Biliary Polycyclic Aromatic Hydrocarbon Metabolite Equivalents Measured in Fish and Subtidal Invertebrates Following the Refugio Beach Oil Spill.

Monterey formation crude oil spilled from an onshore pipeline and entered the surf zone near Refugio State Beach, Santa Barbara County, CA, USA on 19 May 2015. Exposure to nearshore fish was evaluated by measuring biliary polycyclic aromatic hydrocarbon (PAH) metabolite concentrations in surfperch (Embiotocidae), collected near the release point, at a lesser oiled area, Gaviota State Beach, and near the Coal Oil Point oil seep, Campus Point, at both four days and approximately one year after the oil spill. Three to four weeks after the spill, fish, invertebrates, and kelp were collected near the same three sites for PAH analysis of edible tissues to support the fishery closure assessment. Additionally, thirteen days after the spill, vegetation, and invertebrate tissue samples from the seafloor near Refugio State Beach were analyzed for PAHs. In surfperch bile, mean naphthalene, phenanthrene and benzo(a)pyrene metabolite equivalents were significantly higher at Refugio State Beach, compared to Gaviota State Beach or Campus Point in 2015. One year later, there was no significant difference between the three sites. Spatial and depth zone patterns of PAH tissue concentrations from composited invertebrate, vegetation and fish showed highest concentrations were measured in invertebrates collected near Refugio State Beach. Overall, results show elevated PAH levels in nearshore organisms and provide a useful reference for potential PAH exposures in fish and subtidal invertebrates following a large nearshore oil spill.

Invertebrate Gonadotropin-Releasing Hormone Receptor Signaling and Its Relevant Biological Actions.

Gonadotropin-releasing hormones (GnRHs) play pivotal roles in reproduction via the hypothalamus-pituitary-gonad axis (HPG axis) in vertebrates. GnRHs and their receptors (GnRHRs) are also conserved in invertebrates lacking the HPG axis, indicating that invertebrate GnRHs do not serve as "gonadotropin-releasing factors" but, rather, function as neuropeptides that directly regulate target tissues. All vertebrate and urochordate GnRHs comprise 10 amino acids, whereas amphioxus, echinoderm, and protostome GnRH-like peptides are 11- or 12-residue peptides. Intracellular calcium mobilization is the major second messenger for GnRH signaling in cephalochordates, echinoderms, and protostomes, while urochordate GnRHRs also stimulate cAMP production pathways. Moreover, the ligand-specific modulation of signal transduction via heterodimerization between GnRHR paralogs indicates species-specific evolution in Ciona intestinalis. The characterization of authentic or putative invertebrate GnRHRs in various tissues and their in vitro and in vivo activities indicate that invertebrate GnRHs are responsible for the regulation of both reproductive and nonreproductive functions. In this review, we examine our current understanding of and perspectives on the primary sequences, tissue distribution of mRNA expression, signal transduction, and biological functions of invertebrate GnRHs and their receptors.

Comparative dynamics of tropomyosin in vertebrates and invertebrates.

Tropomyosin (Tpm) is an extended α-helical coiled-coil homodimer that regulates actinomyosin interactions in muscle. Molecular simulations of four Tpms, two from the vertebrate class Mammalia (rat and pig), and two from the invertebrate class Malacostraca (shrimp and lobster), showed that despite extensive sequence and structural homology across metazoans, dynamic behavior-particularly long-range structural fluctuations-were clearly distinct. Vertebrate Tpms were more flexible and sampled complex, multi-state conformational landscapes. Invertebrate Tpms were more rigid, sampling a highly constrained harmonic landscape. Filtering of trajectories by principle component analysis into essential subspaces showed significant overlap within but not between phyla. In vertebrate Tpms, hinge-regions decoupled long-range interhelical motions and suggested distinct domains. In contrast, crustacean Tpms did not exhibit long-range dynamic correlations-behaving more like a single rigid rod on the nanosecond time scale. These observations suggest there may be divergent mechanisms for Tpm binding to actin filaments, where conformational flexibility in mammalian Tpm allows a preorganized shape complementary to the filament surface, and where rigidity in the crustacean Tpm requires concerted bending and binding.

Bioaccumulation and Dispersion of Uranium by Freshwater Organisms.

