A smart curtailment approach for reducing bat fatalities and curtailment time at wind energy facilities.

The development and expansion of wind energy is considered a key global threat to bat populations. Bat carcasses are being found underneath wind turbines across North and South America, Eurasia, Africa, and the Austro-Pacific. However, relatively little is known about the comparative impacts of techniques designed to modify turbine operations in ways that reduce bat fatalities associated with wind energy facilities. This study tests a novel approach for reducing bat fatalities and curtailment time at a wind energy facility in the United States, then compares these results to operational mitigation techniques used at other study sites in North America and Europe. The study was conducted in Wisconsin during 2015 using a new system of tools for analyzing bat activity and wind speed data to make near real-time curtailment decisions when bats are detected in the area at control turbines (N = 10) vs. treatment turbines (N = 10). The results show that this smart curtailment approach (referred to as Turbine Integrated Mortality Reduction, TIMR) significantly reduced fatality estimates for treatment turbines relative to control turbines for pooled species data, and for each of five species observed at the study site: pooled data (-84.5%); eastern red bat (Lasiurus borealis, -82.5%); hoary bat (Lasiurus cinereus, -81.4%); silver-haired bat (Lasionycteris noctivagans, -90.9%); big brown bat (Eptesicus fuscus, -74.2%); and little brown bat (Myotis lucifugus, -91.4%). The approach reduced power generation and estimated annual revenue at the wind energy facility by ≤ 3.2% for treatment turbines relative to control turbines, and we estimate that the approach would have reduced curtailment time by 48% relative to turbines operated under a standard curtailment rule used in North America. This approach significantly reduced fatalities associated with all species evaluated, each of which has broad distributions in North America and different ecological affinities, several of which represent species most affected by wind development in North America. While we recognize that this approach needs to be validated in other areas experiencing rapid wind energy development, we anticipate that this approach has the potential to significantly reduce bat fatalities in other ecoregions and with other bat species assemblages in North America and beyond.

Increasing the discrimination power of rapid evaporative ionisation mass spectrometry (REIMS) in analytical control tissue quality screening and cell line sample identification.

RATIONALE: Rapid Evaporative Ionisation Mass Spectrometry (REIMS) has been evaluated as a tool to improve analytical efficiency and add capability in areas within Pharmaceutical Research and Development (Pharma R&D). This article reports the comparison of single MS, and tandem MS/MS REIMS (REIMS and REIMS/MS) methodologies to investigate which mode produces maximum discrimination power for screening applications. METHODS: Control tissue samples and cell line suspension samples were analysed using optimised REIMS and REIMS/MS to evaluate which technique produced optimal discrimination power for control tissue and cell line identification. The iKnife sampling tool and a prototype 'cell sampler' were utilised for tissue and cell analysis, respectively. The REIMS source was coupled to a hybrid Quadrupole-Time Of Flight (QTOF) mass spectrometer. Multivariate Analysis (MVA) was utilised to evaluate the resulting Mass Spectrometry (MS) data and discriminate between sample types. RESULTS: Proof of concept investigations demonstrating that REIMS/MS offered increased MVA discrimination for sample identification, compared with REIMS, is presented for the first time. Control tissue data showed discrimination by timepoint classification over 0-144 h storage after removal from the host. Timepoint discrimination was optimised using REIMS/MS with a collision energy that effectively maximised ion fragmentation. Similar optimisation was observed when REIMS/MS was applied to the identification of cell lines. CONCLUSIONS: The proof of concept results demonstrate that REIMS/MS can offer advantages over REIMS for control tissue quality screening, and cell line identification applications in Pharma R&D. Further work following this proof of concept investigation is being undertaken to implement the technology for these applications, utilising the optimised REIMS/MS methodology. REIMS/MS will also be used as an optimised tool for other applications.

Investigation of different approaches to incorporating internal standard in DBS quantitative bioanalytical workflows and their effect on nullifying hematocrit-based assay bias.

