Ranaviruses in snakes,Ranaviruses in snakes,
lizards, and chelonianslizards, and chelonians
Rachel E. Marschang1
, Anke C....
• Screening of reptiles for ranavirusesScreening of reptiles for ranaviruses
– Conventional PCR targeting a portion of the...
• 2010-2013:2010-2013:
– Screening of samples from:Screening of samples from:
• 834 reptiles, 82 species834 reptiles, 82 s...
• 2010:2010:
– 85 chelonians, 13 species (all negative)85 chelonians, 13 species (all negative)
• 20112011
– 31 chelonians...
• 2010:2010:
– 141 lizards, 19 species (all negative)141 lizards, 19 species (all negative)
• 20112011
– 190 lizards, 18 s...
• 2010:2010:
– 4 snakes, 3 species (all negative)4 snakes, 3 species (all negative)
• 20112011
– 68 snakes, 17 species (al...
• 2010:2010:
– 230 animals, 35 species, all negative230 animals, 35 species, all negative
• 20112011
– 289 animals, 44 spe...
• Species in which ranaviruses have been detectedSpecies in which ranaviruses have been detected
(2010-2013):(2010-2013):
...
Comparison
Virus No. Host Country Year
Short case
history/clinical
signs
Ref.
Testudo
hermanni
ranavirus
CH8/96
Hermann’s
...
Virus No. Host Country Year Case history Ref.
German
Gecko
ranavirus
GGRV
(2000/99)
Leaf-tailed
gecko
Uroplatus
fimbriatus...
Comparison
Virus No. Host Country Year
Short case
history/clinical
signs
Ref.
Python
brongersmai
ranavirus
PBRV
Red blood
...
• Based on:Based on:
– Sequence comparison of partial genes:Sequence comparison of partial genes:
• MCP (1402 bp), DNA pol...
0 .0 0 5
S S T IV ( C h in a )
E S V (G e r )
B IV ( A u s t r a lia )
R m a x - D K
A C R V (U S A - > G e r )
P B R V (A...
100bp50bpTRV1TRV2TRV2ASRV GGRV PBRV ACRV JSRV FV3 LMO DGRV CH8/96 ZPRV1 ZPRV2 NK
Microsatellite PCR
TRV1 TRV2 ACRV GGRV ASRV JSRV10bp
150bp
100bp
50bp
0 .0 0 5
S S T IV (C h in a )
E S V (G e r )
B IV (A u s tr a lia )
R m a x - D K
A C R V (U S A -> G e r )
P B R V (A s i...
0 .0 0 5
S S T IV (C h in a )
E S V (G e r )
B IV (A u s tr a lia )
R m a x - D K
A C R V (U S A -> G e r )
P B R V (A s i...
0 .0 0 5
S S T IV (C h in a )
E S V (G e r )
B IV (A u s tr a lia )
R m a x - D K
A C R V (U S A -> G e r )
P B R V (A s i...
0 .0 0 5
S S T IV (C h in a )
E S V (G e r )
B IV (A u s tr a lia )
R m a x - D K
A C R V (U S A -> G e r )
P B R V (A s i...
• Ranaviruses found in reptiles areRanaviruses found in reptiles are
genetically variablegenetically variable
• More commo...
• American Association of ZooAmerican Association of Zoo
VeterinariansVeterinarians
Helge Behncke, Waldkirch, Jutta Wieche...
Thank you for yourThank you for your
attention!attention!
