Search

1. Conservation managers rely on accurate estimates of disease parameters, such as pathogen prevalence and infection intensity, to assess disease status of a host population. However, these disease metrics may be biased if low-level infection intensities are missed by sampling methods or laboratory diagnostic tests. These false negatives underestimate pathogen prevalence and overestimate mean infection intensity of infected individuals. 2. Our objectives were two-fold. First, we quantified false negative error rates of Batrachochytrium dendrobatidis (Bd) on non-invasive skin swabs collected from an amphibian community in El Copé, Panama. We swabbed amphibians twice in sequence, and we used a recently developed hierarchical Bayesian estimator to assess disease status of the population. Second, we developed a novel hierarchical Bayesian model to simultaneously account for imperfect pathogen detection from field sampling and laboratory diagnostic testing. We evaluated the performance of the model, using simulations and varying sampling design to quantify the magnitude of bias in estimates of pathogen prevalence and infection intensity. 3. We show that Bd detection probability from skin swabs was related to host infection intensity, where Bd infections <10 zoospores have <95% probability of being detected. If imperfect Bd detection was not considered, then Bd prevalence was underestimated by as much as 71%. In the Bd-amphibian system, this indicates a need to correct for imperfect pathogen detection in enzootic host populations persisting with low-level infections. More generally, our results have implications for study designs in other disease systems, particularly those with similar objectives, biology, and sampling decisions. 4. Uncertainty in pathogen detection is an inherent property of most sampling protocols and diagnostic tests, where the magnitude of bias depends on the study system, type of infection, and false negative error rates. Given that it may be difficult to know this information in advance, we advocate that the most cautious approach is to assume all errors are possible and to accommodate them by adjusting sampling designs. The modelling framework presented here improves the accuracy in estimating pathogen prevalence and infection intensity. [ABSTRACT FROM AUTHOR]

1. Epidemiological studies are crucial for understanding the distribution and dynamics of emerging infectious diseases. To accurately assess infection states in wild populations, researchers need to account for observational uncertainty. We focus on two sources of uncertainty when estimating epidemiological parameters: nondetection of infection in sampled individuals and sampling error when quantifying infection intensity for infected individuals. 2. We developed new analytical methods to simultaneously estimate prevalence and the distribution of infection intensities based on repeated sampling of individuals in the wild. The methods are an extension of those used for occupancy estimation and address both sources of observation error. At the same time, we account for heterogeneity in detection probability that results from individual variation in infection intensity. 3. We use two estimation approaches to account for detection. The first is to use the complete likelihood in a hierarchical Bayesian model and fit using Markov chain Monte Carlo sampling. The second is to estimate the detection relationship using a mark-recapture abundance estimator and use those results to calculate weighted estimates for prevalence and mean infection intensities. 4. We use data from a field survey of Batrachochytrium dendrobatidis in Illinois amphibians to test these methods. We show that detection probability using quantitative PCR is strongly related to infection intensity, measured in zoospore equivalents. Sites in the study varied greatly in estimated prevalence and to a lesser extent in mean infection intensities of infected individuals. We did not find evidence of a relationship of snout-vent-length to infection intensity or prevalence. Naïve estimates of prevalence that do not account for detection were less than estimates for either of our methods, which yielded similar prevalence values for most sites. 5. Uncertainty when assessing disease state is a characteristic of most diagnostic tests. The estimators presented here account for this uncertainty and thus can improve accuracy when assessing the relationship of ecological factors to prevalence and infection intensity. [ABSTRACT FROM AUTHOR]

Many declines of amphibian populations have been associated with chytridiomycosis, a disease caused by the aquatic fungus Batracjiochytrium dendrobatidis (Bd). Despite the relatively high prevalence of chytridiomycosis in stream amphibians globally, most surveys in North America have focused primarily on wetland-associated species, which are frequently infected. To better understand the distribution and prevalence of Bd in headwater amphibian communities, we sampled 452 tailed frogs (Ascaphus truei and Aseaphus inontanus) and 304 stream salamanders (seven species in the Dicamptodonti- dae and Plethodontidae) for Bd in 38, first- to third-order streams in five montane areas across the United States. We tested for presence of Bd by using PCR on skin swabs from salamanders and metamorphosed tailed frogs or the oral disc of frog larvae. We detected Bd on only seven individuals (0.93%) in four streams. Based on our study and results from five other studies that have sampled headwater- or seep-associated amphibians in the United States, Bd has been detected on only 3% of 1,322 individuals from 21 species. These results differ strongly from surveys in Central America and Australia, where Bd is more prevalent on stream-breeding species, as well as results from wetland-associated anurans in the same regions of the United States that we sampled. Differences in the prevalence of Bd between stream- and wetland-associated amphibians in the United States may be related to species-specific variation in susceptibility to chytridiomycosis or habitat differences. [ABSTRACT FROM AUTHOR]

The amphibian chytrid fungus Batrachochytrium dendrobatidis, responsible for the potentially fatal amphibian disease chytridiomycosis, is known to occur in a large and ever increasing number of amphibian populations around the world. However, sampling has been biased towards stream- and wetland-breeding anurans, with little attention paid to stream-associated salamanders. We sampled three frog and three salamander species in the Chesapeake and Ohio Canal National Historic Park, Maryland, by swabbing animals for PCR analysis to detect DNA of B. dendrobatidis. Using PCR, we detected B. dendrobatidis DNA in both stream and wetland amphibians, and report here the first occurrence of the pathogen in two species of stream-associated salamanders. Future research should focus on mechanisms within habitats that may affect persistence and dissemination of B. dendrobatidis among stream-associated salamanders [ABSTRACT FROM AUTHOR]