Hematocrit (HCT)-based assay bias (composed of area and recovery bias) is an important contributing factor to the barriers that currently hinder the development and acceptance of dried blood spots (DBS) as a widely used quantitative bioanalytical sampling technique for regulatory studies. This article describes the evaluation of a practical internal standard spray addition technique, used prior to LC-MS/MS analysis, which is demonstrated to nullify the effect of recovery bias. To our knowledge, this is the first time a potential solution to HCT-based recovery bias has been investigated in detail and reported in the literature. This new technique is coupled with accurate volume DBS sampling, whole-spot extraction, and automated direct elution techniques to demonstrate a workflow that both nullifies HCT-based assay bias and the additional manual extraction burden associated with DBS analysis.

Game bird carcasses are less persistent than raptor carcasses, but can predict raptor persistence dynamics.

Researchers conduct post-construction fatality monitoring (PCFM) to determine a wind energy facility's direct impacts on wildlife. Results of PCFM can be used to evaluate compliance with permitted take, potentially triggering adaptive management measures or offsetting mitigation; reducing uncertainty in fatality rates benefits wind companies, wildlife agencies, and other stakeholders. As part of PCFM, investigators conduct carcass persistence trials to account for imperfect detection during carcass surveys. In most PCFM studies, pen-raised game birds and other non-raptor surrogates have been used to estimate persistence of all large birds, including raptors. However, there is a growing body of evidence showing carcass persistence varies by bird type; raptor fatality estimates based on game bird carcass persistence may therefore be biased high. We conducted raptor and game bird carcass persistence field trials for 1 year at 6 wind energy facilities. Raptor carcass persistence varied by habitat and season, whereas the best-supported game bird model only included habitat. Raptor persistence probabilities were higher than corresponding game bird persistence probabilities for 13 of the 16 habitat and season combinations. Analysis of a curated large bird persistence meta-dataset showed that raptor carcass persistence varied by season, habitat, and region. The probability of persisting through a 30-day search interval ranged from 0.44 to 0.99 for raptors and from 0.16 to 0.79 for game birds. Raptor persistence was significantly higher than game bird persistence for 95% of the sampled strata. We used these carcass persistence estimates to develop linear mixed-effects models that predict raptor persistence probabilities based on estimated game bird persistence probabilities. Our scaling model provides an important statistical method to address gaps in raptor persistence data at sites in a broad range of landscape contexts in the continental United States and should be used to inform fatality estimation when site-specific raptor persistence data are limited or absent.

Efficacy and cost of acoustic-informed and wind speed-only turbine curtailment to reduce bat fatalities at a wind energy facility in Wisconsin.

Current research estimates hundreds of thousands of turbine-related bat fatalities in North America annually. In an effort to reduce impacts of wind energy production on bat populations, many facilities implement operational curtailment strategies that limit turbine blade rotation during conditions when nighttime wind speeds are low. Incorporating real-time bat activity data into wind speed-only curtailment (WOC) strategies may increase operational flexibility by allowing turbines to operate normally when bats are not present near turbines. We evaluated costs and benefits of implementing the Turbine Integrated Mortality Reduction (TIMR) system, an approach that informs a curtailment-triggering algorithm based on wind speed and real-time bat acoustic data, compared to a WOC strategy in which turbines were curtailed below 4.5 meters per second (m/s) at a wind energy facility in Fond Du Lac County, Wisconsin. TIMR is a proprietary system and we had no access to the acoustic data or bat call analysis software. Operational parameters for the TIMR system were set to allow curtailment at all wind speeds below 8.0 m/s during the study period when bats were acoustically detected. Overall, the TIMR system reduced fatalities by 75% compared to control turbines, while the WOC strategy reduced fatalities by 47%. An earlier analysis of the same TIMR data neglected to account for carcasses occurring outside the plot boundary and estimated an 84.5% fatality reduction due to the TIMR system. Over the study period, bat activity led to curtailment of TIMR turbines during 39.4% of nighttime hours compared to 31.0% of nighttime hours for WOC turbines, and revenue losses were approximately 280% as great for TIMR turbines as for turbines operated under the WOC strategy. The large cost difference between WOC and TIMR was driven by the 4.5 m/s versus 8.0 m/s wind speed thresholds for curtailment, but our study site has a relatively low average wind speed, which may also have contributed; other wind operators considering the TIMR system will need to consider their ability to absorb production losses in relation to their need to reduce bat fatality rates.

