First recorded outbreak of Veronaea botryosa in North American amphibians: Clinicopathologic features of a rare cause of phaeohyphomycosis in captive White's tree frogs (Litoria caerulea).

We describe fatal phaeohyphomycosis due to Veronaea botryosa in captive White's tree frogs (Litoria caerulea), the first confirmed report in amphibians in North America. Over 15 months, six frogs developed ulcerative dermatitis on distal extremities/ventrum, which in one animal progressed to vasculitis and necrotizing osteomyelitis. All six frogs died. Clinicopathologic findings, diagnostic challenges, and control are discussed. Emerging fungi such as V. botryosa pose serious concerns for zoonosis and potential spread through the pet trade.

Response to Comment on "Amphibian fungal panzootic causes catastrophic and ongoing loss of biodiversity".

Lambert et al question our retrospective and holistic epidemiological assessment of the role of chytridiomycosis in amphibian declines. Their alternative assessment is narrow and provides an incomplete evaluation of evidence. Adopting this approach limits understanding of infectious disease impacts and hampers conservation efforts. We reaffirm that our study provides unambiguous evidence that chytridiomycosis has affected at least 501 amphibian species.

Amphibian fungal panzootic causes catastrophic and ongoing loss of biodiversity.

Anthropogenic trade and development have broken down dispersal barriers, facilitating the spread of diseases that threaten Earth's biodiversity. We present a global, quantitative assessment of the amphibian chytridiomycosis panzootic, one of the most impactful examples of disease spread, and demonstrate its role in the decline of at least 501 amphibian species over the past half-century, including 90 presumed extinctions. The effects of chytridiomycosis have been greatest in large-bodied, range-restricted anurans in wet climates in the Americas and Australia. Declines peaked in the 1980s, and only 12% of declined species show signs of recovery, whereas 39% are experiencing ongoing decline. There is risk of further chytridiomycosis outbreaks in new areas. The chytridiomycosis panzootic represents the greatest recorded loss of biodiversity attributable to a disease.

The Emerging Amphibian Fungal Disease, Chytridiomycosis: A Key Example of the Global Phenomenon of Wildlife Emerging Infectious Diseases.

The spread of amphibian chytrid fungus, Batrachochytrium dendrobatidis, is associated with the emerging infectious wildlife disease chytridiomycosis. This fungus poses an overwhelming threat to global amphibian biodiversity and is contributing toward population declines and extinctions worldwide. Extremely low host-species specificity potentially threatens thousands of the 7,000+ amphibian species with infection, and hosts in additional classes of organisms have now also been identified, including crayfish and nematode worms.Soon after the discovery of B. dendrobatidis in 1999, it became apparent that this pathogen was already pandemic; dozens of countries and hundreds of amphibian species had already been exposed. The timeline of B. dendrobatidis's global emergence still remains a mystery, as does its point of origin. The reason why B. dendrobatidis seems to have only recently increased in virulence to catalyze this global disease event remains unknown, and despite 15 years of investigation, this wildlife pandemic continues primarily uncontrolled. Some disease treatments are effective on animals held in captivity, but there is currently no proven method to eradicate B. dendrobatidis from an affected habitat, nor have we been able to protect new regions from exposure despite knowledge of an approaching "wave" of B. dendrobatidis and ensuing disease.International spread of B. dendrobatidis is largely facilitated by the commercial trade in live amphibians. Chytridiomycosis was recently listed as a globally notifiable disease by the World Organization for Animal Health, but few countries, if any, have formally adopted recommended measures to control its spread. Wildlife diseases continue to emerge as a consequence of globalization, and greater effort is urgently needed to protect global health.

Amphibian Chytrid Fungus in Madagascar neither Shows Widespread Presence nor Signs of Certain Establishment.

