Climate data from the Rocky Mountain Biological Laboratory (1975-2022).

The Rocky Mountain Biological Laboratory (RMBL; Colorado, USA) is the site for many research projects spanning decades, taxa, and research fields from ecology to evolutionary biology to hydrology and beyond. Climate is the focus of much of this work and provides important context for the rest. There are five major sources of data on climate in the RMBL vicinity, each with unique variables, formats, and temporal coverage. These data sources include (1) RMBL resident billy barr, (2) the National Oceanic and Atmospheric Administration (NOAA), (3) the United States Geological Survey (USGS), (4) the United States Department of Agriculture (USDA), and (5) Oregon State University's PRISM Climate Group. Both the NOAA and the USGS have automated meteorological stations in Crested Butte, CO, ~10 km from the RMBL, while the USDA has an automated meteorological station on Snodgrass Mountain, ~2.5 km from the RMBL. Each of these data sets has unique spatial and temporal coverage and formats. Despite the wealth of work on climate-related questions using data from the RMBL, previous researchers have each had to access and format their own climate records, make decisions about handling missing data, and recreate data summaries. Here we provide a single curated climate data set of daily observations covering the years 1975-2022 that blends information from all five sources and includes annotated scripts documenting decisions for handling data. These synthesized climate data will facilitate future research, reduce duplication of effort, and increase our ability to compare results across studies. The data set includes information on precipitation (water and snow), snowmelt date, temperature, wind speed, soil moisture and temperature, and stream flows, all publicly available from a combination of sources. In addition to the formatted raw data, we provide several new variables that are commonly used in ecological analyses, including growing degree days, growing season length, a cold severity index, hard frost days, an index of El Niño-Southern Oscillation, and aridity (standardized precipitation evapotranspiration index). These new variables are calculated from the daily weather records. As appropriate, data are also presented as minima, maxima, means, residuals, and cumulative measures for various time scales including days, months, seasons, and years. The RMBL is a global research hub. Scientists on site at the RMBL come from many countries and produce about 50 peer-reviewed publications each year. Researchers from around the world also routinely use data from the RMBL for synthetic work, and educators around the United States use data from the RMBL for teaching modules. This curated and combined data set will be useful to a wide audience. Along with the synthesized combined data set we include the raw data and the R code for cleaning the raw data and creating the monthly and yearly data sets, which facilitate adding additional years or data using the same standardized protocols. No copyright or proprietary restrictions are associated with using this data set; please cite this data paper when the data are used in publications or scientific events.

Current and lagged climate affects phenology across diverse taxonomic groups.

The timing of life events (phenology) can be influenced by climate. Studies from around the world tell us that climate cues and species' responses can vary greatly. If variation in climate effects on phenology is strong within a single ecosystem, climate change could lead to ecological disruption, but detailed data from diverse taxa within a single ecosystem are rare. We collated first sighting and median activity within a high-elevation environment for plants, insects, birds, mammals and an amphibian across 45 years (1975-2020). We related 10 812 phenological events to climate data to determine the relative importance of climate effects on species' phenologies. We demonstrate significant variation in climate-phenology linkage across taxa in a single ecosystem. Both current and prior climate predicted changes in phenology. Taxa responded to some cues similarly, such as snowmelt date and spring temperatures; other cues affected phenology differently. For example, prior summer precipitation had no effect on most plants, delayed first activity of some insects, but advanced activity of the amphibian, some mammals, and birds. Comparing phenological responses of taxa at a single location, we find that important cues often differ among taxa, suggesting that changes to climate may disrupt synchrony of timing among taxa.

Complex PrEP: the factors requiring consultant-led review of PrEP users.