Uranium is the heaviest naturally occurring element on Earth. Uranium mining may result in ground and surface water contamination with potential bioaccumulation and dispersion by aquatic invertebrates with aerial stages. We investigated the effects of uranium contamination at community level in terms of abundance, richness, the composition of invertebrate communities, and functional traits. We also investigated uranium mobility across aquatic food webs and its transfer to land via the emergence of aquatic insects. We sampled water, sediment, biofilm, macrophytes, aquatic invertebrates, adult insects, and spiders in the riparian zone across sites with a gradient of uranium concentrations in stream water (from 2.1 to 4.7 µg L-1) and sediments (from 10.4 to 41.8 µg g-1). Macroinvertebrate assemblages differed between sites with a higher diversity and predominance of Nemouridae and Baetidae at the reference site and low diversity and predominance of Chironomidae in sites with the highest uranium concentration. Uranium concentrations in producers and consumers increased linearly with uranium concentration in stream water and sediment (p < 0.05). The highest accumulation was found in litter (83.76 ± 5.42 µg g-1) and macrophytes (47.58 ± 6.93 µg g-1) in the most contaminated site. Uranium was highest in scrapers (14.30 ± 0.98 µg g-1), followed by shredders (12.96 ± 0.81 µg g-1) and engulfer predators (7.01 ± 1.3 µg g-1). Uranium in adults of aquatic insects in the riparian zone in all sites ranged from 0.25 to 2.90 µg g-1, whereas in spiders it ranged from 0.96 to 1.73 µg g-1, with no differences between sites (p > 0.05). There was a negative relationship between δ15N and uranium, suggesting there is no biomagnification along food webs. We concluded that uranium is accumulated by producers and consumers but not biomagnified nor dispersed to land with the emergence of aquatic insects.

Convergence of detrital stoichiometry predicts thresholds of nutrient-stimulated breakdown in streams.

Nutrient enrichment of detritus-based streams increases detrital resource quality for consumers and stimulates breakdown rates of particulate organic carbon (C). The relative importance of dissolved inorganic nitrogen (N) vs. phosphorus (P) for detrital quality and their effects on microbial- vs. detritivore-mediated detrital breakdown are poorly understood. We tested effects of experimental N and P additions on detrital stoichiometry (C:N, C:P) and total and microbial breakdown (i.e., with and without detritivorous shredders, respectively) of five detritus types (four leaf litter species and wood) with different initial C : nutrient content. We enriched five headwater streams continuously for two years at different relative availabilities of N and P and compared breakdown rates and detrital stoichiometry to pretreatment conditions. Total breakdown rates increased with nutrient enrichment and were predicted by altered detrital stoichiometry. Streamwater N and P, fungal biomass, and their interactions affected stoichiometry of detritus. Streamwater N and P decreased detrital C:N, whereas streamwater P had stronger negative effects on detrital C:P. Nutrient addition and fungal biomass reduced C:N by 70% and C:P by 83% on average after conditioning, compared to only 26% for C:N and 10% for C:P under pretreatment conditions. Detritus with lowest initial nutrient content changed the most and had greatest increases in total breakdown rates. Detrital stoichiometry was reduced and differences among detritus types were homogenized by nutrient enrichment. With enrichment, detrital nutrient content approached detritivore nutritional requirements and stimulated greater detritivore vs. microbial litter breakdown. We used breakpoint regression to estimate values of detrital stoichiometry that can potentially be used to indicate elevated breakdown rates. Breakpoint ratios for total breakdown were 41 (C:N) and 1518 (C:P), coinciding with total breakdown rates that were ~1.9 times higher when C:N or C:P fell below these breakpoints. Microbial and shredder-mediated breakdown rates both increased when C:N and C:P were reduced, suggesting that detrital stoichiometry is useful for predicting litter breakdown dominated by either microbial or shredder activity. Our results show strong effects of nutrient enrichment on detrital stoichiometry and offer a robust link between a potential holistic nutrient loading metric (decreased and homogenized detrital stoichiometry) and increased C loss from stream ecosystems.

Biomagnification of mercury and selenium in two lakes in southern Norway.

