Aquatic phosphorus behaviour within a UK Ramsar wetland: Impacts of seasonality and hydrology on algal growth and implications for management.

Fundamental to all life, phosphorus is an essential nutrient and, contrastingly, a significant threat to surface water biodiversity globally as one of the most common causes of eutrophication in surface waters worldwide. Freshwater wetland ditches affected by these conditions undergo a conversion from primarily submerged aquatic vegetation to algae or duckweed dominance, leading to anoxic conditions. However, macrophyte biomass harvesting in eutrophic water systems is a promising means of remediation and nutrient recycling. This study seasonally assesses spatial distribution and chemical fractionation of surface water phosphorus, as well as surface biomass abundance and total phosphorus content in the ditch systems at West Sedgemoor (Somerset, UK), a designated site of special scientific interest. Elevated phosphorus concentrations in the surface water were observed across the site, with all sites exceeding e hi the Common Standards Monitoring environmental quality standard value of 0.1 mg L-1 during summer and autumn Sites lacking hydrological flow connectivity with contaminated freshwater inputs, typically had lower surface water phosphorus concentrations than the rest of the moor, with localised hotspots associated with likely cattle related agricultural activity. Summer and autumn were determined as the dominant duckweed growth seasons, in which an estimated 39 kg of phosphorus could be removed via duckweed biomass harvesting, per harvest period.

New Approach Methodologies for the Endocrine Activity Toolbox: Environmental Assessment for Fish and Amphibians.

Multiple in vivo test guidelines focusing on the estrogen, androgen, thyroid, and steroidogenesis pathways have been developed and validated for mammals, amphibians, or fish. However, these tests are resource-intensive and often use a large number of laboratory animals. Developing alternatives for in vivo tests is consistent with the replacement, reduction, and refinement principles for animal welfare considerations, which are supported by increasing mandates to move toward an "animal-free" testing paradigm worldwide. New approach methodologies (NAMs) hold great promise to identify molecular, cellular, and tissue changes that can be used to predict effects reliably and more efficiently at the individual level (and potentially on populations) while reducing the number of animals used in (eco)toxicological testing for endocrine disruption. In a collaborative effort, experts from government, academia, and industry met in 2020 to discuss the current challenges of testing for endocrine activity assessment for fish and amphibians. Continuing this cross-sector initiative, our review focuses on the current state of the science regarding the use of NAMs to identify chemical-induced endocrine effects. The present study highlights the challenges of using NAMs for safety assessment and what work is needed to reduce their uncertainties and increase their acceptance in regulatory processes. We have reviewed the current NAMs available for endocrine activity assessment including in silico, in vitro, and eleutheroembryo models. New approach methodologies can be integrated as part of a weight-of-evidence approach for hazard or risk assessment using the adverse outcome pathway framework. The development and utilization of NAMs not only allows for replacement, reduction, and refinement of animal testing but can also provide robust and fit-for-purpose methods to identify chemicals acting via endocrine mechanisms. Environ Toxicol Chem 2023;42:757-777. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Towards sustainability: Threat of water quality degradation and eutrophication in Usangu agro-ecosystem Tanzania.

The agrochemicals and nutrient losses from farming areas such as paddy farming significantly dictate quality and eutrophication of the freshwater resource. However, how farming and land use pattern affect water qualities and eutrophication remain poorly understood in most African agro-ecosystems. The present study characterized how paddy farming influences water qualities and eutrophication in 10 irrigation schemes in Usangu agro-ecosystem (UA). About 42 water samples were sampled from intakes, channels, paddy fields, and drainages and analyzed for EC, Cl, P, NH4-N, NO3-N, TN, Zn, Cu, Ca, and Mg. We observed water pH ranging from 4.89 to 6.76, which was generally below the acceptable range (6.5-8.4) for irrigation water. NH4-N concentration was in a range of 10.6-70.0 mg/L, NO3-N (8.4-33.9 mg/L), and TN (19.1-21,104 mg/L). NH4-N increased along sampling transect (sampling points) from intakes (5.7-29.1 mg/L), channels (19-20 mg/L), fields (12.9-35.8 mg/L), and outflow (10.6-70.0 mg/L), the same trend were found for NO3-N and TN. The TP determined in water samples were in the range of 0.01 to 1.65 mg/L; where some sites had P > 0.1 mg/L exceeding the allowable P concentration in freshwater resource, thus indicating P enrichment and eutrophication status. The P concentration was observed to increase from intake through paddy fields to drainages, where high P was determined in drainages (0.02-1.65 mg/L) and fields (0.0-0.54 mg/L) compared to channels (0.01-0.13 mg/L) and intakes (0.01-0.04 mg/L). Furthermore, we determined appreciable amount of potentially toxic elements (PTEs) such as Cu, Pb, Cd and Cr in studied water samples. The high N, P, and PTEs in drainages indicate enrichment from agricultural fields leading to water quality degradation and contaminations (eutrophication). The study demonstrates that water quality in UA is degrading potentially due to paddy rice farming and other associated activities in the landscape. Thus, the current study recommends starting initiatives to monitor irrigation water quality in UA for better crop productivity, and improved quality of drainage re-entering downstream through the introduction of mandatory riparian buffer, revising irrigation practices, to include good agronomic practices (GAP) to ensure water quality and sustainability.

