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Thirteen elements including Al, Ag, As, Co, Cu, Cd, Cr, Fe, Mn, Ni, Se, Zn, and Pb were measured in 107 surface grab sediment samples and 175 segments of eight cores from Lakes Superior, Michigan, and Huron, using inductively coupled plasma mass spectrometry (ICP-MS). Concentrations in Ponar grabs vary considerably among metals and among locations, ranging from the highest median for Fe in Lake Superior (42,000 mg/kg) to the lowest median for Ag in the main Lake Huron (0.05 mg/kg). The inventory at coring sites ranged from 7 × 10 6  mg/m 2 of Fe to 3 mg/m 2 of Ag. The background concentrations were estimated from deeper core segments, and enrichment factors (EFs) were calculated with Fe or Al as the reference element. The results show that Al, Fe, Co, Cr, and Mn did not enrich, Ag, Cu, and Ni were present higher than expected from natural sources alone, while Pb, Cd, Se, Zn, and As have been enriched at most sites after European settlement in the region. EFs of most metals are higher for Lake Michigan than the other lakes. However, EF comparison among sampling sites revealed intrinsic problems of this approach for the assessment of human interference. Preliminary risk assessment, conducted by calculating risk quotients, revealed environmental risks of some metals in each lake; however, the results should be interpreted with caution because the approach used is considered to be conservative., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Microplastic risk assessment often characterizes primary plastics, plastics intentionally manufactured at the micro- and nanoscale, or plastics collected within the natural environment, which often lack repeatability and the volume necessary for reliable hazard characterization. There is limited understanding of how environmentally representative plastics prepared at the microscale impact aquatic organisms. The aim of the present study was to create environmentally representative microplastics and characterize their toxicokinetics and hazards. Plastic cups and forks were micronized to <120 μm particles and Daphnia magna were exposed for 48 h at concentrations ranging from 0.01 mg/L to 100 mg/L. Uptake and depuration experiments were conducted at the highest concentration where accumulation was confirmed. Raman spectroscopy identified that both plastics were polystyrene and had similar size distributions. Microplastics were not acutely toxic but accumulated and rapidly depurated. Toxicokinetics demonstrated that cup MPs were consumed at higher rates than fork MPs despite similar physical characteristics. Daphnia magna preferentially selected smaller particles from the heterogenous suspensions. Future research will need to further explore the relationship between physicochemical properties, particularly size, and ecotoxicity. The study focused on mortality as the primary hazard endpoint. However alternative, sublethal biomarkers may be more appropriate in describing the effects of microplastic exposure., Competing Interests: Declaration of competing interest The authors declare no conflict of interest at this time., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study aimed to investigate the impact of Vernonia amygdalina leaf extract on FLT3 regulation. V. amygdalina was extracted with 96% ethanol (VAE-96), and its cytotoxicity against FLT3- cell lines (MOLM-13 and MV-4-11) was evaluated. The results indicated that VAE-96 induced apoptosis in these cells and inhibited the phosphorylation of AKT, MAPK, and FLT3. Additionally, VAE-96 substantially diminished the activity of the FLT3 promoter and the expression of FLT3 mRNA. The extract was found to contain alkaloids, saponin, reduced sugar compounds, and polyphenols, including tannins and flavonoids, as per the phytoconstituents analysis. The potential of alkaloid fractions on MOLM-13 cells was indicated by the robust cytotoxic effect of the alkaloid fractions, which resulted in over 50% cell mortality at 30 µg/ml. Our results suggest that VAE-96 may be a beneficial agent for the prevention and treatment of AML with FLT3-ITD mutation.

It has been demonstrated that Lantana camara possesses several therapeutic properties that can be used to treat various human diseases, including dermatological and gastrointestinal conditions, tetanus, malaria, and tumours. In this investigation, every collected part of L. camara was extracted with absolute methanol to examine its antioxidant capacity using the DPPH assay and its anti-leukemia activity on two AML cell lines, MOLM-13 and MV4-11. In addition, anti-inflammatory effectiveness was evaluated. The results show that extracts from various sections of L. camara have a significant ability to neutralize free radicals, as indicated by their EC50 values. Most of the extracts had values less than 100 μg/ml, with the flower extract having an even lower value of less than 50 μg/ml. Experiments on two AML cell lines showed that the anti-leukemia effects of the extracts were remarkable, with the most potent impact belonging to the root extract (IC50 was 9.78 ± 0.61 and 12.48 ± 1.69 for MOLM-13 and MV4-11 cell lines). The antitumor effect of the extracts was determined to be time- and dose-dependent and did not correlate with antioxidant capacity. Furthermore, when BJ cells were exposed to L. camara root and leaf extracts, their migratory potential was dramatically reduced compared to untreated cells. The extracts demonstrated potential anti-inflammatory capabilities by lowering NO production in LPS-induced BJ cells.

Biological therapies are available for the treatment of the severe allergic asthma (SAA) with blood eosinophil count ≥ 0.3 × 10 9 /L. Several of them also showed benefits on nasal polyps (NP), one of the most frequent comorbidities of the severe asthma, but comparative studies on their effectiveness in the association SAA-NP are currently lacking. The aim of this study is to compare the effectiveness of benralizumab, mepolizumab and omalizumab in patients with SAA-NP in real-life settings. A retrospective, observational, multicenter real-life study was realized including patients with SAA-NP treated by benralizumab, mepolizumab or omalizumab for 6 months. We analysed the nasal and respiratory symptoms, the number of asthma attacks and salbutamol use/week, acute sinusitis and severe exacerbation rates, the asthma control score, the lung function parameters, the NP endoscopic score, the sinus imaging and the blood eosinophil count 6 months before and after treatment. Seventy-two patients with SAA-NP were included: 16 treated by benralizumab, 21 by mepolizumab and 35 by omalizumab. After 6 months of treatment, almost all studied parameters were improved (except sinus imaging) with a greater effect of omalizumab on the nasal pruritus (p = 0.001) and more benefits of benralizumab on exacerbations rate, asthma attacks per week and lung function (all p < 0.05). Benralizumab and mepolizumab were more effective to improve the NP endoscopic score and the blood eosinophil count (both p < 0.001). All three biological therapies showed effectiveness by improving asthma and nasal outcomes in patients with SAA-NP. Several differences have been found that should be confirmed by larger comparative studies., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Poloxamers are negatively temperature-sensitive hydrogels and their hydrophilic groups interact with water molecules at lower temperatures (liquid phase) while their hydrophobic groups interact more strongly with increases in temperature causing gelation. To investigate the factors affecting the rheological properties of poloxamers, various parameters including different poloxamer P407 concentrations, poloxamers P407/P188 blending ratios and additives were examined. The results presented a clear trend of decreasing gelling temperature/time when P407 was at higher concentrations. Moreover, the addition of P188 enhanced the gelling temperature regardless of poloxamer concentration. Polysaccharides and their derivatives have been widely used as components of hydrogel and we found that alginic acid (AA) or carboxymethyl cellulose (CMC) reduced the gelling temperature of poloxamers. In addition, AA-containing poloxamer promoted cell proliferation and both AA -and CMC-containing poloxamer hydrogels reduced cell migration. This study investigated the intriguing characteristics of poloxamer-based hydrogel, providing useful information to compounding an ideal and desired thermo-sensitive hydrogel for further potential clinical applications such as development of sprayable anti-adhesive barrier, wound-healing dressings or injectable drug-delivery system for cartilage repair.