The global spread of amphibian chytrid fungus (Batrachochytrium dendrobatidis, Bd) is associated with amphibian mass mortality, population decline, and extinction. Over the past decade, concern has been expressed for the potential introduction of Bd to Madagascar, a global hotspot of amphibian biodiversity. Following years without detection, widespread Bd presence in Madagascar has now been reported (Bletz et al. 2015a), raising international conservation concern. Before reacting to this finding with a significant management response, the accuracy and context of the data warrant cautious review. Re-examination of a 10-year dataset together with results from more recent surveillance (Kolby et al. 2015) does not yet demonstrate widespread Bd presence. Detection of Bd at "positive" locations in Madagascar has been inconsistent for unknown reasons. Whether Bd is established in Madagascar (i.e. populations are self-sustaining) or instead requires continued introduction to persist also remains uncertain. The deployment of emergency conservation rescue initiatives is expected to target areas where the distribution of Bd and the risk of chytridiomycosis endangering amphibians is believed to overlap. Thus, erroneous description of Bd presence would misdirect limited conservation resources. Standardized surveillance and confirmatory surveys are now imperative to reliably characterize the distribution, potential spread, virulence and overall risk of Bd to amphibians in Madagascar.

Rapid Response to Evaluate the Presence of Amphibian Chytrid Fungus (Batrachochytrium dendrobatidis) and Ranavirus in Wild Amphibian Populations in Madagascar.

We performed a rapid response investigation to evaluate the presence and distribution of amphibian pathogens in Madagascar following our identification of amphibian chytrid fungus (Batrachochytrium dendrobatidis, Bd) and ranavirus in commercially exported amphibians. This targeted risk-based field surveillance program was conducted from February to April 2014 encompassing 12 regions and 47 survey sites. We simultaneously collected amphibian and environmental samples to increase survey sensitivity and performed sampling both in wilderness areas and commercial amphibian trade facilities. Bd was not detected in any of 508 amphibian skin swabs or 68 water filter samples, suggesting pathogen prevalence was below 0.8%, with 95% confidence during our visit. Ranavirus was detected in 5 of 97 amphibians, including one adult Mantidactylus cowanii and three unidentified larvae from Ranomafana National Park, and one adult Mantidactylus mocquardi from Ankaratra. Ranavirus was also detected in water samples collected from two commercial amphibian export facilities. We also provide the first report of an amphibian mass-mortality event observed in wild amphibians in Madagascar. Although neither Bd nor ranavirus appeared widespread in Madagascar during this investigation, additional health surveys are required to disentangle potential seasonal variations in pathogen abundance and detectability from actual changes in pathogen distribution and rates of spread. Accordingly, our results should be conservatively interpreted until a comparable survey effort during winter months has been performed. It is imperative that biosecurity practices be immediately adopted to limit the unintentional increased spread of disease through the movement of contaminated equipment or direct disposal of contaminated material from wildlife trade facilities. The presence of potentially introduced strains of ranaviruses suggests that Madagascar's reptile species might also be threatened by disease. Standardized population monitoring of key amphibian and reptile species should be established with urgency to enable early detection of potential impacts of disease emergence in this global biodiversity hotspot.

Terrestrial Dispersal and Potential Environmental Transmission of the Amphibian Chytrid Fungus (Batrachochytrium dendrobatidis).

Dispersal and exposure to amphibian chytrid fungus (Batrachochytrium dendrobatidis, Bd) is not confined to the aquatic habitat, but little is known about pathways that facilitate exposure to wild terrestrial amphibians that do not typically enter bodies of water. We explored the possible spread of Bd from an aquatic reservoir to terrestrial substrates by the emergence of recently metamorphosed infected amphibians and potential deposition of Bd-positive residue on riparian vegetation in Cusuco National Park, Honduras (CNP). Amphibians and their respective leaf perches were both sampled for Bd presence and the pathogen was detected on 76.1% (35/46) of leaves where a Bd-positive frog had rested. Although the viability of Bd detected on these leaves cannot be discerned from our quantitative PCR results, the cool air temperature, closed canopy, and high humidity of this cloud forest environment in CNP is expected to encourage pathogen persistence. High prevalence of infection (88.5%) detected in the recently metamorphosed amphibians and frequent shedding of Bd-positive residue on foliage demonstrates a pathway of Bd dispersal between aquatic and terrestrial habitats. This pathway provides the opportunity for environmental transmission of Bd among and between amphibian species without direct physical contact or exposure to an aquatic habitat.