Method of applying internal standard to dried matrix spot samples for use in quantitative bioanalysis.

A novel technique is presented that addresses the issue of how to apply internal standard (IS) to dried matrix spot (DMS) samples that allows the IS to integrate with the sample prior to extraction. The TouchSpray, a piezo electric spray system, from The Technology Partnership (TTP), was used to apply methanol containing IS to dried blood spot (DBS) samples. It is demonstrated that this method of IS application has the potential to work in practice, for use in quantitative determination of circulating exposures of pharmaceuticals in toxicokinetic and pharmacokinetic studies. Three different methods of IS application were compared: addition of IS to control blood prior to DBS sample preparation (control 1), incorporation into extraction solvent (control 2), and the novel use of TouchSpray technology (test). It is demonstrated that there was no significant difference in accuracy and precision data using these three techniques obtained using both manual extraction and direct elution.

Direct quantitative bioanalysis of drugs in dried blood spot samples using a thin-layer chromatography mass spectrometer interface.

The CAMAG thin-layer chromatography mass spectrometer (TLC-MS) interface has been assessed as a tool for the direct quantitative bioanalysis of drugs from dried blood spot (DBS) samples, using an MS detector, with or without high-performance liquid chromatography (HPLC) separation. The approach gave acceptable sensitivity, linearity, accuracy, and precision data for bioanalytical validations with and without the inclusion of HPLC separation. In addition, the direct elution technique was shown to increase assay sensitivity for a range of analytes representing a wide "chemical space" for pharmaceutical-type molecules over that obtained by conventional manual extraction of samples (punching of DBS and elution with solvent prior to HPLC-MS analysis). Investigations were performed to optimize extraction time, minimize sample-to-sample carry-over, and compare chromatographic performance. On the basis of this preliminary assessment, it has been demonstrated that the TLC-MS interface has the potential to be an effective tool for the direct analysis of drugs in DBS samples at physiologically relevant concentrations, an approach that could provide significant time and cost savings and greatly simplify bioanalytical procedures compared to current manual practices. Further, the increased sensitivity compared to that of manual extraction may enable the analysis of analytes not currently amenable to DBS sampling due to limitations in assay sensitivity.

Validation of methods for determining pediatric midazolam using wet whole blood and volumetric absorptive microsampling.

Aim: Collection and quantitative analysis in dry blood using volumetric absorptive microsampling (VAMS™) potentially offers significant advantages over conventional wet whole blood analysis. This manuscript explores their use for pediatric sampling and explores additional considerations for the validation of the bioanalytical method. Results: HPLC-MS/MS methods for the determination of midazolam and its major metabolite 1-OH midazolam in both whole wet blood, and dry blood collected on VAMS were developed, validated, and used to support an observational clinical study to compare pharmacokinetic parameters in pediatric patients. Conclusion: Validation data met internationally accepted guideline criteria. A strong correlation was observed in calculated concentrations between wet and dry test samples, indicating that VAMS is a suitable technique for use in pediatric clinical studies.

Developing an efficient protocol for monitoring eagle fatalities at wind energy facilities.

Researchers typically conduct fatality monitoring to determine a wind energy facility's direct impacts on wildlife. In the United States, wind energy impacts on eagles have received increased attention in recent years because eagle incidental take permits became available. Permit holders are required to conduct fatality monitoring to evaluate compliance with permitted eagle take. Our objective was to develop an efficient eagle fatality monitoring protocol with a quantifiable detection probability based on a stationary scanning search method. We conducted scanning searches for eagle carcasses at four wind energy facilities. We estimated searcher efficiency of the scanning search method using feathered turkey decoys as eagle carcass surrogates, used publicly available data on large raptor carcass distances from turbines to evaluate the proportion of carcasses expected to occur in searched areas, and estimated carcass persistence rates for game birds and raptors. These three bias adjustments were combined to estimate the overall probability of detection for the scanning search method. We found generally high searcher efficiency for the scanning search method, with 76% of decoys detected; however, detection decreased with distance and difficulty of visibility class. Mean carcass persistence time varied between 28 and 76 days for raptors and between three and nine days for game birds, showing that game birds do not persist as long as raptors. We estimated that 95% of large avian carcasses fall within 100 m of turbine bases, and 99% fall within 150 m. Using these estimates and assuming a 30-day search interval for all facility turbines, we estimated that the probability of detecting a large raptor carcass using the scanning search method at a wind facility ranged from 0.50 to 0.69. Our research suggests a monitoring program that uses scanning searches can be a cost-effective approach for gathering data necessary to meet incidental eagle take permit requirements.