Freshwater organisms are often exposed to contaminants such as heavy metals from stormwater discharges, which are dependent on rainfall duration and intensity. Therefore, standardized (48- or 96-h) continuous exposure methods developed for whole effluent toxicity (WET) testing might not always accurately convey the effects of stormwater and runoff contaminants. The present study characterized the acute toxicity of copper (Cu), zinc (Zn), and cadmium (Cd) to freshwater amphipods (Hyalella azteca) and cadmium (Cd) to water fleas (Ceriodaphnia dubia) using a modified exposure design that integrated relevant pulsed durations and included post-exposure monitoring. Less than 24-h-old C. dubia and 7 to 8-day-old H. azteca were exposed to water spiked with Cu, Zn, or Cd using 6-, 12-, 26-, or 96-h durations under standard laboratory conditions and monitored for cumulative mortality and reproduction (C. dubia only). Lethal effect (LC10s, LC25s, LC50s) and reproductive effect (EC25s, EC50s) were determined based on either mortality or reproduction of organisms at the end of each pulse (6, 12, or 26 h) and at the end of their respective tests (96 h). For all metals exposed to each organism, acute toxicity was found to be highest for the (96 h) continuous exposures. For pulsed exposures, mortality continued to increase following transfer to clean water for post-exposure monitoring. These results indicate a latent effect of Cu, Zn, and Cd to H. azteca and Cd to C. dubia. The present study concluded that using the continuous (48- or 96-h) WET exposure method overestimates the effects of stormwater and runoff contaminants. However, pulsed exposures without post-exposure monitoring also underestimate the toxicity of contaminants. The proposed pulsed exposure design provides a compromise that is more realistic than current WET methods to assess impacts from episodic events and accounts for potential latent effects that may be overlooked without monitoring post-exposure. Environ Toxicol Chem 2022;41:2488-2499. © 2022 SETAC., (© 2022 SETAC.)

This study examines the abundance and types of microplastic (MP) content in the digestive system of different bird species to help us better understand MP transfer to birds from their environments. The lower GI tracts of six bird species (Tree Swallow (Tachycineta bicolor), Ovenbird (Seiurus aurocapilla), Dark-eyed Junco (Junco hyemalis), Hermit Thrush (Catharus guttatus), Tennessee Warbler (Leiothlypis peregrine), White-throated Sparrow (Zonotrichia albicollis)) were collected for MP analysis. Tree Swallows were collected from nine locations along three drainages in the Milwaukee area and at a remote lake in Wisconsin, USA and consisted of nestlings not yet fledged. The five other bird species were adult migratory bird fatalities from window strikes at the Loyola University Chicago's Lake Shore Campus. Results of plastic analysis showed that the lower GI tracts of all bird species contained different types of MPs (i.e., fibers, fragment, beads). Fiber was the most dominant, followed by fragment and beads. Microplastics were polyethylene, polyethylene terephthalate, nylon, and polyvinyl base materials. There was no statistically significant difference in the site average MP concentration for nestling Tree Swallows or in the species average concentration for adult migratory birds. However, except for Ovenbird, species of adult migratory birds had a species average MP concentration (20.1) that was statistically higher than the site-average MP concentration for Tree Swallow nestlings (5.9). The presence of MPs in the lower GI tracts of unfledged swallows could suggest that MPs had been transferred to the birds from the environment via their diet, such as flying insects-the food source of the birds. Further study should be conducted to investigate the potential transfer of MPs from aquatic insects to birds., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Mercury (Hg) is a ubiquitous contaminant in the environment and its methylated form, methylmercury (MeHg), poses a worldwide health concern for humans and wildlife, primarily through fish consumption. Global production of forest fire ash, derived from wildfires and prescribed burns, is rapidly increasing due to a warming climate, but their interactions with aqueous and sedimentary Hg are poorly understood. Herein, we compared the differences of wildfire ash with activated carbon and biochar on the sorption of aqueous inorganic Hg and sedimentary Hg methylation. Sorption of aqueous inorganic Hg was greatest for wildfire ash materials (up to 0.21 μg g -1 or 2.2 μg g -1 C) among all of the solid sorbents evaluated. A similar Hg adsorption mechanism for activated carbon, biochar made of walnut, and wildfire ash was found that involves the formation of complexes between Hg and oxygen-containing functional groups, especially the -COO group. Notably, increasing dissolved organic matter from 2.4 to 70 mg C L -1 remarkably reduced Hg sorption (up to 40% reduction) and increased the time required to reach Hg-sorbent pseudo-equilibrium. Surprisingly, biochar and wildfire ash, but not activated carbon, stimulated MeHg production during anoxic sediment incubation, possibly due to the release of labile organic matter. Overall, our study indicates that while wildfire ash can sequester aqueous Hg, the leaching of its labile organic matter may promote production of toxic MeHg in anoxic sediments, which has an important implication for potential MeHg contamination in downstream aquatic ecosystems after wildfires.

A long-term exposure outdoor microcosm study was conducted to evaluate the effects of zinc (Zn) on zooplankton, phytoplankton, and periphyton in a freshwater system. Five Zn treatment concentrations (nominal: 8, 20, 40, 80, and 160 μg/L Zn) and an untreated control with 3 replicates each were used. Various physical and chemical characteristics of the microcosms and biological assessment endpoints (e.g., total abundance, group abundance, species richness, chlorophyll a, etc.) were measured to determine the effects of Zn over time. In general, physical and chemical characteristics (e.g., total dissolved solids, total suspended solids, dissolved oxygen, pH, dissolved organic carbon) of water fluctuated over time, but they were not significantly different within treatments and controls during the study. Zinc significantly affected the population dynamics and community structure of plankton. The effects occurred 7 d after initial treatment exposures began and continued to the end of the treatment phase, especially at the high treatment concentrations. Total and group abundance, species richness, the Shannon index, and chlorophyll a concentrations for high Zn treatment concentrations were significantly lower than the controls during the treatment phase. The no-observed-effect, lowest-observed-effect, and median effect concentrations were generally lower than the literature-reported results from single-species toxicity tests for fish and invertebrates, suggesting that plankton are more sensitive to Zn than planktivores. Although primary producers play an important role in the ecosystem, they have not been consistently incorporated into numerical environmental quality criteria for freshwater organisms, at least in the United States. The results of the present study are useful for development of environmental quality guidelines for freshwater ecosystems and ecological risk assessment. Environ Toxicol Chem 2021;40:2053-2072. © 2021 SETAC., (© 2021 SETAC.)

Background: Viral infections are known to exacerbate asthma in adults. Previous studies have found few patients with asthma among severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pneumonia cases. However, the relationship between SARS-CoV-2 infection and severe asthma exacerbation is not known., Objective: To assess the frequency of asthma exacerbation in patients with asthma hospitalized for SARS-CoV-2 pneumonia and compare symptoms and laboratory and radiological findings in patients with and without asthma with SARS-CoV-2 pneumonia., Methods: We included 106 patients between March 4 and April 6, 2020, who were hospitalized in the Chest Diseases Department of Strasbourg University Hospital; 23 had asthma. To assess the patients' asthma status, 3 periods were defined: the last month before the onset of COVID-19 symptoms (p1), prehospitalization (p2), and during hospitalization (p3). Severe asthma exacerbations were defined according to Global INitiative for Asthma guidelines during p1 and p2. During p3, we defined severe asthma deterioration as the onset of breathlessness and wheezing requiring systemic corticosteroids and inhaled β2 agonist., Results: We found no significant difference between patients with and without asthma in terms of severity (length of stay, maximal oxygen flow needed, noninvasive ventilation requirement, and intensive care unit transfer); 52.2% of the patients with asthma had Global INitiative for Asthma step 1 asthma. One patient had a severe exacerbation during p1, 2 patients during p2, and 5 patients were treated with systemic corticosteroids and inhaled β2 agonist during p3., Conclusions: Our results demonstrate that patients with asthma appeared not to be at risk for severe SARS-CoV-2 pneumonia. Moreover, SARS-CoV-2 pneumonia did not induce severe asthma exacerbation., (Copyright © 2020 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.)

