Native Amphibians Research Articles

Ranavirosis in Invasive Bullfrogs, Belgium

To the Editor: Massive global declines in amphibians have been attributed to various causes, including infectious diseases such as chytridiomycosis and ranavirosis. Chytridiomycosis and ranaviral disease are international notifiable diseases because they have been listed by the World Organisation for Animal Health in its Animal Health Code. Ranavirosis is caused by icosahedral cytoplasmic DNA viruses that belong to the family Iridoviridae, in particular by 4 species of Ranavirus: Frog Virus 3 (FV3), Bohle iridovirus, Ambystoma tigrinum virus, and a possible species Rana catesbeiana virus Z. In Europe, FV3 has been identified in several outbreaks of ranavirosis, characterized by mass deaths, notably in green frogs (Pelophylax sp.) in Denmark, Croatia, and the Netherlands (1,2); Rana temporaria and Bufo bufo in the United Kingdom (3,4); and Alytes obstetricans and Ichthyosaura alpestris in Spain (5). The invasive exotic bullfrog (Lithobates catesbeianus) has been introduced in several European countries and has established large breeding populations in France, Italy, Germany, Greece, and Belgium (6). In addition to their direct effect on native amphibians through competition and predation, bullfrogs are thought to be carriers of chytridiomycosis (7,8) and, possibly, ranaviruses. Although mass deaths of L. catesbeianus tadpoles has been reported in aquaculture facilities, L. catesbeianus tadpoles are generally considered a subclinical reservoir of ranaviruses in the United States (9). To assess the role of bullfrogs as carriers of ranaviruses in Europe, we collected 400 clinically healthy tadpoles of L. catesbeianus from 3 invasive bullfrog populations at Hoogstraten, Belgium (51°47′Ν, 4°75′E) during May–June 2010. All larvae were euthanized as part of an invasive species eradication project and stored at –20°C until further use. At necropsy, liver tissues were collected, and DNA was extracted by using the Genomic DNA Mini Kit (BIOLINE, London, UK). PCR to detect ranavirus was performed as described by Mao et al. (10). Three samples showed positive results with this PCR. These samples were sequenced by using primers M4 and M5 described by Mao et al. (10) and blasted in GenBank. A 100% homology with the common midwife toad (A. obstetricans) ranavirus partial major capsid protein gene (GenBank accession no. {type:entrez-nucleotide,attrs:{text:FM213466.1,term_id:209869735,term_text:FM213466.1}}FM213466.1) was found (5). Despite the low prevalence of Ranavirus infection (0.75%) in the bullfrog tadpoles examined, this study shows that invasive bullfrogs, a known reservoir of chytridiomycosis, are also a likely carrier of ranaviral disease in Europe.

Invasive shrub alters native forest amphibian communities

Although invasive plants can have transformative effects on native plant communities, studies of the consequences of plant invasion for native fauna are generally restricted to primary consumers. Here we investigate whether an invasive shrub, Lonicera maackii, impacts native amphibians and evaluate evidence for the role of invasive plant-induced alteration of forest understory microclimate as a mechanism driving amphibian responses to L. maackii invasion. We sampled amphibian communities in forest plots with high or low density of L. maackii, and monitored microclimate (temperature and humidity at ground level) in the same forest plots. Amphibian species richness and evenness were lower in invaded plots. Invasion also resulted in shifts in amphibian species composition. Mean daily maximum temperature and mean daily temperature were lower in invaded plots, and counts of the Green frog Lithobates clamitans were marginally negatively related to mean daily temperature. Our work illustrates how an invasive ecosystem engineer may affect native organisms with which it shares no trophic connection, and suggests that changes in microclimate may be one mechanism by which alien plants affect communities where they invade.

