Targeted and non-targeted analysis of young-of-year smallmouth bass using comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry.

Smallmouth bass in the Susquehanna River Basin, Chesapeake Bay Watershed, USA, have been exhibiting clinical signs of disease and reproductive endocrine disruption (e.g., intersex, male plasma vitellogenin) for over fifteen years. Previous histological and targeted chemical analyses have identified infectious agents and pollutants in fish tissues including organic contaminants, mercury, and perfluorinated compounds, but a common causative link for the observed signs of disease across this widespread area has not been determined. This study examines 146 young-of-year smallmouth bass collected from 14 sampling sites in the Susquehanna River Basin, Pennsylvania, USA with varying levels of disease prevalence. Whole fish were extracted by a recently developed modification to the quick, easy, cheap, effective, rugged, and safe extraction method and analyzed by comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry. A targeted analysis was conducted to identify the presence and quantity of 127 known contaminants, including polychlorinated biphenyls, brominated diphenyl ethers, organochlorinated pesticides, and pharmaceutical and personal care products. A non-targeted analysis was conducted on the same data set to identify analytes of interest not included on routine target compound lists. Chromatographic alignment through Statistical Compare (ChromaTOF GC) was followed by Fisher ratio and principal component analysis to reduce the data set from thousands of peaks per sample to a final data set of 65 analytes of interest. Comparisons of these 65 compounds between Normal (no observed health anomalies) and Lesioned (observed health anomaly at time of collection) fish revealed increased levels of three chemical families in Lesioned fish including esters, ketones, and nitrogen containing compounds.

Optimization of a suite of flathead catfish (Pylodictis olivaris) microsatellite markers for understanding the population genetics of introduced populations in the northeast United States.

OBJECTIVE: Flathead catfish are rapidly expanding into nonnative waterways throughout the United States. Once established, flathead catfish may cause disruptions to the local ecosystem through consumption and competition with native fishes, including species of conservation concern. Flathead catfish often become a popular sport fish in their introduced range, and so management strategies must frequently balance the need to protect native and naturalized fauna while meeting the desire to maintain or enhance fisheries. However, there are currently few tools available to inform management of invasive flathead catfish (Pylodictis olivaris). We describe a suite of microsatellite loci that can be used to characterize population structure, predict invasion history, and assess potential mitigation strategies for flathead catfish. RESULTS: Our panel of 13 microsatellite loci were polymorphic and appear to be informative for population genetic studies of flathead catfish. We found moderate levels of diversity in four nonnative collections of flathead catfish in the Pennsylvania and Maryland sections of the Susquehanna River and the Schuylkill River, Pennsylvania. Analyses suggested patterns of genetic differentiation within- and among-rivers, highlighting the utility of this marker panel for understanding the structure and assessing the degree of connectivity among flathead catfish populations.

Modified QuEChERS extraction for the analysis of young-of-year smallmouth bass using GC × GC-TOFMS.

Signs of disease, such as external lesions, have been prevalent in smallmouth bass throughout the Susquehanna River Basin, USA. Previous targeted chemical studies in this system have identified known persistent organic pollutants, but a common explanatory link across multiple affected sites remains undetermined. A fast and robust extraction method that can be applied to young-of-year fish is needed to effectively screen for target and non-target compounds that may be impacting organism health. The quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction methodology was optimized to perform both targeted and non-targeted chemical analyses from a single extraction of whole young-of-year fish. Comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC × GC-TOFMS) was used for extract analysis. Sample extraction was performed using the solvent ethyl acetate, followed by a two-step cleanup in which samples were frozen for lipid removal and subjected to dispersive solid phase extraction using Florisil. A sample of 21 young-of-year smallmouth bass collected from areas with disease and exhibiting different types of external lesions were evaluated for 233 target compounds. A total of 34 organic contaminants, including polychlorinated biphenyls, brominated diphenyl ethers, organochlorinated pesticides, and personal care products, were detected. Data from this sample set was then analyzed for non-targets. Using the Fisher ratio method and multivariate analysis, an additional 10 significant features were identified specific to either fish with visible lesions or with no visible disease characteristics.

Spatiotemporal variation in occurrence and co-occurrence of pesticides, hormones, and other organic contaminants in rivers in the Chesapeake Bay Watershed, United States.

