Fast quantitative PCR, locked nucleic acid probes and reduced volume reactions are effective tools for detecting Batrachochytrium dendrobatidis DNA

The fungal pathogen Batrachochytrium dendrobatidis threatens amphibian populations around the world. The ability to detect this pathogen on infected animals and in the environment is critical for understanding and controlling this pandemic. We tested several advances in quantitative PCR (qPCR) techniques to detect B. dendrobatidis DNA. We used a fast PCR thermocycler and enzymes that reduced the volume and the duration of the reaction. We also compared a conventional TaqMan minor groove binding (MGB) probe to an identical locked nucleic acid (LNA) counterpart. The fast qPCR reaction had a high degree of sensitivity to B. dendrobatidis DNA. The LNA probe was effective for detecting B. dendrobatidis DNA and produced results -similar to those of the MGB probe. The modifications that we tested can improve the cost, time efficiency and specificity of quantitative PCR as a tool for detecting pathogen DNA.     

Reptiles as potential vectors and hosts of the amphibian pathogen Batrachochytrium dendrobatidis in Panama

Chytridiomycosis, the disease caused by Batrachochytrium dendrobatidis, is considered to be a disease exclusively of amphibians. However, B. dendrobatidis may also be capable of persisting in the environment, and non-amphibian vectors or hosts may contribute to disease transmission. Reptiles living in close proximity to amphibians and sharing similar ecological traits could serve as vectors or reservoir hosts for B. dendrobatidis, harbouring the organism on their skin without succumbing to disease. We surveyed for the presence of B. dendrobatidis DNA among 211 lizards and 8 snakes at 8 sites at varying elevations in Panama where the syntopic amphibians were at pre-epizootic, epizootic or post-epizootic stages of chytridiomycosis. Detection of B. dendrobatidis DNA was done using qPCR analysis. Evidence of the amphibian pathogen was present at varying intensities in 29 of 79 examined Anolis humilis lizards (32%) and 9 of 101 A. lionotus lizards (9%), and in one individual each of the snakes Pliocercus euryzonus, Imantodes cenchoa, and Nothopsis rugosus. In general, B. dendrobatidis DNA prevalence among reptiles was positively correlated with the infection prevalence among co-occurring anuran amphibians at any particular site (r = 0.88, p = 0.004). These reptiles, therefore, may likely be vectors or reservoir hosts for B. dendrobatidis and could serve as disease transmission agents. Although there is no evidence of B. dendrobatidis disease-induced declines in reptiles, cases of coincidence of reptile and amphibian declines suggest this potentiality. Our study is the first to provide evidence of non-amphibian carriers for B. dendrobatidis in a natural Neotropical environment.     

Transmission of Batrachochytrium dendrobatidis to wood frogs (Lithobates sylvaticus) via a bullfrog (L. catesbeianus) vector

Chytridiomycosis, an emerging infectious disease caused by the chytrid fungus Batrachochytrium dendrobatidis, threatens anuran populations worldwide. Effects of B. dendrobatidis on frog species are variable. Some species typically develop nonlethal infections and may function as carriers; others typically develop lethal infections that can lead to population declines. Nonlethal infections in the bullfrog (Lithobates catesbeianus) are well-documented. In contrast, recently metamorphosed wood frogs (L. sylvaticus) can die from chytridiomycosis. We conducted an ex-situ experiment between May and July 2010 to determine whether B. dendrobatidis-infected bullfrogs could transmit the fungus to wood frog tadpoles when the two species shared a body of water. We tested for B. dendrobatidis infections with quantitative polymerase chain reactions (qPCR) in a subsample of the wood frog tadpoles and in all metamorphosed wood frogs and compared risk of death of froglets exposed and unexposed to infected bullfrogs. We detected B. dendrobatidis sporadically in subsampled treatment tadpoles (nine of 90, 10%) and frequently in treatment froglets (112 of 113, 99.1%). Pooled risk of froglet death was higher (P<0.001) in treatment enclosures than in control enclosures. Our results indicate that, at the low infection loads bullfrogs tend to carry, swabbing for PCR analyses may underestimate prevalence of B. dendrobatidis in this species. We highlight bullfrog disease screening as a management challenge, especially in light of exotic bullfrog colonies on multiple continents and large-scale global trade in this species. We document the importance of quantifying lethal and sublethal effects of bullfrog vectors on B. dendrobatidis-susceptible species.     

