Evan H Campbell Grant Northeast Amphibian Research and Monitoring Initiative US Geological Survey Patuxent Wildlife Research Center

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Evan H Campbell Grant Northeast Amphibian Research and Monitoring Initiative US Geological Survey Patuxent Wildlife Research Center New England Chapter of the Wildlife Society Spring Workshop 2015

Color slides of 5 common lethal infectious diseases of amphibians Considerations for surveillance and monitoring of diseases at multiple scales

5 Common Lethal Infectious Diseases of Amphibians - Chytrid Fungus - Ranaviruses - Perkinsus-like organism - Ichthyophonus sp. - Ribeiroia ondatrae (fluke)

1) Pathogenic Chytrid Fungus: Batrachochytrium dendrobatidis ( Bd ) Major cause of amphibian population declines worldwide Kills only frogs & toads after metamorphosis; occasionally lethal in adult newts Die-offs are insidious; rarely are more than 2 dead frogs or toads found at a site Mixed effect of disease on populations in northeastern US

Chytridiomycosis: Gross Findings: Tadpoles Larvae: non-lethal oral infection of tadpoles

Chytridiomycosis: Gross Findings: Adult Frogs Lethal in majority of spp. within 2-3 wks of end of metamorphosis Harlequin frog (Panama) Chiricahua leopard frog (Rana chiricahuensis)

Bd: Salamanders Pathogenicity largely unknown; non-lethal to aquatic life-stages, occasionally lethal to red-spotted newts. Susceptibility of terrestrial salamanders is largely unstudied Stream salamanders susceptible; no die-offs observed.

Chytridiomycosis in Amphibians Year of First Detection Museum specimens 02 01 01 c. 95 04 76 01 74 c. 74 02 03 99 00 74 99 06 02 03 00 01 Revised Aug 2010 00 00 01 01 95 97 00 02 Puerto Rico 77

Salamander chytrid fungus Recent discovery of a second chytrid pathogen, Batrachochytrium salamandrivorans sp. nov., that causes lethal skin infections in salamanders, which has resulted in steep declines in salamander populations in northwestern Europe. Several species of US amphibians (including red-spotted newts in NE) are susceptible. Risk of introduction Richgels et al., in prep

2) Ranaviruses Agent: Ranavirus; Family: Iridoviridae - 5 or 6 distinct strains or species in USA - Some strains are host-specific, other strains may be panzootic Epizootiology: - Kills larvae & larvae in metamorphosis - Seldom affects adult amphibians in USA - Onset is sudden (explosive) - Mortality rate often >95% of larvae

Ranavirus Infection Gross Findings: 1. Petechia or ulcers in skin

Ranavirus Infection Gross Findings: 2. Effusions under skin and into body cavity

Bunch of dead decaying tadpoles Matt Gray, UT

Ranavirus Die-offs: Frogs and Toads (tadpoles) Revised Aug 2010 Alaska Puerto Rico Hawai i

Ranavirus Die-offs: Salamanders (larvae) Revised Aug 2010 Alaska Puerto Rico Hawai i

Source: Global Ranavirus reporting system (2013)

3) Perkinsus-like Organism Kills ranid tadpoles only, rarely other spp. Die-offs may continue for weeks with bloated weak tadpoles floating on surface Often causes >90% mortality of ranid tadpoles in a pond (hence, little recruitment) Mississippi gopher frog (Rana sevosa)

Perkinsus-like Organism: Gross Findings - Mildly to massively enlarged light yellow liver, spleen, pronephroi & mesonephroi - Abdominal distention

Perkinsus-like Organism: Distribution Revised Aug 2010 Alaska Puerto Rico Hawai i

4) Ichthyophoniasis Eastern red-spotted newt 1. Definition: A slight to severe (and lethal) infection of amphibian skeletal muscle Swollen rump in recently metamorphosed frogs Lethal Infections occur in Eastern red-spotted newts Larval Rana spp. Adult Rana spp. Green frog

Amphibian Ichthyophoniasis: Distribution Revised Aug 2010 Puerto Rico

5) Ribeiroia ondatrae-induced Malformations: Infection by metacercariae must occur at very specific tadpole ages to induce malformation (Gosner 26-28) Polymelia Polypodia Polydactyly & Skin Web Ectromelia

Now that we know what we re looking for And where it has been found

Sampling design considerations for amphibian diseases Interest in diseases at multiple levels Mapping detections/dieoffs Number of ponds affected Prevalence at an infected pond Temporal and spatial dynamics Population-level effects

Two primary challenges (1) addressing uncertainty with (only partially) observed disease dynamics in the wild (2) determining how environmental, spatial, and community processes interact to cause outbreaks and disease persistence.

Challenge 1: Uncertainty (False positives and/or negatives) McClintock et al. 2010. Ecol Lett. 13:659

Uncertainty at each level: Pr(Refuge contains disease)

Pr(Refuge contains disease) Pr(Pond diseased Refuge diseased)

Pr(Refuge contains disease) Pr(Pond diseased Refuge diseased) Pr(Frog diseased Pond diseased)

Pr(Refuge contains disease) Pr(Pond diseased Refuge diseased) Pr(Frog diseased Pond diseased) Pr(Detect disease Frog diseased)

Ignoring uncertainty False + and/or errors Results in biased estimates of presence, prevalence and dynamics. We need to design methods and apply sampling to efficiently address these sources of uncertainty. Probabilistic choice of sampling units Repeat sampling in space and time How to allocate effort across the hierarchy?

2) Determining causes of outbreaks and disease persistence Matt Gray, UT Sampling design considerations (what, when, where) Statistical methods which account for imperfect detection Combinations of field and laboratory studies Research needs: Pathogen ecology Reservoirs/vectors - persistence Mechanism(s) for outbreaks Influence of spatial structure of habitats

Recommendations and challenges Consider hierarchical nature of disease question (presence vs. prevalence; scale)? Dynamics in presence and prevalence annual allocation of effort Uncertainty in ecology of many amphibian diseases. Management? Developing amphibian disease monitoring program for northeast - ehgrant@usgs.gov