The present study characterizes the dependence of microplastic consumption and excretion on particle size and body shape of fathead minnow (Pimephales promelas) over time that has not been studied. Specifically, the study is to answer four important questions: 1) how do P. promelas consume microplastic particles at different size ranges over time? 2) how long does it take for P. promelas to excrete microplastic particles after consumption? 3) do P. promelas reconsume microplastic particles after excretion? 4) are microplastic consumption and excretion by P. promelas dependent on the body shape? To answer these questions, larval P. promelas were exposed to polyethylene microbeads (PMBs) at two different consumable size ranges of 63-75 µm and 125-150 µm in moderately hard water. The experiments were designed to allow and to not allow fish to reconsume the particles they excreted. Results of the present study showed that P. promelas consumed significant amount of PMBs after 1 h of exposure to PMBs regardless particle size. The number of consumed PMBs per fish at smaller size range was up to 10 times higher than that at larger size range. When expressing the consumption in µg PMBs/fish, this difference was approximately 1.3 times, suggesting the importance of the measurement unit. After consuming, fish excreted PMBs over time and reconsumed excreted PMBs if reconsumption was allowed. Interestingly, it took longer for bent body fish to excrete PMBs than regular straight body fish. Our observation showed that excreted PMBs were likely coated with intestinal fluid that is denser than water, resulting in aggregation and deposition of PMBs. This result suggests that in the natural environment, the consumption and excretion of plastics by fish would enhance the movement of plastics from the water column to the waterbed and make it available for benthic organisms., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Wildfires are expected to become more frequent and intensive at the global scale due to climate change. Many studies have focused on the loss of mercury (Hg) from burned forests; however, little is known about the origins, concentration, reactivity, and bioavailability of Hg in residual ash materials in postfire landscapes. We examine Hg levels and reactivity in black ash (BA, low burn intensity) and white ash (WA, high burn intensity) generated from two recent northern California wildfires and document that all ash samples contained measurable, but highly variable, Hg levels ranging from 4 to 125 ng/g dry wt. ( n = 28). Stable Hg isotopic compositions measured in select ash samples suggest that most Hg in wildfire ash is derived from vegetation. Ash samples had a highly variable fraction of Hg in recalcitrant forms (0-75%), and this recalcitrant Hg pool appears to be associated with the black carbon fraction in ash. Both BA and WA were found to strongly sequester aqueous inorganic Hg but not gaseous elemental Hg under controlled conditions. During anoxic ash incubation with natural surface water, we find that Hg in most ash samples had a minimal release and low methylation potential. Thus, the formation of wildfire ash can sequester Hg into relatively nonbioavailable forms, attenuating the potentially adverse effects of Hg erosion and transport to aquatic environments along with eroded wildfire ash.

The present study assessed the chronic toxicity of cadmium (Cd) and nickel (Ni) mixtures to Daphnia magna. Using a titration design, Ni concentrations of 20, 40, 80, 100, 120, 140, and 160 μg/L were tested alone and simultaneously titrated in increments against a constant concentration of 1.5 μg/L Cd. The results demonstrated that Cd at 1.5 μg/L was highly toxic to D. magna, and Ni alone concentrations ≥80 μg/L were toxic to D. magna survival, reproduction, and growth. No Ni alone concentration was found to induce a toxic effect on undeveloped embryos and the time to first brood. Only the Ni alone treatment containing 200 μg/L affected the reproductive rates of D. magna. For CdNi mixtures, Ni concentrations of 20, 40, and 80 μg/L were found to strongly protect D. magna from Cd toxicity at the survival and growth endpoints, resulting in less-than-additive effects, but not on the reproductive endpoint. At higher concentrations, Ni exceeded the necessary concentration needed to protect D. magna, and appeared to contribute to the toxicity. Overall, the results of metal uptake support the competitive binding mechanism at the biotic ligand and explain the less-than-additive effects observed in the CdNi mixtures concentration. The embryonic effects of CdNi mixtures are not explained by the competitive binding mechanism at the biotic ligand. More research is needed to determine the mechanisms that produce embryonic impairment when cellular metals interact. Overall, the results of the present study are relevant for the development of improved environmental quality guidelines for metal mixtures., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The aim of the present study was to determine the influence of salinity and organism age on the chronic toxicity of waterborne lead (Pb) to Atherinops affinis and to compare the relative Pb sensitivity of A. affinis with other marine species. Chronic Pb exposure experiments were conducted in a water flow-through testing system. Survival, standard length, dry weight, and tissue Pb concentration were measured and lethal concentrations (LCs), effect concentrations (ECs), and bioconcentration factors (BCFs) were calculated. In general, increasing salinity and organism age decreased Pb toxicity. The LC50s for larval fish at 14 and 28 ppt salinity were 15.1 and 79.8 μg/L dissolved Pb, respectively; whereas, the LC50 for juvenile fish was 167.6 μg/L dissolved Pb at 28 ppt salinity. Using standard length data, the EC10 values for larval fish were 16.4 and 82.4 μg/L dissolved Pb at 14 and 28 ppt salinity, respectively. The dry weight EC25s for low and high salinity were 15.6 and 61.84 μg/L dissolved Pb, respectively. The BCFs were higher with the lower salinity study (1703) in comparison to the higher salinity study (654). Results of Pb speciation calculation showed higher fraction of Pb 2+ in water with lower salinity, explaining the higher observed toxicity of Pb in lower salinity water than higher salinity water. Atherinops affinis is more sensitive to Pb than several other marine species. Evidence of abnormal swimming and skeletal deformities were observed in Pb exposure treatments. Results of the present study are useful for marine biotic ligand modeling and support ecological risk assessment and deriving Pb environmental quality criteria for marine environments. Environ Toxicol Chem 2018;37:2705-2713. © 2018 SETAC., (© 2018 SETAC.)

The rapid increase in plastic use over the last few decades has resulted in plastic pollution in freshwater and marine ecosystems. However, more attention has been paid to plastic pollution in marine ecosystems than to freshwater ecosystems. This research determined microplastic ingestion by Daphnia magna and the potential effect of microplastics on the organism's survival and reproduction. The study also examined the potential of microplastics to enhance algal growth in support of understanding effects of microplastic ingestion on the organism. When exposed to 25, 50, and 100mg/L fluorescent green polyethylene microbeads at size of 63-75μm, D. magna ingested significant amount of plastic microbeads. The number of ingested beads increased with increasing particle concentration and exposure time. However, no significant effect on survival and reproduction was observed although the gut of D. magna was filled with plastic microbeads. In the algal experiment, Raphidocelis subcapitata grew more in the exposure media with the present of plastic microbeads than without plastic microbeads. This result suggests that plastic microbeads could serve as substrates for R. subcapitata to grow. Raphidocelis subcapitata then could be transferred to the organism's gut and provided energy for survival and reproduction. Results of the present study add to the literature of microplastic ingestion by aquatic organisms. Caution should be taken when interpreting hazards of microplastics based on ingestion, such as the measurement unit and the presence of algae in the environment., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The present study characterized the chronic effect of binary-metal mixtures of cadmium (Cd) and zinc (Zn) on Daphnia magna. The titration design was chosen to characterize the 21-d chronic effects of the binary-metal mixtures on survival, growth, reproduction, and metal accumulation in D. magna. Using this design, increasing concentrations of Zn (10, 20, 40, 80, 120, 160, and 200 μg/L) were titrated against a constant concentration of 1.5 μg/L Cd. The results demonstrated that Cd was highly toxic to D. magna. In a mixture with Cd and Zn, sublethal concentrations of 10 and 20 μg/L Zn were insufficient to protect D. magna from chronic Cd toxicity, whereas mixtures containing 40, 80, and 120 μg/L Zn provided strong protective effects to D. magna at all endpoints and resulted in less-than-additive effects. At higher Zn concentrations, such as 160 and 200 μg/L, Zn appeared to contribute to the toxicity. The less-than-additive effects observed in the Cd-Zn mixture can be explained by the decrease in body Cd concentration when the Zn concentration was increased in the exposure media. Embryos analyzed for morphological alterations in the Cd-Zn mixtures demonstrated severe developmental defects. The effect of Cd on undeveloped embryos while both Zn and Cd are present in the organisms raises a question of whether the competitive binding mechanism of Zn and Cd is still happening at the cellular level in the organisms. The results of the present study are useful for development of the biotic ligand model and environmental quality guidelines for metal mixtures. Environ Toxicol Chem 2017;36:2739-2749. © 2017 SETAC., (© 2017 SETAC.)