We have investigated bioaccumulation and trophic transfer of both mercury (Hg) and selenium (Se) in two lakes in southern Norway to reveal a suggested mitigating effect of Se on Hg biota accumulation. The study included analysis of total Se (Se), total Hg (Hg), and methyl-mercury (MeHg) in water, littoral and pelagic invertebrates and perch (Perca fluviatilis), together with stable isotope analysis (δ(15)N and δ(13)C) in biota. Mean dissolved Se ranged from 22 to 59ngL(-1), while Hg and MeHg in lake water ranged from 1 to 3ngL(-1) and 0.01 to 0.06ngL(-1). Biota Se and Hg concentrations (dry weight) ranged from 0.41mgSekg(-1) and 0.06mgHgkg(-1) in primary littoral invertebrates and up to 2.9mg Sekg(-1) and 3.6mgHgkg(-1) in perch. Both Hg and Se biomagnified in the food web, with a trophic magnification factor (TMF) of 4.64 for Hg and 1.29 for Se. The reported positive transfer of Se in the food web, despite the low measured dissolved Se, suggest that a major proportion of the Se in these lakes are both highly bioavailable and bioaccumulative. However, we did not find support for a Se-facilitated inhibition in the accumulation of Hg in perch, as Se and Hg concentrations in perch muscle correlated positively and Se did not explain any variations in Hg after we controlled for the effects of other important covariates. We postulate that this may be a result of insufficient concentrations of dissolved Se and subsequently in biota in our studied lakes for an efficient Hg sequestration up the food web.

Using molecular biomarkers and traditional morphometric measurements to assess the health of slimy sculpin (Cottus cognatus) from streams with elevated selenium in North-Eastern British Columbia.

Canadian fish-based environmental effects monitoring programs use individual and population-level endpoints to assess aquatic health. Impacts of coal mining and selenium (Se) exposure were assessed in slimy sculpin (Cottus cognatus) from reference streams located both inside and outside of a coal zone, and from 1 stream with a history of coal mining, using traditional environmental effects monitoring endpoints. In addition, physical characteristics of the streams and benthic macro-invertebrate communities were assessed. To determine whether the assessment of effects could be improved by including molecular markers, real-time polymerase chain reaction assays were optimized for genes associated with reproduction (vtg, esr1, star, cyp19a1, and gys2), and oxidative and cellular stress (sod1, gpx, gsr, cat, and hsp 90). Water Se levels exceeded guidelines in the stream with historical mining (4 µg/L), but benthic macroinvertebrates did not exceed dietary thresholds (2-3 µg/g dry wt). Whole-body Se levels were above British Columbia's tissue guideline in fish from all streams, but only above the draft US Environmental Protection Agency (USEPA) criterion (7.91 µg/g dry wt) at the reference stream inside the coal zone. Some markers of cellular and oxidative stress were elevated in fish liver at the exposed site (sod1, gpx), but some were lower (cat, sod1, gpx, gsr, hsp90) in the gonads of fish inside the coal zone. Some of the differences in gene expression levels between the reference and impacted sites were sex dependent. Based on benthic macroinvertebrate assessments, the authors hypothesize that traditional and molecular differences in slimy sculpin at impacted sites may be driven by food availability rather than Se exposure. The present study is the first to adapt molecular endpoints in the slimy sculpin for aquatic health assessments.

Chemoinformatic analysis as a tool for prioritization of trypanocidal marine derived lead compounds.

Marine trypanocidal natural products are, most often, reported with trypanocidal activity and selectivity against human cell lines. The triaging of hits requires a consideration of chemical tractability for drug development. We utilized a combined Lipinski's rule-of-five, chemical clustering and ChemGPS-NP principle analysis to analyze a set of 40 antitrypanosomal natural products for their drug like properties and chemical space. The analyses identified 16 chemical clusters with 11 well positioned within drug-like chemical space. This study demonstrated that our combined analysis can be used as an important strategy for prioritization of active marine natural products for further investigation.

Marine natural products: bryostatins in preclinical and clinical studies.