Early assessment of the impact of alien species: differential consequences of an invasive crayfish on adult and larval amphibians

Aim Early assessment of the impact of invasive alien species is crucial to set up timely management, but often the impact is evident when it is too late for action. We evaluated relationships between the alien crayfish, Procambarus clarkii, the distribution of native amphibians, and the abundance of their larvae. We assessed whether considering measures of reproductive success provide a more prompt measure of impact than considering just species distribution. Location One hundred and twenty-five wetlands in Northern Italy, in an area recently invaded by P. clarkii. Methods We surveyed wetlands to assess the presence of breeding activity of amphibians and the distribution of P. clarkii. We measured the abundance of amphibian larvae before metamorphosis through pipe sampling. We built models analysing the relationships between amphibian and crayfish distribution, while taking into account spatial autocorrelation and environmental features. Analyses were performed at both the species level (generalized linear models and spatial eigenvector mapping) and community level (constrained redundancy analysis). Results In terms of breeding site distribution, only two amphibians (Lissotriton vulgaris and Hyla intermedia) were negatively associated with P. clarkii, while the relationships between other the species and P. clarkii were positive or not significant. However, larval abundance for all amphibian species was negatively associated with the alien crayfish. Analyses performed at community and single species levels yielded consistent results. Main conclusions Procambarus clarkii impacts amphibians through different processes. Newts probably avoid invaded wetlands for breeding. Other species attempt breeding in wetlands with crayfish, but suffer very low success. Considering distribution data alone would not provide a correct picture of the impact of this alien species; measures of reproductive success may allow a more accurate assessment of the impact.

Disease dynamics vary spatially and temporally in a North American amphibian

Local environmental conditions are a primary factor influencing chytridiomycosis, an emerging disease caused by the fungus Batrachochytrium dendrobatidis ( Bd) that has affected over 200 amphibian species worldwide. In North America, seasonality and regional habitat differences predict considerable spatial and temporal disease variability, yet the sparse and opportunistic nature of most studies have provided insufficient data for understanding regional Bd epidemiology. We present a five-year field study that reveals spatial and temporal Bd dynamics across Lithobates yavapaiensis populations in Arizona, USA. Two populations showed no Bd infection or mortality, ten populations showed winter Bd infection, and five populations experienced winter mortality. Infection intensity decreased over winter sampling seasons, whereas mortality and infection prevalence did not change over time. Frogs dying from chytridiomycosis were significantly larger and had significantly higher infection intensities than survivors. We conclude that conserving L. yavapaiensis and other native amphibians requires protection and management of riparian ecosystems to promote populations large enough to survive winter cycles of chytridiomycosis.

Linking the Distribution of an Invasive Amphibian (Rana catesbeiana) to Habitat Conditions in a Managed River System in Northern California

Extensive modifications of river systems have left floodplains some of the most endangered ecosystems in the world and made restoration of these systems a priority. Modified river ecosystems frequently support invasive species to the detriment of native species. Rana catesbeiana (American bullfrog) is an invasive amphibian that thrives in modified aquatic habitats. In 2004–2005 we studied the distribution of bullfrogs along a 98‐km reach of the Trinity River below the Lewiston Dam to identify habitat characteristics associated with bullfrogs and to recommend actions to reduce their prevalence in the system. We also examined native amphibian distributions relative to bullfrogs and disturbance regimes. We used regression techniques to model the distribution of bullfrogs in relation to environmental conditions. Models assessing breeding habitat outperformed models assessing bullfrog presence. Top‐ranked predictor variables of bullfrog distribution included water depth, percent rooted floating vegetation, and river km. Most breeding sites of bullfrogs were relict mine tailing ponds or inactive side channels created during restoration activities in the 1990s. Native species were more common in the lower reach where habitats were less modified, in contrast to the distribution of bullfrogs that dominated the upper, more modified reach. To control bullfrogs along a managed river, we suggest reducing the suitability of breeding sites by decreasing depth or reducing hydroperiod and increasing connection with the active river channel. Current management goals of restoring salmonid habitat and returning the river to a more natural hydrologic condition should aid in control of bullfrogs and improve conditions for native amphibians.

