Circulating Tumor DNA as a Predictive Biomarker for Clinical Outcomes With Margetuximab and Pembrolizumab in Pretreated HER2-Positive Gastric/ Gastroesophageal Adenocarcinoma.

PURPOSE: To assess the ability of circulating tumor DNA (ctDNA)-based testing to identify patients with HER2 (encoded by ERBB2)-positive gastric/gastroesophageal adenocarcinoma (GEA) who progressed on or after trastuzumab-containing treatments were treated with combination therapy of anti-HER2 and anti-PD-1 agents. METHODS: ctDNA analysis was performed retrospectively using plasma samples collected at study entry from 86 patients participating in the phase 1/2 CP-MGAH22-05 study (NCT02689284). RESULTS: Objective response rate (ORR) was significantly higher in evaluable ERBB2 amplification-positive vs - negative patients based on ctDNA analysis at study entry (37% vs 6%, respectively; P = .00094). ORR was 23% across all patients who were evaluable for response. ERBB2 amplification was detected at study entry in 57% of patients (all HER2 positive at diagnosis), and detection was higher (88%) when HER2 status was determined by immunohistochemistry fewer than 6 months before study entry. ctDNA was detected in 98% (84/86) of patients tested at study entry. Codetected ERBB2-activating mutations were not associated with response. CONCLUSIONS: Current ERBB2 status may be more effective than archival status at predicting clinical benefit from margetuximab plus pembrolizumab therapy. ctDNA testing for ERBB2 status prior to treatment will spare patients from repeat tissue biopsies, which may be reserved for reflex testing when ctDNA is not detected.

Differing behavioral changes in crayfish and bluegill under short- and long-chain PFAS exposures: Field study in Northern Michigan, USA.

The emergent contaminant family, per- and poly-fluorinated alkyl substances (PFAS) has gained research attention due to their widespread detection and stability within the environment. Despite the growing amount of research on perfluorooctanesulfonic acid (PFOS) and perfluoro-n-octanoic acid (PFOA) in aquatic organisms, investigations detailing behavioral and physiological effects of aquatic organisms exposed to a mixture of PFAS analytes in the wild have been limited. The objective of this study was to evaluate the potential behavioral and histological effects of environmental exposure to PFAS compounds within multiple trophic levels of aquatic ecosystems. The current study investigates effects of environmentally relevant PFAS concentration exposures in crayfish (Faxonius immunis, F. rusticus, F. virilis) and bluegill (Lepomis macrochirus) sourced from four water bodies in Northern Michigan. Antipredator response and foraging behavioral assays were used to investigate potential effects on crayfish; a swimming speed behavioral assay and liver and gill histology analysis were used to investigate potential effects on fish. Linear mixed model and multiple regression analyses resulted in significant relationships between tissue accumulation levels of long chain PFAS compounds and crayfish foraging and fish critical swimming speed responses. Crayfish foraging decreased and fish critical swim speeds increased with PFAS exposure which may lead to energetic and population concerns. Antipredator response in crayfish and liver and gill histology in fish were not significantly related to PFAS tissue or water concentrations. The sensitivity of crayfish and bluegill behavior contributes to the growing body of research regarding the differential toxicity of short-chain and long-chain PFAS compounds. The sensitivity of some aquatic organism behaviors to PFAS accumulated in tissue may have implications for PFAS transfer and alterations to ecosystem functioning; based on the results of this field study, further laboratory research is recommended to further evaluate these relationships.

Running away or running to? Do prey make decisions solely based on the landscape of fear or do they also include stimuli from a landscape of safety?

