Reduced Freshwater Mussel Juvenile Production as a Result of Agricultural and Urban Contaminant Mixture Exposures.

Freshwater mussels provide invaluable ecological services but are threatened by habitat alteration, poor water quality, invasive species, climate change, and contaminants, including contaminants of emerging concern (CECs). Contaminants of emerging concerns are well documented in aquatic environments, including the Great Lakes Basin, but limited information is available on how environmentally relevant mixtures affect freshwater mussel biology throughout their varied life stages. Our main goal was to assess mussels' reproductive output in response to exposure to agricultural and urban CEC mixtures during glochidial development through juvenile transformation and excystment focusing on how exposure duration and treatment affect: (1) the number of glochidia prematurely released by brooding females, (2) glochidial transformation through host-fish excystment, and (3) the number of fully metamorphosed juveniles able to continue the lifecycle. Mussels and host fish were exposed to either a control water (CW), control ethanol (CE), agriculture CEC mixture (AM), or urban CEC mixture (UM) for 40 and 100 days. We found no effect from treatment or exposure duration on the number of glochidia prematurely released. Fewer partially and fully metamorphosed AM juveniles were observed during the 100-day exposure, compared with the 40-day. During the 40-day exposure, CW produced more fully metamorphosed individuals compared with CE and UM, but during the 100-day exposure AM produced more fully metamorphosed individuals compared with the CW. There was reduction in fully metamorphosed juveniles compared with partially metamorphosed for CE and UM during the 40-day exposure, as well as in the CW during the 100-day exposure. These results will be important for understanding how mussel populations are affected by CEC exposure. The experiments also yielded many insights for laboratory toxicology exposure studies. Environ Toxicol Chem 2024;43:1112-1125. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Assessing Contaminants of Emerging Concern in the Great Lakes Ecosystem: A Decade of Method Development and Practical Application.

Assessing the ecological risk of contaminants in the field typically involves consideration of a complex mixture of compounds which may or may not be detected via instrumental analyses. Further, there are insufficient data to predict the potential biological effects of many detected compounds, leading to their being characterized as contaminants of emerging concern (CECs). Over the past several years, advances in chemistry, toxicology, and bioinformatics have resulted in a variety of concepts and tools that can enhance the pragmatic assessment of the ecological risk of CECs. The present Focus article describes a 10+- year multiagency effort supported through the U.S. Great Lakes Restoration Initiative to assess the occurrence and implications of CECs in the North American Great Lakes. State-of-the-science methods and models were used to evaluate more than 700 sites in about approximately 200 tributaries across lakes Ontario, Erie, Huron, Michigan, and Superior, sometimes on multiple occasions. Studies featured measurement of up to 500 different target analytes in different environmental matrices, coupled with evaluation of biological effects in resident species, animals from in situ and laboratory exposures, and in vitro systems. Experimental taxa included birds, fish, and a variety of invertebrates, and measured endpoints ranged from molecular to apical responses. Data were integrated and evaluated using a diversity of curated knowledgebases and models with the goal of producing actionable insights for risk assessors and managers charged with evaluating and mitigating the effects of CECs in the Great Lakes. This overview is based on research and data captured in approximately about 90 peer-reviewed journal articles and reports, including approximately about 30 appearing in a virtual issue comprised of highlighted papers published in Environmental Toxicology and Chemistry or Integrated Environmental Assessment and Management. Environ Toxicol Chem 2023;42:2506-2518. © 2023 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Validation of a vulnerability index of exposure to chemicals of emerging concern in surface water and sediment of Great Lakes tributaries of the United States.

Widespread occurrence of emerging contaminants in Great Lakes tributaries led to the development and publication of a vulnerability index (VI) to assess the potential exposure of aquatic communities to chemicals of emerging concern (CEC) in the Great Lakes basin. The robust nature of the VI was tested to evaluate the underlying statistical model and expand the spatial domain of the index. Data collected at 131 new sampling sites (Test 1) and published data from independent studies (Test 2) were used to test the model predictions. Test 1 water and sediment samples were analyzed for the same classes of CEC chemicals and compared to the predictions for the original VI. Concentrations and numbers of unique CECs detected in water and sediment samples were similar between the original data and the two test datasets, although CECs tended to have higher detection frequencies in the original dataset compared to the Test 1 and Test 2 datasets. For example, 69 CECs were detected in ≥30% of water samples in the original dataset compared with 17 CECs in the Test 1 data and 59 in the Test 2 data. Predicted vulnerability for test sites agreed with actual vulnerability 64% of the time for water and 71% of the time for sediment. Agreement percentage results were greater when individual sites were grouped by river, with 82% agreement between predictions and actual vulnerability for water and 78% agreement for sediment. For the entire dataset, the VI ranks correlated with an independent estimate of potential biological impact. Agreement percentage was the greatest for low or high vulnerability index values but highly variable for sites that are classified as having medium vulnerability. Despite the underlying variability, there is a significant correlation (R2 = 0.26; p < 0.01) between the VI ranking of tributaries and the independent ranking of potential negative biological impact.

