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Ranavirus outbreak in captive eastern box turtles
- 1. TURTLE (TERRAPENE CAROLINA CAROLINA) POPULATION WITH MYCOPLASMA AND HERPESVIRUS CO-INFECTION: MANAGEMENT AND MONITORING RR Sim, AN Wack, MC Allender, KJ Murphy, E Bronson
- 2. Start of the outbreak July 13: Presumed index case Found dead in naturalized exhibit, too autolyzed for diagnostics July 19 Second autolyzed case Three moribund turtles: oral swabs from 2 of 3 PCR for Ranavirus (FV-3): 2 of 2 positive PCR for Herpesvirus (TerHV1): 1 of 2 positive All three died July 20-22
- 3. Ambient temperature 20 25 30 35 40 45 7/5/2011 7/7/2011 7/9/2011 7/11/2011 7/13/2011 7/15/2011 7/17/2011 7/19/2011 7/21/2011 7/23/2011 7/25/2011 7/27/2011 7/29/2011 7/31/2011 8/2/2011 8/4/2011 Max. Temp. Mean Temp. Min. Temp. Temp (ºC)
- 4. Eastern Box Turtles severe diffuse fibrinonecrotic glossitis mild to moderate multifocal fibrinonecrotic gastritis
- 5. Management for 22 EBTs Quarantine Sorted & segregated by level of disease: No lesions, mild, or severe Individuals moved up severity scale as needed
- 6. Treatment De-wormed: Pyrantel Pamoate 5 mg/kg PO once Antibiotics: Ceftazidime: 20 mg/kg IM q72h Enrofloxacin: 10 mg/kg IM q48h for severe group Famciclovir: 10, 20, or 30 mg/kg PO q24h Random assignment of dosage level within each severity group Fluid & Nutritional support
- 7. Duration of treatment Continuation of treatment based on clinical signs No lesion group: 34 d Other severity groups: Treatment stopped after 3 checks q3d without lesions Median duration of treatment All survivors: 34 d (range 34 – 80 d) Ranavirus-pos. survivors: 49 d (range 34 – 80 d) day 80, the surviving 14 turtles were returned to the outdoor exhibit for brumation
- 8. Molecular Monitoring Oral swabs days 0, 10, 34, and 60 were tested for Ranavirus by quantitative real-time PCR day 0 for Herpesvirus and Mycoplasma by conventional PCR Overall prevalence of viral shedding Ranavirus-positive: 86.4% (19/22) 6 asymptomatic, positive at start of treatment period 2 of 6 developed clinical signs Herpesvirus-positive: 54.5% (12/22) Mycoplasma-positive: 68.2% (15/22)
- 9. qPCR for Ranavirus 0.0% 20.0% 40.0% 60.0% 80.0% 0 10 34 60 16/22 9/18 5/16 0/14 0.00E+00 2.50E+07 5.00E+07 7.50E+07 1.00E+08 0 10 34 60 Prevalence of viral shedding Median viral copies Day Day
- 10. Characterizing co-infection Ranavirus- negative Ranavirus- positive Infection None HV & M RV alone RV & M RV & HV RV, HV & M Number 1 2 4 5 2 8 3 19 Prevalenc e 4.5% 9.1% 18.2% 22.7% 9.1% 36.4% 13.6% 86.4% Survival 100% 100% 50% 40% 50% 62.5%
- 11. Post-brumation follow-up All 14 turtles survived brumation 5-months after emergence: no clinical signs and all Ranavirus- negative Herpesvirus- positive at Courtesy of in.gov
- 12. Discussion Origin of the outbreak and Transmission Treatment choices: Famciclovir, Esophagostomy tubes Characterization of Ranavirus infection Asymptomatic infection Relationship of severity and survival? Role of co-infection with Mycoplasma and Herpesvirus Immune response?
- 13. Conclusion Ranavirus infection in this captive population was managed in a way that increased survival as compared to previous reports Co-infection of Herpesvirus and Mycoplasma does not seem to influence survival Level of intensive intervention impractical for free-ranging population Nutritional and fluid support is felt to be a critical component of care Famciclovir is of uncertain utility, but should be further investigated
Editor's Notes
- Ranavirus: Frog Virus-3Signs in cheloniansTan fibrinonecrotic plaques associated with necrotizing and ulcerative stomatitis, esophagitis, gastritis, and enterocolitisConjunctivitis and blepharoedemaFibrinoidvasculitis mostly in the spleenDegenerative hepatitis with necrosis Hyperemic edematous lungsEffect of concurrent herpesvirus (Stomatitis, rhinitis, conjunctivitis, CNS disease)? Mycoplasma?
- To monitor the progression of disease on a molecular level. Oral swabs were collected on days 0, 10, 34, and 60. They were frozen at -80 degrees Celsius until testing. Real time TaqManPCR was done to quantify the number of Ranavirusviral copies being orally shed by individual Box turtles and conventional PCR was done at day 0 to get a baseline of Herpesvirus in the population.77% were shedding Ranavirus particles at some point in the disease course. Of note 6 Box turtles were asymptomatic shedders at the beginning of the treatment period and only 2 of these 6 went on to develop clinical signs. 54.5% of the Box turtles were Herpesvirus-positive at the start of the study.- Ranavirus -positive: 86.4% (19/22) - Herpesvirus-positive: 54.5% (12/22) - Mycoplasma-positive: 68.2% (15/22)