Predator-prey interactions are a key part of ecosystem function, and non-consumptive effects fall under the landscape of fear theory. Under the landscape of fear, the antipredator responses of prey are based on the spatial and temporal distribution of predatory cues in the environment. However, the aversive stimuli (fear) are not the only stimuli prey can utilize when making behavioral decisions. Prey might also be using attractive stimuli that represent safety to guide decision making. Using a novel, orthogonal design, we were able to spatially separate aversive and attractive stimuli to determine whether prey are utilizing safety cues to navigate their environment. Crayfish Faxonius rusticus were placed in the center of a behavioral arena. Aversive stimuli of either predatory bass Micropterus salmoides cues or conspecific alarm cues increased along the x-axis of the behavioral arena. Safety cues (shelters) increased along the y-axis by decreasing the number of shelter openings in this direction. Crayfish were allowed two phases to explore the arena: one without the fearful stimuli and one with the stimuli. Linear mixed models were conducted to determine whether movement behaviors and habitat utilization were affected by the phase of the trial and the type of aversive stimuli. Crayfish responded more strongly to alarm cues than to fear cues, with only alarm cues significantly impacting habitat utilization. When responding to alarm cues, crayfish used safety cues as well as fear cues to relocate themselves within the arena. Based on these results, we argue that crayfish utilize a landscape of safety in conjunction with a landscape of fear when navigating their environment.

Spatial, but not temporal, aspects of orientation are controlled by the fine-scale distribution of chemical cues in turbulent odor plumes.

Orientation within turbulent odor plumes occurs across a vast range of spatial and temporal scales. From salmon homing across featureless oceans to microbes forming reproductive spores, the extraction of spatial and temporal information from chemical cues is a common sensory phenomenon. Yet, given the difficulty of quantifying chemical cues at the spatial and temporal scales used by organisms, discovering what aspects of chemical cues control orientation behavior has remained elusive. In this study, we placed electrochemical sensors on the carapace of orienting crayfish and measured, with fast temporal rates and small spatial scales, the concentration fluctuations arriving at the olfactory appendages during orientation. Our results show that the spatial aspects of orientation (turning and heading angles) are controlled by the temporal aspects of odor cues.

Mapping Dynamic Exposure: Constructing GIS Models of Spatiotemporal Heterogeneity in Artificial Stream Systems.

In flowing environments, the degree of turbulent flow determines the movement and distribution of chemicals. Variation in flow alters the patchiness of toxicant plumes within a stream ecosystem. This patchiness translates into variability in exposure pulses for organisms encountering the toxic plume. Throughout a stream, the processes that give rise to chemical plume structure will vary as a function of local flow characteristics. This research examines the influence of toxicant mode of entry and stream flow velocity on the spatiotemporal patterning of exposure. Two introduction treatments were evaluated: one mimicking groundwater and the other mimicking runoff. The influence of flow regime was examined through the comparison of models constructed under two stream flow velocities. Concentrations of a tracer molecule were recorded using an electrochemical monitoring system. From these localized, direct measurements, geographic information systems (GIS) were used to model exposure throughout the stream. Conceptualizing exposure as a series of toxicant pulses, exposure can be defined using a variety of chemical peak characteristics. Three-dimensional, layered maps were constructed defining exposure as the integrated area of toxicant peaks, the magnitude of peaks, and peak frequency. Differences in the spatial and temporal patterning of exposure were apparent both within treatments and between treatments. No two definitions of exposure yielded the same exposure distributions for any treatment. These models demonstrate that distribution of chemical exposure throughout a stream ecosystem is linked to both toxicant mode of introduction and stream hydrodynamics. Furthermore, these results demonstrate that optimal exposure modeling relies on first defining exposure.

Express yourself: Individuals with bold personalities exhibit increased behavioral sensitivity to dynamic herbicide exposure.

The majority of ecotoxicological studies performed measure average responses from individuals which do not account for the inter-individual variation in the responses of animals to environmental stimuli (i.e. the personality of individuals). Thus, these designs assume that all individuals will respond to contaminant exposure in a similar manner. Additionally, commonly used constant, static exposure regime designs neglect to recognize the spatial and temporal variation in contaminant plume structures as they move throughout fluid environments. The purpose of this study was to understand the effects of the structural characteristics (concentration, duration, and frequency) of temporally and spatially variant contaminant plumes on the personality of individuals. This experimental design aimed to construct a sensitive definition of exposure by connecting sublethal effects of toxicants and realistic exposure regimes. This study used escape response of Faxonius virilis crayfish from the predatory odor of Micropterus salmoides prior to and following exposure to the herbicide, atrazine. Atrazine was delivered in pulses to flow through exposure arenas for a total of 47 h while manipulating the concentration, frequency, and duration of the herbicide pulses. Escape response of crayfish prior to exposure was used to categorize animals into bold and shy personalities. The change in escape response was analyzed and resulted in a personality-dependent behavioral sensitivity to the polluted environment. Individuals classified as bold showed increased change in response to predatory odor relative to shy animals. Bold animals exhibited decreased activity after exposure where no change was presented in shy individuals. Shifts in individual behavior have impacts on the population level (e.g. resource acquisition/value; interspecies competition) and the ecosystem level (e.g. food web dynamics; trophic cascades). This study demonstrates the importance of sensitive measures in ecological risk assessment methods.