Population modeling to inform management and recovery efforts for lake sturgeon, Acipenser fulvescens.

Lake sturgeon (Acipenser fulvescens) populations have significantly declined across their historic range, in large part due to anthropogenic impacts that have likely been exacerbated by the life-history traits of this slow-growing and long-lived species. We developed a population model to explore how Contaminants of Emerging Concern (CECs) impact lake sturgeon populations. We explored how different physiological modes of action (pMoAs) of CECs impacted population abundance and recovery and how different simulated management actions could enable recovery. We first estimated the impacts on population abundance and recovery by comparing the trajectory of an unexposed population to a population that had been exposed to a CEC with a specific pMoA after the end of the exposure. We then predicted how different management actions would impact population recovery by comparing the trajectories of an unexposed population to an exposed population for which a management action started at a fixed time without discontinuation of the exposure. Our results predicted that the individual-level pMoA of CECs has an important impact on population-level effects because different stressor's pMoA impacts the life-history traits of sturgeon differently. For example, the feeding and reproduction pMoAs caused the strongest and weakest population declines, respectively. For the same reason, pMoA also impacted recovery. For example, recovery was delayed when the pMoA was growth, maintenance, or feeding, but it was immediate when the pMoA was reproduction. We found that management actions that increased the egg survival rate or the stocking of fingerlings resulted in faster and stronger recovery than management actions that increased the juvenile or adult survival rate. This result occurred because the first two management actions immediately impacted recruitment, whereas the impact was delayed for the last two. Finally, there was greater potential for recovery when management action targeted eggs and fingerlings because these life stages have lower natural survival rates. Integr Environ Assess Manag 2022;18:1597-1608. © 2022 Society of Environmental Toxicology & Chemistry (SETAC). This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Multiple lines of evidence for identifying potential hazards to fish from contaminants of emerging concern in Great Lakes tributaries.

Contaminants of emerging concern (CECs; e.g., pharmaceuticals, flame retardants, pesticides, and industrial chemicals) are omnipresent throughout tributaries to the Great Lakes. Furthermore, CECs are often present at concentrations that are potentially hazardous to aquatic species. Since 2010, we characterized the presence of CECs at 309 sites within 47 Great Lakes tributaries and characterized responses of fathead minnow (Pimephales promelas) exposed to river water at a subset of 26 sites within four tributaries. Our work resulted in three independent lines of evidence related to the potential hazards of CEC exposure to fish. First, vulnerability (where vulnerability refers to likelihood) of surface waters to CEC presence was predicted using select watershed characteristics. Second, hazard to fish (where hazard means the potential for adverse biological responses) was predicted using screening values for a subset of CECs. Third, biological responses of fathead minnow exposed to river water in streamside exposures were measured. We assessed the congruence of these three lines of evidence for identifying sites with elevated hazards to CEC exposure. Predicted vulnerability and hazards agreed at 66% of all sites. Where the two indices did not agree, vulnerability often underestimated predicted hazard. When compared with measured biological responses from streamside exposures, predicted hazards agreed for 42% of samples. Furthermore, when predicted hazards for specific effect categories were compared with similar measured biomarkers, 26% and 46% of samples agreed for reproductive and physiological effect categories, respectively. Overall, vulnerability and hazard predictions tended to overestimate the measured biological responses, providing a protective estimate of the potential hazards of CEC exposure to fish. When used together, these three approaches can help resource managers prioritize management activities in minimizing hazards of CEC exposure and can be used by researchers to prioritize studies focused on understanding the hazards of CEC exposure to fish. Integr Environ Assess Manag 2022;18:1246-1259. © 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC). This article has been contributed to by US Government employees and their work is in the public domain in the USA.

A simulation-based evaluation of management actions to reduce the risk of contaminants of emerging concern (CECs) to walleye in the Great Lakes Basin.

