Comparing modern identification methods for wild bees: Metabarcoding and image-based morphological taxonomic assignment.

With the decline of bee populations worldwide, studies determining current wild bee distributions and diversity are increasingly important. Wild bee identification is often completed by experienced taxonomists or by genetic analysis. The current study was designed to compare two methods of identification including: (1) morphological identification by experienced taxonomists using images of field-collected wild bees and (2) genetic analysis of composite bee legs (multiple taxa) using metabarcoding. Bees were collected from conservation grasslands in eastern Iowa in summer 2019 and identified to the lowest taxonomic unit using both methods. Sanger sequencing of individual wild bee legs was used as a positive control for metabarcoding. Morphological identification of bees using images resulted in 36 unique taxa among 22 genera, and >80% of Bombus specimens were identified to species. Metabarcoding was limited to genus-level assignments among 18 genera but resolved some morphologically similar genera. Metabarcoding did not consistently detect all genera in the composite samples, including kleptoparasitic bees. Sanger sequencing showed similar presence or absence detection results as metabarcoding but provided species-level identifications for cryptic species (i.e., Lasioglossum). Genus-specific detections were more frequent with morphological identification than metabarcoding, but certain genera such as Ceratina and Halictus were identified equally well with metabarcoding and morphology. Genera with proportionately less tissue in a composite sample were less likely to be detected using metabarcoding. Image-based methods were limited by image quality and visible morphological features, while genetic methods were limited by databases, primers, and amplification at target loci. This study shows how an image-based identification method compares with genetic techniques, and how in combination, the methods provide valuable genus- and species-level information for wild bees while preserving tissue for other analyses. These methods could be improved and transferred to a field setting to advance our understanding of wild bee distributions and to expedite conservation research.

Conserved grasslands support similar pollinator diversity as pollinator-specific practice regardless of proximal cropland and pesticide exposure.

Pollinator diversity and abundance are declining globally. Cropland agriculture and the corresponding use of agricultural pesticides may contribute to these declines, while increased pollinator habitat (flowering plants) can help mitigate them. Here we tested whether the relative effect of wildflower plantings on pollinator diversity and counts were modified by proportion of nearby agricultural land cover and pesticide exposure in 24 conserved grasslands in Iowa, USA. Compared with general grassland conservation practices, wildflower plantings led to only a 5% increase in pollinator diversity and no change in counts regardless of the proportion of cropland agriculture within a 1 km radius. Pollinator diversity increased earlier in the growing season and with per cent flower cover. Unexpectedly, neither insecticide nor total pesticide concentrations on above-ground passive samplers were related to pollinator diversity. However, pollinator community composition was most strongly related to date of sampling, total pesticide concentration, and forb or flower cover. Our results indicate very little difference in pollinator diversity between grassland conservation practices with and without wildflower plantings. Given the relatively high economic costs of wildflower plantings, our research provides initial evidence that investment in general grassland conservation may efficiently conserve pollinator diversity in temperate regions of intensive cropland agriculture.

Organohalogenated contaminants in multiple life stages of the Pacific lamprey (Entosphenus tridentatus), Oregon, USA.

Pacific lamprey (Entosphenus tridentatus) are ecologically and culturally important anadromous animals native to the West Coast of the United States. Pacific lamprey populations are in decline, and contaminants may be a contributing factor. Between 2017 and 2021, three life stages of Pacific lamprey and collocated sediment samples were collected in Oregon (larval lamprey, sediment, and returning adult lamprey) and off the coast of Oregon and Washington (ocean juvenile lamprey). Tissue and sediment samples were analyzed for 56 organohalogenated compounds that included legacy pesticides, current use pesticides, polybrominated diphenyl ether congeners, and polychlorinated biphenyl congeners. Organohalogenated compounds were detected in all three Pacific lamprey life stages. The organohalogenated compounds detected in collocated sediment and larval lamprey samples were generally dissimilar, and compounds detected in larval lamprey indicate potential point sources along the rivers. Ocean-caught juvenile lamprey had significantly higher lipid contents than returning adult lamprey, but lipid content and concentrations of select compounds were not strongly correlated. Concentrations of select compounds detected in both ocean juvenile and returning adult lamprey were either not significantly different or were higher in returning adult lamprey. Concentrations of some compounds in returning adult lamprey-which are consumed by Indigenous peoples-exceeded state and national human health consumption thresholds. Collaboration among Tribes and public-sector agencies helped make this study successful.

Field Assessment of Naled and Its Primary Degradation Product (Dichlorvos) in Aquatic Ecosystems Following Aerial Ultra-low Volume Application for Mosquito Control.

Naled, an organophosphate insecticide, is applied aerially at ultra-low volumes over aquatic ecosystems near Sacramento, California, USA, during summer months for mosquito control. Two ecosystem types (rice fields and a flowing canal) were sampled in 2020 and 2021. Naled and its primary degradation product (dichlorvos) were measured in water, biofilm, grazer macroinvertebrates, and omnivore/predator macroinvertebrates (predominantly crayfish). Maximum naled and dichlorvos concentrations detected in water samples one day after naled application were 287.3 and 5647.5 ng/L, respectively, which were above the U.S. Environmental Protection Agency's aquatic life benchmarks for invertebrates. Neither compound was detected in water more than one day after the application. Dichlorvos, but not naled, was detected in composite crayfish samples up to 10 days after the last aerial application. Detections in water from the canal showed that the compounds were transported downstream of the target application area. Factors such as vector control flight paths, dilution, and transport through air and water likely affected concentrations of naled and dichlorvos in water and organisms from these aquatic ecosystems.