INTRODUCTION: HIV-1 pre-exposure prophylaxis (PrEP) has been available in England since March 2020 on the National Health Service using generic emtricitabine and tenofovir disoproxil. 56 Dean Street (56DS) provided PrEP through (1) additional private care from September 2015, estimated to be providing 11% of England's PrEP in 2019; and (2) the IMPACT trial, as well as monitoring those self-sourcing PrEP. Providing PrEP at scale through a nurse-led service required a safety net for complex individuals. 56DS introduced a consultant-led PrEP outpatient service, the PrEP review clinic, in January 2018 and we report the outcomes of this service. METHODS: We present a retrospective case note review of the PrEP review clinic with descriptive outcomes from 26 January 2018 to 20 December 2019. Reason for referral, demographics, PrEP management and PrEP discontinuations were recorded. RESULTS: 13 980 unique users accessed PrEP from 56DS during the two year evaluation period. 220 individuals were seen in the PrEP review clinic. Majority of patients were referred for renal issues (114 of 220, 51.8%), followed by side effects (59 of 220, 26.8%) and comorbidities (38 of 220, 17.2%). Of those with renal issues, 89 (out of 114, 78.1%) users were referred for an abnormal estimated glomerular filtration rate (eGFR). 35 (out of 114, 30.7%) PrEP users had an eGFR between 45 and 59 mL/min/1.73 m2, of whom 2 (5.7%) discontinued PrEP. Majority of users were advised to stop supplements±switch to event-based dosing (24 of 35, 68.6%). Ten PrEP users were referred with an eGFR between 30 and 44 mL/min/1.73 m2; 4 (40%) stopped or did not start PrEP and 6 (60%) were asked to stop supplements±switch to event-based dosing. DISCUSSION: A small proportion of PrEP users have complex PrEP issues. Methods to manage renal dysfunction with PrEP included stopping supplements and switching to event-based dosing. Those with side effects were managed with an array of options, with only modest effectiveness. Other PrEP options are needed to support those with toxicities or intolerances.

Effects of two pesticides on northern leopard frog (Lithobates pipiens) stress metrics: Blood cell profiles and corticosterone concentrations.

Amphibians are declining globally. Exposure to pesticides has been implicated in decreasing amphibian immune function, thus increasing their susceptibility to parasites and disease and thereby negatively affecting individuals and populations. Amphibians are likely exposed to neonicotinoids because these widely used insecticides are highly soluble in water and because amphibian freshwater habitats are often embedded in agroecosystems. Herein, we investigate the effects of long-term exposure to two individual neonicotinoids (clothianidin or thiamethoxam) at either low or high concentrations (2.5 or 250 µg/L) on northern leopard frog (Lithobates pipiens) blood cell profiles and concentrations of corticosterone, an energy-mediating hormone associated with stress. Larval frogs from Gosner stage 25 to 46 were exposed to pesticide and control treatments in outdoor mesocosms. Corticosterone concentrations were measured after 6 d of exposure, and blood cell profiles were assessed once frogs reached Gosner stage 46 (following 8 w of exposure). No significant changes were found in erythrocyte counts, leukocyte counts, monocyte to leukocyte ratios or corticosterone concentrations between treatments. However, exposure to either 2.5 or 250 µg/L of clothianidin, or 250 µg/L of thiamethoxam decreased neutrophil to lymphocyte ratios and neutrophil to leukocyte ratios, and exposure to 2.5 µg/L of clothianidin or 250 µg/L of thiamethoxam decreased eosinophil to leukocyte ratios. Our results indicate that long-term exposure to neonicotinoids can alter leukocyte profiles, indicative of a stress response. Future studies should investigate whether chronic exposure to neonicotinoids affect multiple measures of stress differently or influences the susceptibility of amphibians to parasites and pathogens. Our work underscores the importance of continued use of multiple measures of stress for different amphibian species when undertaking ecotoxicological assessments.

Clothianidin alters leukocyte profiles and elevates measures of oxidative stress in tadpoles of the amphibian, Rana pipiens.

