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Identification and Epidemiology of Streptoccocus iniae and S. agalactiae

Identification and Epidemiology of Streptoccocus iniae and S. agalactiae. Joyce J. Evans, Phillip H. Klesius, Craig A. Shoemaker and David J. Pasnik USDA/ARS Aquatic Animal Health Research Laboratory Chestertown, MD and Auburn, AL. Streptococcus iniae and S. agalactiae Overview.

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Identification and Epidemiology of Streptoccocus iniae and S. agalactiae

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  1. Identification and Epidemiology of Streptoccocus iniae and S. agalactiae Joyce J. Evans, Phillip H. Klesius, Craig A. Shoemaker and David J. Pasnik USDA/ARS Aquatic Animal Health Research Laboratory Chestertown, MD and Auburn, AL

  2. Streptococcus iniae and S. agalactiaeOverview • Global Distribution • Clinical Disease Signs • Fish host susceptibility & alternative hosts • Sample Collection, Transport & Storage • Characterization • Conventional , automated & molecular diagnostic techniques • Environmental Influences or parasitism on disease susceptibility

  3. Streptococcus iniae and S. agalactiae • Highly fatal bacterial fish pathogens implicated also in human & bovine infections • Major disease problem in food fish production and feral fish worldwide • Neurotropic disease with high mortality • More than 40 species of fish affected • Affects fish from diverse habitats

  4. Bottlenose dolphin(Tursiops truncatus)* Gulf menhaden (Brevoortia patronus)* Spot (Leiostomus xanthurus)* Atlantic croaker (Micropogon undulatus)* Sea catfish (Arius felix)* Striped mullet (Mugil cephalus)* Pinfish (Lagodon rhomboids)* Stingray (Dasyatis sp)* Silver trout (Cynoscion nothus)* Sea trout (Cynoscion regalis)* Gulf killifish (Fundulus grandis)* Bluefish (Pomatomus saltatrix)* Klunzingeri mullet (Liza klunzingeri)* Gilthead sea bream (Sparus auratus)* European sea bass (Dicentrarchus labrax) Silver pomfret (Pampus argenteus) Barramundi (Lates calcarifer) Yellowtail (Seriola quinqueradita) Snapper (Ocyurus chrysurus) Grunt (Haemulidae sp) Brown spot grouper (Epinephelus coioides) Borneo grouper (Liza macrolepis) Lined piggy (Pomadasys stridens) Parrot fish (Sparisoma aurofrenatum)/ (Sparisoma viride) Black margate (Anisotremus sp) Chubb (Scaridae sp.) Common mackerel (Scomber japonicus) Japanese/ Olive flounder (Paralicthys olivaceus) Whitespotted spinefoot “ rabbitfish” (Siganus canaliculatus) Marbled spinefoot (Siganus rivulatus) Dusky spinefoot (Siganus fuscescens) Lizard fish (Synodus variegates) Estuarine & Marine fish speciesaffected by S.iniae & S. agalactiae

  5. Amazon dolphin (Inia geoffrensis) Mossambique tilapia (Oreochromis mossambicus) Blue tilapia (Oreochromis aureus) Nile tilapia (Oreochromis niloticus)* Tilapia hybrid (Oreochromis niloticus x O. mossambicus)* Red tilapia (Oreochromis niloticus x O. aureus)* Tilapia spp unspecified (Oreochromis spp.)* Golden shiners (Notemigonus crysoleucas) Rainbow trout (Oncorhynchus mykiss) Coho salmon (Oncorhynchus kisutch) Amago (Oncorhynchus rhodurus var. macrostomus) Ayu (Plecoglossus altivelis) Gray mullet (Mugil cephalus) Hybrid striped bass/ Sunshine bass (Morone chrysops x M. saxatilis) Striped bass (M. saxatilis) Freshwater fish speciesaffected by S. iniae &S. agalactiae

  6. Streptococcusiniae & S. agalactiae in tilapia Countries where Streptococcus iniae has been reported ( 9 ) Countries where Streptococcus agalactiae has been reported ( 3 ) Countries where both Streptococcus iniae and Streptococcus agalactiae have been reported (4 countries, 3 continents)

  7. S.iniae & S. agalactiaein tilapia species Oreochromis niloticus F / N & E Japan, U.S.A. F / N Thailand, Indonesia, Venezuela, Ecuador, Honduras F / N Thailand Brazil, U.S.A (E) SA Oreochromis mossambicus M / N Israel F / N Taiwan Oreochromis aureus F / E U.S.A. Oreochromis niloticus x O. aureus F / E U.S.A.   F / N Saudi Arabia, IsraelSA Oreochromis spp. F / N Israel, U.S.A, Canada, Columbia, Philippines, China Oreochromis niloticus x O. mossambicus F / N ThailandSA