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Ranaviruses in snakes, lizards and chelonians

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2013 International Symposium on Ranaviruses
By Rachel Marschang

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Ranaviruses in snakes, lizards and chelonians

  1. 1. Ranaviruses in snakes,Ranaviruses in snakes, lizards, and chelonianslizards, and chelonians Rachel E. Marschang1 , Anke C. Stöhr1 , Silvia Blahak2 , Maria Filomena Alcobia da Silva3,4 , Antonio Pedro Alves de Matos4,5 , Tibor Papp6 1 Fachgebiet für Umwelt- und Tierhygiene, University of Hohenheim, Germany 2 Chemisches und Veterinäruntersuchungsamt OWL, Detmold, Germany 3 University of Lisbon, Faculty of Sciences, Department of Plant Biology, Lisbon, Portugal 4 CESAM – Centre for Environmental and Marine Studies, Aveiro University, Aveiro, Portugal 5 Anatomic Pathology Department, Curry Cabral Hospital, Lisbon, Portugal 6 Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences, Budapest, Hungary
  2. 2. • Screening of reptiles for ranavirusesScreening of reptiles for ranaviruses – Conventional PCR targeting a portion of the MCPConventional PCR targeting a portion of the MCP genegene – Virus isolation in cell culture (TH-1, IgH2, VH2)Virus isolation in cell culture (TH-1, IgH2, VH2) Diagnosis
  3. 3. • 2010-2013:2010-2013: – Screening of samples from:Screening of samples from: • 834 reptiles, 82 species834 reptiles, 82 species • Chelonians: 232 animals, at least 24 speciesChelonians: 232 animals, at least 24 species • Lizards: 423 animals, at least 35 speciesLizards: 423 animals, at least 35 species • Snakes: 179 animals, at least 23 speciesSnakes: 179 animals, at least 23 species Diagnosis
  4. 4. • 2010:2010: – 85 chelonians, 13 species (all negative)85 chelonians, 13 species (all negative) • 20112011 – 31 chelonians, 9 species (all negative)31 chelonians, 9 species (all negative) • 20122012 – 64 chelonians, 14 species (5 positive from 4 different64 chelonians, 14 species (5 positive from 4 different owners)owners) • 20132013 – 52 chelonians, 8 species (3 positive from 3 different52 chelonians, 8 species (3 positive from 3 different owners)owners) Diagnosis: chelonians
  5. 5. • 2010:2010: – 141 lizards, 19 species (all negative)141 lizards, 19 species (all negative) • 20112011 – 190 lizards, 18 species (1 group positive)190 lizards, 18 species (1 group positive) • 20122012 – 60 lizards, 13 species (6 positive)60 lizards, 13 species (6 positive) • 20132013 – 32 lizards, 7 species (1 positive)32 lizards, 7 species (1 positive) Diagnosis: lizards
  6. 6. • 2010:2010: – 4 snakes, 3 species (all negative)4 snakes, 3 species (all negative) • 20112011 – 68 snakes, 17 species (all negative)68 snakes, 17 species (all negative) • 20122012 – 77 snakes, 13 species (4 positive from 2 different owners)77 snakes, 13 species (4 positive from 2 different owners) • 20132013 – 30 snakes, 7 species (1 positive)30 snakes, 7 species (1 positive) Diagnosis: snakes
  7. 7. • 2010:2010: – 230 animals, 35 species, all negative230 animals, 35 species, all negative • 20112011 – 289 animals, 44 species, positive in one group of lizards289 animals, 44 species, positive in one group of lizards • 20122012 – 201 animals, 40 species, 15 positive from 12 owners201 animals, 40 species, 15 positive from 12 owners • 20132013 – 114 animals, 22 species, 5 positive from 5 different114 animals, 22 species, 5 positive from 5 different ownersowners Diagnosis: positives