Bt Proteins Exacerbate Negative Growth Effects in Juvenile Rusty (F. rusticus) Crayfish Fed Corn Diet.

The adoption of genetically modified (GM) crops has occurred rapidly in the United States. The transfer of GM corn byproducts from agricultural fields to nearby streams after harvest is significant and occurs well into the post-harvest year. These corn leaves, stems, and cobs then become a detrital food source for organisms, such as shredders in the stream ecosystem. Considering that the nontarget effects of Bt corn have been observed in some terrestrial organisms, we assessed whether Bt toxins affect an important aquatic organism, juvenile F. rusticus crayfish. Juvenile crayfish were fed six distinct diet treatments: two varieties of Bt corn, two non-Bt controls of herbicide tolerant corn, and two controls: fish gelatin and river detritus. Juveniles were fed these diets while housed in flow-through artificial streams that received natural stream water from a local source. Specific growth rate and survivorship of the crayfish were measured throughout the study. Juveniles fed corn diets grew significantly less and had reduced survival compared with juveniles fed fish gelatin or river detritus diets. Furthermore, juveniles fed one Bt variety of corn (VT Triple Pro®) exhibited significantly less growth than those fed one of the herbicide tolerant varieties (Roundup Ready 2®). Our study shows that corn inputs to streams may be detrimental to the growth and survivorship of juvenile crayfish and that certain Bt varieties may exacerbate these negative effects. These effects on crayfish will have repercussions for the entire ecosystem, because crayfish are conduits of energy between many trophic levels.

Variable Background Flow on Aquatic Toxicant Exposure Alters Foraging Patterns on Crayfish.

Climate change is expected to alter hydrological cycles on global and regional scales, impacting groundwater and surface water inputs to stream habitats. In the midwestern United States, the volume and frequency of inputs are expected to become increasingly variable. This region has a high incidence of agriculture, creating enormous potential for transport of pesticides and herbicides into aquatic ecosystems. Metolachlor, an herbicide for corn and soybean crops, has been demonstrated to contaminate surface water and groundwater in the region. This study examines the impact of variable flow conditions on the toxicity of environmentally relevant concentrations of metolachlor in a macroinvertebrate found in midwestern streams, the rusty crayfish (Faxonius rusticus). Changes in crayfish foraging behavior were analyzed using a Mixed Model ANCOVA. Under toxicant exposure, crayfish significantly increased their consumption of macrophytes, but only under the variable flow regime. Thus, the increased variability in toxicant exposure impacted crayfish foraging behavior more than other flow regimes. This significant interaction between flow regime and metolachlor exposure suggests that the greater variability in toxicant inputs to streams may lead to more severe changes in behavior for exposed organisms.

Targeting CD123 in acute myeloid leukemia using a T-cell-directed dual-affinity retargeting platform.

T-cell-directed killing of tumor cells using bispecific antibodies is a promising approach for the treatment of hematologic malignancies. Here we describe our preclinical work with a dual-affinity retargeting (DART) molecule generated from antibodies to CD3 and CD123, designed to redirect T cells against acute myeloid leukemia blasts. The CD3×CD123 DART (also referred to as MGD006/S80880) consists of 2 independent polypeptides, each composed of the VH of 1 antibody in tandem with the VL of the other antibody. The target antigen CD123 (interleukin 3RA) is highly and differentially expressed in acute myeloid leukemia (AML) blasts compared with normal hematopoietic stem and progenitor cells. In this study we demonstrate that the CD3×CD123 DART binds to both human CD3 and CD123 to mediate target-effector cell association, T-cell activation, proliferation, and receptor diversification. The CD3×CD123 DART also induces a dose-dependent killing of AML cell lines and primary AML blasts in vitro and in vivo. These results provide the basis for testing the CD3×CD123 DART in the treatment of patients with CD123(+) AML.