Impact of the small Indian mongoose on native amphibians and reptiles of the Adriatic islands, Croatia

AbstractWe studied impacts of the introduced small Indian mongoose Herpestes auropunctatus on the herpetofauna on six islands in the Adriatic Sea, Croatia, comparing abundances of reptiles and amphibians on three islands with the mongoose to those on three islands without the mongoose. We used four types of sampling surveys: distance‐constrained surveys, visual encounter surveys, special searches and accidental trapping. The horned viper Vipera ammodytes and Balkan green lizard Lacerta trilineata were absent from two mongoose‐infested islands (Korčula and Mljet) and rare on the third (Hvar); they were common only on the mongoose‐free island where they had historically been present (Brač). The European green toad was absent from one mongoose‐infested island, where it had historically been present and rare on the other two. It was common on two of the three mongoose‐free islands. Other herpetofaunal species were either very scarce or completely absent on the three mongoose‐infested islands. Most of these species also occur on the mainland but are already scarce there; some are strictly protected under Appendix II of the Berne Convention. The recent spread of the mongoose to the European mainland suggests the need for urgent control to protect vulnerable herpetofauna.

Can the introduction of Xenopus laevis affect native amphibian populations? Reduction of reproductive occurrence in presence of the invasive species

Biological invasions are regarded as a form of global change and potential cause of biodiversity loss. Xenopus laevis is an anuran amphibian native to sub-Saharan Africa with strong invasive capacity, especially in geographic regions with a Mediterranean climate. In spite of the worldwide diffusion of X. laevis, the effective impact on local ecosystems and native amphibian populations is poorly quantified. A large population of X. laevis occurs in Sicily and our main aim of this work was to assess the consequences of introduction of this alien species on local amphibian populations. In this study we compare the occurrence of reproduction of native amphibians in ponds with and without X. laevis, and before and after the alien colonization. The results of our study shows that, when X. laevis establishes a conspicuous population in a pond system, the populations of Discoglossus pictus, Hyla intermedia and Pelophylax synklepton esculentus show clear signs of distress and the occurrence of reproduction of these native amphibians collapses. In contrast, the populations of Bufo bufo do not appear to be affected by the alien species. Since the Sicilian population of X. laevis shows a strong dispersal capacity, proportionate and quick interventions become necessary to bound the detriment to the Sicilian amphibians populations.

Indirect effects of introduced trout on Cascades frogs (Rana cascadae) via shared aquatic prey

Summary 1. The introduction of trout to montane lakes has negatively affected amphibian populations across the western United States. In northern California’s Klamath–Siskiyou Mountains, introduced trout have diminished the distribution and abundance of a native ranid frog, Rana (=Lithobates) cascadae. This is primarily thought to be the result of predation on frog larvae. However, if trout feed on larval aquatic insects that are available to R. cascadae only after emergence, then resource competition may also affect this declining native amphibian. 2. Stomach contents of R. cascadae were compared between lakes that contained trout and those from which introduced trout were removed. Total prey mass in stomach contents relative to frog body mass was not significantly different between lakes with fish and fish-removal lakes, but in the former R. cascadae consumed a smaller proportion of adult aquatic insects. The stomach contents of fish included larvae of aquatic insects that are, as adults, eaten by R. cascadae. 3. Rana cascadae consumed fewer caddisflies (Trichoptera) and more grasshoppers (Orthoptera) at lakes with higher densities of fish. At lakes with greater aquatic habitat complexity, R. cascadae consumed more water striders (Hemiptera: Gerridae) and terrestrial spiders (Araneae). 4. We suggest that reductions in the availability of emerging aquatic insects cause R. cascadae to consume more terrestrial prey where trout are present. Thus, introduced trout may influence native amphibians directly through predation and, indirectly, through pre-emptive resource competition.