Neonicotinoid pesticide use is widespread and highly debated, as evidenced by recent attention received from the public, academics and pesticide regulatory agencies. However, relatively little is known about the physiological effects of neonicotinoid insecticides on aquatic vertebrates. Amphibians (larval stages in particular) are excellent vertebrate bioindicators in aquatic systems due to their risk of exposure and sensitivity to environmental stressors. Previous work with wood frog (Rana sylvatica) tadpoles exposed to formulated products containing thiamethoxam or clothianidin in outdoor mesocosms found significant shifts in leukocyte profiles, suggesting the tadpoles were physiologically stressed. The main objective of the present study was to characterize this stress response further using complementary measures of stress after exposure to clothianidin on northern leopard frogs (Rana pipiens) during their aquatic larval stages. Laboratory static-renewal exposures were conducted over eight weeks with the technical product clothianidin at 0, 0.23, 1, 10 and 100 µg/L, and diquat dibromide at 532 µg/L was used as a positive control. We assessed tadpole leukocyte profiles and measures of oxidative stress as these sub-lethal alterations could affect amphibian fitness. We found changes in several types of leukocytes at 1 and 10 µg/L, suggesting that these tadpoles exhibited signs of mild physiological stress. Clothianidin also induced an oxidative stress response at 0.23, 1 and 100 µg/L. However, we found no differences in survival, growth, development time or hepatosomatic index in frogs exposed to clothianidin. Our study indicates that tadpoles chronically exposed to clothianidin have increased stress responses, but in the absence of concentration-response relationships and effects on whole-organism endpoints, the implications on the overall health and fitness of these changes are unclear.

Effects of 2 Neonicotinoid Insecticides on Blood Cell Profiles and Corticosterone Concentrations of Wood Frogs (Lithobates sylvaticus).

Neonicotinoids are widely used insecticides that are detectable in agricultural waterways. These insecticides are of concern due to their potential impacts on nontarget organisms. Pesticides can affect development of amphibians and suppress the immune system, which could impact disease susceptibility and tolerance. No previous studies on amphibians have examined the effects of these insecticides on differential blood cell proportions or concentrations of corticosterone (a general stress hormone). We investigated the effects of chronic exposure to 2 neonicotinoids, thiamethoxam and clothianidin, on immunometrics of wood frogs (Lithobates sylvaticus). Frogs were exposed to single, chronic treatments of 2.5 or 250 µg/L of clothianidin or thiamethoxam for 7 wk from Gosner stages 25 to 46. The juvenile frogs were then maintained for 3 wk post metamorphosis without exposure to neonicotinoids. We measured water-borne corticosterone twice: at 6 d and 8 wk after exposure in larval and juvenile frogs, respectively. We assessed differential blood cell profiles from juvenile frogs. Corticosterone was significantly lower in tadpoles exposed to 250 µg/L of thiamethoxam compared with other tadpole treatments, but no significant differences in corticosterone concentrations were found in treatments using juvenile frogs. Anemia was detected in all treatments compared with controls with the exception of tadpoles exposed to 2.5 µg/L of clothianidin. Neutrophil-to-leukocyte and neutrophil-to-lymphocyte ratios were elevated in frogs exposed to 250 µg/L of thiamethoxam. Collectively, these results indicate that chronic exposure to neonicotinoids has varied impacts on blood cell profiles and corticosterone concentrations of developing wood frogs, which are indicative of stress. Future studies should investigate whether exposure to neonicotinoids increases susceptibility to infection by parasites in both larval and adult wood frogs. Environ Toxicol Chem 2019;38:1273-1284. © 2019 Crown in the right of Canada. Published by Wiley Periodicals Inc. on behalf of SETAC.

Sublethal effects on wood frogs chronically exposed to environmentally relevant concentrations of two neonicotinoid insecticides.