We identified and evaluated differentially expressed genes (DEGs) by RNA-Sequencing (RNA-Seq) in the intestinal mucosa of two Fayoumi chicken lines, M5.1 and M15.2, that are affected by necrotic enteritis (NE); these chicken lines share the same genetic background but have different major histocompatibility complexes. RNA-Seq generated over 49 and 40 million reads for lines M5.1 and M15.2, respectively. The alignment of these sequences with the Gallus gallus genome database revealed the expression of more than 14,500 genes in two lines, among which 581, 1270, and 1140 DEGs were detected when lines M15.2 and M5.1 were compared with the control and compared between each other. The analysis of all DEGs using the gene ontology database revealed annotations for 111 biological processes, 32 cellular components, and 17 molecular functions, and KEEG pathway mapping indicated that the DEGs were primarily involved in immunity, responses to various stimuli, and signal transduction. In addition, we analyzed 183 innate immune genes that were differentially expressed in NE-induced chicken lines, including 46 CD molecular genes, 89 immune-related genes, and 13 β -defensin genes with 3 lineage-specific duplications. Taken together, the transcriptional profiles showed that line M5.1 was more resistant to NE than line M15.2 and that differential gene expression patterns were associated with host genetic differences in resistance to NE. qRT-PCR and RNA-Seq analyses showed that all the genes examined had similar responses to NE (correlation coefficient R=0.84 to 0.88, p <0.01) in both lines. To the best of our knowledge, this is the first study that describes NE-induced DEGs using RNA-seq in two lines with different levels of susceptibility to NE. These results will lead to increased insights on NE disease resistance mechanisms and the role of host genes in the control of the host immune response., Competing Interests: The authors have declared that no competing interests exist., (2017, Japan Poultry Science Association.)

The present study investigated copper (Cu) toxicity and the influence of water quality characteristics of Dongnai River and Mekong River (Vietnam) surface waters to three tropical species; Daphnia lumholtzi, Ceriodaphnia cornuta, and Danio rerio. The river waters had a range of water quality parameters that modify Cu bioavailability and toxicity. The range of total hardness, alkalinity, pH and dissolved organic carbon were 15-64 mg/L as CaCO3, 18-58 mg/L as CaCO3, 6.62-7.88, and 6.9-14.7 mg/l, respectively. The US EPA acute toxicity test method with a modification to the light photoperiod and temperature for tropical organisms was used to investigate Cu toxicity. Result of the present study found that Cu produced toxic effect to the studied organisms at low concentrations. The 48-h LC50 ranged from 3.92 to 8.61 µg/l, 2.92-9.56 µg/l, and 15.71-68.69 µg/l dissolved Cu for D. lumholtzi, C. cornuta, and D. rerio, respectively. In general, water quality had an influence on Cu bioavailability and toxicity to the studied organisms. The toxicity of Cu was higher in water with lower hardness, DOC, and/or pH. The present study indicates a contribution of Cu hydroxide and carbonate to Cu bioavailability to Mekong organisms. Results of the present study will be used for calibrating the US Cu Biotic Ligand Model (BLM) to Mekong River water and organisms in support of application of the BLM for setting site-specific Cu water quality guidelines in the ecosystem of the Lower Mekong River Basin., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Metal mixture toxicity has been studied for decades. However, the results are not consistent, and thus ecological risk assessment and regulation of mixtures has been difficult. The objective of the present study was to use a systematic experimental design to characterize the toxicity of binary-metal mixture of Cu, Zn, and Ni to Pimephales promelas, typically to determine whether the effect of these binary-metal mixtures on P. promelas is more-than-additive. Standard 96-h toxicity tests were conducted with larval P. promelas based on US Environmental and Protection Agency methods to determine metal mixture effects. All experiments were conducted in synthetic moderately hard water with no addition of dissolved organic matter. Three different effect analysis approaches, the MixTox model, the Finney model, and the toxic unit method, were used for comparison. The results indicate that the toxicity of Cu+Zn, Cu+Ni, and Zn+Ni mixtures to P. promelas was more-than-additive. Among the 3 mixtures, the effect of the Cu+Ni mixture was the most profound. The results of the present study are useful for applications to models such as the metal mixture biotic ligand model. More research should be conducted to determine the mechanisms of acute and chronic toxicity of metal mixtures., (© 2015 SETAC.)

The present study characterized the influence of water-quality characteristics on zinc (Zn) toxicity to the Florida apple snail (Pomacea paludosa) and the sensitivity of freshwater snails to Zn. Standard 96-h renewal acute toxicity tests were conducted with Zn and juvenile P. paludosa under 3 conditions of pH and alkalinity, water hardness, and dissolved organic carbon (DOC). Median lethal effect concentrations (96-h LC50s), no-observed- effect concentrations, lowest-observed-effect concentrations, LC10s, and LC20s were determined for each test. The results showed that Zn toxicity to P. paludosa decreased linearly with increasing hardness, pH, and DOC. A multiple linear regression model based on pH, hardness, and DOC was able to explain 99% of the observed variability in LC50s. These results are useful for the development of a biotic ligand model (BLM) for P. paludosa and Zn. Zinc acute toxicity data were collected from the literature for 12 freshwater snail species in a wide range of water-quality characteristics for species sensitivity distribution analysis. The results showed that P. paludosa is the second most sensitive to Zn. The present study also suggested that aqueous ZnCO3 and ZnHCO3 (-) can be bioavailable to P. paludosa. Therefore, bioavailability models (e.g., BLM) should take these Zn species into consideration for bioavailability when applied to snails., (© 2014 SETAC.)

Three Florida native larval butterflies (Junonia coenia, Anartia jatrophae, Eumaeus atala) were used in the present study to determine the acute toxicity, hazard, and risk of a 24h ingestion of leaves contaminated with the adult mosquito control insecticides permethrin, naled, and dichlorvos to late 4th and early 5th in-star caterpillars. Based on 24-h LD50s for ingestion, naled was more acutely toxic than permethrin and dichlorvos to caterpillars. Hazard quotients using the ratio of the highest doses and the 90th percentile doses from field measurements in host plant foliage following actual mosquito control applications to the toxicological benchmarks from laboratory toxicity tests indicate potential high acute hazard for naled compared to permethrin and dichlorvos. Based on probabilistic ecological risk methods, naled exposure doses in the environment also presented a higher acute risk to caterpillars than permethrin and dichlorvos. The acute toxicity laboratory results and ecological risk assessment are based only on dietary ingestion and single chemical doses. It does not include other typical exposure scenarios that may occur in the environment. It is thus plausible to state that the ecological risk assessment presented here underestimates the potential risks in the field to caterpillars. However, one assumption that is scientifically feasible and certainly real from the results - if the environmental exposure doses of mosquito control operations are similar or higher to those presented here in leaves from the field, after applications, there will likely be significant mortalities and other adverse effects on caterpillar populations., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A comprehensive probabilistic terrestrial ecological risk assessment (ERA) was conducted to characterize the potential risk of mosquito control insecticide (i.e., naled, it's metabolite dichlorvos, and permethrin) usage to adult butterflies in south Florida by comparing the probability distributions of environmental exposure concentrations following actual mosquito control applications at labeled rates from ten field monitoring studies with the probability distributions of butterfly species response (effects) data from our laboratory acute toxicity studies. The overlap of these distributions was used as a measure of risk to butterflies. The long-term viability (survival) of adult butterflies, following topical (thorax/wings) exposures was the environmental value we wanted to protect. Laboratory acute toxicity studies (24-h LD50) included topical exposures (thorax and wings) to five adult butterfly species and preparation of species sensitivity distributions (SSDs). The ERA indicated that the assessment endpoint of protection, of at least 90% of the species, 90% of the time (or the 10th percentile from the acute SSDs) from acute naled and permethrin exposures, is most likely not occurring when considering topical exposures to adults. Although the surface areas for adulticide exposures are greater for the wings, exposures to the thorax provide the highest potential for risk (i.e., SSD 10th percentile is lowest) for adult butterflies. Dichlorvos appeared to present no risk. The results of this ERA can be applied to other areas of the world, where these insecticides are used and where butterflies may be exposed. Since there are other sources (e.g., agriculture) of pesticides in the environment, where butterfly exposures will occur, the ERA may under-estimate the potential risks under real-world conditions., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The St. Lucie estuary (SLE) ecosystem in South Florida has been shown to be contaminated with metals and pesticides. Our earlier studies also showed that aquatic organisms, especially benthic species in the SLE ecosystem, might be potentially at high risk from copper (Cu) exposure. The objectives of this study were to conduct studies with separate groups of organisms exposed to seven field-collected sediment samples from the St. Lucie River according to standard procedures to evaluate toxicity and tissue concentrations of Cu and zinc (Zn). Short term and longer term whole sediment acute toxicity studies were performed with Ampelisca abdita and Mercenaria mercenaria. Analysis of sediment chemical characteristics showed that Cu and Zn are of most concern because their concentrations in 86 % of the sediments were higher than the threshold effect concentrations for Florida sediment quality criteria and the National Oceanic and Atmospheric Administration Screening Quick Reference Tables (SQuiRTs) sediment values. There was no significant effect on survival of the tested organisms. However, increased Cu and Zn concentrations in the test organisms were found. Dry weight of the tested organisms was also inversely related to Cu and Zn concentrations in sediments and organisms. The effects on organism weight and Cu and Zn uptake raise concerns about the organism population dynamics of the ecosystem because benthic organisms are primary food sources in the SLE system and are continuously exposed to Cu- and Zn-contaminated sediments throughout their life cycle. The results of the present study also indicate that Cu and Zn exposures by way of sediment ingestion are important routes of exposure.