Predator–prey interactions between Synbranchus marmoratus (Teleostei: Synbranchidae) and Hypsiboas pulchellus tadpoles (Amphibia: Hylidae): Importance of lateral line in nocturnal predation and effects of fenitrothion exposure

Environmental contaminants can disrupt interactions between aquatic species by altering community structure. We explored predator–prey interactions between marbled swamp juvenile eels ( Synbranchus marmoratus; predator) and anuran tadpoles ( Hypsiboas pulchellus; prey) in relation to two aspects: the importance of lateral line in the predator and whether the absence of light modifies predation rates; and the effect of a sub-lethal concentration of fenitrothion on both predator and prey. Eels were tested under two sensory conditions (lateral line intact and lateral line blocked by cobalt chloride) in dark conditions. Predation rates were evaluated using different treatments that combined predator and prey exposed or not to insecticide. Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities were also measured in muscle samples of eels and tadpoles to explore whether fenitrothion affects predator and prey differentially. Marbled swamp eels were more efficient in feeding on tadpoles during the night than during the day, showing that lateral line makes an important contribution to prey detection and capture. Regarding pesticide effects, short-term (6 h) exposure to an ecologically relevant fenitrothion dose of 2.5 mg L −1 altered the predator–prey relationship by changing prey behaviour, reducing prey detection and therefore increasing tadpole survival. At this concentration, the outcome of the predator–prey relationship appears biased in favor of the exposed tadpoles, which were released from predation risk, despite their altered behaviour and the higher inhibition percentages of tail BChE (70%) and AChE (51%) than in control individuals. Our study involving these model species and agrochemicals demonstrates that fenitrothion affected the outcome of a predator–prey relationship. Further studies are needed, in these species and other native amphibians, to investigate the nature of the mechanisms responsible for the adverse effects of pesticides on antipredator behaviour and predation efficiency.

Non‐native salmonids affect amphibian occupancy at multiple spatial scales

AbstractAim The introduction of non‐native species into aquatic environments has been linked with local extinctions and altered distributions of native species. We investigated the effect of non‐native salmonids on the occupancy of two native amphibians, the long‐toed salamander (Ambystoma macrodactylum) and Columbia spotted frog (Rana luteiventris), across three spatial scales: water bodies, small catchments and large catchments.Location Mountain lakes at ≥ 1500 m elevation were surveyed across the northern Rocky Mountains, USA.Methods We surveyed 2267 water bodies for amphibian occupancy (based on evidence of reproduction) and fish presence between 1986 and 2002 and modelled the probability of amphibian occupancy at each spatial scale in relation to habitat availability and quality and fish presence.Results After accounting for habitat features, we estimated that A. macrodactylum was 2.3 times more likely to breed in fishless water bodies than in water bodies with fish. Ambystoma macrodactylum also was more likely to occupy small catchments where none of the water bodies contained fish than in catchments where at least one water body contained fish. However, the probability of salamander occupancy in small catchments was also influenced by habitat availability (i.e. the number of water bodies within a catchment) and suitability of remaining fishless water bodies. We found no relationship between fish presence and salamander occupancy at the large‐catchment scale, probably because of increased habitat availability. In contrast to A. macrodactylum, we found no relationship between fish presence and R. luteiventris occupancy at any scale.Main conclusions Our results suggest that the negative effects of non‐native salmonids can extend beyond the boundaries of individual water bodies and increase A. macrodactylum extinction risk at landscape scales. We suspect that niche overlap between non‐native fish and A. macrodactylum at higher elevations in the northern Rocky Mountains may lead to extinction in catchments with limited suitable habitat.

First Evidence of Batrachochytrium dendrobatidis in China: Discovery of Chytridiomycosis in Introduced American Bullfrogs and Native Amphibians in the Yunnan Province, China