First evidence of amphibian chytrid fungus (Batrachochytrium dendrobatidis) and ranavirus in Hong Kong amphibian trade.

The emerging infectious amphibian diseases caused by amphibian chytrid fungus (Batrachochytrium dendrobatidis, Bd) and ranaviruses are responsible for global amphibian population declines and extinctions. Although likely to have been spread by a variety of activities, transcontinental dispersal appears closely associated with the international trade in live amphibians. The territory of Hong Kong reports frequent, high volume trade in amphibians, and yet the presence of Bd and ranavirus have not previously been detected in either traded or free-ranging amphibians. In 2012, a prospective surveillance project was conducted to investigate the presence of these pathogens in commercial shipments of live amphibians exported from Hong Kong International Airport. Analysis of skin (Bd) and cloacal (ranavirus) swabs by quantitative PCR detected pathogen presence in 31/265 (11.7%) and in 105/185 (56.8%) of amphibians, respectively. In addition, the water in which animals were transported tested positive for Bd, demonstrating the risk of pathogen pollution by the disposal of untreated wastewater. It is uncertain whether Bd and ranavirus remain contained within Hong Kong's trade sector, or if native amphibians have already been exposed. Rapid response efforts are now urgently needed to determine current pathogen distribution in Hong Kong, evaluate potential trade-associated exposure to free-ranging amphibians, and identify opportunities to prevent disease establishment.

Presence of the amphibian chytrid fungus Batrachochytrium dendrobatidis in native amphibians exported from Madagascar.

The emerging infectious disease chytridiomycosis is driven by the spread of amphibian chytrid fungus (Batrachochytrium dendrobatidis, Bd), a highly virulent pathogen threatening global amphibian biodiversity. Although pandemic in distribution, previous intensive field surveys have failed to detect Bd in Madagascar, a biodiversity hotspot home to hundreds of endemic amphibian species. Due to the presence of Bd in nearby continental Africa and the ecological crisis that can be expected following establishment in Madagascar, enhanced surveillance is imperative. I sampled 565 amphibians commercially exported from Madagascar for the presence of Bd upon importation to the USA, both to assist early detection efforts and demonstrate the conservation potential of wildlife trade disease surveillance. Bd was detected in three animals via quantitative PCR: a single Heterixalus alboguttatus, Heterixalus betsileo, and Scaphiophryne spinosa. This is the first time Bd has been confirmed in amphibians from Madagascar and presents an urgent call to action. Our early identification of pathogen presence prior to widespread infection provides the necessary tools and encouragement to catalyze a swift, targeted response to isolate and eradicate Bd from Madagascar. If implemented before establishment occurs, an otherwise likely catastrophic decline in amphibian biodiversity may be prevented.

Amphibian chytrid fungus Batrachochytrium dendrobatidis in Cusuco National Park, Honduras.

Amphibian population declines in Honduras have long been attributed to habitat degradation and pollution, but an increasing number of declines are now being observed from within the boundaries of national parks in pristine montane environments. The amphibian chytrid fungus Batrachochytrium dendrobatidis has been implicated in these declines and was recently documented in Honduras from samples collected in Pico Bonito National Park in 2003. This report now confirms Cusuco National Park, a protected cloud forest reserve with reported amphibian declines, to be the second known site of infection for Honduras. B. dendrobatidis infection was detected in 5 amphibian species: Craugastor rostralis, Duellmanohyla soralia, Lithobates maculata, Plectrohyla dasypus, and Ptychohyla hypomykter. D. soralia, P. dasypus, and P. hypomykter are listed as critically endangered in the IUCN Red List of Threatened Species and have severely fragmented or restricted distributions. Further investigations are necessary to determine whether observed infection levels indicate an active B. dendrobatidis epizootic with the potential to cause further population declines and extinction.