Ranavirus associated Ranavirus associated Ranavirus associated Ranavirus associated dermatitis in lizardsdermatitis in lizards

Anke C StöhrAnke C Stöhr11 Kim O Heckers Kim O Heckers22 Jutta Wiechert Jutta Wiechert3†3† Helge Behncke Helge Behncke44 Anke 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, Germany22Laboklin 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, Germany44ImportImport--Export Peter Hoch GmbH, Waldkirch, Germany Export 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

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%)

First case: First case: J l l didJ l l didJapalura splendidaJapalura splendida

•• Green Green stripedstriped treetree dragonsdragons importedimported fromfromsouthwesternsouthwestern China via Florida China via Florida toto Germany in Germany in spring 2011spring 2011spring 2011spring 2011

•• Clinical Clinical signssigns: : dehydratationdehydratation, , apathyapathy, , skinskin lesionslesions, , centralcentral nervousnervous disordersdisorders highhigh mortalitymortalitycentralcentral nervousnervous disordersdisorders, , highhigh mortalitymortality

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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 in in 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, hyperemiaand necrosis in the liver, basophilic intracytoplasmatic inclusion bodies

First case: First case: J l l didJ l l didJapalura splendidaJapalura splendida

•• Virological examination (5 animals):Virological examination (5 animals):–– Detection of Detection of ranavirus (RV), adenovirus (AdV), invertebrate ranavirus (RV), adenovirus (AdV), invertebrate

i id ii id i (IIV) i PCRi PCRiridovirusiridovirus (IIV) via PCR via 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)

Second case: Second case: D i iliD i iliDopasia gracilisDopasia gracilis

•• 2012: Asian glass lizards illegally imported into 2012: 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 virus Detection of RV via PCR (skin + mixed organs) and virus isolation (skin, IgHisolation (skin, IgH--2)2)

Reptilia

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 repeatedly Green 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

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)

Fourth case: Fourth case: I iI iIguana iguanaIguana iguana

•• Adult male and female green iguanas from different origins Adult 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 GermanyO 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 developed Over 5 years, individual animals repeatedly developed hyperkeratotic skin lesionshyperkeratotic skin lesions interrupted by asymptomatic interrupted by asymptomatic periodsperiodspp

• Further examination: bacterial infection, dermatomycosis• No treatment effective• May 2012: last iguana developed disease

-> skin biopsy for virological testingRV iti (PCR)-> RV positive (PCR)

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

->

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

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

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) - - -

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

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 lizard Asian 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?

DiscussionDiscussion

•• Origin: Asia, USA, GermanyOrigin: Asia, USA, Germany--> global distribution> global distribution global distribution global 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 other Coinfections with other

viruses/parasites/bacteria/fungiviruses/parasites/bacteria/fungi•• disease after transport disease 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…)?

DiscussionDiscussion

•• Contact with other reptiles/amphibians Contact 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 ulcerativenecrotic dermatitis and hyperkeratosis)necrotic dermatitis and hyperkeratosis) in all lizardsin all lizards--> DD mortality + skin lesions> DD mortality + skin lesions DD mortality skin lesions DD mortality skin lesions

AcknowledgementsAcknowledgements

Christa Schäfer and Silvia Speck:F h bi U l d Ti h iFachgebiet Umwelt und TierhygieneUniversity of HohenheimStuttgart, Germany

ThankThank youyou forfor youryouryy yyattentionattention!!