Although the chytrid fungus Batrachochytrium dendrobatidis (Bd), the etiological agent of amphibian chytridiomycosis, has been implicated in mass mortality and population declines on several continents around the world, there have been no reports on the presence of Bd infections in amphibians in China. We employed quantitative PCR and histological techniques to investigate the presence of Bd in introduced North American bullfrogs (Rana catesbeiana) (referred to hereafter as bullfrog) and native amphibians in bullfrog-invaded areas of the Yunnan Province, China. A total of 259 samples at five wild sites were collected between June and September in 2007 and 2008, including bullfrogs and four native amphibian species (Rana pleuraden, Rana chaochiaoensis, Odorrana andersonii, and Bombina maxima). In addition, 37 samples of adult bullfrogs were obtained from a food market. Bd infections were discovered in bullfrogs and three native amphibian species from all of the surveyed sites. Of the 39 Bd-positive samples, 35 were from wild-caught bullfrog tadpoles, postmetamorphic bullfrogs, R. pleuraden, R. chaochiaoensis, and O. andersonii, and four were from adult bullfrogs from the market. Our results provide the first evidence of the presence of Bd in Chinese amphibians, suggesting that native amphibian diversity in China is at risk from Bd. There is an urgent need to monitor the distribution of Bd in amphibians in China and understand the susceptibility of native amphibian species to chytridiomycosis. Strict regulations on the transportation of bullfrogs and the breeding of bullfrogs in markets and farms should be drafted in order to stop the spread of Bd by bullfrogs.

Lethal Effect and In Vivo Genotoxicity of Profenofos to Chinese Native Amphibian (Rana spinosa) Tadpoles

Amphibian populations are decreasing in size due to environmental stressors in most areas of southern China. Pesticides are known to be a group of potential stressors to amphibians, especially in agricultural ecosystems. Profenofos, an organophosphate insecticide and acaricide, is widely used for controlling insect pests in China. The aim of this study is to evaluate the acute lethality and genotoxicity of profenofos to amphibian under controlled conditions. Results showed that profenofos was highly lethal to tadpoles of Rana spinosa, with 50% lethal concentration (LC(50)) values of 1.59, 1.14, 0.77, and 0.58 mg l(-1) at 24, 48, 72, and 96 h, respectively. DNA damage of erythrocytes was observed by alkaline single-cell gel electrophoresis assay at all tested sublethal concentrations. The study also showed, by micronucleus test, that profenofos at moderate to high sublethal concentration might have genotoxicity to the tadpole after 96 h exposure. Furthermore, based on our results, it is suggested that the alkaline single-cell gel electrophoresis assay could be used as a screening tool for biomonitoring of pesticide contamination in aquatic systems or agricultural ecosystems.

Using occupancy models to understand the distribution of an amphibian pathogen, Batrachochytrium dendrobatidis

Batrachochytrium dendrobatidis is a fungal pathogen that is receiving attention around the world for its role in amphibian declines. Study of its occurrence patterns is hampered by false negatives: the failure to detect the pathogen when it is present. Occupancy models are a useful but currently underutilized tool for analyzing detection data when the probability of detecting a species is <1. We use occupancy models to evaluate hypotheses concerning the occurrence and prevalence of B. dendrobatidis and discuss how this application differs from a conventional occupancy approach. We found that the probability of detecting the pathogen, conditional on presence of the pathogen in the anuran population, was related to amphibian development stage, day of the year, elevation, and human activities. Batrachochytrium dendrobatidis was found throughout our study area but was only estimated to occur in 53.4% of 78 populations of native amphibians and 66.4% of 40 populations of nonnative Rana catesbeiana tested. We found little evidence to support any spatial hypotheses concerning the probability that the pathogen occurs in a population, but did find evidence of some taxonomic variation. We discuss the interpretation of occupancy model parameters, when, unlike a conventional occupancy application, the number of potential samples or observations is finite.