Environmental detection of Batrachochytrium dendrobatidis in a temperate climate

The aetiological agent of amphibian chytridiomycosis Batrachochytrium dendrobatidis is a primary cause of amphibian population declines. Current surveillance is based on the detection of B. dendrobatidis in its host but in vitro work suggests infective stages may survive in the abiotic environment for at least 3 mo. We describe here a surveillance system using filtration and quantitative PCR that can detect B. dendrobatidis in small (< 1 l) volumes of water. After assessing the analytical sensitivity of the protocol for both water and sediment samples in the laboratory, we analyzed environmental samples from the Sierra de Guadarrama mountain range in Spain at locations associated with chytrid-related die-offs and at other sites across Spain. B. dendrobatidis was detected in samples from 64% of the ponds in the Sierra de Guadarrama and at 2 sites outside this region, showing that levels of amphibian exposure to B. dendrobatidis are spatially heterogeneous. In experimental microcosms, we detected B. dendrobatidis for up to 12 wk, though we found no evidence for an overall increase in biomass. Our results emphasise the need to further investigate the life cycle of B. dendrobatidis to more completely understand the epidemiology of this emerging pathogen.     

鸿雁迁徙对停歇地的选择特征

建立合理的湿地水鸟保护地是缓解水鸟栖息地丧失和种群下降的重要手段。以往的保护地设计中,由于越冬地和和繁殖地水鸟停留时间长、种群数量大,受到较多的关注。分析湿地水鸟对停歇地的选择偏好,掌握停歇地的自然条件和人类活动特征可为水鸟保护网络优化和保护地管理提供决策依据。而在水鸟迁徙过程中,停歇地作为保护网络的重要节点也发挥了重要作用。因此选择鸿雁为伞护种,获取了29只鸿雁项圈追踪数据,分析蒙古国Khukh湖-中国东北鸭绿江口秋季迁徙路线对停歇地生境选择偏好,识别了鸿雁在湿地周边不同距离梯度下的活动频率变化。根据鸿雁停歇数据共识别停歇地63处,以此为基础分析停歇的自然条件和人类活动因素特征。结果表明,鸿雁除选择湖泊和沼泽为停歇地外,周边250m内的裸地和草地也是重要栖息地;当鸿雁停歇地人类活动较少时,鸿雁倾向于选择土壤肥沃、食物丰富区,而人类活动强度加大时,栖息地植被条件提高能够为鸿雁提供遮避条件,也吸引了更多鸿雁停歇。研究建议,在水鸟迁徙重要廊道区应增加水鸟停歇地保护区,保护区的设计应根据关键保护对象活动频率加强对湿地周边的栖息地保护,减少水滨人类活动对鸿雁停歇的负面影响;在人类活动强度较大的地区设立水鸟保护地,还应加强对植被的保护,在水鸟利用频率较高的停歇地周边进行植被修复。     

Fungicidal effects of chemical disinfectants, UV light, desiccation and heat on the amphibian chytrid Batrachochytrium dendrobatidis