Wild Bee Exposure to Pesticides in Conservation Grasslands Increases along an Agricultural Gradient: A Tale of Two Sample Types.

Conservation efforts have been implemented in agroecosystems to enhance pollinator diversity by creating grassland habitat, but little is known about the exposure of bees to pesticides while foraging in these grassland fields. Pesticide exposure was assessed in 24 conservation grassland fields along an agricultural gradient at two time points (July and August) using silicone band passive samplers (nonlethal) and bee tissues (lethal). Overall, 46 pesticides were detected including 9 herbicides, 19 insecticides, 17 fungicides, and a plant growth regulator. For the bands, there were more frequent/higher concentrations of herbicides in July (maximum: 1600 ng/band in July; 570 ng/band in August), while insecticides and fungicides had more frequent/higher concentrations in August (maximum: 110 and 65 ng/band in July; 1500 and 1700 ng/band in August). Pesticide concentrations in bands increased 16% with every 10% increase in cultivated crops. The bee tissues showed no difference in detection frequency, and concentrations were similar among months; maximum concentrations of herbicides, insecticides, and fungicides in July and August were 17, 27, and 180 and 19, 120, and 170 ng/g, respectively. Pesticide residues in bands and bee tissues did not always show the same patterns; of the 20 compounds observed in both media, six (primarily fungicides) showed a detection-concentration relationship between the two media. Together, the band and bee residue data can provide a more complete understanding of pesticide exposure and accumulation in conserved grasslands.

Contaminant Concentrations in Sediments, Aquatic Invertebrates, and Fish in Proximity to Rail Tracks Used for Coal Transport in the Pacific Northwest (USA): A Baseline Assessment.

Railway transport of coal poses an environmental risk, because coal dust contains polycyclic aromatic hydrocarbons (PAHs), mercury, and other trace metals. In the Pacific Northwest of the United States, proposed infrastructure projects could result in an increase in coal transport by train through the Columbia River corridor. Baseline information is needed on current distributions, levels, and spatial patterns of coal dust-derived contaminants in habitats and organisms adjacent to existing coal transport lines. To that end, we collected aquatic surface sediments, aquatic insects, and juvenile fish in 2014 and 2015 from Horsethief Lake State Park and Steigerwald National Wildlife Refuge, both located in Washington state close to the rail line and within the Columbia River Gorge National Scenic Area. Two subsites in each area were selected: one close to the rail line and one far from the rail line. Detected PAH concentrations were relatively low compared with those measured at more urbanized areas. Some contaminants were measured at higher concentrations at the subsites close to the rail line, but it was not possible to link the contaminants to a definitive source. Trace metal concentrations were only slightly higher than background concentrations, but a few of the more sensitive benchmarks were exceeded, including those for arsenic, lead, and selenium in fish tissue and fluoranthene, cadmium, copper, manganese, nickel, zinc, iron, and arsenic in sediments. At Horsethief Lake, Chinook salmon and yellow perch showed lower total mercury body burdens than other species, but PAH body burdens did not differ significantly among species. Differences in the species caught among subsites and the low number of invertebrate samples rendered food web comparisons difficult, but these data show that the PAHs and trace metals, including mercury, are accumulating in these wetland sites and in some resident organisms.

Prior Hydrologic Disturbance Affects Competition between Aedes Mosquitoes via Changes in Leaf Litter.

Allochthonous leaf litter is often the main resource base for invertebrate communities in ephemeral water-filled containers, and detritus quality can be affected by hydrologic conditions. The invasive mosquito Aedes albopictus utilizes container habitats for its development where it competes as larvae for detritus and associated microorganisms with the native Aedes triseriatus. Different hydrologic conditions that containers are exposed to prior to mosquito utilization affect litter decay and associated water quality. We tested the hypothesis that larval competition between A. albopictus and A. triseriatus would be differentially affected by prior hydrologic conditions. Experimental microcosms provisioned with Quercus alba L. litter were subjected to one of three different hydrologic treatments prior to the addition of water and mosquito larvae: dry, flooded, and a wet/dry cycle. Interspecific competition between A. albopictus and A. triseriatus was mediated by hydrologic treatment, and was strongest in the dry treatment vs. the flooded or wet/dry treatments. Aedes triseriatus estimated rate of population change (λ') was lowest in the dry treatment. Aedes albopictus λ' was unaffected by hydrologic treatment, and was on average always increasing (i.e., > 1). Aedes triseriatus λ' was affected by the interaction of hydrologic treatment with interspecific competition, and was on average declining (i.e., < 1.0), at the highest interspecific densities in the dry treatment. Dry treatment litter had the slowest decay rate and leached the highest concentration of tannin-lignin, but supported more total bacteria than the other treatments. These results suggest that dry conditions negatively impact A. triseriatus population performance and may result in the competitive exclusion of A. triseriatus by A. albopictus, possibly by reducing microbial taxa that Aedes species browse. Changing rainfall patterns with climate change are likely to affect competition between A. triseriatus and A. albopictus, probably enhancing negative competitive effects of A. albopictus on A. triseriatus in areas that experience drought.