Amphibian chytridiomycosis in Japan: distribution, haplotypes and possible route of entry into Japan

A serious disease of amphibians caused by the chytrid fungus Batrachochytrium dendrobatidis was first found in Japan in December 2006 in imported pet frogs. This was the first report of chytridiomycosis in Asia. To assess the risk of pandemic chytridiomycosis to Japanese frogs, we surveyed the distribution of the fungus among captive and wild frog populations. We established a nested PCR assay that uses two pairs of PCR primers to amplify the internal transcribed spacer (ITS) region of a ribosomal RNA cassette to detect mild fungal infections from as little as 0.001 pg (1 fg) of B. dendrobatidis DNA. We collected swab samples from 265 amphibians sold at pet shops, 294 bred at institutes and 2103 collected at field sites from northern to southwestern Japan. We detected infections in native and exotic species, both in captivity and in the field. Sequencing of PCR products revealed 26 haplotypes of the B. dendrobatidis ITS region. Phylogenetic analysis showed that three of these haplotypes were specific to the Japanese giant salamander (Andrias japonicus) and appeared to have established a commensal relationship with this native amphibian. Many other haplotypes were carried by alien amphibians. The highest genetic diversity of B. dendrobatidis was found in the American bullfrog (Rana catesbeiana). Some strains of B. dendrobatidis appeared to be endemic to Japanese native amphibians, but many alien strains are being introduced into Japan via imported amphibians. To improve chytridiomycosis risk management, we must consider the risk of B. dendrobatidis changing hosts as a result of anthropogenic disturbance of the host-specific distribution of the fungus.

Chytridiomycosis in frogs from Uruguay

Amphibian chytridiomycosis caused by Batrachochytrium dendrobatidis is reported in Uruguayan native amphibians for the first time. Histological evidence of infection was observed in tadpoles of Hypsiboas pulchellus, Odontophrynus maisuma, Physalaemus henselii, and Scinax squalirostris. The effects of chytridiomycosis on these species are still unknown. However, the disease is of potential concern for the conservation of the apparently declining species P. henselii and also for O. maisuma, given its restricted distribution in habitats which are being increasingly disturbed.

Western Mosquitofish Predation on Native Amphibian Eggs and Larvae

ABSTRACT The western mosquitofish (Gambusia affinis) is commonly stocked for mosquito control, but this species has the potential to negatively impact native amphibians through predation on early life stages. We examined predation by western mosquitofish on eggs and larvae of wood frog (Rana sylvatica), American toad (Bufo americanus), eastern tiger salamander (Ambystoma tigrinum tigrinum), and western chorus fiog (Pseudacris triseriata triseriata) using a series of laboratory microcosm experiments. There were no eggs of any amphibian species consumed by western mosquitofish, and only one predation attempt was observed on a western chorus frog egg. No predation was observed on larvae of wood frog or American toad, and there was no significant difference in the percentage of larvae of these two species missing at the end of the experimental trials (11% and 6%, respectively). However, eastern tiger salamander and western chorus fiog larvae experienced significantly higher predation rates than the other two amphibian species, with approximately 59% of tiger salamander and 92% of chorus frog larvae missing at the end of the experiments. Based on these results, we do not recommend stocking western mosquitofish into waterways that contain early life stages of native amphibian species.

Distribution of long-toed salamanders and introduced trout in high- and low-elevation wetlands in southwestern Alberta, Canada

Several species of trout have been introduced into most wetlands within the mountain and foothill regions of southwestern Alberta, Canada. In this study, we used standard survey methods to evaluate the distribution of long-toed salamander (Ambystoma macrodactylum) larvae relative to introduced trout within 27 low-elevation wetlands and 30 high-elevation wetlands in Waterton Lakes National Park and the adjacent Castle Special Management Area. Results provided indirect evidence of salamander displacement by trout. At high elevations, salamander larvae were not detected in any of the sites that contained trout. The complete allotopy between trout and long-toed salamanders within high-elevation sites supports previous studies completed within the core of this species' range, further documenting the widespread negative impacts of introduced trout on native amphibians in upper basins. Similarly, salamander larvae were present in each of the low-elevation wetlands that lacked trout; they co-occurred in only 4 of 27 low-elevation wetlands. Allotopy within lower elevation sites is particularly important from a conservation perspective because, relative to populations in high-elevation wetlands, these populations tend to contain individuals with rapid growth and high dispersal rates.