The efficacy of a number of disinfection treatments was tested on in vitro cultures of the fungus Batrachochytrium dendrobatidis, the causative agent of chytridiomycosis in amphibians. The aim was to evaluate the fungicidal effects of chemical disinfectants, sterilising ultraviolet (UV) light, heat and desiccation, using methods that were feasible for either disinfection in the field, in amphibian husbandry or in the laboratory. The chemical disinfectants tested were: sodium chloride, household bleach (active ingredient: sodium hypochlorite), potassium permanganate, formaldehyde solution, Path-X agricultural disinfectant (active ingredient: didecyl dimethyl ammonium chloride, DDAC), quaternary ammonium compound 128 (DDAC), Dithane, Virkon, ethanol and benzalkonium chloride. In 2 series of experiments using separate isolates of B. dendrobatidis, the fungicidal effect was evaluated for various time periods and at a range of chemical concentrations. The end point measured was death of 100% of zoospores and zoosporangia. Nearly all chemical disinfectants resulted in 100%, mortality for at least one of the concentrations tested. However, concentration and time of exposure was critical for most chemicals. Exposure to 70% ethanol, 1 mg Virkon ml(-1) or 1 mg benzalkonium chloride ml(-1) resulted in death of all zoosporangia after 20 s. The most effective products for field use were Path-X and the quaternary ammonium compound 128, which can be used at dilutions containing low levels (e.g. 0.012 or 0.008%, respectively) of the active compound didecyl dimethyl ammonium chloride. Bleach, containing the active ingredient sodium hypochlorite, was effective at concentrations of 1% sodium hypochlorite and above. Cultures did not survive complete drying, which occurred after <3 h at room temperature. B. dendrobatidis was sensitive to heating, and within 4 h at 37 degrees C, 30 min at 47 degrees C and 5 min at 60 degrees C, 100% mortality occurred. UV light (at 1000 mW m(-2) with a wavelength of 254 nm) was ineffective at killing B. dendrobatidis in culture.     

Disturbance Caused by Aerial Waterfowl Surveys During the Nonbreeding Season

Aerial surveys flown at low altitudes allow detection, identification, and enumeration of waterfowl and other waterbirds, but few studies have assessed disturbance to these guilds during the nonbreeding period. Excessive disturbance can potentially increase energy expenditure and exposure to hunting mortality contrary to objectives of many waterfowl sanctuaries where surveys are conducted. We used concurrent ground and aerial surveys to estimate the proportion of waterfowl and other waterbirds that exhibited a noticeable reaction (i.e., disturbance) or left the survey area entirely (i.e., abandonment) during low-altitude (i.e., 60–90 m above ground level) aerial surveys during September through January 2014–2017 in Illinois, USA. Overall, disturbance and abandonment probabilities of waterfowl (x̄ = 14 ± 2% [SE] and x̄ = 3 ± 1%, respectively) during aerial surveys were low. However, disturbance and abandonment probabilities varied considerably among taxa (e.g., American coot [Fulica americana] x̄ = 2 ± 1% and x̄ = 0 ± 0%, respectively; killdeer [Charadrius vociferus] x̄ = 92 ± 8% and x̄ = 17 ± 17%, respectively). Additionally, disturbance and abandonment probabilities of light geese (i.e., snow goose [Chen caerulescens] and Ross's goose [C. rossii]) and greater white-fronted geese (Anser albifrons) were relatively high, and nearly all light geese abandoned a survey location as a result of the aerial survey. Among waterfowl taxa, the odds of disturbance from the survey aircraft were 2.2–6.2 times greater at locations closed to waterfowl hunting than locations open to waterfowl hunting. Temperature, wind speed, and cloud cover were not important predictors of disturbance for most guilds, except for a negative influence of temperature on disturbance of geese. Low-altitude aerial surveys were not a significant source of disturbance for many taxa and abandonment events were rare, except events involving light geese. Periodic low-altitude aerial surveys appear to be compatible with objectives of providing sanctuary conditions for most waterfowl and other waterbirds. © 2020 The Wildlife Society.     

Possible modes of dissemination of the amphibian chytrid Batrachochytrium dendrobatidis in the environment

Amphibian chytridiomycosis caused by Batrachochytrium dendrobatidis has spread at an alarming rate over large distances throughout sensitive frog populations in eastern Australia, Central America and New Zealand. Infected amphibians and contaminated water are implicated in translocation, but other vectors are unknown. Through in vitro studies we show that potential means of translocation may be moist soil and bird feathers. B. dendrobatidis survived for up to 3 mo in sterile, moist river sand with no other nutrients added. B. dendrobatidis attached to and grew on sterile feathers and were able to be transported by feathers to establish new cultures in media, surviving between 1 and 3 h of drying between transfers. If these in vitro results are valid in the natural environment, the findings raise the possibilities that B. dendrobatidis may be translocated by movement of moist river sand and that birds may carry the amphibian chytrid between frog habitats. However, further studies using sand and feathers containing normal microflora are essential.     