  8. 8. • Species in which ranaviruses have been detectedSpecies in which ranaviruses have been detected (2010-2013):(2010-2013): – Chelonians:Chelonians: Stigmochelys pardalisStigmochelys pardalis,, T. graecaT. graeca,, T.T. hermannihermanni (n=2),(n=2), T. horsfieldiiT. horsfieldii,, Emys orbicularisEmys orbicularis – Lizards:Lizards: Japalura splendidaJapalura splendida,, Dopasia gracilisDopasia gracilis,, VaranusVaranus macraeimacraei,, Anolis carolinensisAnolis carolinensis,, Iguana iguanaIguana iguana,, PogonaPogona vitticepsvitticeps (n=2),(n=2), Lacerta agilisLacerta agilis – Snakes: 2Snakes: 2 Python regiusPython regius, 2, 2 Python molurusPython molurus + 1+ 1 EunectesEunectes spp.spp. Diagnosis: positives
  9. 9. Comparison Virus No. Host Country Year Short case history/clinical signs Ref. Testudo hermanni ranavirus CH8/96 Hermann’s tortoise Testudo hermanni Switz. 1996 Stomatitis, death of several tortoises. Marschang et al., 1999 Testudo ranavirus 1 TRV1 (882/96) Egyptian tortoise Testudo kleinmanni Germany 1996 Rhinitis, stomatitis, hepatitis. 2nd animal in collection survived. Blahak and Uhlenbrok, 2010 Testudo ranavirus 2 TRV 2 (5187/97) Hermann’s tortoise Testudo hermanni Germany 2007 Stomatitis, emaciation. Blahak and Uhlenbrok, 2010 TRV 2 (CU60/09) Marginated tortoise Testudo marginata Germany 2009 Stomatitis, necrosis in the trachea. Blahak and Uhlenbrok, 2010
  10. 10. Virus No. Host Country Year Case history Ref. German Gecko ranavirus GGRV (2000/99) Leaf-tailed gecko Uroplatus fimbriatus Germany 2001 Granulomatous lesions on the tongue, hepatitis Marschang et al., 2005 Lacerta monticola ranavirus LMRV Iberian mountain lizard Lacerta monticola Portugal 2003/ 2004 Wild-caught, co-inf. with erythrocytic necrosis virus Alves de Matos et al., 2011 Japalura splendida ranavirus JSRV Green striped tree dragon Japalura splendida Germany (imported from Asia via Florida) 2011 Skin lesions, systemic haemorrhages, liver necrosis, large number of animals died. AdV / IIV in the same group Behncke et al., 2013 Anolis sagrei ranavirus ASRV Brown anole Anolis sagrei Germany (imported from Florida) 2008 2011 RV found repeatedly in different imported groups during 3 years. Low to high mortality, apathy, skin lesions. Coinfection with reovirus in one animal. Stöhr et al., submitted for publication Dopasia gracilis ranavirus DGRV Asian glass lizard Dopasia gracilis Germany (imported from Asia) 2012 Illegally imported animals, a number of animals died. Skin lesions. IIV in the same animal. Stöhr et al., submitted for publication Anolis carolinensis ranavirus ACRV Green anole Anolis carolinensis Germany (imported from Florida) 2012 Several animals in poor body condition separated, high mortality, skin lesions. AdV and IIV in the same animal. Stöhr et al., submitted for publication
  11. 11. Comparison Virus No. Host Country Year Short case history/clinical signs Ref. Python brongersmai ranavirus PBRV Red blood python Python brongersmai Germany (imported from Indonesia) 2007 100 animals imported, 30 % developed severe diphteroid stomatitis and hepatitis. An unknown number of snakes died. Blahak, unpub. • Total: 10 different reptilian ranavirusTotal: 10 different reptilian ranavirus isolatesisolates