Scaling to the Organism: An Innovative Model of Dynamic Exposure Hotspots in Stream Systems.

In flowing systems, fluctuations in the frequency, magnitude, and duration of exposure occurs due to turbulence and geomorphology, causing spatial and temporal variations in chemical exposure at the scale of the organism. Spatial models representing toxicant distribution at the appropriate scales of stream organisms are noticeably missing from the literature. To characterize the fine scale distribution of pollutants in freshwater streams at the scale of a benthic organism, nine artificial stream habitats were created (riffle, pool, run, bend, woody debris) with either sand or gravel substrate. Dopamine was released as a chemical tracer, mimicking a groundwater source, and measurements were recorded with a microelectrode and Epsilon electrochemical recording system. Proxies for the frequency, magnitude, and duration of chemical exposure were extracted. Geographic information systems and an inverse distance weight interpolation technique were used to predict spatially the chemical distribution throughout the habitats. Spatial and temporal variations of exposure were exhibited within and across habitats, indicating that the frequency, magnitude, and duration of exposure is influenced by the organism's location within a habitat and the habitat it resides in. The run and pool with sand substrate contained the greatest frequency, magnitude, and duration of exposure, suggesting a more detrimental exposure compared to other habitats. Differences in peak heights within and across habitats are orders of magnitude in value. Spatial and temporal fluctuations of fine scale exposure need to be considered in both ecotoxicology and water quality modeling to represent and understand the exposure of pollutants impacting benthic organisms.

Exposure Through Runoff and Ground Water Contamination Differentially Impact Behavior and Physiology of Crustaceans in Fluvial Systems.

Chemical pollutants enter aquatic systems through numerous pathways (e.g., surface runoff and ground water contamination), thus associating these contaminant sources with varying hydrodynamic environments. The hydrodynamic environment shapes the temporal and spatial distribution of chemical contaminants through turbulent mixing. The differential dispersal of contaminants is not commonly addressed in ecotoxicological studies and may have varying implications for organism health. The purpose of this study is to understand how differing routes of exposure to atrazine alter social behaviors and physiological responses of aquatic organisms. This study used agonistic encounters in crayfish Orconectes virilis as a behavioral assay to investigate impact of sublethal concentrations of atrazine (0, 40, 80, and 160 µg/L) delivered by methods mimicking ground water and surface runoff influx into flow-through exposure arenas for a total of 23 h. Each experimental animal participated in a dyadic fight trial with an unexposed opponent. Fight duration and intensity were analyzed. Experimental crayfish hepatopancreas and abdominal muscle tissue samples were analyzed for cytochrome P450 and acetylcholinesterase levels to discern mechanism of detoxification and mode of action of atrazine. Atrazine delivered via runoff decreased crayfish overall fight intensity and contrastingly ground water delivery increased overall fight intensity. The behavioral differences were mirrored by increases in cytochrome P450 activity, whereas no differences were found in acetylcholinesterase activity. This study demonstrates that method of delivery into fluvial systems has differential effects on both behavior and physiology of organisms and emphasizes the need for the consideration of delivery pathway in ecotoxicological studies and water-impairment standards.

Exposure to Sublethal Ammonia Concentrations Alters the Duration and Intensity of Agonistic Interactions in the Crayfish, Orconectes rusticus.

Crayfish extract information from chemical stimuli during social interactions. Commercial fertilizers increase background ammonia concentrations which may interfere with chemical communication. Background pollution can disrupt perception of chemical stimuli in three ways: masking, sensory impairment, physiological impairment or in combination. We investigated whether exposure to ammonia alters agonistic behavior. Crayfish pairs exposed to 0.9 mg/L ammonia fought for a longer duration, while crayfish exposed to 9.0 mg/L ammonia fought for a shorter duration. Altering activity patterns of crayfish may alter crayfish populations leading to a nonproportional impact because of their importance to the structure and function of aquatic ecosystems.

Targeting HIV Reservoir in Infected CD4 T Cells by Dual-Affinity Re-targeting Molecules (DARTs) that Bind HIV Envelope and Recruit Cytotoxic T Cells.