Impacts of disturbance on migratory waterfowl

It is well known that disturbance from human activities can cause temporary changes in behaviour and locally affect temporal and spatial distribution of migratory and wintering waterfowl. But it is also known that, to some extent, birds can compensate for disturbance by altering their behaviour or habituating to human activities. Comparatively little is known about how these reactions to disturbance may impact on the large-scale dispersion of waterfowl and, ultimately, on their population dynamics. To be able to answer these questions, a better theoretical framework, based on optimal foraging theory incorporating predation risk, and field experiments are required. Furthermore, we need to study the waterfowl throughout their winter ranges to interpret the overall impacts of disturbance. This paper examines two cases where the impacts of disturbance have been assessed from field experiments. In one study, disturbance effects of shooting were tested by setting up experimental reserves in two Danish coastal wetlands. Over a 5-year period, these became two of the most important staging areas for coastal waterfowl, and the national totals of key species were significantly increased. A national management plan which will establish more than 50 new shooting-free refuges on Danish coastal areas within the next 5 years is likely to boost waterfowl numbers even more. Such retention of birds at more northerly sites on the fiyway should result in a more efficient resource utilization and may positively affect the population dynamics where numbers are affected by winter resources. In a second study, the impacts of disturbance by farmers on spring fattening of Pink-footed Geese Anser brachyrhynchus were analysed. In undisturbed areas in northern Norway, abdominal profiles of the geese increased rapidly, whereas in disturbed sites they did not. Subsequently, geese that had used undisturbed sites reproduced better than geese from disturbed sites.     

Microsatellites identify depredated waterfowl remains from glaucous gull stomachs

Prey remains can provide valuable sources of information regarding causes of predation and the species composition of a predator's diet. Unfortunately, the highly degraded state of many prey samples from gastrointestinal tracts often precludes unambiguous identification. We describe a procedure by which PCR amplification of taxonomically informative microsatellite loci were used to identify species of waterfowl predated by glaucous gulls ( Larus hyperboreus ). We found that one microsatellite locus unambiguously distinguished between species of the subfamily Anserinae (whistling ducks, geese and swans) and those of the subfamily Anatidae (all other ducks). An additional locus distinguished the remains of all geese and swan species known to nest on the Yukon–Kuskokwim delta in western Alaska. The study focused on two waterfowl species which have experienced precipitous declines in population numbers: emperor geese ( Chen canagica ) and spectacled eiders ( Somateria fischeri ). No evidence of predation on spectacled eiders was observed. Twenty-six percent of all glaucous gull stomachs examined contained the remains of juvenile emperor geese.     

Multi-species patterns of avian cholera mortality in Nebraska's Rainwater Basin

Nebraska's Rainwater Basin (RWB) is a key spring migration area for millions of waterfowl and other avian species. Avian cholera has been endemic in the RWB since the 1970s and in some years tens of thousands of waterfowl have died from the disease. We evaluated patterns of avian cholera mortality in waterfowl species using the RWB during the last quarter of the 20th century. Mortality patterns changed between the years before (1976-1988) and coincident with (1989-1999) the dramatic increases in lesser snow goose abundance and mortality. Lesser snow geese (Chen caerulescens caerulescens) have commonly been associated with mortality events in the RWB and are known to carry virulent strains of Pasteurella multocida, the agent causing avian cholera. Lesser snow geese appeared to be the species most affected by avian cholera during 1989-1999; however, mortality in several other waterfowl species was positively correlated with lesser snow goose mortality. Coincident with increased lesser snow goose mortality, spring avian cholera outbreaks were detected earlier and ended earlier compared to 1976-1988. Dense concentrations of lesser snow geese may facilitate intraspecific disease transmission through bird-to-bird contact and wetland contamination. Rates of interspecific avian cholera transmission within the waterfowl community, however, are difficult to determine.     