Neonicotinoids are prophylactically used globally on a variety of crops, and there is concern for the potential impacts of neonicotinoids on aquatic ecosystems. The intensive use of pesticides on crops has been identified as a contributor to population declines of amphibians, but currently little is known regarding the sublethal effects of chronic neonicotinoid exposure on amphibians. The objective of the present study was to characterize the sublethal effect(s) of exposure to 3 environmentally relevant concentrations (1 µg/L, 10 µg/L, and 100 µg/L) of 2 neonicotinoids on larval wood frogs (Lithobates sylvaticus) using outdoor mesocosms. We exposed tadpoles to solutions of 2 commercial formulations containing imidacloprid and thiamethoxam, and assessed survival, growth, and development. Exposure to imidacloprid at 10 µg/L and 100 µg/L increased survival and delayed completion of metamorphosis compared with controls. Exposure to thiamethoxam did not influence amphibian responses. There was no significant effect of any treatment on body mass or size of the metamorphs. The results suggest that current usage of imidacloprid and thiamethoxam does not pose a threat to wood frogs. However, further assessment of both direct and indirect effects on subtle sublethal endpoints, and the influence of multiple interacting stressors at various life stages, is needed to fully understand the effects of neonicotinoids on amphibians. Environ Toxicol Chem 2017;36:1101-1109. © 2017 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

Spatial patterning in PM2.5 constituents under an inversion-focused sampling design across an urban area of complex terrain.

Health effects of fine particulate matter (PM2.5) vary by chemical composition, and composition can help to identify key PM2.5 sources across urban areas. Further, this intra-urban spatial variation in concentrations and composition may vary with meteorological conditions (e.g., mixing height). Accordingly, we hypothesized that spatial sampling during atmospheric inversions would help to better identify localized source effects, and reveal more distinct spatial patterns in key constituents. We designed a 2-year monitoring campaign to capture fine-scale intra-urban variability in PM2.5 composition across Pittsburgh, PA, and compared both spatial patterns and source effects during "frequent inversion" hours vs 24-h weeklong averages. Using spatially distributed programmable monitors, and a geographic information systems (GIS)-based design, we collected PM2.5 samples across 37 sampling locations per year to capture variation in local pollution sources (e.g., proximity to industry, traffic density) and terrain (e.g., elevation). We used inductively coupled plasma mass spectrometry (ICP-MS) to determine elemental composition, and unconstrained factor analysis to identify source suites by sampling scheme and season. We examined spatial patterning in source factors using land use regression (LUR), wherein GIS-based source indicators served to corroborate factor interpretations. Under both summer sampling regimes, and for winter inversion-focused sampling, we identified six source factors, characterized by tracers associated with brake and tire wear, steel-making, soil and road dust, coal, diesel exhaust, and vehicular emissions. For winter 24-h samples, four factors suggested traffic/fuel oil, traffic emissions, coal/industry, and steel-making sources. In LURs, as hypothesized, GIS-based source terms better explained spatial variability in inversion-focused samples, including a greater contribution from roadway, steel, and coal-related sources. Factor analysis produced source-related constituent suites under both sampling designs, though factors were more distinct under inversion-focused sampling.

Multilocus Sex Determination Revealed in Two Populations of Gynodioecious Wild Strawberry, Fragaria vesca subsp. bracteata.

Gynodioecy, the coexistence of females and hermaphrodites, occurs in 20% of angiosperm families and often enables transitions between hermaphroditism and dioecy. Clarifying mechanisms of sex determination in gynodioecious species can thus illuminate sexual system evolution. Genetic determination of gynodioecy, however, can be complex and is not fully characterized in any wild species. We used targeted sequence capture to genetically map a novel nuclear contributor to male sterility in a self-pollinated hermaphrodite of Fragaria vesca subsp. bracteata from the southern portion of its range. To understand its interaction with another identified locus and possibly additional loci, we performed crosses within and between two populations separated by 2000 km, phenotyped the progeny and sequenced candidate markers at both sex-determining loci. The newly mapped locus contains a high density of pentatricopeptide repeat genes, a class commonly involved in restoration of fertility caused by cytoplasmic male sterility. Examination of all crosses revealed three unlinked epistatically interacting loci that determine sexual phenotype and vary in frequency between populations. Fragaria vesca subsp. bracteata represents the first wild gynodioecious species with genomic evidence of both cytoplasmic and nuclear genes in sex determination. We propose a model for the interactions between these loci and new hypotheses for the evolution of sex determining chromosomes in the subdioecious and dioecious Fragaria.

