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Abstract The use of quantitative real-time PCR (qPCR) to monitor pathogens is common; however, quantitative frameworks that consider the observation process, dynamics in pathogen presence, and pathogen load are lacking. This can be problematic in the early stages of disease progression, where low level detections may be treated as ‘inconclusive’ and excluded from analyses. Alternatively, a framework that accounts for imperfect detection would provide more robust inferences. To better estimate pathogen dynamics, we developed a hierarchical multi-scale dynamic occupancy hurdle model (MS-DOHM). The model used data gathered during sampling forPseudogymnoascus destructans (Pd), the causative agent of white-nose syndrome, a fungal disease that has cause severe declines in several species of hibernating bats in North America. The model allowed us to estimate initial occupancy, colonization, persistence and prevalence ofPdat bat hibernacula. Additionally, utilizing the relationship between cycle threshold and pathogen load, we estimated pathogen detectability and modeled expected colony and bat pathogen loads. To assess the ability of MS-DOHM to estimate pathogen dynamics, we compared MS-DOHM’s results to those of a dynamic occupancy model and naïve detection/non-detection. MS-DOHM’s estimates of site-level pathogen presence were up to 11.9% higher than estimates from the dynamic occupancy model and 35.7% higher than naïve occupancy. Including prevalence and load in our modeling framework resulted in estimates of pathogen arrival that were two to three years earlier compared to the dynamic occupancy and naïve detection/non-detection, respectively. Compared to naïve values, MS-DOHM predicted greater pathogen loads on colonies; however, we found no difference between model estimates and naïve values of prevalence. While the model predicted no declines in site-level prevalence, there were instances where pathogen load decreased in colonies that had beenPdpositive for longer periods of time. Our findings demonstrate that accounting for pathogen load and prevalence at multiple scales changes our understanding ofPddynamics, potentially allowing earlier conservation intervention. Additionally, we found that accounting for pathogen load and prevalence within hibernacula and among individuals resulted in a better fitting model with greater predictive ability. 

Avian malaria is expanding upslope with warmer temperatures and driving multiple species of Hawaiian birds towards extinction. Methods to reduce malaria transmission are urgently needed to prevent further declines. ReleasingWolbachia-infected incompatible male mosquitoes could suppress mosquito populations and releasingWolbachia-infected female mosquitoes (or both sexes) could reduce pathogen transmission if theWolbachiastrain reduced vector competence. We clearedCulex quinquefasciatusof their naturalWolbachia pipientis wPip infection and transinfected them withWolbachia wAlbB isolated fromAedes albopictus. We show thatwAlbB infection was transmitted transovarially, and demonstrate cytoplasmic incompatibility with wild-type mosquitoes infected withwPip from Oahu and Maui, Hawaii. We measured vector competence for avian malaria,Plasmodium relictum, lineage GRW4, of seven mosquito lines (two withwAlbB; three with naturalwPip infection, and two cleared ofWolbachiainfection) by allowing them to feed on canaries infected with recently collected field isolates of HawaiianP.relictum. We tested 73 groups (N_total= 1176) of mosquitoes forP.relictuminfection in abdomens and thoraxes 6–14 days after feeding on a range of parasitemias from 0.028% to 2.49%, as well as a smaller subset of salivary glands. We found no measurable effect ofWolbachiaon any endpoint, but strong effects of parasitemia, days post feeding, and mosquito strain on both abdomen and thorax infection prevalence. These results suggest that releasing malewAlbB-infectedC.quinquefasciatusmosquitoes could suppresswPip-infected mosquito populations, but would have little positive or negative impact on mosquito vector competence forP.relictumifwAlbB became established in local mosquito populations. More broadly, the lack ofWolbachiaeffects on vector competence we observed highlights the variable impacts of both native and transinfectedWolbachiainfections in mosquitoes. 

The Arctic is undergoing rapid changes in climate, altering the status and functioning of high-latitude soils and permafrost. The vast majority of studies on Arctic soils and permafrost are conducted during the summer period due to ease of accessibility, sampling, instrument operation, and making measurements, in comparison to during winter and transition seasons. However, there is increasing evidence that microbial activity continues in Arctic soils outside of the summer period. Moreover, it is becoming clear that understanding the seasonal dynamics of Arctic soils is of critical importance, especially considering that the under-studied winter is the period that is most sensitive to climate warming. Soil biogeochemical models have advanced our understanding of the functioning and fate of soils in the Arctic, however it is vital that seasonality in biotic and abiotic processes is accurately captured in these models. Here we synthesize recent investigations and observations of the year-round functioning of Arctic soils, review soil biogeochemical modelling frameworks, and highlight certain processes and behaviors that are shaped by seasonality and thus warrant particular consideration within these models. More attention to seasonal processes will be critical to improving datasets and soil biogeochemical models that can be used to understand the year-round functioning of soils and the fate of the soil carbon reservoir in the Arctic. 

Understanding experimental design (e.g. control of variable strategy or CVS) is foundational for scientific reasoning. Previous research has demonstrated that demonstrations with cognitive conflict (e.g. asking students to evaluate and explain different experimental designs) are effective in promoting children’s scientific reasoning, however, the implementation of this approach often requires significant instructional time and resources. This study reports the impact of a brief, scalable intervention on one component of scientific reasoning, understanding experimental design, by providing brief instruction on the control-of-variable strategy (CVS), embedded in a food science activity (popping popcorn). Threehundred and seven (307) 3rd-5th graders in the midwestern US participated in either a CVS intervention or a demonstration on the science of popcorn without a CVS intervention. Performance on a pre-activity test (involving identification of good and bad experiments) did not differ between conditions. By contrast, postactivity performance was significantly greater for classes who received the CVS intervention. Thus, a brief discussion of the CVS embedded within a food-science demonstration can have a meaningful impact on children’s understanding of conducting a quality experiment. Our results demonstrate the efficacy of a simple, low-cost intervention for CVS that is potentially scalable. 

Molecular phylogenetic analyses were conducted to infer relationships between the eastern and western Nearctic Androprosopa Mik and amongst the considerably more diverse western Nearctic species. Fresh, molecular-grade material was obtained for all Nearctic Androprosopa species except two Mexican species, An. sonorensis (Arnaud & Boussy) and An. zempoala Sinclair & Huerta, that eluded capture. Molecular sequences from two nuclear proteincoding genes, big zinc finger (BZF) and molybdenum cofactor sulfurase (MCS), were sampled from representatives of several outgroup and ingroup taxa and analyzed phylogenetically using maximum likelihood criteria to confirm identifications of females and immatures using a barcoding approach, test species boundaries among morphologically similar species, and infer relationships among more morphologically disparate groups. Resulting phylogenies suggest the following with significant node (bootstrap) support: (1) the eastern Nearctic Androprosopa species form the sister group to the lineage comprised of all sampled Palearctic thaumaleids, i.e., An. larvata (Mik), An. striata (Okada), and Thaumalea testacea Ruthe; (2) the aforementioned lineage is the sister group to the clade comprised of western Nearctic Androprosopa species; (3) the western Nearctic Androprosopa species form three multispecies lineages, two of which can be further divided into three or more well founded species groups. Our results suggest that Androprosopa as currently defined is paraphyletic. Additionally, we propose several new species groups within the western Nearctic Androprosopa based on molecular and morphological data.