Survey for the pathogenic chytrid fungus Batrachochytrium dendrobatidis in southwestern North Carolina salamander populations

Batrachochytrium dendrobatidis is a fungal pathogen responsible for a potentially fatal disease of amphibians. We conducted a survey for B. dendrobatidis in the Appalachian Mountains of southwestern North Carolina, USA, from 10 June to 23 July 23 2009. Ventral skin swabs were collected from plethodontid salamanders (n=278) and real-time PCR was performed to test for the presence of B. dendrobatidis. We found no evidence of B. dendrobatidis, suggesting that B. dendrobatidis is absent or present in such low levels that it was undetected. If B. dendrobatidis was present at the time of our sampling, this survey supports evidence of low prevalence of B. dendrobatidis in North American headwater stream salamander populations.     

New species records and changes in abundance of waterfowl in northwest Greenland

Breeding populations of Nearctic and Palearctic waterfowl have undergone significant changes in abundance and distribution over the past 50 years. The Avanersuaq District in northwest Greenland is home to an assemblage of waterfowl from both geographic areas; however, minimal historic or current information is available on species abundance. In 2008 and 2009, we conducted field surveys in Greenland from 76.00° to 77.35°N for breeding and non-breeding waterfowl and have collected anecdotal field notes of avian observations over a 20-year period (1993–2012). During these periods, we documented the first observation of a Ross’s goose (Chen rossii) and the first confirmed breeding by lesser snow geese (Chen caerulescens caerulescens) in Greenland. Northern pintails (Anas acuta) were observed for the first time in northwest Greenland, and a previously unknown breeding location for brent geese (Branta bernicla hrota) was also identified. Local populations of greater snow (C. c.) and Canada geese (B. canadensis) have increased in size. The Booth Sound and Drown Bay wetland areas and many islands throughout the Avanersuaq District were identified as critical habitat for both breeding and non-breeding waterfowl. Further increases in waterfowl abundance, including more frequent rare and new visitors, are likely in the study area as breeding populations further south continue to increase and an ameliorating climate allows for a longer breeding season. These results will prove useful as a baseline for comparisons with future surveys.     

Mercury in waterfowl from a contaminated river in Virginia

Many bodies of water around the world are contaminated with mercury from historic industrial and mining activities or ongoing atmospheric deposition, resulting in numerous fish consumption advisories. However, concerns about mercury have only rarely led to consumption advisories on waterfowl. In contrast with fish, waterfowl frequently disperse long distances to new watersheds, so hunters and wildlife managers do not know whether waterfowl at a pristine site have spent time at a contaminated site elsewhere. We sampled tissue mercury concentrations of mallards (Anas platyrhynchos), wood ducks (Aix sponsa), and Canada geese (Branta canadensis) at a site contaminated with mercury, during the breeding and hunting seasons. We found that many mallards had bioaccumulated mercury to levels that had the potential to produce reproductive effects and exceeded consumption advisories set for fish by regulatory agencies, whereas this was true for only a few wood ducks and Canada geese. We also documented that mercury-exposed waterfowl from this contaminated site were harvested by hunters as far as 1,054 km away. Our results suggest the need for more proactive sampling of waterfowl for mercury, and likely other bioaccumulating contaminants, in order to allow hunters to make more informed choices about consumption of their harvest. © 2012 The Wildlife Society.     

Avian cholera in waterfowl in Saskatchewan, spring 1977.

Avian cholera was diagnosed in lesser snow geese (Anser c. caerulescens), Ross' geese (Anser rossii) and individuals of several other waterfowl species in a small area of south-western Saskatchewan over a 1 month period during the 1977 spring migration. Approximately 250 dead birds were found. This is apparently the first time avian cholera has been reported in migrating waterfowl in Canada. The site of the mortality was midway between the wintering and nesting areas of the two principal species, and the significance of the occurrence of the disease this far north is discussed.     

