/bacteria/fungiviruses/parasites/bacteria/fungi
•• disease after transportdisease after transport
--> clinical outcome of disease multifactorial> clinical outcome of disease multifactorial
( i f ti t i i )?( i f ti t i i )?(coinfections, stress, immunosuppression…)?(coinfections, stress, immunosuppression…)?

17. DiscussionDiscussion
•• Contact with other reptiles/amphibiansContact with other reptiles/amphibians
--> Role of reptiles in the epidemiology?> Role of reptiles in the epidemiology?Role of reptiles in the epidemiology?Role of reptiles in the epidemiology?
•• Skin lesions (Skin lesions (ppurulent to granulomatous ulcerativeurulent to granulomatous ulcerative--Skin lesions (Skin lesions (ppurulent to granulomatous ulcerativeurulent to granulomatous ulcerative
necrotic dermatitis and hyperkeratosis)necrotic dermatitis and hyperkeratosis) in all lizardsin all lizards
--> DD mortality + skin lesions> DD mortality + skin lesionsDD mortality skin lesionsDD mortality skin lesions

18. AcknowledgementsAcknowledgements
Christa Schäfer and Silvia Speck:
F h bi U l d Ti h iFachgebiet Umwelt und Tierhygiene
University of Hohenheim
Stuttgart, Germany

19. ThankThank youyou forfor youryouryy yy
attentionattention!!