Background: Food fortification began in Ghana in 1996 when legislation was passed to enforce the iodization of salt. This paper describes the development of the Ghanaian fortification program and identifies lessons learned in implementing fortification initiatives (universal salt iodization, fortification of vegetable oil and wheat flour) from 1996 to date., Objective: This paper identifies achievements, challenges, and lessons learned in implementing large scale food fortification in Ghana., Methodology: Primary data was collected through interviews with key members of the National Food Fortification Alliance (NFFA), implementation staff of the Food Fortification Project, and staff of GAIN. Secondary data was collected through desk review of documentation from the project offices of the National Food Fortification Project and the National Secretariat for the Implementation of the National Salt Iodization in Ghana., Results: Reduction of the prevalence of goiter has been observed, and coverage of households with adequately iodized salt increased between 1996 and 2006. Two models were designed to increase production of adequately iodized salt: one to procure and distribute potassium iodate (KIO3) locally, and the second, the salt bank cooperative (SBC) model, specifically designed for small-scale artisanal salt farmers. This resulted in the establishment of a centralized potassium iodate procurement and distribution system, tailored to local needs and ensuring competitive and stable prices. The SBC model allowed for nearly 157 MT of adequately iodized salt to be produced in 2011 in a region where adequately iodized salt was initially not available. For vegetable oil fortification, implementing quantitative analysis methods for accurate control of added fortificant proved challenging but was overcome with the use of a rapid test device, confirming that 95% of vegetable oil is adequately fortified in Ghana. However, appropriate compliance with national standards on wheat flour continues to pose challenges due to adverse sensory effects, which have led producers to reduce the dosage of premix in wheat flour., Conclusions: Challenges to access to premix experienced by small producers can be overcome with a central procurement model in which the distributor leverages the overall volume by tendering for a consolidated order. The SBC model has the potential to be expanded and to considerably increase the coverage of the population consuming iodized salt in Ghana. Successful implementation of the cost-effective iCheck CHROMA rapid test device should be replicated in other countries where quality control of fortified vegetable oil is a challenge, and extended to additional food vehicles, such as wheat flour and salt. Only a reduced impact on iron deficiency in Ghana can be expected, given the low level of fortificant added to the wheat flour. An integrated approach, with complementary programs including additional iron-fortified food vehicles, should be explored to maximize health impact.

Endosulfan is an insecticide which has been widely used in agriculture. The technical grade material consists of two isomers (alpha and beta). Under natural environmental conditions, endosulfan is metabolized through oxidation and the main metabolite in the environment is endosulfan sulfate. Most ecotoxicology research has been conducted with technical grade endosulfan to determine effects on non-target aquatic organisms. Little data on the effects of endosulfan sulfate on aquatic organisms are available in the literature. This study characterizes endosulfan sulfate bioconcentration and depuration in mosquito fish (Gambusia affinis). During the study, G. affinis was exposed to an environmentally relevant endosulfan sulfate concentration of 0.25 μg L(-1) for 5 weeks (uptake phase) followed by a 3-week period (depuration phase) in clean water. This study found that G. affinis bioconcentrated endosulfan sulfate. During the exposure phase, fish tissue concentrations of endosulfan sulfate increased with time up to 730 μg kg(-1) dw or 215 μg kg(-1) ww. The bioconcentration data followed Michaelis-Menten kinetics better than the one-compartment first order kinetics (1-CFOK). Using these models, the bioconcentration factors for endosulfan sulfate-exposed G. affinis were from 687 to 888 L kg(-1) in wet weight or 2263 to 2936 L kg(-1) in dry weight. During the depuration phase, endosulfan sulfate concentrations in tissue significantly decreased and the data followed first order kinetics. The half-life of endosulfan sulfate in G. affinis was about 9 d. There was no significant difference in standard length or weight between control and exposed fish. The growth data followed the von Bertalanffy growth model. However, the condition factor of exposed fish increased with time during the exposure phase., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

This study characterizes the effects of copper (Cu) on Florida apple snails (Pomacea paludosa) and mosquito fish (Gambusia affinis) using a replicated outdoor microcosm design. Soils used in this study were collected from two Cu-enriched citrus agricultural sites in South Florida (Agler property (AGLR) in St. Lucie County and Sunrise Boys property (SRB) in Palm Beach County) and a reference site (Equus property) in St. Lucie County. The study included a 5-week aging phase, an 11 month exposure phase, and a 3 month post-treatment (exposure) phase. The aging phase was initiated by flooding agricultural soils with rainwater in 4 m(3) fiberglass microcosm tanks. Introducing juvenile apple snails (≤7 d old) and mosquito fish (2-3 cm) into the microcosm tanks initiated the exposure phase. Survival, growth, and reproduction of apple snails and fish, and Cu uptake in apple snails, fish, and periphyton were determined in this study. Water chemistry (e.g., dissolved Cu concentration, dissolved organic carbon and dissolved oxygen concentrations, pH, hardness, alkalinity, etc.) was measured daily or weekly during the study. Initial soil Cu concentrations in Equus, SRB, and AGLR microcosms were 7, 55, and 99 mg/kg dw, respectively. Dissolved Cu concentrations in Equus, SRB and AGLR microcosms at the beginning of the study were 3, 82, and 43 μg/L, respectively and decreased to low saturation levels of about ≤9 μg/L Cu after the first 3 months of the study. The decrease of dissolved Cu concentrations was likely due to the dilution of rainwater. Snail and fish mortality appeared to be higher in SRB microcosms than in Equus and AGLR microcosms. There was no significant difference in growth of the snails between treatments. Snail growth data followed the von Bertalanffy Model. The maximum shell length, shell height, and shell width of the snails calculated by the von Bertalanffy Model (L(∞)) were 2.76, 2.05, and 2.18 cm, respectively. The maximum wet weight was 9.38 g. Growth rate (k) of the snails increased in order of shell height (0.459), shell length (0.550), and shell weight (0.598). There was no reproduction in the snails in any treatments including the reference during the exposure phase. However, Cu did not affect reproduction of fish during this period. Copper concentrations in periphyton from Equus, SRB, and AGLR microcosms ranged from 2 to 62, 31 to 371, and 13 to 478 mg/kg, respectively. Copper concentrations in fish at the beginning, days 30 and 150 of the study ranged from 3.19 to 7.53 mg/kg and were not significantly different from the different treatments. Average Cu concentrations in the soft tissue of dead snails from SRB and AGLR microcosms were 4602 mg/kg dw (ranged from 2913 to 8370 mg/kg dw) and 2824 mg/kg dw (ranged from 2118 to 3600 mg/kg dw), respectively. The Cu concentrations in the soft tissue of dead snails found in this study were higher than the tissue Cu concentrations in live aquatic organisms reported in the literature. These high Cu concentrations in edible apple snail soft tissue might pose a risk to Florida apple snail predators, including the snail kite. The post-exposure phase, with snails exposed to only water (i.e., no soils) showed depuration of copper from apple snails and reproduction in all treatments., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Honeybees are the standard insect test species used for toxicity testing of pesticides on nontarget insects for the U.S. Environmental Protection Agency (U.S. EPA) under the Federal Insecticide Fungicide and Rodenticide Act (FIFRA). Butterflies are another important insect order and a valued ecological resource in pollination. The current study conducted acute toxicity tests with naled, permethrin, and dichlorvos on fifth larval instar (caterpillars) and adults of different native Florida, USA, butterfly species to determine median lethal doses (24-h LD50), because limited acute toxicity data are available with this major insect group. Thorax- and wing-only applications of each insecticide were conducted. Based on LD50s, thorax and wing application exposures were acutely toxic to both caterpillars and adults. Permethrin was the most acutely toxic insecticide after thorax exposure to fifth instars and adult butterflies. However, no generalization on acute toxicity (sensitivity) of the insecticides could be concluded based on exposures to fifth instars versus adult butterflies or on thorax versus wing exposures of adult butterflies. A comparison of LD50s of the butterflies from this study (caterpillars and adults) with honeybee LD50s for the adult mosquito insecticides on a µg/organism or µg/g basis indicates that several butterfly species are more sensitive to these insecticides than are honeybees. A comparison of species sensitivity distributions for all three insecticides shows that permethrin had the lowest 10th percentile. Using a hazard quotient approach indicates that both permethrin and naled applications in the field may present potential acute hazards to butterflies, whereas no acute hazard of dichlorvos is apparent in butterflies. Butterflies should be considered as potential test organisms when nontarget insect testing of pesticides is suggested under FIFRA., (Copyright © 2011 SETAC.)