Condition Bias of Decoy-Harvested Light Geese During the Conservation Order

Evidence that decoy harvest techniques primarily remove individuals of poorer body condition is well established in short-lived duck species; however, there is limited support for condition bias in longer-lived waterfowl species, such as geese, where decoy harvest is considered primarily additive because of their high natural survival rates. We evaluated support for the harvest condition bias hypothesis of 2 long-lived waterfowl species, the lesser snow goose (Anser caerulescens caerulescens) and Ross's goose (Anser rossii). We used proximate analysis to quantify lipid and protein content of lesser snow and Ross's geese collected during the Light Goose Conservation Order (LGCO) in 2015 and 2016 during spring migration in Arkansas, Missouri, Nebraska, and South Dakota, USA. In each state, LGCO participants collected birds using traditional decoy techniques and we collected birds from the general population using jump-shooting tactics. Total body lipid content in both lesser snow and Ross's geese varied with age, region of harvest, and harvest type (decoy or jump-shooting). On average, adult lesser snow and Ross's geese harvested over decoys had 60 g and 41 g, respectively, fewer lipids than conspecifics collected using jump-shooting. We observed lower lipid reserves in decoy-shot geese in all 4 states sampled despite general gains in lipid reserves as migration chronology progressed. Our data support that the harvest condition bias extends to longer-lived waterfowl species and during a life-history event (spring migration) in which harvest is not normally observed. In the case of overabundant light geese, the disproportionate harvest of poorer-conditioned lesser snow and Ross's geese may serve as an additional challenge against any realized effects of harvest to reduce the population, in addition to extremely low harvest rates. © 2019 The Wildlife Society.     

Antimicrobial peptide defenses of the Tarahumara frog,Rana tarahumarae

Populations of the Tarahumara frog Rana tarahumarae have decreased markedly in recent years in the northern part of their range. Infection by the chytrid fungus Batrachochytrium dendrobatidis has been implicated in these declines. To determine whether antimicrobial peptides in the skin provide protection against this pathogen, norepinephrine-stimulated skin secretions were tested for their ability to inhibit growth of B. dendrobatidis in vitro. After concentration, crude mixtures of skin peptides inhibited the growth of the chytrid in a concentration-dependent manner. Proteomic analysis led to the identification and characterization of three peptides belonging to the brevinin-1 family of antimicrobial peptides and three belonging to the ranatuerin-2 family. The two most abundant peptides, ranatuerin-2TRa (GIMDSIKGAAKEIAGHLLDNLKCKITGC) and brevinin-1TRa (FLPVIAGIAANVLPKLFCKLTKRC), were active against B. dendrobatidis (MIC of 50 microM for ranatuerin-2TRa and 12.5 microM for brevinin-1TRa against zoospores). These data clearly show that antimicrobial peptides in the skin secretions of the Tarahumara frog are active against B. dendrobatidis and should provide some protection against infection. Therefore, the observed susceptibility of these frogs to this pathogen in the wild may be due to the effects of additional environmental factors that impair this innate defense mechanism, leading to the observed population declines.     

Surveillance for batrachochytrium dendrobatidis using Mixophyes (Anura: Myobatrachidae) larvae