HIV reservoirs and production of viral antigens are not eliminated in chronically infected participants treated with combination antiretroviral therapy (cART). Novel therapeutic strategies aiming at viral reservoir elimination are needed to address chronic immune dysfunction and non-AIDS morbidities that exist despite effective cART. The HIV envelope protein (Env) is emerging as a highly specific viral target for therapeutic elimination of the persistent HIV-infected reservoirs via antibody-mediated cell killing. Dual-Affinity Re-Targeting (DART) molecules exhibit a distinct mechanism of action via binding the cell surface target antigen and simultaneously engaging CD3 on cytotoxic T lymphocytes (CTLs). We designed and evaluated Env-specific DARTs (HIVxCD3 DARTs) derived from known antibodies recognizing diverse Env epitopes with or without broadly neutralizing activity. HIVxCD3 DARTs derived from PGT121, PGT145, A32, and 7B2, but not VRC01 or 10E8 antibodies, mediated potent CTL-dependent killing of quiescent primary CD4 T cells infected with diverse HIV isolates. Similar killing activity was also observed with DARTs structurally modified for in vivo half-life extension. In an ex vivo model using cells isolated from HIV-infected participants on cART, combinations of the most potent HIVxCD3 DARTs reduced HIV expression both in quiescent and activated peripheral blood mononuclear cell cultures isolated from HIV-infected participants on suppressive cART. Importantly, HIVxCD3 DARTs did not induce cell-to-cell virus spread in resting or activated CD4 T cell cultures. Collectively, these results provide support for further development of HIVxCD3 DARTs as a promising therapeutic strategy for targeting HIV reservoirs.

Mimicking natural systems: Changes in behavior as a result of dynamic exposure to naproxen.

Animals living in aquatic habitats regularly encounter anthropogenic chemical pollution. Typically, the toxicity of a chemical toxicant is determined by the median lethal concentration (LC50) through a static exposure test. However, LC50 values and static tests do not provide an accurate representation of exposure to pollutants within natural stream systems. In their native habitats, animals experience exposure as a fluctuating concentration due to turbulent mixing, temporal variations of contamination (seasonal inputs), and contaminant input type (point vs. non-point). Research has shown that turbulent environments produce exposures with a high degree of fluctuation in frequency, duration, and intensity. In order to more effectively evaluate the effects of pollutants, we created a dynamic exposure paradigm, utilizing both flow and substrate within a small mesocosm. A commonly used pharmaceutical, naproxen, was used as the toxicant and female crayfish (Orconectes virilis) as the target organism to investigate changes in fighting behavior as a result of dynamic exposure. Crayfish underwent either a 23h long static or a dynamic exposure to naproxen. Following exposure, the target crayfish and an unexposed size matched opponent underwent a 15min fight trial. These fight trials were recorded and later analyzed using a standard ethogram. Results indicate that exposure to sublethal concentrations of naproxen, in both static and flowing conditions, negatively impact aggressive behavior. Results also indicate that a dynamic exposure paradigm has a greater negative impact on behavior than a static exposure. Turbulence and habitat structure play important roles in shaping chemical exposure. Future research should incorporate features of dynamic chemical exposure in order to form a more comprehensive image of chemical exposure and predict the resulting sublethal effects from exposure. Possible techniques for assessment include utilizing flow-through experimental set-ups in tandem with behavioral or physiological endpoints as opposed to acute toxicity. Other possibilities of assessment could involve utilizing fine-scale chemical measurements of pollutants to determine the actual concentrations animals encounter during an exposure event.

The Degree of Impairment of Foraging in Crayfish (Orconectes virilis) due to Insecticide Exposure is Dependent upon Turbulence Dispersion.

