A probabilistic model for designing and assessing the performance of eDNA sampling protocols.

Environmental DNA (eDNA) sampling, the detection of species-specific genetic material in water samples, is an emerging tool for monitoring aquatic invasive species. Optimizing eDNA sampling protocols can be challenging because there is imperfect understanding of how each step of the protocol influences its sensitivity. This paper develops a probabilistic model that characterizes each step of an eDNA sampling protocol to evaluate the protocol's overall detection sensitivity for one sample. The model is then applied to analyse how changes over time made to the eDNA sampling protocol to detect bighead (BH) and silver carp (SC) eDNA have influenced its sensitivity, and hence interpretation of the results. The model shows that changes to the protocol have caused the sensitivity of the protocol to fluctuate. A more efficient extraction method in 2013, new species-specific markers with a qPCR assay in 2014, and a more efficient capture method in 2015 have improved the sensitivity, while switching to a larger elution volume in 2013 and a smaller sample volume in 2015 have reduced the sensitivity. Overall, the sensitivity of the current protocol is higher for BH eDNA detection and SC eDNA detection compared to the original protocol used from 2009 to 2012. The paper shows how this model of eDNA sampling can be used to evaluate the effect of proposed changes in an eDNA sampling and analysis protocol on the sensitivity of that protocol to help researchers optimize their design.

Sampling Designs for Landscape-level eDNA Monitoring Programs.

Effective natural resources management requires accurate information about species distributions. Environmental DNA (eDNA) analysis is a commonly used method to determine species presence and distribution. However, when understanding eDNA-based distribution data, managers must contend with imperfect detection in collection samples and subsamples (i.e., molecular analyses) impacting their ability to detect species and estimate occurrence. Occurrence models can estimate 3 probabilities: occurrence, capture, and eDNA detection. However, most occurrence models do not. To quantify imperfect detection in rare versus common species, we examined multiple field capture and detection probabilities. We studied this with 3 objectives: Determine sample sizes required to detect eDNA given imperfect detection, determine sample sizes required to estimate eDNA capture parameters, and examine performance of a 3-level occurrence model. We found detecting eDNA in ≥1 sample at a site required ≤15 samples per site for common species, but detecting eDNA when looking for rare species required 45 to 90 samples per site. Our occurrence model recovered known parameters unless capture and detection probabilities were <0.2 where >100 samples per site and ≥8 molecular replicates were required. Our findings illustrate the importance of sample size and molecular replication for eDNA-based work. Integr Environ Assess Manag 2019;15:760-771. Published 2019. This article is a US Government work and is in the public domain in the USA.

FLEAS OF BLACK-FOOTED FERRETS (MUSTELA NIGRIPES) AND THEIR POTENTIAL ROLE IN THE MOVEMENT OF PLAGUE.

Sylvatic plague is one of the major impediments to the recovery of the black-footed ferret ( Mustela nigripes ) because it decimates their primary prey species, prairie dogs ( Cynomys spp.), and directly causes mortality in ferrets. Fleas are the primary vector of Yersinia pestis , the causative agent of sylvatic plague. The goal of this research was to better understand the flea fauna of ferrets and the factors that might influence flea abundance on ferrets. Fleas from ferrets were tested for Y. pestis in a post hoc assessment to investigate the plausibility that some ferrets could act as incidental transporter hosts of fleas infected with Y. pestis . Fleas were collected from ferrets captured on the Lower Brule Indian Reservation in central South Dakota, US from 2009 to 2012. A total of 528 fleas collected from 67 individual ferrets were identified and tested for the presence of Y. pestis with a nested PCR assay. The predominant flea recovered from ferrets was Oropsylla hirsuta , a species that comprises 70-100% of the fleas recovered from prairie dogs and their burrows in the study area. Yersinia pestis was detected at low levels in fleas collected from ferrets with prevalence ranging from 0% to 2.9%; male ferrets harbored significantly more fleas than female ferrets. Six of 67 ferrets vaccinated against plague carried fleas that tested positive for Y. pestis , which suggests ferrets vaccinated against plague could inadvertently act as incidental transporter hosts of Y. pestis -positive fleas.

Detections of Yersinia pestis East of the Known Distribution of Active Plague in the United States.

We examined fleas collected from black-tailed prairie dog (Cynomys ludovicianus) burrows from 2009 through 2011 in five national park units east of the known distribution of active plague across the northern Great Plains for the presence of Yersinia pestis. Across all national park units, Oropsylla tuberculata and Oropsylla hirsuta were the most common fleas collected from prairie dog burrows, 42.4% and 56.9%, respectively, of the 3964 fleas collected from burrow swabbing. Using a nested PCR assay, we detected 200 Y. pestis-positive fleas from 3117 assays. In total, 6.4% of assayed fleas were Y. pestis positive and 13.9% of prairie dog burrows swabbed contained Y. pestis-positive fleas. Evidence of the presence of Y. pestis was observed at all national park units except Devils Tower National Monument in Wyoming. We detected the presence of Y. pestis without large die-offs, i.e., enzootic sylvatic plague, east of the known distribution of active plague and near the eastern edge of the present distribution of black-tailed prairie dogs. This study, in combination with previous work suggests that sylvatic plague likely occurs across the range of black-tailed prairie dogs and should now be treated as endemic across this range.

Habitat correlates with the spatial distribution of ectoparasites on Peromyscus leucopus in southern Michigan.

The goal of this study was to evaluate the role of habitat in determining ectoparasite distribution of Peromyscus leucopus. We tested the hypothesis that ectoparasite occurrence is associated with particular host environments and this association is stronger for ectoparasites with limited interactions (i.e., ticks) than those with frequent interactions (i.e., lice). Ectoparasites from three different groups (Acari, Siphonaptera, and Phthiraptera) were collected from P. leucopus inhabiting a number of forested habitats in southern Michigan. Measurements of plant species structure and composition were collected and models were developed using quadratic discriminant function analysis to determine if habitats associated with ectoparasite presence were different from those associated with their absence. Mice parasitized by ticks were more likely to be found in areas having undergone a recent disturbance. Mice parasitized by ticks, fleas, and lice were more likely to be found in areas having tree species associated with dry soils. Our results show there is a distinct difference in habitats associated with the presence of ectoparasites, though we did not observe a stronger association of host habitat for ticks than for fleas or lice. This implies habitat should be included as an important component of assessments of the spatial distribution of ectoparasites.