Determining in situ periphyton community responses to nutrient and atrazine gradients via pigment analysis.

Agrochemicals, including fertilizers and herbicides, are significant contributors of non-point source pollution to surface waters and have the potential to negatively affect periphyton. We characterized periphyton communities using pigment markers to assess the effects of nutrient enrichment and the herbicide atrazine with in situ experimental manipulations and by examining changes in community structure along existing agrochemical gradients. In 2008, the addition of nutrients (20 mg/L nitrate and 1.25 mg/L reactive phosphate), atrazine (20 µg/L) and a combination of both nutrients and atrazine had no significant effect on periphyton biomass or community structure in a stream periphytometer experiment. In 2009, similar experiments with higher concentrations of atrazine (200 µg/L) at two stream sites led to some minor effects. In contrast, at the watershed scale (2010) periphyton biomass (mg/m(2) chlorophyll a) increased significantly along correlated gradients of nitrate and atrazine but no direct effects of reactive phosphate were observed. Across the watershed, the average periphyton community was composed of Bacillariophyceae (60.9%), Chlorophyceae (28.1%), Cryptophyceae (6.9%) and Euglenophyceae (4.1%), with the Bacillariophyceae associated with high turbidity and the Chlorophyceae with nitrate enrichment. Overall, effects of nitrate on periphyton biomass and community structure superseded effects of reactive phosphate and atrazine.

Atrazine contamination at the watershed scale and environmental factors affecting sampling rates of the polar organic chemical integrative sampler (POCIS).

Polar organic chemical integrative samplers (POCIS) were used to estimate atrazine contamination at 24 stream/river sites located across a watershed with land use ranging from 6.7 to 97.4% annual crops and surface water nitrate concentrations ranging from 3 to 5404 µg/L. A gradient of atrazine contamination spanning two orders of magnitude was observed over two POCIS deployments of 28 d and was positively correlated with measures of agricultural intensity. The metabolite desisopropyl atrazine was used as a performance reference compound in field calibration studies. Sampling rates were similar between field sites but differed seasonally. Temperature had a significant effect on sampling rates while other environmental variables, including water velocity, appeared to have no effect on sampling rates. A performance reference compound approach showed potential in evaluating spatial and temporal differences in field sampling rates and as a tool for further understanding processes governing uptake of polar compounds by POCIS.

Maternal sex effects and inbreeding depression under varied environmental conditions in gynodioecious Fragaria vesca subsp. bracteata.

BACKGROUND AND AIMS: Gynodioecy (coexistence of females and hermaphrodites) is a sexual system that occurs in numerous flowering plant lineages. Thus, understanding the features that affect its maintenance has wide importance. Models predict that females must have a seed fitness advantage over hermaphrodites, and this may be achieved via seed quality or quantity. Females in a population of Fragaria vesca subsp. bracteata, a long-lived gynodioecious perennial, do not demonstrate a seed quantity advantage, so this study explored whether females produced better quality seed via maternal sex effects or avoidance of inbreeding depression (IBD). METHODS: Families of selfed and outcrossed seed were created using hermaphrodite mothers and families of outcrossed seed were created using female mothers. The effects of these pollination treatments were assessed under benign conditions early in life and under varied conditions later in life. To test for an effect of maternal sex, fitness components and traits associated with acclimation to variable environments of progeny of outbred hermaphrodites and females were compared. To test for expression of IBD, fitness parameters between inbred and outbred progeny of hermaphrodites were compared. KEY RESULTS: Offspring of females were more likely to germinate in benign conditions and survive in harsh resource environments than outbred progeny of hermaphrodites. IBD was low across most life stages, and both the effect of maternal sex on progeny quality and the expression of IBD depended on both maternal family and resource condition of the progeny. CONCLUSIONS: The effect of maternal sex and IBD on progeny quality depended on resource conditions, maternal lineage and progeny life stage. In conjunction with known lack of differences in seed quantity, the quality advantages and IBD observed here are still unlikely to be sufficient for maintenance of gynodioecy under nuclear inheritance of male sterility.