Endosulfan sulfate is a persistent environmental metabolite of endosulfan, an organochlorine insecticide-acaricide presently registered by the United States Environmental Protection Agency. There is, however, limited acute fish toxicity data for endosulfan sulfate. This study determines the acute toxicity (LC₅₀s and LC₁₀s) of endosulfan sulfate to three inland Florida native fish species (mosquitofish [Gambusia affinis]; least killifish [Heterandria formosa]; and sailfin mollies [Poecilia latipinna]) as well as fathead minnows (Pimephales promelas). Ninety-six-h acute toxicity tests were conducted with each fish species under flow-through conditions. For all of the above-mentioned fish species, 96-h LC₅₀ estimates ranged from 2.1 to 3.5 μg/L endosulfan sulfate. The 96-h LC₁₀ estimates ranged from 0.8 to 2.1 μg/L endosulfan sulfate. Of all of the fish tested, the least killifish appeared to be the most sensitive to endosulfan sulfate exposure. The above-mentioned data were combined with previous acute toxicity data for endosulfan sulfate and freshwater fish for an effects analysis. The effects analysis estimated hazardous concentrations expected to exceed 5, 10, and 50% of the fish species' acute LC₅₀ or LC₁₀ values (HC₅, HC₁₀, and HC₅₀). The endosulfan sulfate freshwater-fish acute tests were also compared with the available freshwater-fish acute toxicity data for technical endosulfan. Technical endosulfan is a mixture of α- and β-endosulfan. The LC₅₀s had a wider range for technical endosulfan, and their distribution produced a lower HC₁₀ than for endosulfan sulfate. The number of freshwater-fish LC₅₀s for endosulfan sulfate is much smaller than the number available for technical endosulfan, reflecting priorities in examining the toxicity of the parent compounds of pesticides. The toxicity test results and effects analyses provided acute effect values for endosulfan sulfate and freshwater fish that might be applied in future screening level ecologic risk assessments. The effects analyses also discussed several deficiencies in conventional methods for setting water-quality criteria and determining ecologic effects from acute toxicity tests.

Background: Imatinib, a selective tyrosine kinase inhibitor, has been used as a standard first-line therapy for irresectable and metastasized gastrointestinal stromal tumor (GIST) patients. Unfortunately, most patients responding to imatinib will eventually exhibit imatinib-resistance, the cause of which is not fully understood. The serious clinical problem of imatinib-resistance demands alternative therapeutic strategy. This study was conducted to investigate the effect of all-trans retinoic acid (ATRA) on GIST cell lines., Methods: Cell proliferation was determined by trypan blue dye exclusion test. Western blot analysis was performed to test the expression of activated KIT, its downstream proteins, and apoptosis associated proteins. The cytotoxic interactions of imatinib with ATRA were evaluated using the isobologram of Steel and Peckham., Results and Conclusion: In this work, for the first time we have demonstrated that ATRA affected on cell proliferation of GIST-T1 and GIST-882 cell line through inhibition of cell growth in a dose dependent manner and induced apoptosis. High dose of ATRA induced morphologic change in GIST-T1 cells, rounded-up cells, and activated the caspase-3 protein. In further examination, we found that the ATRA-induced apoptosis in GIST-T1 cells was accompanied by the down-regulated expression of survivin and up-regulated expression of Bax protein. Moreover, ATRA suppressed the activity of KIT protein in GIST-T1 cells and its downstream signal, AKT activity, but not MAPK activity. We also have demonstrated that combination of ATRA with imatinib showed additive effect by isobologram, suggesting that the combination of ATRA and imatinib may be a novel potential therapeutic option for GIST treatment. Furthermore, the scracht assay result suggested that ATRA was a potential reagent to prevent the invasion or metastasis of GIST cells.

Single-species flow-through toxicity tests were conducted to determine the times-to-death of two indigenous fish to South Florida--least killifish (Heterandria formosa) and mosquitofish (Gambusia affinis)--from acute exposure to endosulfan sulfate. Mortalities were recorded within 8-h periods from test initiation to termination at 96 h. The 96-h LC(50)s for least killifish and mosquitofish estimated using the trimmed-Spearman-Karber method were 2.0 and 2.3 microg/l, respectively. An accelerated failure time model was used to estimate times to death at selected concentrations. Data were fit to log-normal, log-logistic, and Weibull distributions. Acute toxicity data fit to the Weibull distribution produced a better relative fit than log-normal or log-logistic distributions for both toxicity tests. The survival-time profiles and associated statistics illustrate the benefit of considering exposure duration as well as concentration when predicting acute risk to species' populations. Both toxicity tests had similar outcomes from exposure to endosulfan sulfate, with least killifish being slightly more likely to die at lower concentrations and shorter time periods than mosquitofish. From the models generated by the toxicity tests, times-to-death for least killifish and mosquitofish were estimated for environmentally relevant concentrations of total endosulfan at a site of concern in South Florida. When the results from the current toxicity tests were compared to environmental concentrations from previous screening-level ecological risk assessments, the durations necessary to potentially kill 10% or more of the populations of the two native south Florida fish species were estimated to be 77 and 96 h for least killifish and mosquitofish, respectively. However, the exposure values included the alpha and beta isomers as well as endosulfan sulfate; therefore, an understanding of their toxicity might be important in understanding the survival dynamics of fish species in endosulfan sulfate-contaminated sites.