The frog-killing chytrid fungus Batrachochytrium dendrobatidis (Bd) is decimating amphibian populations around the world. 1–4 Bd has a biphasic life cycle, alternating between motile zoospores that disperse within aquatic environments and sessile sporangia that grow within the mucus-coated skin of amphibians. 5,6 Zoospores lack cell walls and swim rapidly through aquatic environments using a posterior flagellum and crawl across solid surfaces using actin structures similar to those of human cells. 7,8 Bd transitions from this motile dispersal form to its reproductive form by absorbing its flagellum, rearranging its actin cytoskeleton, and rapidly building a chitin-based cell wall—a process called "encystation." 5–7 The resulting sporangium increases in volume by two or three orders of magnitude while undergoing rounds of mitosis without cytokinesis to form a large ceonocyte. The sporangium then cellurizes by dividing its cytoplasm into dozens of new zoospores. After exiting the sporangium through a discharge tube onto the amphibian skin, daughter zoospores can then reinfect the same individual or find a new host. 5 Although encystation is critical to Bd growth, whether and how this developmental transition is triggered by external signals was previously unknown. We discovered that exposure to amphibian mucus triggers rapid and reproducible encystation within minutes. This response can be recapitulated with purified mucin, the bulk component of mucus, but not by similarly viscous methylcellulose or simple sugars. Mucin-induced encystation does not require gene expression but does require surface adhesion, calcium signaling, and modulation of the actin cytoskeleton. Mucus-induced encystation may represent a key mechanism for synchronizing Bd development with the arrival at the host. • Bd encysts within 5 min of exposure to amphibian mucus or purified mucin • Mucin-induced encystation includes ciliary retraction and de novo cell wall assembly • Triggered encystation requires surface adhesion but not gene expression • Calcium flux appears necessary and sufficient to induce encystation The chytrid fungus Batrachochytrium dendrobatidis (Bd) causes a skin infection that is decimating global amphibian populations. Robinson et al. report here that exposure to amphibian mucus causes Bd to rapidly transition from its dispersal to its growth form and identify key molecular requirements of this essential developmental transition. [ABSTRACT FROM AUTHOR]

Diverse bacteria inhabit amphibian skin; some of those bacteria inhibit growth of the fungal pathogen Batrachochytrium dendrobatidis. Yet there has been no systematic survey of anti-B. dendrobatidis bacteria across localities, species, and elevations. This is important given geographic and taxonomic variations in amphibian susceptibility to B. dendrobatidis. Our collection sites were at locations within the Appalachian Mountains where previous sampling had indicated low B. dendrobatidis prevalence. We determined the numbers and identities of anti-B. dendrobatidis bacteria on 61 Plethodon salamanders (37 P. cinereus, 15 P. glutinosus, 9 P. cylindraceus) via culturing methods and 16S rRNA gene sequencing. We sampled co-occurring species at three localities and sampled P. cinereus along an elevational gradient (700 to 1,000 meters above sea level [masl]) at one locality. We identified 50 anti-B. dendrobatidis bacterial operational taxonomic units (OTUs) and found that the degree of B. dendrobatidis inhibition was not correlated with relatedness. Five anti-B. dendrobatidis bacterial strains occurred on multiple amphibian species at multiple localities, but none were shared among all species and localities. The prevalence of anti-B. dendrobatidis bacteria was higher at Shenandoah National Park (NP), VA, with 96% (25/26) of salamanders hosting at least one anti-B. dendrobatidis bacterial species compared to 50% (7/14) at Catoctin Mountain Park (MP), MD, and 38% (8/21) at Mt. Rogers National Recreation Area (NRA), VA. At the individual level, salamanders at Shenandoah NP had more anti-B. dendrobatidis bacteria per individual (μ = 3.3) than those at Catoctin MP (μ = 0.8) and at Mt. Rogers NRA (μ= 0.4). All salamanders tested negative for B. dendrobatidis. Anti-B. dendrobatidis bacterial species are diverse in central Appalachian Plethodon salamanders, and their distribution varied geographically. The antifungal bacterial species that we identified may play a protective role for these salamanders. IMPORTANCE Amphibians harbor skin bacteria that can kill an amphibian fungal pathogen, Batrachochytrium dendrobatidis. Some amphibians die from B. dendrobatidis infection, whereas others do not. The bacteria that can kill B. dendrobatidis, called anti-B. dendrobatidis bacteria, are thought to influence the B. dendrobatidis infection outcome for the amphibian. Yet how anti-B. dendrobatidis bacterial species vary among amphibian species and populations is unknown. We determined the distribution of anti-B. dendrobatidis bacterial species among three salamander species (n = 61) sampled at three localities. We identified 50 unique anti-B. dendrobatidis bacterial species and found that all of the tested salamanders were negative for B. dendrobatidis. Five anti-B. dendrobatidis bacterial species were commonly detected, suggesting a stable, functional association with these salamanders. The number of anti-B. dendrobatidis bacteria per individual varied among localities but not among co-occurring salamander species, demonstrating that environment is more influential than host factors in structuring the anti-B. dendrobatidis bacterial community. These anti-B. dendrobatidis bacteria may serve a protective function for their salamander hosts. [ABSTRACT FROM AUTHOR]