Investigating the spatiotemporal dynamics of contaminants in surface water is crucial to better understand how introduced chemicals are interacting with and potentially influencing aquatic organisms and environments. Within the Chesapeake Bay Watershed, United States, there are concerns about the potential role of contaminant exposure on fish health. Evidence suggests that exposure to contaminants in surface water is causing immunosuppression and intersex in freshwater fish species. Despite these concerns, there is a paucity of information regarding the complex dynamics of contaminant occurrence and co-occurrence in surface water across both space and time. To address these concerns, we applied a Bayesian hierarchical joint-contaminant model to describe the occurrence and co-occurrence patterns of 28 contaminants and total estrogenicity across six river sites and over three years. We found that seasonal occurrence patterns varied by contaminant, with the highest occurrence probabilities during the spring and summer months. Additionally, we found that the proportion of agricultural landcover in the immediate catchment, as well as stream discharge, did not have a significant effect on the occurrence probabilities of most compounds. Four pesticides (atrazine, metolachlor, fipronil and simazine) co-occurred across sites after accounting for environmental covariates. These results provide baseline information on the contaminant occurrence patterns of several classes of compounds within the Chesapeake Bay Watershed. Understanding the spatiotemporal dynamics of contaminants in surface water is the first step in investigating the effects of contaminant exposure on fisheries and aquatic environments.

Spatial and temporal variability of myxozoan parasite, Myxobolus inornatus, prevalence in young of the year smallmouth bass in the Susquehanna River Basin, Pennsylvania.

A myxozoan parasite, Myxobolus inornatus, is one disease agent identified in young of the year (YOY) smallmouth bass in the Susquehanna River Basin, Pennsylvania. We investigated spatial and temporal variability in M. Inornatus prevalence across the Susquehanna River Basin and at several out-of-basin sites. We examined potential land use drivers of M. Inornatus prevalence including agricultural and developed land use. In 1,267 YOY smallmouth bass collected from 32 sites during 2013-2016, M. Inornatus was documented in 43.6% of samples. Among-site variability in parasite prevalence was greater than among-year variability. The effect of agricultural land use on M. Inornatus prevalence had a high probability of being positively correlated at multiple spatial scales (probability of positive effect > 0.80). The effect of developed land use on M. Inornatus prevalence had a relatively high probability of being negatively correlated at multiple spatial scales (probability of negative effect > 0.70). Our results suggest that land use practices could be related to M. Inornatus infection of smallmouth bass. Further study will be necessary to determine whether disease dynamics are a consequence of effects on the host, alterations of instream habitat mediating invertebrate host dynamics and/or survival and dispersal of the parasite infective stage.

Accurate reading comprehension rate as an indicator of broad reading in students in first, second, and third grades.

The relationship between reading comprehension rate measures and broad reading skill development was examined using data from approximately 1425 students (grades 1-3). Students read 3 passages, from a pool of 30, and answered open-ended comprehension questions. Accurate reading comprehension rate (ARCR) was calculated by dividing the percentage of questions answered correctly (%QC) by seconds required to read the passage. Across all 30 passages, ARCR and its two components, %QC correct and time spent reading (1/seconds spent reading the passage), were significantly correlated with broad reading scores, with %QC resulting in the lowest correlations. Two sequential regressions supported previous findings which suggest that ARCR measures consistently produced meaningful incremental increases beyond %QC in the amount of variance explained in broad reading skill; however, ARCR produced small or no incremental increases beyond reading time. Discussion focuses on the importance of the measure of reading time embedded in brief accurate reading rate measures and directions for future research.

Selective serotonin receptor stimulation of the medial nucleus accumbens differentially affects appetitive motivation for food on a progressive ratio schedule of reinforcement.

Previously, we reported that stimulation of selective serotonin (5-HT) receptor subtypes in the nucleus accumbens shell differentially affected consumption of freely available food. Specifically, activation of 5-HT(6) receptors caused a dose-dependent increase in food intake, while the stimulation of 5-HT(1/7) receptor subtypes decreased feeding [34]. The current experiments tested whether similar pharmacological activation of nucleus accumbens serotonin receptors would also affect appetitive motivation, as measured by the amount of effort non-deprived rats exerted to earn sugar reinforcement. Rats were trained to lever press for sugar pellets on a progressive ratio 2 schedule of reinforcement. Across multiple treatment days, three separate groups (N=8-10) received bilateral infusions of the 5-HT(6) agonist EMD 386088 (at 0.0, 1.0 and 4.0 µg/0.5 µl/side), the 5-HT(1/7) agonist 5-CT (at 0, 0.5, 1.0, or 4.0 µg/0.5 µl/side), or the 5-HT(2C) agonist RO 60-0175 fumarate (at 0, 2.0, or 5.0 µg/0.5 µl/side) into the anterior medial nucleus accumbens prior to a 1-h progressive ratio session. Stimulation of 5-HT(6) receptors caused a dose-dependent increase in motivation as assessed by break point, reinforcers earned, and total active lever presses. Stimulation of 5-HT(1/7) receptors increased lever pressing at the 0.5 µg dose of 5-CT, but inhibited lever presses and break point at 4.0 µg/side. Injection of the 5-HT(2C) agonist had no effect on motivation within the task. Collectively, these experiments suggest that, in addition to their role in modulating food consumption, nucleus accumbens 5-HT(6) and 5-HT(1/7) receptors also differentially regulate the appetitive components of food-directed motivation.