Exotic Plant Infestation Is Associated with Decreased Modularity and Increased Numbers of Connectors in Mixed-Grass Prairie Pollination Networks.

The majority of pollinating insects are generalists whose lifetimes overlap flowering periods of many potentially suitable plant species. Such generality is instrumental in allowing exotic plant species to invade pollination networks. The particulars of how existing networks change in response to an invasive plant over the course of its phenology are not well characterized, but may shed light on the probability of long-term effects on plant-pollinator interactions and the stability of network structure. Here we describe changes in network topology and modular structure of infested and non-infested networks during the flowering season of the generalist non-native flowering plant, Cirsium arvense in mixed-grass prairie at Badlands National Park, South Dakota, USA. Objectives were to compare network-level effects of infestation as they propagate over the season in infested and non-infested (with respect to C. arvense) networks. We characterized plant-pollinator networks on 5 non-infested and 7 infested 1-ha plots during 4 sample periods that collectively covered the length of C. arvense flowering period. Two other abundantly-flowering invasive plants were present during this time: Melilotus officinalis had highly variable floral abundance in both C. arvense-infested and non-infested plots and Convolvulus arvensis, which occurred almost exclusively in infested plots and peaked early in the season. Modularity, including roles of individual species, and network topology were assessed for each sample period as well as in pooled infested and non-infested networks. Differences in modularity and network metrics between infested and non-infested networks were limited to the third and fourth sample periods, during flower senescence of C. arvense and the other invasive species; generality of pollinators rose concurrently, suggesting rewiring of the network and a lag effect of earlier floral abundance. Modularity was lower and number of connectors higher in infested networks, whether they were assessed in individual sample periods or pooled into infested and non-infested networks over the entire blooming period of C. arvense. Connectors typically did not reside within the same modules as C. arvense, suggesting that effects of the other invasive plants may also influence the modularity results, and that effects of infestation extend to co-flowering native plants. We conclude that the presence of abundantly flowering invasive species is associated with greater network stability due to decreased modularity, but whether this is advantageous for the associated native plant-pollinator communities depends on the nature of perturbations they experience.

Optimization of an automated IS addition system for use in high-throughput quantitative DBS analysis.

BACKGROUND: Automated DBS direct elution systems are available that incorporate IS spray modules which, unlike conventional IS addition via the extraction solvent, apply IS prior to DBS samples prior to extraction, allowing analyte and IS to be coextracted. RESULTS: IS spray system parameters were optimized to identify the conditions that produced the best analytical performance in quantitative bioanalytical assays, without interfering with the integrity of the DBS sample prior to extraction. CONCLUSION: LC-MS/MS method validations across four representative small molecule assays using the optimized IS spray conditions were demonstrated to produce analytical performance comparable to conventional methods of IS addition, demonstrating that the spray technique is a viable alternative.

DBS direct elution: optimizing performance in high-throughput quantitative LC-MS/MS analysis.

BACKGROUND: Automated DBS direct elution techniques eliminate the manual extraction burden of DBS bioanalysis, offer good quantitative performance, the ability to eliminate hematocrit-based assay bias, and, previous reports have demonstrated that significant increases in assay sensitivity compared with manual DBS extraction are possible. RESULTS: An investigation into elucidating parameters for optimized generic DBS direct elution for high sample throughput quantitative bioanalytical applications is presented for the first time. Generic direct elution conditions were identified that enabled LC-MS/MS assay sensitivity to be maximized while retaining acceptable chromatographic performance. CONCLUSION: Compared with generic conventional DBS manual extraction, assay sensitivity was demonstrated to be increased up to 33-fold across four representative small molecule compounds, using the recommended direct elution conditions.

Multiplexed extraction and quantitative analysis of pharmaceuticals from DBS samples using digital microfluidics.