CONTEXT: Bryostatins represent an important group of pharmaceutically promising substances. These compounds are produced by commensal microorganisms naturally occurring in marine invertebrates, mainly in bryozoans. The most frequently investigated substance is bryostatin-1, which is a highly oxygenated macrolide with a polyacetate backbone. OBJECTIVE: The aim of this work was to summarize documented preclinical and clinical effects of bryostatin-class compounds. METHODS: A literature search was made of Medline and Web of Science databases in 2012. RESULTS AND CONCLUSION: Our review showed that bryostatins are potent agonists of protein kinase C. In addition to this, their significant antineoplastic activity against several tumor types has also been established and described. Bryostatin's anticancer activity has been proved against various cancer types. Moreover, significant results have been achieved by using bryostatin-1 in combination with other therapies, including combination with vaccine testing. Concerning other important properties that bryostatins possess, their ability to sensitize some resistant cells to chemotherapy agents, or immunoactivity and further stimulating growth of new neural connections, and enhancing effect on long-term memory are worth mentioning. In particular, some new bryostatin analogs could represent potential therapeutic agent for the treatment of cancer and other diseases in future.

[Body mass and excretion of phosphorus in aqueous invertebrates].

Empirical materials on the dependence of the intensity of mineral phosphorus excretion (Ex) on the body mass (W) in invertebrates are summarized. The parameters of the average dependence of Ex on Win animals at 20 degrees C are determined. The main factors that influence the parameters ofthis dependence are discussed. It is shown that the decrease of Ex with an increase of Win animals is, as a rule, more significant compared with the decrease in the intensity of metabolism.

Distribution of polychlorinated biphenyls in an urban riparian zone affected by wastewater treatment plant effluent and the transfer to terrestrial compartment by invertebrates.

In this study, we investigated the distribution of polychlorinated biphenyls (PCBs) in a riparian zone affected by the effluent from a wastewater treatment plant (WWTP). River water, sediment, aquatic invertebrates and samples from the surrounding terrestrial compartment such as soil, reed plants and several land based invertebrates were collected. A relatively narrow range of δ(13)C values was found among most invertebrates (except butterflies, grasshoppers), indicating a similar energy source. The highest concentration of total PCBs was observed in zooplankton (151.1 ng/g lipid weight), and soil dwelling invertebrates showed higher concentrations than phytophagous insects at the riparian zone. The endobenthic oligochaete Tubifex tubifex (54.28 ng/g lw) might be a useful bioindicator of WWTP derived PCBs contamination. High bioaccumulation factors (BAFs) were observed in collected aquatic invertebrates, although the biota-sediment/soil accumulation factors (BSAF) remained relatively low. Emerging aquatic insects such as chironomids could carry waterborne PCBs to the terrestrial compartment via their lifecycles. The estimated annual flux of PCBs for chironomids ranged from 0.66 to 265 ng⋅m(-2)⋅y(-1). Although a high prevalence of PCB-11 and PCB-28 was found for most aquatic based samples in this riparian zone, the mid-chlorinated congeners (e.g. PCB-153 and PCB-138) became predominant among chironomids and dragonflies as well as soil dwelling invertebrates, which might suggest a selective biodriven transfer of different PCB congeners.

Predators and resources influence phosphorus transfer along an invertebrate food web through changes in prey behaviour.

Predators play a fundamental role in prey trophic behaviour, with indirect consequences for species coexistence and ecosystem functioning. Resource quality and availability also influence prey trophic behaviour, with potential effects on predator-prey dynamics. Although many studies have addressed these topics, little attention has been paid to the combined effects of predators and resources on prey species coexistence and nutrient transfer along food chains, especially in detritus-based systems. To determine the influence of predators and resource quality on the movement and P uptake of detritivores, we carried out a field experiment on the River Kelvin (Scotland) using (32)P to test the hypothesis of reduced prey vagility among resource patches as a strategy to avoid predation. Thirty leaf sacks containing alder leaves and two detritivore prey populations (Asellus aquaticus and Lymnaea peregra) were placed in cages, half of them with two predator species (Dendrocoelum lacteum and Erpobdella octoculata) and the other half without predators. Five alder leaf bags, each individually inoculated with a different fungus strain to simulate a patchy habitat, were placed inside each leaf sack. One bag in each sack was labelled with (32)P, in order to assess the proportion of detritivores using it as food and thus their movement among the five resource patches. Three replicates for each labelled fungus and each predation treatment (i.e. with and without predators) were left on the riverbed for 7 days. The presence of predators had negligible effects on the number of detritivores in the leaf bags, but it did reduce the proportion of (32)P-labelled detritivores and their P uptake. The most strongly affected species was A. aquaticus, whose vagility, trophic overlap with L. peregra and P uptake were all reduced. The results confirm the importance of bottom-up and top-down forces acting simultaneously to regulate nutrient transfer along food chains in patchy habitats.