As toxicants move into aquatic systems, the concentration at any point in space or time is heavily influenced by the flow dynamics. The dispersion of these chemicals creates a toxicant concentration that fluctuates widely in time and is highly dependent on the spatial heterogeneity of turbulence. Despite this knowledge on the movement of toxicants in natural systems, most ecotoxicological studies use static exposure paradigms that ignore the spatio-temporal dynamics of toxicants in aquatic systems. Although recent studies have begun to use pulsed paradigms in an attempt to mimic natural conditions, the heterogeneity of real concentrations in natural systems rarely is considered for use in these tests. Thus, understanding how organisms are impaired by naturally distributed toxicants is relatively unknown. The purpose of this experiment was to determine how turbulent dispersion of a toxicant negatively impacts a behavioral task and if altering the nature of turbulence will change the negative impact of the toxicant. Crayfish were exposed to a turbulent plume of carbaryl, an insecticide, under two different turbulent conditions and two different spatial conditions. Turbulence was altered by placing an obstruction within the flow which mimics a natural obstruction in lentic systems. Crayfish were exposed to sublethal concentrations of carbaryl for 48 h under these different dynamic conditions. After toxicant exposure, crayfish foraging ability was measured in a flow-through Y maze. We hypothesized that crayfish exposed to the toxicant under more turbulent conditions would exhibit more detrimental responses due to the increased variation in chemical fluctuations. The fine-scale chemical distribution of the toxicant and the three-dimensional velocity profile were characterized for each of the turbulent conditions and each of the spatial locations. Analyses of these data showed that changes in turbulence or spatial location created a unique exposure condition. Particularly, significant variations in the rise time, intermittency, and slope of toxicant pulses were quantified, whereas average concentration of the peaks remained constant across locations. Deficits in the foraging ability of crayfish exposed under these dynamic conditions paralleled the differences quantified in parameters of the turbulent toxicant plume. Given these results, the concept of toxicant exposure needs revision and needs to incorporate the more temporally based measures of toxicant dispersion. In addition, static and pulsed exposure models do not duplicate natural exposure and may not reflect behavioral or physiological impairments that occur under more realistic exposure conditions.

An Increased Rate for Major Birth Anomalies was not Found Following Dental Treatment Requiring Local Anesthesia During Pregnancy.

ARTICLE TITLE AND BIBLIOGRAPHIC INFORMATION: Pregnancy outcome after in utero exposure to local anesthetics as part of dental treatment: a prospective comparative cohort study. Hagai A, Diav-Citrin O, Shechtman S, Ornoy A. J Am Dent Assoc 2015;146(8):572-80. SOURCE OF FUNDING: Not reported TYPE OF STUDY/DESIGN: Comparative cohort study.

Blood-based lung cancer biomarkers identified through proteomic discovery in cancer tissues, cell lines and conditioned medium.

BACKGROUND: Support for early detection of lung cancer has emerged from the National Lung Screening Trial (NLST), in which low-dose computed tomography (LDCT) screening reduced lung cancer mortality by 20 % relative to chest x-ray. The US Preventive Services Task Force (USPSTF) recently recommended annual screening for the high-risk population, concluding that the benefits (life years gained) outweighed harms (false positive findings, abortive biopsy/surgery, radiation exposure). In making their recommendation, the USPSTF noted that the moderate net benefit of screening was dependent on the resolution of most false-positive results without invasive procedures. Circulating biomarkers may serve as a valuable adjunctive tool to imaging. RESULTS: We developed a broad-based proteomics discovery program, integrating liquid chromatography/mass spectrometry (LC/MS) analyses of freshly resected lung tumor specimens (n = 13), lung cancer cell lines (n = 17), and conditioned media collected from tumor cell lines (n = 7). To enrich for biomarkers likely to be found at elevated levels in the peripheral circulation of lung cancer patients, proteins were prioritized based on predicted subcellular localization (secreted, cell-membrane associated) and differential expression in disease samples. 179 candidate biomarkers were identified. Several markers selected for further validation showed elevated levels in serum collected from subjects with stage I NSCLC (n = 94), relative to healthy smoker controls (n = 189). An 8-marker model was developed (TFPI, MDK, OPN, MMP2, TIMP1, CEA, CYFRA 21-1, SCC) which accurately distinguished subjects with lung cancer (n = 50) from high risk smokers (n = 50) in an independent validation study (AUC = 0.775). CONCLUSIONS: Integrating biomarker discovery from multiple sample types (fresh tissue, cell lines and conditioned medium) has resulted in a diverse repertoire of candidate biomarkers. This unique collection of biomarkers may have clinical utility in lung cancer detection and diagnoses.

Sublethal copper toxicity impairs chemical orientation in the crayfish, Orconectes rusticus.