  8. S. iniae and S. agalactiaeClinical & Behavioral Signs of Disease are similar • Erratic swimming behavior • “C-shaped” body posture • Exophthalmia, multiple ocular lesions • Lethargy, Off feed • Fecal cast & operculum clear-S. agalactiae

  9. S. iniae/ S. agalactiae experimental infections Nile tilapia behavioral signs “C” shape/ erratic swimming Upside down/side to side Head up Head down

  10. Nile tilapia S. iniae/ S. agalactiae disease signs

  11. Ocular and operculaSAlesions cloudy eye hemorrhagic eye opaque eye Popeye missing eye collapsed eye

  12. Body and fin hemorrhagic lesions

  13. Streptococcus iniae & S. agalactiae Isolation Microbiological Collection, Transport & Storage • Culture sites-lethal (brain, HK)/ non-lethal (Nares) • Evans et al., 2000,2001 • Transport system for maintaining viability • Evans et al., 2002 • Survival in frozen fish (archival, retrospective) • Evans et al., 2004

  14. Genus Identification of catalase negative Gram positive cocci in chainsBasis of confusion Genera Hemolysis Lancefield Starch Growth Hydolysis @10 45 6.5% NaCl Streptococcusα/β/n A-H, L, K-N, +/- v  v  - O,U,V, ng Enterococcus α/β/n  D  +/-  +  +  + Lactococcus   α/n  N   -  +  -c  v

  15. Hemolysis on 5% SBA Motility Growth @10°C 45°C in 6.5% NaClbroth Reaction on Bile-Esculinmedia Production of: Pyrroliydonyl arylamidase PYR Leucine aminopeptidase LAP Gas from glucose in MRS broth Susceptibility to Vancomycin (30 ug) Vogas-Prokauer (VP) reaction CAMP test Si Sa + +/- - - + - - - - - - - + - + + - nr + + - - + + Conventional tests to confirm Streptococcus genus and/or species

  16. S. iniae No Lancefield group –non groupable Starch hydrolysis + S. agalactiae Lancefield group B Starch hydrolysis – Conventional techniques to differentiate between S. iniae & S.agalactiae

  17. S. iniae API rapid strep 32- No S.iniae ID-Not in database Biolog- excellent ID Monoclonal antibody-based indirect fluorescent antibody technique- Klesius et al., 2006 S. agalactiae API rapid strep 32- S. agalactiae ID Biolog- excellent ID Commercial multi-test systems & Molecular techniques

  18. Streptococcus iniae research Pathogenesis • S. iniae infectivity and distribution after eye, nare & gill inoculation in Nile tilapia Evans et al., 2000;2001; McNulty et al., 2003 • Effect of density on infection- densities of 11.2 g/L and above sig effect on S. iniae mortality Shoemaker et al, 2000 • Amoxicillin oral feed treatment prior to and during S. iniae infection in Nile tilapia no sig differences in mortality-Evans unpublished

  19. Streptococcus agalactiae research Stress & Disease Susceptibility in Nile tilapia • Environmental influences (DO, UIA) reported as key components in epizootics • Sublethal D0 stress-Evans et al.,2003 • Sublethal Unionized ammonia-(0.35 mg/L) Evans et al.,2006 • Increased susceptibility to S. agalactiae at elevated salinities (30 ppt) at 20 and 30°C Chang and Plumb,1996

  20. S. Iniae & S. agalactiae research Disease Susceptibility in non-traditional parasitized & non parasitized host-Channel catfish Reduced survival among Trichodina sp parasitized channel catfish fry challenged with either S. iniae or S. agalactiaeEvans et al in press Challenged, parasitized fish: Challenged, non-parasitized fish: • Clinical signs No clinical signs • Streptococcus sp. cultured No Streptococcus sp. cultured • 0 to 73% survival 88 to 100% survival

  21. Streptococcus iniae and S. agalactiae Vaccine development Early indicators of need U.S. wide epidemiology survey of the incidence of Streptococcus iniae in catfish, tilapia and hybrid striped bass production farms indicated prevalence Shoemaker, Klesius, Evans, 2001 Worldwide epizootics Fish epizootics in Gulf of Mexico (Plumb et al 1974), Chesapeake Bay (Baya et al 1990) & Kuwait Bay (Evans et al., 2002; Glibert et al., 2002), Thailand (Suanyuk et al 2005) and Brazil (Salvadore et al 2005) due to S. agalactiae.

  22. Acknowledgments • Dr. Richard Shelby- USDA/ARS, Auburn, AL • Crystal Braden- USDA/ARS, Auburn, AL • Daniel Brougher- USDA/ARS, Chestertown, MD • Lisa Biggar- USDA/ARS, Chestertown, MD

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