Mazzoni

566 views

Published on

Surveillance of ranavirus in frog farms and surrounding environments in Brazil

Published in: Education, Technology, Lifestyle
0 Comments
0 Likes
Statistics
Notes
  • Be the first to comment

  • Be the first to like this

No Downloads
Views
Total views
566
On SlideShare
0
From Embeds
0
Number of Embeds
1
Actions
Shares
0
Downloads
5
Comments
0
Likes
0
Embeds 0
No embeds

No notes for slide

Mazzoni

  1. 1. Surveillance onSurveillance on RANAVIRUSESRANAVIRUSES 2nd. INTERNATIONAL SYMPOSIUM ON RANAVIRUSES Knoxville, TN – 27-28 July 2013 RANAVIRUSESRANAVIRUSES inin Farmed frogs and SympatricFarmed frogs and Sympatric Amphibia, Fish & ReptilesAmphibia, Fish & Reptiles Rolando Mazzoni & Adriano Q. Mesquita CPA/EVZ/UFG BRAZIL
  2. 2. BrazilBrazil American bullfrogsAmerican bullfrogs BACKGROUND Virus isolation from sick tadpolesVirus isolation from sick tadpoles (farmed)(farmed)
  3. 3. Presence ofPresence of “FV3 like” viruses“FV3 like” viruses BACKGROUND “FV3 like” viruses“FV3 like” viruses
  4. 4. OLDER TADPOLES & FROGSOLDER TADPOLES & FROGS BACKGROUND 10% Positive10% Positive (conventional PCR)(conventional PCR) No virus associated to lessionsNo virus associated to lessions (TEM)(TEM)
  5. 5. Bullfrogs are TEVBullfrogs are TEV -- FV3 carriersFV3 carriers BACKGROUND WOLF et al., 1968; WILLIAMS et al., 2005; WHITTINGTON et al., 2010 PRESENCE # DISEASE !!!!
  6. 6. FV3 in American BullfrogsFV3 in American Bullfrogs •• INTRODUCEDINTRODUCED oror •• SouthSouth--American VirusAmerican Virus BACKGROUND Frog farms shed virus to the environment?Frog farms shed virus to the environment? Pathogenic to other aquatic organisms?Pathogenic to other aquatic organisms?
  7. 7. OBJECTIVESOBJECTIVES Ranavirus surveillanceRanavirus surveillance:: •• Farmed American bullfrogsFarmed American bullfrogs •• Surrounding areasSurrounding areas •• Other areasOther areas•• Other areasOther areas Disease reportsDisease reports
  8. 8. MATERIAL & METHODSMATERIAL & METHODS Samples were obtained from Am. bullfrogsSamples were obtained from Am. bullfrogs •• EggsEggs •• TadpolesTadpoles •• FrogsFrogs Sympatric amphibian, fish & reptilesSympatric amphibian, fish & reptiles •• Frogs (Frogs (Leptodactylus ocellatusLeptodactylus ocellatus;; TrachycephalusTrachycephalus typhoniustyphonius;; •• Toads (Toads (BufoBufo marinusmarinus)) •• Lizards (Lizards (PhyllopezusPhyllopezus pollicarispollicaris)) •• Fish (Fish (Oreochromis niloticusOreochromis niloticus)) •• Snakes (Snakes (BothropsBothrops jararacussujararacussu))
  9. 9. GOIAS BRAZIL SAO PAULO SAMPLED AREAS
  10. 10. GOIAS FARMS LOCATION AND SAMPLING SITES Farm 3 Farm 2
  11. 11. SAMPLED ORGANSSAMPLED ORGANS •• LIVERLIVER •• KIDNEYKIDNEY •• SPLEENSPLEEN MATERIAL & METHODS •• SPLEENSPLEEN •• BLOODBLOOD •• GONADSGONADS •• SKINSKIN •• MUSCLEMUSCLE
  12. 12. METHODMETHOD •• REAL TIME PCRREAL TIME PCR MATERIAL & METHODS •• REAL TIME PCRREAL TIME PCR •• TARGET SEQUENCE = MCPTARGET SEQUENCE = MCP
  13. 13. Positions: 97689 to 97751 Frog virus 3, complete genome GenBank: AY548484.1 Tan et al. (2004) MATERIAL & METHODS RCVBRRCVBR--F = GAGCGTCACCCTCTCATTCF = GAGCGTCACCCTCTCATTC RCVBRRCVBR--R = GCGTCCAGGTATGCCGTGR = GCGTCCAGGTATGCCGTG RCVBRRCVBR--S = CGACATCAGCGCCCAGTCS = CGACATCAGCGCCCAGTC Tm = 61°C
  14. 14. RESULTSRESULTS •• Positive percentage for RVs in farmed bullfrogsPositive percentage for RVs in farmed bullfrogs Farm % positive 01 1501 (40 samples) 15 02 (45 samples) 35 03 (150 samples) 100
  15. 15. RESULTS Positive percentage for RVs in sympatric speciesPositive percentage for RVs in sympatric species from Farm 03from Farm 03 Bullfrogs (150 samples) Amphibian (12 samples) Reptiles (10 samples) Fish (20 samples) 100 10 10 10
  16. 16. RESULTS Amphibian Reptiles Fish Results for RVs in sympatric species from regionsResults for RVs in sympatric species from regions where bullfrogs were not introducedwhere bullfrogs were not introduced Amphibian (12 samples) Reptiles (8 samples) Fish (12 samples) Negative
  17. 17. RESULTS DISEASE REPORTS IN FARMED FROGSDISEASE REPORTS IN FARMED FROGS Farm 01 Sporadic outbreak (once)(once) 02 No outbreak 03 100% mortality in tadpoles
  18. 18. RESULTS DISEASE REPORTS IN OTHER SPECIESDISEASE REPORTS IN OTHER SPECIES NO MORTALITIES OR DISEASES REGISTERED
  19. 19. CONCLUSIONS Farmed FrogsFarmed Frogs •• High to moderate presence of RVsHigh to moderate presence of RVs Farms with High RV prevalenceFarms with High RV prevalenceFarms with High RV prevalenceFarms with High RV prevalence •• Poor managementPoor management •• Whole cycleWhole cycle •• High densitiesHigh densities •• No biosecurity measuresNo biosecurity measures
  20. 20. CONCLUSIONS Sympatric speciesSympatric species •• Low detectionLow detection •• No disease reportedNo disease reported Other areasOther areas = No detection= No detection
  21. 21. CONCLUSIONS Ranavirus in Frog FarmsRanavirus in Frog Farms •• IntroducedIntroduced withwith AmericanAmerican bullfrogsbullfrogs •• ContaminatingContaminating sympatricsympatric speciesspecies Sympatric speciesSympatric species •• NoNo diseasedisease yetyet reportedreported•• NoNo diseasedisease yetyet reportedreported ResistanceResistance//lowlow susceptibilitysusceptibility ?? Low virulence ?Low virulence ? Higher sensitivity with RTHigher sensitivity with RT--PCRPCR
  22. 22. FUTURE DIRECTIONS Experimental ChallengesExperimental Challenges SequencingSequencing Compare RCVCompare RCV--BR with RCVBR with RCV--Z and FV3Z and FV3 •• Virulence genes id.Virulence genes id.
  23. 23. THANKS !THANKS !

×