Contaminants of emerging concern (CECs) are ubiquitous, present in complex chemical mixtures, and represent a threat to the Great Lake ecosystem. Mitigation strategies are needed to protect populations of key species, but knowledge about ecological and biological effects of CECs at the population level are limited. In this study, we combined laboratory data on CEC effects at the individual-level with in-situ CEC concentration data in a walleye (Sander vitreus) population model to simulate the effectiveness of different CEC mitigation strategies in the Maumee River and Lake Erie. We compared the effectiveness of moderate mitigation (50% reduction in exposure level) of an entire watershed versus intensive mitigation (reduction of exposure to a level that does not affect walleye) of single river sites for three CEC mixture scenarios (agricultural, urban, and combined). We also explored the impact of hypothetical chemical toxicokinetics (the time course of chemicals in walleye) on the relative effectiveness of the mitigation strategies. Our results suggest that when CECs impact fecundity, single-site mitigation is more effective when it focuses on spawning sites and nearby downstream sites that are substantially impaired. Our simulations also suggest that chemical toxicokinetics are important when evaluating single-site mitigation strategies, but that population characteristics, such as stage-specific mortality rate, are more important when evaluating watershed mitigation strategies. Results can be used to guide fisheries management, such as choosing habitat restoration sites, and identify key knowledge gaps that direct future research and monitoring.

Characterization of pharmaceuticals, personal care products, and polybrominated diphenyl ethers in lake sturgeon serum and gametes.

Recent research suggests contaminants of emerging concern (CECs) are widespread and environmentally relevant concentrations can impact fishes. However, little is known about impacts of CECs to long-lived or rare species. The objective of this study was to characterize CEC concentrations in lake sturgeon (Acipenser fulvescens) serum and gametes. Blood serum was collected non-lethally from lake sturgeon at four lower Great Lakes basin sites: Detroit, upper Niagara, lower Niagara, and St. Lawrence rivers; additionally, gametes were collected from lake sturgeon in the St. Lawrence River. Samples were analyzed for pharmaceuticals and personal care products (PPCPs) and polybrominated diphenyl ethers (PBDEs). Overall, 44 different PPCPs were identified in serum and gamete samples across sites, with 22 PPCPs identified in at least 25% of serum samples and three PPCPs identified in 25% of gamete samples. PPCP concentrations in serum and gametes ranged from 0.00208 to 130 ppb and 0.00538-190 ppb, respectively. NMDS ordination revealed differences in the presence and concentrations of PPCPs in lake sturgeon serum across sites, however, N,N-diethyl-meta-toluamide (DEET), hydrocortisone, benztropine, and amitriptyline were detected in at least one serum sample at all sites. Additionally, DEET, 10-hydroxy-amitriptyline, and sertraline were detected in ≥25% of gamete samples collected from the St. Lawrence River. Twenty-six PBDE congeners were identified in 25% of serum samples and 24 were identified in 25% of gamete samples. PBDEs in serum were present across all sites and in gametes of St. Lawrence River lake sturgeon, and total PBDE concentrations in serum and gametes ranged from 0.184 to 12.7 ppb and 0.0826-0.44 ppb, respectively. Managers of lake sturgeon populations may need to consider the impacts of CECs if reproductive, developmental, behavioral, growth effects, or mortality are observed in the Great Lakes basin or other areas that are impacted by increased exposures to PPCPs and PBDEs.

Land Use Contributions to Adverse Biological Effects in a Complex Agricultural and Urban Watershed: A Case Study of the Maumee River.

Agricultural and urban contaminants are an environmental concern because runoff may contaminate aquatic ecosystems, resulting in stress for exposed fish. The objective of the present controlled, field-based study was to assess the impacts of high-intensity agriculture and urban land use on multiple life stages of the fathead minnow (Pimephales promelas), using the Maumee River (Toledo, OH, USA) as a case study. Laboratory cultured adult and larval fathead minnows were exposed for 21 d, and embryos were exposed until hatching to site-specific water along the lower reach of the Maumee River. Adult minnows were analyzed for reproduction and alterations to hematologic characteristics (vitellogenin, glucose, estradiol, 11-ketotestosterone). Water and fish tissue samples were analyzed for a suite of multiresidue pesticides, hormones, and pharmaceuticals. Contaminants were detected in every water and tissue sample, with 6 pesticides and 8 pharmaceuticals detected in at least 82% of water samples and at least half of tissue samples. Effects differed by exposed life stage and year of exposure. Fecundity was the most sensitive endpoint measured and was altered by water from multiple sites in both years. Physiological parameters associated with fecundity, such as plasma vitellogenin and steroid hormone concentrations, were seldom impacted. Larval fathead minnows appeared to be unaffected. Embryonic morphological development was delayed in embryos exposed to site waters collected in 2016 but not in 2017. A distinction between agricultural and urban influences in the Maumee River was not realized due to the great overlap in contaminant presence and biological effects. Differences in precipitation patterns between study years likely contributed to the observed biological differences and highlight the need for environmental exposure studies to assess the environmental risk of contaminants. Environ Toxicol Chem 2019;00:1-17. © 2019 SETAC.