Selenium bioaccumulation in stocked fish as an indicator of fishery potential in pit lakes on reclaimed coal mines in Alberta, Canada.

Pit lakes are a common reclamation strategy for open pit mines; however, there is a concern about their water quality and suitability as fish habitat because they are often contaminated by metals or metalloids. This study assessed the exposure of fish and invertebrates to selenium (Se) and other metals and metalloids in pit lakes formed by open pit coal mining in Tertiary (thermal coal) and in Cretaceous (metallurgical coal) bedrock. Juvenile hatchery rainbow trout, Oncorhynchus mykiss, and brook trout, Salvelinus fontinalis, were stocked into two thermal coal pit lakes (water Se < 2 µg/L, low water Se) and two metallurgical coal pit lakes (water Se > 15 µg/L, high water Se). Se accumulation in stocked fish and concentrations in invertebrates were characterized over a period of 2 years. In the metallurgical pits, invertebrates had higher Se concentrations and fish accumulated Se to higher levels (exceeding USEPA tissue Se guidelines) than biota in the thermal pits. Rainbow and brook trout accumulated similar concentrations of Se in their muscle and exhibited a similar relationship between whole-body and muscle Se concentrations. These results may be used by resource managers to assess compliance with whole-body tissue Se guidelines and to determine if pit lakes in coal mining areas pose a significant Se risk to wildlife or human health. The high Se exposure in metallurgical coal pits indicates that under the current mining and reclamation strategy, these lakes are not suitable for management as recreational "put and take" fisheries.

Direct soil contact values for ecological receptors exposed to weathered petroleum hydrocarbon (PHC) fraction 2.

Ecological tier 1 Canada-wide standards (CWS) for petroleum hydrocarbon (PHC) fraction 2 (F2; >nC10-C16) in soil were derived using ecotoxicological assessment endpoints (effective concentrations [ECs]/lethal concentrations [LCs]/inhibitory concentrations, 25% [IC25s]) with freshly spiked (fresh) fine- and coarse-grained soils. These soil standards might be needlessly conservative when applied to field samples with weathered hydrocarbons. The purpose of the present study was to assess the degradation and toxicity of weathered PHC F2 in a fine-grained soil and to derive direct soil contact values for ecological receptors. Fine-grained reference soils were spiked with distilled F2 and weathered for 183 d. Toxicity tests using plants and invertebrates were conducted with the weathered F2-spiked soils. Endpoint EC/IC25s were calculated and used to derive soil standards for weathered F2 in fine-grained soil protective of ecological receptors exposed via direct soil contact. The values derived for weathered F2 were less restrictive than current ecological tier 1 CWS for F2 in soil.

Does equilibrium passive sampling reflect actual in situ bioaccumulation of PAHs and petroleum hydrocarbon mixtures in aquatic worms?

Over the past couple of years, several analytical methods have been developed for assessing the bioavailability of environmental contaminants in sediments and soils. Comparison studies suggest that equilibrium passive sampling methods generally provide the better estimates of internal concentrations in organisms and thus of subsequent risks. However, field studies to validate the potential of passive sampling to predict actual in situ bioaccumulation are scarce and limited information only exists on selected, individual compounds. The present study investigated whether bioaccumulation of PAH and complex petroleum hydrocarbon mixtures in field-exposed aquatic worms could be predicted properly with passive samplers. To this end, in situ bioaccumulation in aquatic worms at 6 PAH-contaminated locations and 8 petroleum hydrocarbon (oil)-contaminated locations was compared with the results of in situ solid phase micro extraction (SPME) applications. For the oil-contaminated sediments, bioaccumulation was also assessed in the lab with polyoxymethylene solid phase extraction (POM-SPE). Actual PAH bioaccumulation was generally predicted within a factor of 4 with in situ SPME, using temperature-adjusted SPME fiber-water partition coefficients and lab-derived bioaccumulation factors (BAFs) for the worm species used, demonstrating the method's potential under field conditions. In situ SPME appeared to be less suitable for predicting bioaccumulation of oil however, in contrast to POM-SPE in the lab, which assessed in situ oil bioaccumulation within a factor of 3, while also closely reflecting the actual distribution of oil boiling point fractions (the hydrocarbon block profile) as accumulated by the worms. All in all, the results indicated that (specific) equilibrium passive samplers, either applied in the field or the lab, have great potential for assessing bioaccumulation of environmental contaminant mixtures from field-contaminated sediments.