Fourteen populations of anuran larvae (tadpoles), including three populations of the endangered Fleay's Barred Frog (Mixophyes fleayi) and 11 populations of the common Great Barred Frog (Mixophyes fasciolatus), in creek sites in the southeast region of Queensland were selected. Site selection was based on a history (within the district) of adult frog population declines and/or disappearances or records of infection of adult frogs or larvae by Batrachochytrium dendrobatidis. Larvae were collected once from each creek site between October 2002 and October 2004, and were between Gosner developmental stages 25 and 40. Total body length ranged from 18 mm to 100 mm. Mouthparts were examined under a dissecting microscope for grossly visible abnormalities, and then examined for histologic evidence of B. dendrobatidis. The most consistent mouthpart abnormalities found were multifocal depigmentation of the jaw sheaths and loss or shortening of the tooth rows. At the individual larva level, presence of mouthpart abnormalities was strongly associated with histologic diagnosis of B. dendrobatidis (93%). At least one larva with abnormal mouthparts was detected at 12 of the 14 sites and histologic evidence of B. dendrobatidis was detected at 13 of the 14 sites. These findings suggest that larvae of Mixophyes species are suitable for surveillance for B. dendrobatidis. We conclude that surveillance of B. dendrobatidis where individual larva prevalences of mouthpart abnormalities and histologic evidence of B. dendrobatidis are as high as those observed in this study (66% and 78%, respectively), relatively small numbers of larvae are required to detect these infections. Medium to large larvae (body length >30 mm) were much more likely to be affected than small larvae (body length < or =30 mm), suggesting that larger individuals should be targeted for surveillance.     

Evaluation of tadpole mouthpart depigmentation as a diagnostic test for infection by Batrachochytrium dendrobatidis for four California anurans

The objective of this study was to evaluate the utility of gross morphologic examination of larval mouthpart defects as a diagnostic screening test to detect Batrachochytrium dendrobatidis infection in four California, USA, anuran species. We examined mouthparts of 2,034 tadpoles of Bufo boreas, Pseudacris regilla, and Rana catesbeiana collected in 2003 and 2004 and Bufo canorus collected in 2004. Data were recorded for three morphologic features: upper toothrows, lower toothrows, and combined jaw sheaths. Mouthpart defects were observed in all four species (n=757), but only two species were infected with B. dendrobatidis (n=84). Sensitivity and specificity of the mouthparts test were 76% and 58%, respectively. Forty-two percent of B. dendrobatidis-negative animals would have been designated positive based on mouthpart defects. Observed prevalence was 43%, and true prevalence was 3.0%. Tests of the null hypothesis using logistic regression analysis showed that anuran larval mouthpart defects were not associated with B. dendrobatidis infection whether mouthparts scores were tested by individual morphologic feature or in combination (P=0.37). We conclude that B. dendrobatidis infection and anuran larval mouthpart defects are two separate processes that may occur concurrently and that evaluation of tadpole oral morphology is neither an accurate nor a reliable diagnostic test for B. dendrobatidis infection for the four species tested.     

Biogeographic models of gene flow in two waterfowl of the Australo‐Papuan tropics

There are many large, easy‐to‐observe anseriform birds (ducks, geese, and swans) in northern Australia and New Guinea and they often gather in large numbers. Yet, the structure of their populations and their regional movements are poorly understood. Lack of understanding of population structure limits our capacity to understand source‐sink dynamics relevant to their conservation or assess risks associated with avian‐borne pathogens, in particular, avian influenza for which waterfowl are the main reservoir species. We set out to assess present‐day genetic connectivity between populations of two widely distributed waterfowl in the Australo‐Papuan tropics, magpie goose Anseranas semipalmata (Latham, 1798) and wandering whistling‐duck Dendrocygna arcuata (Horsfield, 1824). Microsatellite data were obtained from 237 magpie geese and 64 wandering whistling‐duck. Samples were collected across northern Australia, and at one site each in New Guinea and Timor Leste. In the wandering whistling‐duck, genetic diversity was significantly apportioned by region and sampling location. For this species, the best model of population structure was New Guinea as the source population for all other populations. One remarkable result for this species was genetic separation of two flocks sampled contemporaneously on Cape York Peninsula only a few kilometers apart. In contrast, evidence for population structure was much weaker in the magpie goose, and Cape York as the source population provided the best fit to the observed structure. The fine scale genetic structure observed in wandering whistling‐duck and magpie goose is consistent with earlier suggestions that the west‐coast of Cape York Peninsula is a flyway for Australo‐Papuan anseriforms between Australia and New Guinea across Torres Strait.