Predicting the occurrence of chemicals of emerging concern in surface water and sediment across the U.S. portion of the Great Lakes Basin.

Chemicals of emerging concern (CECs) are introduced into the aquatic environment via various sources, posing a potential risk to aquatic organisms. Previous studies have identified relationships between the presence of CECs in water and broad-scale watershed characteristics. However, relationships between the presence of CECs and source-related watershed characteristics have not been explored across the Great Lakes basin. Boosted regression tree (BRT) analyses were used to develop predictive models of CEC occurrence in water and sediment throughout 24 U.S. tributaries to the Great Lakes. Models were based on the distribution of both broad-scale and source-related watershed characteristics. Twenty-one upstream watershed characteristics, including land cover, number of permitted point sources, and distance to point sources were used to develop models predicting the probability of CEC occurrence in surface water and bottom sediment. Total accuracy of BRT models ranged from 66% to 94% for both matrices. All 21 watershed characteristics were important predictor variables in at least one surface-water model; twenty were important in at least one bottom-sediment model. Among the model variables, developed land use and distance to point sources were important predictors of the presence of CEC classes in both water and sediment. Although limitations exist, BRT models are one tool available for assessing vulnerability of fisheries and aquatic resources to CEC occurrences.

On the Cutting Edge of Research to Conserve At-Risk Species: Maximizing Impact through Partnerships.

Today's conservation challenges are complex. Solving these challenges often requires scientific collaborations that extend beyond the scope, expertise, and capacity of any single agency, organization, or institution. Conservation efforts can benefit from interdisciplinary collaboration, scientific and technological innovations, and the leveraging of capacity and resources among partners. Here we explore a series of case studies demonstrating how collaborative scientific partnerships are furthering the mission of the US Fish and Wildlife Service (USFWS), including: (1) contaminants of emerging concern in the Great Lakes Basin, (2) Poweshiek skipperling conservation, (3) using technology to improve population survey methods for bats and monarch butterfly, and (4) Big River restoration in the Southeast Missouri lead mining district. These case studies illustrate how strategic and effective scientific collaboration is a multi-stage process that requires investment of time and resources by all participants. Early coordination and communication is crucial to aligning planned work with scientific and decision-making needs. Collaborations between USFWS and external scientists can be mutually beneficial by supporting the agency mission while also providing an avenue for innovative research to be directly applied in conservation decisions and management actions.

Is Angiostatin Involved in Physiological Foveal Avascularity?

Purpose: The primate central retina is characterized by an avascular fovea and well-defined perifoveal capillary plexus. Neither blood vessels nor their accompanying astrocytes enter the fovea during any stage of retinal development; a balance of angiogenic and angiostatic factors probably maintains foveal avascularity throughout life. The aim of this study was to identify potentially angiorepulsive factors involved in the development of the avascular primate retinal fovea. Methods: Retinas of newborn, juvenile, and adult Callithrix jacchus and Macaca fascicularis monkeys and control human retinas were studied to determine the localization of angiostatin relative to III ß-tubulin, glial fibrillary acidic protein, vascular endothelial growth factor (VEGF), platelet endothelial cell adhesion molecule-1 (PECAM), and the angiostatin receptor αvß3-integrin in the foveal, macular, and peripheral retina. Expression studies were performed using immunohistochemistry (IHC) on retinal whole-mount and paraffin sections, and Western blotting on frozen material. The complex network of the main retinal cell types was identified by IHC of retinal whole mounts. Results: In general, lifetime expression of angiostatin was found in all retinas. Colabeling with different markers revealed retinal ganglion cells as the main source of angiostatin expression in the primate retina, whereas PECAM-immunopositive blood capillaries expressed the angiostatin receptor αvß3-integrin, and capillary-associated astrocytes expressed VEGF. Conclusions: This study provides the first evidence of angiostatin expression in the primate retina; the expression of angiostatin in the avascular foveal region and the peripheral retina suggests that angiostatin may play a role in the regulation of retinal vascularization, providing a possible explanation for the development and persistence of an avascular fovea.