Evidence of indirect impacts of introduced trout on native amphibians via facilitation of a shared predator

Hyperpredation occurs when non-native prey facilitate invasive predators, which then suppress native prey. Direct impacts of introduced fish on amphibians are well studied, but the role of fish in supporting shared predators has not been considered. We present evidence for indirect effects of trout on amphibians through snake predation. Analyses of the diet, distribution and density of the Pacific coast aquatic garter snake (Thamnophis atratus) relative to the sympatric common garter snake (Thamnophis sirtalis) in the Klamath Mountains of California suggest that trout introductions facilitated expansion of T. atratus by providing alternative prey. T. atratus diet included trout and amphibians whereas T. sirtalis preyed solely upon amphibians. The distribution and density of T. atratus matched that of introduced trout instead of native amphibians. Populations of T. atratus could reach high densities in the absence of high densities of amphibians. When the snakes opportunistically prey upon amphibians whose numbers are already directly impacted by trout, they can cause significant additional declines. When T. atratus was present in lake basins, native Cascades frogs (Rana cascadae) were rarer than in basins without T. atratus. This case differs from other hyperpredation studies because the two prey species also interact via intraguild predation. Given the worldwide practice of stocking fish into aquatic habitats, it is important to understand the consequences of the practice on food-web structure and ecosystem functioning. Bottom-up impacts of introduced predators should be considered as well as top-down so that managers can incorporate the range of ecosystem-level effects into conservation goals and decisions.

Population genetics reveals origin and number of founders in a biological invasion

Propagule pressure is considered the main determinant of success of biological invasions: when a large number of individuals are introduced into an area, the species is more likely to establish and become invasive. Nevertheless, precise data on propagule pressure exist only for a small sample of invasive species, usually voluntarily introduced. We studied the invasion of the American bullfrog, Rana catesbeiana, into Europe, a species that is considered a major cause of decline for native amphibians. For this major invader with scarce historical data, we used population genetics data (a partial sequence of the mitochondrial cytochrome b gene) to infer the invasion history and to estimate the number of founders of non-native populations. Based on differences between populations, at least six independent introductions from the native range occurred in Europe, followed by secondary translocations. Genetic diversity was strongly reduced in non-native populations, indicating a very strong bottleneck during colonization. We used simulations to estimate the precise number of founders and found that most non-native populations derive from less than six females. This capability of invasion from a very small number of propagules challenges usual management strategies; species with such ability should be identified at an early stage of introduction.

Survey for the amphibian chytrid Batrachochytrium dendrobatidis in Hong Kong in native amphibians and in the international amphibian trade

Chytridiomycosis, caused by the pathogen Batrachochytrium dendrobatidis, is responsible for many amphibian declines and has been identified in wild amphibian populations on all continents where they exist, except for Asia. In order to assess whether B. dendrobatidis is present on the native amphibians of Hong Kong, we sampled wild populations of Amolops hongkongensis, Paa exilispinosa, P. spinosa and Rana chloronota during 2005-2006. Amphibians infected with B. dendrobatidis have been found in the international trade, so we also examined the extent and nature of the amphibian trade in Hong Kong during 2005-2006, and assessed whether B. dendrobatidis was present in imported amphibians. All 274 individuals of 4 native amphibian species sampled tested negative for B. dendrobatidis, giving an upper 95% confidence limit for prevalence of 1.3%. Approximately 4.3 million amphibians of 45 species from 11 countries were imported into Hong Kong via air over 12 mo; we did not detect B. dendrobatidis on any of 137 imported amphibians sampled. As B. dendrobatidis generally occurs at greater than 5% prevalence in infected populations during favorable environmental conditions, native amphibians in Hong Kong appear free of B. dendrobatidis, and may be at severe risk of impact if it is introduced. Until it is established that the pathogen is present in Hong Kong, management strategies should focus on preventing it from being imported and decreasing the risk of it escaping into the wild amphibian populations if imported. Further research is needed to determine the status of B. dendrobatidis in Hong Kong with greater certainty.