  12. 12. • Based on:Based on: – Sequence comparison of partial genes:Sequence comparison of partial genes: • MCP (1402 bp), DNA polymerase(560 bp), RNR-MCP (1402 bp), DNA polymerase(560 bp), RNR-αα (806 bp), RNR-(806 bp), RNR-ββ (646 bp)(646 bp) (Mao et al., 1997; Marschang(Mao et al., 1997; Marschang et al., 1999; Ariel et al., 2010; Hyatt et al., 2000; Holopainen etet al., 1999; Ariel et al., 2010; Hyatt et al., 2000; Holopainen et al., 2009)al., 2009) – Length and sequences of vIF-Length and sequences of vIF-22αα (247 or 1050(247 or 1050 bp)bp) (Essbauer et al., 2001; Chinchar, personal comm.)(Essbauer et al., 2001; Chinchar, personal comm.) – Length of microsatellite regionLength of microsatellite region (Mavian et al., 2012)(Mavian et al., 2012) Comparison
  13. 13. 0 .0 0 5 S S T IV ( C h in a ) E S V (G e r ) B IV ( A u s t r a lia ) R m a x - D K A C R V (U S A - > G e r ) P B R V (A s ia -> G e r ) C M T V ( S p a in + N e t h .) J S R V (C h in a -> U S A - > G e r ) T R V 2 (G e r ) E H N V (A u s t r a lia ) T F V (C h in a ) G G R V ( 2 0 0 0 /9 9 ) (G e r ) T R V 1 (G e r ) L M R V (P o r t) E C V ( F r a n c e ) A S R V ( U S A - > G e r ) C H 8 /9 6 (S w itz e r la n d ) Z P R V 2 (E u r o p e ) R E V ( Ita l) Z P R V 1 ( G e r ) C o d V (D K ) A T S V (U S A ) D G R V (A s ia - > G e r ) F V 3 (U S A ) S E R V P P IV ( F in n l) N C R V ( Ir a q - > G e r ) R g r y l (C h in a ) 1 0 0 /1 0 0 /1 6 9 /5 7 /0 .9 4 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 8 2 /6 4 /0 .9 7 6 2 /4 8 /0 .9 1 0 0 /8 4 /1 8 3 /7 2 /0 . 9 7 1 0 0 /1 0 0 /1 1 0 0 /8 6 /1 7 8 /5 9 /0 .7 4 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 8 7 /6 8 /0 .8 6 8 /5 9 /0 .8 1 1 0 0 /1 0 0 /1 9 8 /9 3 /1 8 5 /n .a ./0 .9 9 1 0 0 /9 9 /1 7 4 /7 3 /0 .9 6 Concatenated tree 3223 bp Red=full length vIF-2a Green=truncated vIF2a Black=no data available
  14. 14. 100bp50bpTRV1TRV2TRV2ASRV GGRV PBRV ACRV JSRV FV3 LMO DGRV CH8/96 ZPRV1 ZPRV2 NK Microsatellite PCR
  15. 15. TRV1 TRV2 ACRV GGRV ASRV JSRV10bp 150bp 100bp 50bp
  16. 16. 0 .0 0 5 S S T IV (C h in a ) E S V (G e r ) B IV (A u s tr a lia ) R m a x - D K A C R V (U S A -> G e r ) P B R V (A s ia -> G e r ) C M T V (S p a in + N e th .) J S R V ( C h in a -> U S A -> G e r ) T R V 2 (G e r ) E H N V (A u s tr a lia ) T F V (C h in a ) G G R V (2 0 0 0 /9 9 ) (G e r ) T R V 1 (G e r ) L M R V (P o r t ) E C V (F r a n c e ) A S R V (U S A -> G e r ) C H 8 /9 6 (S w itz e r la n d ) Z P R V 2 (E u r o p e ) R E V (Ita l) Z P R V 1 (G e r ) C o d V (D K ) A T S V (U S A ) D G R V (A s ia -> G e r ) F V 3 ( U S A ) S E R V P P IV ( F in n l) N C R V ( Ir a q -> G e r ) R g r y l (C h in a ) 1 0 0 /1 0 0 /1 6 9 /5 7 /0 .9 4 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 8 2 /6 4 /0 .9 7 6 2 /4 8 /0 . 9 1 0 0 /8 4 /1 8 3 /7 2 /0 . 9 7 1 0 0 /1 0 0 /1 1 0 0 /8 6 /1 7 8 /5 9 /0 .7 4 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 8 7 /6 8 /0 .8 6 8 /5 9 /0 .8 1 1 0 0 /1 0 0 /1 9 8 /9 3 /1 8 5 /n .a ./0 .9 9 1 0 0 /9 9 /1 7 4 /7 3 /0 .9 6 Russian tort, 2012, D Retic. python, 2012, D Retic. python, 2012, D Anaconda, 2012, D Bearded dragon, 2012, D Hermann‘s tort, 2012, D Ball python, 2013, D Bearded dragon, 2013, D Red-eared slider, 2013, D Hermann‘s tort, 2013, UK
  17. 17. 0 .0 0 5 S S T IV (C h in a ) E S V (G e r ) B IV (A u s tr a lia ) R m a x - D K A C R V (U S A -> G e r ) P B R V (A s ia -> G e r ) C M T V (S p a in + N e th .) J S R V ( C h in a -> U S A -> G e r ) T R V 2 (G e r ) E H N V (A u s tr a lia ) T F V (C h in a ) G G R V (2 0 0 0 /9 9 ) (G e r ) T R V 1 (G e r ) L M R V (P o r t ) E C V (F r a n c e ) A S R V (U S A -> G e r ) C H 8 /9 6 (S w itz e r la n d ) Z P R V 2 (E u r o p e ) R E V (Ita l) Z P R V 1 (G e r ) C o d V (D K ) A T S V (U S A ) D G R V (A s ia -> G e r ) F V 3 ( U S A ) S E R V P P IV ( F in n l) N C R V ( Ir a q -> G e r ) R g r y l (C h in a ) 1 0 0 /1 0 0 /1 6 9 /5 7 /0 .9 4 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 8 2 /6 4 /0 .9 7 6 2 /4 8 /0 . 