Inhibitors of CXC chemokine receptor type 4: putative therapeutic approaches in inflammatory diseases.

PURPOSE OF REVIEW: The CXC chemokine receptor type 4 (CXCR4), which is a G-protein coupled receptor, and its ligand CXCL12 play an important role in neutrophil homeostasis and inflammation. This review focuses on involvement of the CXCL12/CXCR4 axis in inflammation and different inflammatory diseases and depicts that blocking CXCR4 is an attractive therapeutic strategy. RECENT FINDINGS: Binding of CXCL12 to CXCR4 retains immature neutrophils in the bone marrow and also participates in leukocyte recruitment into inflamed tissue. The CXCL12/CXCR4 axis is also involved in several inflammatory processes and diseases including the WHIM (warts, hypogammaglobulinemia, infections and myelokathexis) syndrome, HIV, autoimmune disorders, ischemic injury, and pulmonary fibrosis. SUMMARY: Based on these findings, blocking CXCR4 seems to be a therapeutic strategy in inflammatory diseases. Several promising CXCR4 antagonists are in different stages of development and clinical trials. Currently, only plerixafor (AMD3100) has been approved for short-term application.

Identification of specific miRNAs targeting proteins of the apical junctional complex that simulate the probiotic effect of E. coli Nissle 1917 on T84 epithelial cells.

In the intestine, dysregulation of miRNA is associated with inflammation, disruption of the gastrointestinal barrier, and the onset of gastrointestinal disorders. This study identifies miRNAs involved in the maintenance of intercellular junctions and barrier integrity. For the functional identification of barrier affecting miRNAs, we took advantage of the barrier-enforcing effects of the probiotic bacterium Escherichia coli Nissle 1917 (EcN) which can be monitored by enhanced transepithelial resistance (TER). miRNA-profiling of T84 monolayers prior and after co-incubation with EcN revealed for the first time differentially regulated miRNAs (miR-203, miR-483-3p, miR-595) targeting tight junction (TJ) proteins. Using real-time PCR, Western blotting and specific miRNA mimics, we showed that these miRNAs are involved in the regulation of barrier function by modulating the expression of regulatory and structural components of tight junctional complexes. Furthermore, specific inhibitors directed at these miRNA abrogated the disturbance of tight junctions induced by enteropathogenic E. coli (EPEC). The half-maximal inhibitory concentration (IC(50)) was determined to 340 nM by monitoring inhibitor kinetics. In summary, we conclude that specific miRNAs effect regulatory as well as structural proteins of the junctional complex which in turn are involved in the barrier enhancing effect of EcN. Hence, we suggest that the application of miRNAs might be refined and further developed as a novel supportive strategy for the treatment of gastrointestinal disorders.

Differential targeting of the E-Cadherin/ß-Catenin complex by gram-positive probiotic lactobacilli improves epithelial barrier function.

The intestinal ecosystem is balanced by dynamic interactions between resident and incoming microbes, the gastrointestinal barrier, and the mucosal immune system. However, in the context of inflammatory bowel diseases (IBD), where the integrity of the gastrointestinal barrier is compromised, resident microbes contribute to the development and perpetuation of inflammation and disease. Probiotic bacteria have been shown to exert beneficial effects, e.g., enhancing epithelial barrier integrity. However, the mechanisms underlying these beneficial effects are only poorly understood. Here, we comparatively investigated the effects of four probiotic lactobacilli, namely, Lactobacillus acidophilus, L. fermentum, L. gasseri, and L. rhamnosus, in a T84 cell epithelial barrier model. Results of DNA microarray experiments indicating that lactobacilli modulate the regulation of genes encoding in particular adherence junction proteins such as E-cadherin and ß-catenin were confirmed by quantitative reverse transcription-PCR (qRT-PCR). Furthermore, we show that epithelial barrier function is modulated by Gram-positive probiotic lactobacilli via their effect on adherence junction protein expression and complex formation. In addition, incubation with lactobacilli differentially influences the phosphorylation of adherence junction proteins and the abundance of protein kinase C (PKC) isoforms such as PKCδ that thereby positively modulates epithelial barrier function. Further insight into the underlying molecular mechanisms triggered by these probiotics might also foster the development of novel strategies for the treatment of gastrointestinal diseases (e.g., IBD).