Food chain model based on field data to predict westslope cutthroat trout (Oncorhynchus clarkii lewisi) ovary selenium concentrations from water selenium concentrations in the Elk Valley, British Columbia.

Previous studies conducted in the Elk River watershed showed that selenium concentrations are higher in aquatic biota in lentic compared to lotic habitats of the system having similar water selenium concentrations. Studies have also shown that water selenium concentrations have increased over time (~10% per year) and recent annual average concentrations have ranged up to 0.044 mg/L in areas downstream from mine discharges. For the present study, trophic transfer of selenium was characterized in lotic versus lentic habitats using concentrations measured in field-collected samples and assuming a three-step food chain of water to the base of the food web (biofilm), to benthic invertebrates, and then to westslope cutthroat trout (WCT) ovaries. Food chain models were developed for each habitat type (lotic and lentic) by combining linear regression equations for the three transfer relationships, allowing for prediction of fish ovary concentrations from water concentrations. Greater accumulation of selenium in lentic areas was mostly attributable to greater uptake at the base of the food chain compared to lotic areas. Enrichment/trophic transfer factors for selenium at all levels of the lotic and lentic food chains decreased and then became near constant as exposure concentrations increased. The lotic model predicted little increase in WCT ovary selenium concentrations over an eightfold increase in water concentrations (~0.005-0.040 mg/L), accounting for the lack of observed increase in within-area fish tissue concentrations over time despite increasing trends in water concentrations.

Low concentrations of selenium in stream food webs of eastern Canada.

Herbivorous and predatory invertebrates and two species of fish (brook trout and blacknose dace) were collected from 49 streams in New Brunswick, Canada to determine whether concentrations of selenium (Se) in the biota were affected by a point source (a coal-fired power plant), and stream water chemistry (pH, sulphate, conductivity, and total organic carbon), and to determine the trophic transfer of Se through these food webs. Total Se concentrations in the biota were generally low (0.2 to 4.8 µg g(-1) dry weight) across sites and there was no relationship between distance from the coal-fired power plant and Se concentrations in invertebrates or fishes. Water chemistry was an equally poor predictor of Se concentrations in invertebrates and fish. Trophic position (determined using δ(15)N) was a significant predictor of Se concentrations in only five of the stream food webs, and two of these had negative slopes, indicating little or no trophic magnification across most systems; many fishes had lower concentrations than their invertebrate prey and trophic transfer was higher at sites with low invertebrate Se concentrations. Variability in Se concentrations in fishes was explained more by site of capture than microhabitat use within the site (as measured with δ(13)C), suggesting among-site differences in geological sources of Se. Because concentrations were below known toxicity thresholds for fish and other consumers, these results suggest that Se is not an environmental issue in New Brunswick streams that do not receive direct inputs from mining activities.

New methods to explore marine resources for Alzheimer's therapeutics.

Despite the long history of drug discovery from natural sources, the marine environment, which covers 70% of the Earth's surface, is still relatively unexplored. Intense competition for limited resources drives the evolution of specific and potent chemical defenses distinct from their terrestrial counterparts. Based on this rationale, we recently began screening extracts derived from marine invertebrate and cyanobacterial samples for BACE-1 inhibitors in a chemiluminescent enzyme-fragment complementation (EFC) assay. The results of this broad screening are presented here, along with our progress towards the development of a secondary LC-MS homogeneous affinity assay. Incubation of the extracts active in the EFC assay with BACE1, subsequent isolation of the enzyme-inhibitor complex and then analysis of the small molecule inhibitor by LC-MS rapidly links a chemical structure to biological activity. This approach enables the rapid target-orientated discovery of BACE-1 inhibitors from marine sources.