9 1 0 0 /8 4 /1 8 3 /7 2 /0 . 9 7 1 0 0 /1 0 0 /1 1 0 0 /8 6 /1 7 8 /5 9 /0 .7 4 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 8 7 /6 8 /0 .8 6 8 /5 9 /0 .8 1 1 0 0 /1 0 0 /1 9 8 /9 3 /1 8 5 /n .a ./0 .9 9 1 0 0 /9 9 /1 7 4 /7 3 /0 .9 6 Leopard tort, 2012, D Hermann‘s tort, 2012, D Ball python, 2012, D Monitor lizard, 2012, D Hermann‘s tort, 2012, I Iguana, 2012, D
  18. 18. 0 .0 0 5 S S T IV (C h in a ) E S V (G e r ) B IV (A u s tr a lia ) R m a x - D K A C R V (U S A -> G e r ) P B R V (A s ia -> G e r ) C M T V (S p a in + N e th .) J S R V ( C h in a -> U S A -> G e r ) T R V 2 (G e r ) E H N V (A u s tr a lia ) T F V (C h in a ) G G R V (2 0 0 0 /9 9 ) (G e r ) T R V 1 (G e r ) L M R V (P o r t ) E C V (F r a n c e ) A S R V (U S A -> G e r ) C H 8 /9 6 (S w itz e r la n d ) Z P R V 2 (E u r o p e ) R E V (Ita l) Z P R V 1 (G e r ) C o d V (D K ) A T S V (U S A ) D G R V (A s ia -> G e r ) F V 3 ( U S A ) S E R V P P IV ( F in n l) N C R V ( Ir a q -> G e r ) R g r y l (C h in a ) 1 0 0 /1 0 0 /1 6 9 /5 7 /0 .9 4 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 8 2 /6 4 /0 .9 7 6 2 /4 8 /0 . 9 1 0 0 /8 4 /1 8 3 /7 2 /0 . 9 7 1 0 0 /1 0 0 /1 1 0 0 /8 6 /1 7 8 /5 9 /0 .7 4 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 8 7 /6 8 /0 .8 6 8 /5 9 /0 .8 1 1 0 0 /1 0 0 /1 9 8 /9 3 /1 8 5 /n .a ./0 .9 9 1 0 0 /9 9 /1 7 4 /7 3 /0 .9 6 Sand lizard, 2012, UK
  19. 19. 0 .0 0 5 S S T IV (C h in a ) E S V (G e r ) B IV (A u s tr a lia ) R m a x - D K A C R V (U S A -> G e r ) P B R V (A s ia -> G e r ) C M T V (S p a in + N e th .) J S R V ( C h in a -> U S A -> G e r ) T R V 2 (G e r ) E H N V (A u s tr a lia ) T F V (C h in a ) G G R V (2 0 0 0 /9 9 ) (G e r ) T R V 1 (G e r ) L M R V (P o r t ) E C V (F r a n c e ) A S R V (U S A -> G e r ) C H 8 /9 6 (S w itz e r la n d ) Z P R V 2 (E u r o p e ) R E V (Ita l) Z P R V 1 (G e r ) C o d V (D K ) A T S V (U S A ) D G R V (A s ia -> G e r ) F V 3 ( U S A ) S E R V P P IV ( F in n l) N C R V ( Ir a q -> G e r ) R g r y l (C h in a ) 1 0 0 /1 0 0 /1 6 9 /5 7 /0 .9 4 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 8 2 /6 4 /0 .9 7 6 2 /4 8 /0 . 9 1 0 0 /8 4 /1 8 3 /7 2 /0 . 9 7 1 0 0 /1 0 0 /1 1 0 0 /8 6 /1 7 8 /5 9 /0 .7 4 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 1 0 0 /1 0 0 /1 8 7 /6 8 /0 .8 6 8 /5 9 /0 .8 1 1 0 0 /1 0 0 /1 9 8 /9 3 /1 8 5 /n .a ./0 .9 9 1 0 0 /9 9 /1 7 4 /7 3 /0 .9 6 Europ. pond turtle, 2012, D
  20. 20. • Ranaviruses found in reptiles areRanaviruses found in reptiles are genetically variablegenetically variable • More common than we realizedMore common than we realized • Outbreaks?Outbreaks? • Pet trade?Pet trade? • Pathogenicity?Pathogenicity? Summary
  21. 21. • American Association of ZooAmerican Association of Zoo VeterinariansVeterinarians Helge Behncke, Waldkirch, Jutta Wiechert, Mainz, TonyHelge Behncke, Waldkirch, Jutta Wiechert, Mainz, Tony Sainesbury, London, Ross Martin, Leeds, SabineSainesbury, London, Ross Martin, Leeds, Sabine Öfner, Munich, Kim Heckers, Bad Kissingen, JürgenÖfner, Munich, Kim Heckers, Bad Kissingen, Jürgen Seybold, Baiersbronn, Karina Mathes, HannoverSeybold, Baiersbronn, Karina Mathes, Hannover Acknowledgements
  22. 22. Thank you for yourThank you for your attention!attention!

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