Comparing the sensitivity of geographically distinct Lemna minor populations to atrazine.

The objectives of this study were to compare the sensitivities of field populations and a laboratory culture of a duckweed species (Lemna minor) to the herbicide atrazine using three different endpoints and to determine whether sensitivity to atrazine was affected by past exposure to the herbicide. L. minor cultures were purchased commercially or collected from field sites within an agricultural watershed and exposed to atrazine for 7 days under greenhouse conditions. Populations differed significantly in their sensitivity to atrazine. Biomass was more sensitive than frond number, while chlorophyll fluorescence was not a sensitive endpoint. Overall, the sensitivity of the various populations to atrazine was not strongly related to measures of past exposure to agriculture stressors. Positive correlations between biomass twenty-five percent inhibition concentrations (IC25s), biomass estimated marginal means and in-stream atrazine concentrations were observed, providing evidence that atrazine exposure is linked to a decrease in sensitivity to atrazine. However, IC25s generated for each population were similar, ranging from 19 to 40 and 57 to 92 µg/L atrazine for biomass and frond data respectively, and likely do not represent biologically significant differences in atrazine sensitivity. Given the small range in sensitivity observed between populations, commercial laboratory cultures appear to provide a good estimate of the sensitivity of field populations of L. minor to atrazine and should continue to be used in regulatory phytotoxicity testing.

Comparison of the effects of glyphosate and atrazine herbicides on nontarget plants grown singly and in microcosms.

Intensification of agriculture and the corresponding increase in herbicide use has led to concern regarding the effects these chemicals may have on nontarget plants of agroecosystems. Current pesticide registration guidelines are focused on testing crop species grown singly in pots under greenhouse conditions and may not provide adequate measures of protection to noncrop species. The objective of the present study was to compare the response of terrestrial and wetland plants to the herbicides glyphosate and atrazine when grown singly in pots versus under different microcosm conditions. Greenhouse microcosms were generally more sensitive than single-species tests. Plants grown for an extended test period or in seminatural field conditions were generally less sensitive to herbicides. Sensitivity was found to be dependent on interactions between species and test conditions. Changes in community structure were observed in herbicide-treated microcosms that would not be predicted from single-species testing. Single-species tests are useful because they are inexpensive, can demonstrate clear dose-response patterns uncomplicated by other factors influencing growth, and are able to provide a measure of the sensitivity of a given species to glyphosate and atrazine. However, they are unable to predict subtle changes in community structure that may have important long-term consequences.

Germination requirements for 29 terrestrial and wetland wild plant species appropriate for phytotoxicity testing.

BACKGROUND: Species selected for phytotoxicity testing have been limited to a few standard crop species owing to restrictive recommendations at the regulatory level. However, guidelines by the Organisation for Economic Development and Cooperation (OECD) were recently amended in 2006 to include a list of herbaceous non-crop plant species suitable for testing. The objective of this study was to outline the optimum germination requirements for a selection of wild species for which seeds were readily available from commercial suppliers. RESULTS: Of the 29 herbaceous terrestrial and wetland species included in this study, all achieved 50% germination and 23 reached > 70% germination to meet the criterion outlined in the OECD guidelines. Most species attained their maximum germination within 14 days or less. Cold stratification of imbibed seeds improved germination for 14 species. Increasing sowing soil depth did not improve seed germination. The variance attained in this experiment between replicates was low, especially for species with > 70% germination (standard error approximately 5%). CONCLUSION: The present study showed that 23 of the 29 species tested required minimal pretreatments and produced consistent, reliable and uniform germination reaching at least 70%. The inclusion of wild plant species in regulatory testing should be given real consideration.