The uptake and effects (survival, weight) of copper (Cu) on Florida apple snails (Pomacea paludosa) via exposures to copper-enriched agricultural soil-water and water-only treatments were investigated. Soils were collected from citrus sites in south Florida and flooded with laboratory freshwater for 14d. Neonate apple snails (96-h-old) were then exposed to either Cu from a soil-overlying water (i.e., flooded agricultural soils) treatment or overlying water-only (i.e., equilibrated overlying water produced from 14d flooding of agriculture soils) treatment for 14d under standard laboratory conditions. Survival, weight (dry, wet), and whole body Cu uptake were measured. Copper exposure via soil-water exposures resulted in higher mortality and whole body Cu uptake than water-only exposures, indicating Cu uptake from soils. However, snail wet and dry weights were higher in soil-water treatments than in water-only treatments. Micronutrients from soils may be consumed by snails increasing weights. Survival, apple snail dry weight, and whole body Cu concentrations were significantly correlated with soil and water Cu concentrations in soil-water treatments. Survival was significantly correlated with the concentration of Cu(CO3)2(2-) in water-only treatments. This suggests that Cu(CO3)2(2-) is toxic to apple snails. Whole body Cu concentrations were higher in surviving snails than dead snails, suggesting that apple snails have the ability to detoxify accumulated Cu (e.g., through metallothionein induction, granules).

Florida apple snails (Pomacea paludosa) were exposed to three concentrations of copper (Cu), in water (8 microg/L, 16 microg/L, 24 microg/L), for one generation to examine uptake and the effects on survival, growth, and reproduction of the F(0) generation and survival, growth, and whole body Cu of the F(1) generation. During a 9-month Cu exposure, apple snails exposed to 8-16 microg/L Cu had high Cu accumulation (whole body, foot, viscera, and shell) and significantly reduced clutch production (8-16 microg/L) and egg hatching (16 microg/L). Apple snails exposed to the 24 microg/L Cu had low survival and the treatment was therefore terminated. Concentrations of minerals (Na(+), K(+), Mg(2+), Ca(2+)) in tissues were maintained regardless of Cu exposure, but the distribution of Cu in the body of snails differed, depending on exposure concentrations. Higher exposure concentrations resulted in a greater percentage of Cu accumulated in the viscera of the snail. Copper exposure to the F(0) generation did not affect the survival, growth, or whole body Cu concentrations in the F(1) generation. These finding are significant, given the importance of the Florida apple snail in the Everglades food chain. Changes in the abundance of apple snail populations, as a result of Cu exposure, could ultimately affect foraging success of predators.

This study examined the uptake and effects of copper (Cu) from flooded agricultural soils to epibenthic amphipods (Hyalella azteca) using 10-day sediment toxicity tests. Soils were collected from 10 citrus agricultural sites in South Florida. One sediment toxicity test was conducted with one flooding of the 10 soils, and based on the results of this test a second sediment toxicity test was conducted with 4 of the soils, after four 14-day flooding and four 14-day drying intervals over 4 months. Sediment toxicity tests were conducted under flow-through conditions using U.S. EPA methodology. Effects on survival, dry weight, and whole-body Cu concentrations of H. azteca were determined. Cu concentrations in overlying water and sediment of both sediment toxicity tests exceeded regulatory criteria for aquatic organisms. Although survival of H. azteca was not consistently affected from the first to the second sediment toxicity tests, dry weight was consistently reduced and related to Cu concentrations in soil, overlying water, and pore water. Furthermore, whole-body tissue Cu concentrations were significantly higher in H. azteca in all 10 soil-water treatments in the first sediment toxicity test and in all 4 soil-water treatments in the second sediment toxicity test compared to controls. Whole-body tissue concentrations and effects on dry weight were related to Cu exposures in soil, overlying water, and pore water. In these managed soil-water systems, small fish consuming H. azteca with high concentrations of Cu may be at risk.

The present study characterized copper (Cu) uptake and depuration by juvenile and adult Florida apple snails (Pomacea paludosa) from water, soil, and diet. During a 28-day uptake period, juvenile apple snails were exposed to aqueous Cu and adult apple snails were exposed to Cu-contaminated soil, water, and food. In the follow-up 14-day depuration period, both juvenile and adult apple snails were held in laboratory freshwater with background Cu concentrations<4 microg/l. For juvenile apple snails, whole body Cu concentrations increased with time and reached a plateau after 14 days. The data followed Michaelis-Menten kinetics rather than a one compartment first order kinetics model. The mean Cu bioconcentration factor (BCF) for juvenile apple snails was 1493 and the depuration half-life was 10.5-13.8 days. For adult snails, dietary uptake of Cu resulted in higher bioaccumulation factors (BAFs) compared to uptake from soil. Most of the accumulated Cu was located in soft tissue (about 60% in the viscera and 40% in the foot). The shell contained <1% of the total accumulated copper. Soft tissue is usually consumed by predators of the apple snail. Therefore, the results of the present study show that Cu transfer through the food chain to the apple snail may lead to potential risk to its predators.

A screening-level aquatic risk assessment was conducted for copper in south Florida's freshwater and saltwater environments. Risk was quantified by comparing the overlap between the probability distributions of copper exposure from surface water and sediment with the probability distributions of effects data obtained from laboratory studies. Copper concentrations in surface water and sediment in south Florida were summarized by county. For surface water, the highest concentrations of copper were found in Martin and St. Lucie counties for freshwater and saltwater, respectively. From the exposure probability distributions, the 90th centile values were estimated at 14.0 microg/L and 15.4 microg/L in freshwater and saltwater, respectively. Copper concentrations in sediment were evaluated from a probability distribution of predicted pore water concentrations. The 90th centile values of pore water concentrations from freshwater sediments ranged from 5.0 microg/L in Palm Beach County to 71.7 microg/L in Broward County. In saltwater sediments, the 90th centile values for pore water ranged from 26.1 microg/L in St. Lucie County to 27.3 microg/L in Miami-Dade County. Ecological effects data were obtained for acute and chronic copper effects in freshwater and saltwater. The 10th centile values for acute effects data were 21.2 microg/L and 9.8 microg/L for freshwater and saltwater species, respectively. For chronic effects, the 10th centile values were 3.8 microg/L and 3.9 microg/L for freshwater and saltwater species, respectively. The risk of acute copper exposure in surface water was generally low; however, the potential for ecological risk from chronic copper exposure was low to high in several counties including Lee, Martin, and St. Lucie counties. The risk of acute copper exposure in porewater from freshwater sediments also was low with the exception of St. Lucie and Broward counties. However, porewater from saltwater sediments posed a significant acute risk in Miami-Dade and St. Lucie counties. In porewater from freshwater and saltwater sediments chronic risk was high in counties with sufficient data available to calculate risk estimates.

Copper (Cu) desorption and toxicity to the Florida apple snail were investigated from soils obtained from agricultural sites acquired under the Comprehensive Everglades Restoration Plan. Copper concentrations in 11 flooded soils ranged from 5 to 234 mg/kg on day 0 and from 6.2 to 204 mg/kg on day 28 (steady-state). The steady-state Cu concentration in overlying water ranged from 9.1 to 308.2 microg/L. In a 28-d growth study, high mortality in snails occurred within 9 to 16 d in two of three soil treatments tested. Growth of apple snails over 28 d was affected by Cu in these two treatments. Tissue Cu concentrations by day 14 were 12-23-fold higher in snails exposed to the three soil treatments compared to controls. The endangered Florida snail kite and its main food source, the Florida apple snail, may be at risk from Cu exposure in these managed agricultural soil-water ecosystems.

Copper (Cu)-containing compounds have been used in Florida as fungicides, herbicides, and soil amendments, resulting in elevated Cu in the aquatic ecosystem. The Florida apple snail (Pomacea paludosa), a key species in south Florida, may be adversely affected by Cu. Water-quality parameters, such as hardness, dissolved organic carbon (DOC), pH, and alkalinity, affect metal bioavailability and toxicity in aquatic organisms; however, it is uncertain to what extent these factors affect Cu toxicity in the Florida apple snail. The research presented here characterized the acute (96-hour) toxicity of Cu in water to the Florida apple snail at various life stages and under different water-quality parameters. Cu was more toxic to juvenile than adult apple snails. There was no difference between the 96-hour LC(50) at pH 5.5 and 6.5; however, the 96-hour LC(50 )values at pH 7.5 and 8.5 were greater than at lower pHs. The decrease in Cu(2+) above pH 7, as predicted by the MINTEQ model, accounted for the pH effect. Cu toxicity decreased as DOC increased from 0.2 to 30 mg/L. Unlike other aquatic organisms, hardness had no effect on Cu toxicity to the Florida apple snail, suggesting another mechanism of toxicity. Whole-body tissue analysis indicated that the lethal body burden of 120-day-old snails exposed to Cu for 4 days was 30 mg/kg Cu dry weight. Multiple regression analysis indicated that Cu toxicity was a function of organism age, DOC, and pH.