Before reaching concentrations that are high enough to cause mortality, elevated levels of chemical pollution can significantly alter a keystone indicator species' ability to extract sensory information. To organisms that rely on chemical signals to make crucial ecological decisions, increased amounts of a pollutant may impact chemoreceptive abilities by altering the perception of the sensory landscape or impairing the functioning of sensory organs. Heavy metal pollutants entering an aquatic ecosystem are of increasing concern due to discernible effects on chemoreception in many ecologically and economically important species. In order to determine the effects of sublethal copper toxicity on chemically mediated behavior, male and female rusty crayfish, Orconectes rusticus, were exposed to ecologically relevant concentrations of copper (4.5, 45, and 450 µg/l) for 120 h. Following exposure, crayfish were allowed to orient toward a food odor stimulus. During orientation trials, select crayfish oriented under a point or nonpoint source copper background pollutant at the same concentration as the exposure period. Orientation trials were videotaped and analyzed using EthoVision XT 8.5 (Noldus Information Technology, The Netherlands) for differences in overall success in locating the food source and orienting parameters. Significant differences were found in the overall orientation ability of O. rusticus to locate an odor source when previously exposed to copper in combination with a source of pollution in the background of orientation trials. Crayfish exposed to copper in any capacity during the experiment (regardless of concentration or background during trials) showed slower walking speeds toward the source, decreased turning angles, increased heading angles toward the source, and decreased upstream heading angles. Results from this experiment support that copper impairs the ability of crayfish to detect, process, and/or respond appropriately to chemosensory information in order to successfully localize a food odor source.

Fine-scale chemical exposure differs in point and nonpoint source plumes.

Increasing influxes of anthropogenic chemicals into aquatic ecosystems has led to growing global concern surrounding human and ecosystem health. As more freshwater systems are deemed not potable or usable for agricultural purposes, more attention is being paid to remediation and mitigation efforts. Predicting and preventing the impacts of the chemical inputs first requires a thorough understanding of the spatio-temporal distribution of chemical plumes in natural habitats. Plume dispersion is intimately tied to fluid mechanics; therefore, alterations in the way that chemical plumes are introduced to habitats can have profound effects on chemical distribution. Such alterations can subsequently alter the exposure to which organisms are subjected. This study examined the influence of point versus nonpoint sources in structuring the distribution of chemicals in a simulated flowing freshwater habitat. The fine-scale (molecular) spatio-temporal distribution of chemicals was measured in situ using an electrochemical detector. Molecular concentration at varying distance and height from the source was quantified using dopamine coupled with an electrochemical detection system. The fine-scale distribution of chemical plumes from point and nonpoint sources showed significant differences in how organisms will be exposed to chemicals. Overall, this study characterized plumes from nonpoint sources as having significantly longer peak lengths and rise times as well as greater peak heights and maximum slopes than plumes from point sources, thus providing a significantly different exposure paradigm. This quantification of how chemicals move differently throughout a fluid medium when introduced from point and nonpoint sources allows a greater understanding of how chemical plumes can potentially affect aquatic ecosystems.

The Effects of Biodiesel and Crude Oil on the Foraging Behavior of Rusty Crayfish, Orconectes rusticus.

Environmental pollutants, such as crude oil and other petroleum-based fuels, inhibit and limit an organism's ability to perceive a chemical stimulus. Despite the increased use of alternative fuels, such as biodiesel, there have been few studies investigating the impact of these chemicals on the behavior of aquatic organisms. The purpose of this study was to compare the sublethal effects of biodiesel and crude oil exposure on chemically mediated behaviors in a freshwater keystone species. Crayfish (Orconectes rusticus) were tested on their ability to respond appropriately to a positive chemical stimulus within a Y-maze choice paradigm. Behavior was quantified by measuring time spent finding an odor source, duration of time spent at the odor source, percentage of crayfish that found the odor source, and percentage of crayfish that chose the correct arm of the arena. Results indicated negative impacts of both biodiesel and crude oil on the ability of crayfish to locate the food source. However, there were no significant differences between behavioral performances when crayfish were exposed to crude oil compared with biodiesel. Thus, biodiesel and crude oil have equally negative effects on the chemosensory behavior of crayfish. These findings indicate that biodiesel has the potential to have similar negative ecological impacts as other fuel source toxins.