BACKGROUND: Dried blood spot (DBS) sampling is emerging as a valuable technique in a variety of fields, including clinical and preclinical testing of pharmaceuticals. Despite this popularity, current DBS sampling and analysis processes remain laborious and time consuming. Digital microfluidics, a microscale liquid-handling technique, characterized by the manipulation of discrete droplets on open electrode arrays, offers a potential solution to these problems. RESULTS: We report a new digital microfluidic method for multiplexed extraction and analysis of pharmaceuticals in DBS samples. In the new method, four DBS samples are extracted in microliter-sized droplets containing internal standard, and the extract is delivered to dedicated nanoelectrospray ionization emitters for direct analysis by tandem mass spectometry and selected reaction monitoring. CONCLUSION: The new method allows for an order of magnitude reduction in processing time and approximately three-times reduction in extraction solvent relative to conventional techniques, while maintaining acceptable analytical performance for most drugs tested.

Dried matrix spot direct analysis: evaluating the robustness of a direct elution technique for use in quantitative bioanalysis.

BACKGROUND: The surge in interest in switching from traditionally used wet plasma to dried matrix spot (DMS) sampling and analysis to support pharmaceutical drug development is due to the significant ethical, financial and data quality advantages on offer. Unfortunately these advantages do not extend to sample bioanalysis, as DMS extraction is more complex than the protein precipitation method typically used for wet plasma analysis. Direct elution techniques coupled to HPLC-MS/MS have been identified as a potential means to counter this additional complexity. RESULTS: The robustness and reproducibility of DMS HPLC-MS/MS data generated using a CAMAG DBS-MS 16 prototype automated direct elution instrument has been demonstrated to meet or exceed results obtained using a conventional manual extraction methodology. CONCLUSION: The data generated suggest that a simple and fast direct elution method of DMS samples that does not require additional sample or extract clean-up, offers sufficiently robust performance to be compatible with high-sample-throughput quantitative analysis. Further evaluation of the technique and the development of more advanced fully automated direct elution instrumentation is fully warranted.

Quantitative analysis of therapeutic drugs in dried blood spot samples by paper spray mass spectrometry: an avenue to therapeutic drug monitoring.

A method is presented for the direct quantitative analysis of therapeutic drugs from dried blood spot samples by mass spectrometry. The method, paper spray mass spectrometry, generates gas phase ions directly from the blood card paper used to store dried blood samples without the need for complex sample preparation and separation; the entire time for preparation and analysis of blood samples is around 30 s. Limits of detection were investigated for a chemically diverse set of some 15 therapeutic drugs; hydrophobic and weakly basic drugs, such as sunitinib, citalopram, and verapamil, were found to be routinely detectable at approximately 1 ng/mL. Samples were prepared by addition of the drug to whole blood. Drug concentrations were measured quantitatively over several orders of magnitude, with accuracies within 10% of the expected value and relative standard deviation (RSD) of around 10% by prespotting an internal standard solution onto the paper prior to application of the blood sample. We have demonstrated that paper spray mass spectrometry can be used to quantitatively measure drug concentrations over the entire therapeutic range for a wide variety of drugs. The high quality analytical data obtained indicate that the technique may be a viable option for therapeutic drug monitoring.

Study to assess the effect of age of control human and animal blood on its suitability for use in quantitative bioanalytical DBS methods.

BACKGROUND: A study was performed to evaluate the suitability of stored EDTA-treated control whole blood for use in the preparation of calibration standards and quality control samples for quantitative bioanalytical methods employing dried blood spot (DBS) samples to support pharmaceutical exposure studies. RESULTS: It has been demonstrated that a storage time of 14 days for control human and animal blood is suitable for producing quantitative analytical results within internationally recognised acceptance criteria for two analytes. Furthermore, blood hemolysis and chill-thaw cycles have been evaluated and shown not to affect bioanalytical results notably. Aggressive mixing techniques can result in rat blood coagulation; however, this does not occur with other species tested and can be affected by the method of blood collection. CONCLUSION: Control whole blood handled and stored using the recommendations generated from this study will not notably affect quantitative bioanalytical results when used for the preparation of calibration standards and quality control samples for DBS assays. It was demonstrated that control human and animal blood can be stored for periods long enough to effectively eliminate wastage.