The acute toxicity of selenium (Se) to aquatic biota has been studied extensively for decades. However, most studies have used a constant concentration aqueous exposure of Se to an invertebrate species. Since constant concentration exposure of toxicants to invertebrates is unusual in the environment, episodic exposure or pulsed exposures may represent true risk to aquatic biota more accurately. This research was designed to characterize the toxicity effects of pulsed Se exposure to Daphnia magna. Selenium exposure was varied during a 21-d chronic toxicity test to examine the effects of exposure concentration, duration, and recovery on survival, growth, and reproduction of D. magna. While D. magna did not die during exposures, latent mortality was observed. Latent mortality increased with exposure concentration and duration. Hence, standard toxicity test using continuous exposures would underestimate Se toxicity. Risk assessment method using results of continuous exposure would underestimate risk of Se to biota. For double-pulse exposures, cumulative mortality on day 21 was higher when time interval between pulses was shorter. With the same total exposure time, continuous exposure caused higher toxicity than did pulsed exposures due to recovery and tolerance development in D. magna after earlier pulses. Growth and reproduction of surviving D. magna were not affected by pulsed Se exposure due to recovery of D. magna after removal of the pulses. Based on these results, risk assessment for Se should take latent effects and the effect of recovery in to account.

Aquatic organisms living in surface waters experience fluctuating contaminant exposures that vary in concentration, duration, and frequency. This study characterized the role of pulsed concentration, pulsed duration, and the interval between pulses on the toxicity of four metals (Cu, Zn, Se, and As) to Daphnia magna. During 21-d toxicity tests, neonatal D. magna were exposed to single or double pulses. Pulsed concentrations and durations ranged from 32 to 6000 microg/L and 8 to 96 h, respectively. Intervals between two pulses ranged from 24 to 288 h. Mortality, growth, and reproduction were characterized for exposures. For single-pulse exposures of Cu and As, metal concentration had a stronger effect on survival of D. magna than did pulsed duration: pulses with 2X concentration and 1Y duration resulted in more mortality than did pulses with 1X concentration and 2Y duration. In contrast, effects of pulsed duration were stronger than metal concentration for Zn. However, the effects of duration and concentration were similar for Se. The relative effects of pulsed concentration and duration found in the present study revealed that the common method using area under the curve (AUC = concentration x duration) may not always accurately estimate environmental risk from metals (e.g., for Cu, Zn, As). In addition, the occurrence of delayed mortality in the present study revealed that using continuous exposure bioassays might underestimate metal toxicity to aquatic biota. For double-pulse exposures, the toxicity of the second pulse was influenced by the first pulse for all four metals. This influence was dependent on the pulsed concentration and duration and the interval between pulses. Further, toxicity caused by the second pulse decreased as the time between the exposures increased. For all four metals, there existed an interval great enough that the toxicity of the two pulses was independent. This would result in less toxicity for multiple exposures than continuous exposures with the same total exposure duration. The interval time at which the effects of the two pulses were independent increased with increasing concentration. Growth and cumulative reproduction of D. magna over 21 d were not significantly affected by pulsed exposures examined in the present study, indicating recovery of the organisms.

Research on the toxicity of arsenic has focused on sublethal effects that do not provide sufficient information for risk estimation. While most studies have focused on organism response to constant arsenic exposures, organisms in nature are exposed to fluctuating As concentrations. Consequently, results obtained from standardized bioassays with constant exposures may not adequately characterize risk to indigenous biota. This research was designed to characterize the response of Daphnia magna to fluctuating arsenic exposures during 21-day experiments. At concentrations > or =3000 microg/L As, 21-day pulsed exposure mortality increased as a function of exposure concentration and duration. In addition, 21-day pulsed exposure mortality increased with increasing recovery time. Pulsed As exposure did not affect the growth of D. magna over 21 days. Twenty-one day accumulative reproduction of D. magna was only affected by pulsed exposures of high As concentration and long durations.

Aquatic organisms living in surface water experience contaminant exposure at different life stages. While some investigators have examined the influence of organism age on the toxicity of pollutants, the general assumption in toxicology has been that young organisms were more sensitive than older organisms. In fact, some standardized toxicity tests call for the use of organisms less than 24 h old. This research characterized the age sensitivity of the water flea Daphnia magna to copper, zinc, selenium, and arsenic. During 21-d toxicity tests, organisms were exposed to a single 12-h pulse of either 70 microg/L Cu, 750 microg/L Zn, 1000 microg/L Se, or 5000 microg/L As at different ages ranging from 3 h to 10 d old. Mortality and reproduction were compiled over 21 d. During the juvenile stage, mortality increased and cumulative reproduction decreased with age, respectively. However, mortality decreased and cumulative reproduction increased with age when organisms became adult. Peak sensitivity occurred in 4-d-old organisms exposed to Cu and Zn, while 2- to 3-d-old organisms were most sensitive to As and Se. Growth of D. magna over 21 d was not affected by the 12-h pulse of Cu, Zn, Se, or As given at any organism age. This indicates the recovery of the organisms after exposure termination.

Some toxicology research in which toxicant exposures are continual (pulsed) rather than continuous have been reported. A number of toxicity models have been developed for pulsed and continuous exposures. Most of these models were developed based on one- or two-compartment, first-order toxicokinetics and were calibrated with organic compounds. In the present study, the relationship between mortality (after 21 d) of Daphnia magna in response to pulsed and continuous exposures to Cu, Zn, and Se was used to develop a model that integrated the effects of single and multiple pulsed metal exposures based on first-order uptake and depuration kinetics. Mortality was a function of exposure concentration, duration, and recovery time between exposures. The model was successfully validated using an independent data set. It is applicable to risk assessment and, potentially, may be incorporated with other models (e.g., the biotic ligand model) to predict the toxicity of pulsed metal exposures under a range of environmental conditions.

This research characterized the effects of water quality and organism age on the toxicity of nickel (Ni) to fathead minnows (Pimephales promelas) to facilitate the accurate development of site-specific water-quality criteria. Nickel sulfate hexahydrate (NiSO4 x 6H2O) was used as the Ni source for performing acute toxicity tests (median lethal concentration after 96-h exposure [96-h LC50]) with < 1-d-old and 28-d-old P. promelas under varying regimes of hardness, pH, alkalinity, and natural organic matter (NOM). The toxicity of Ni was inversely related to water hardness between hardness values of 20 and 150 mg/L (as CaCO3). Below 30 mg/L alkalinity, Ni toxicity was related to alkalinity. The effect of pH was confounded by hardness and the presence of NOM. In the absence of NOM, the toxicity of Ni increased as pH increased at high hardness and alkalinity. In general, 28-d-old fish were less sensitive than < 1-d-old fish to Ni. This lower sensitivity ranged from 12-fold at low hardness and alkalinity (20 and 4 mg/L, respectively) to 5-fold at high hardness and alkalinity (100 and 400 mg/L, respectively). The presence of NOM (10 mg/L as dissolved organic carbon [DOC]) reduced Ni toxicity by up to 50%, but this effect appeared to be saturated above DOC at 5 mg/L. Incubating Ni with the NOM solution from 1 to 17 days had no effect on Ni toxicity. When using multivariate analysis, the 96-h LC50 for Ni was a function of fish age, alkalinity, hardness, and NOM (96-h LC50 = -0.642 + 0.270(fish age) + 0.005(alkalinity) + 0.018(hardness) + 0.138(DOC)). When using this model, we found a strong relationship between measured and predicted 96-h LC50 values (r2 = 0.94) throughout the treatment water qualities. The biotic ligand model (BLM) did not accurately predict Ni toxicity at high or low levels of alkalinity. Results of our research suggest that the BLM could be improved by considering NiCO3 to be bioavailable.