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
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)
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
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)