Investigating endocrine-disrupting properties of chemicals in fish and amphibians: Opportunities to apply the 3Rs.

Many regulations are beginning to explicitly require investigation of a chemical's endocrine-disrupting properties as a part of the safety assessment process for substances already on or about to be placed on the market. Different jurisdictions are applying distinct approaches. However, all share a common theme requiring testing for endocrine activity and adverse effects, typically involving in vitro and in vivo assays on selected endocrine pathways. For ecotoxicological evaluation, in vivo assays can be performed across various animal species, including mammals, amphibians, and fish. Results indicating activity (i.e., that a test substance may interact with the endocrine system) from in vivo screens usually trigger further higher-tier in vivo assays. Higher-tier assays provide data on adverse effects on relevant endpoints over more extensive parts of the organism's life cycle. Both in vivo screening and higher-tier assays are animal- and resource-intensive and can be technically challenging to conduct. Testing large numbers of chemicals will inevitably result in the use of large numbers of animals, contradicting stipulations set out within many regulatory frameworks that animal studies be conducted as a last resort. Improved strategies are urgently required. In February 2020, the UK's National Centre for the 3Rs and the Health and Environmental Sciences Institute hosted a workshop ("Investigating Endocrine Disrupting Properties in Fish and Amphibians: Opportunities to Apply the 3Rs"). Over 50 delegates attended from North America and Europe, across academia, laboratories, and consultancies, regulatory agencies, and industry. Challenges and opportunities in applying refinement and reduction approaches within the current animal test guidelines were discussed, and utilization of replacement and/or new approach methodologies, including in silico, in vitro, and embryo models, was explored. Efforts and activities needed to enable application of 3Rs approaches in practice were also identified. This article provides an overview of the workshop discussions and sets priority areas for follow-up. Integr Environ Assess Manag 2022;18:442-458. © 2021 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Soil fertility and land sustainability in Usangu Basin-Tanzania.

Soil fertility determines crop growth, productivity and consequently determines land productivity and sustainability. Continuous crop production exploits plant nutrients from soils leading to plant nutrient imbalance, thus affecting soil productivity. This study was conducted to monitor soil fertility status in soils of Usangu agro-ecosystem to establish management strategies. To assess soil fertility status in Usangu agro-ecosystem in Southern Highland Tanzania; 0-30 cm depth soil samples were taken for organic carbon, soil pH, N, P, Ca, K, Mg, S, Al, and micronutrients such as Zn, Mn, Cu, Fe, and Cr analyses by various established standard analytical methods. The results indicated most micronutrients were available in the deficient amount in many studied sites except for Fe and Mn, which were observed to be above optimum requirement. Based on critical levels established in other areas, 90 % of the soils were ranked as N, P, K, and Mg deficient. The micronutrients (Cu, Fe, and Zn) were inadequate in all soils resulting in limited crop growth and productivity. A high concentration of trace metals was detected in agricultural soils, this might affect plant nutrients availability and leading to environmental contamination affecting land productivity and sustainability. The study found that Usangu agro-ecosystem has deprived of soil fertility leading to poor crop growth and productivity. The authors recommend the addition of supplemental materials rich in plant nutrients such as inorganic fertilizer, manure, crop residues, and treated wastes to improve soil fertility for improved productivity and land sustainability.

Accumulation and bioconcentration of heavy metals in two phases from agricultural soil to plants in Usangu agroecosystem-Tanzania.

The build-up of heavy metals (HM) in agricultural soils accelerates the HM uptake by plants, which could potentially affect food quality and food safety. Here we studied the status and bioaccumulation of HM from soils to plant parts (roots, stem, and grains) in Usangu agro-ecosystem-Tanzania. In total 68 soil samples and 42 rice plant samples from six irrigation schemes were studied. The concentrations of cadmium-Cd, chromium-Cr, copper-Cu, lead-Pb, zinc-Zn, nickel-Ni, and iron-Fe were determined to estimate accumulation, distribution, bioconcentration. Total soil HM concentration in soil and plant samples was determined by acid digestion. The concentration of HM in soils samples (in mg/kg) were Cr (4.58-42.76), Co (1.486-6.12), Fe (3513.56-12593.99), Zn (7.89-29.17), Cd (0.008-0.073), Cu (0.84-9.25), Ni (0.92-7.98), and Pb (1.82-18.86). The total HM concentration in plant samples were (in mg/kg) were Cu (5.18-33.56), Zn (57.03-120.88), Fe (963.51-27918.95), Mn (613.15-2280.98), Cd (4.3-17.46), Pb (0.01-28.25), Cr (12.88-57.34) and Ni (9.65-103.33). The concentration of HM in soil and plant parts was observed to vary among locations where high concentrations of HM were detected in stems and roots compared to grains. The ratio HM in plants and soil samples (bioconcentration) was higher than one for some sites indicating higher HM uptakes by plants leading to possible health risk to soil invertebrates, animals, and humans. The bioconcentration factor varied among schemes, with the highest values at Igalako and Mahongole, which could be caused by artisanal gold mining and mining quarry existed in the area. Therefore, steps are needed to reverse the situation to balance the HM in agricultural soils and plant tissues to be within acceptable limits.

Land use patterns influence the distribution of potentially toxic elements in soils of the Usangu Basin, Tanzania.

Spatial distribution of Potentially Toxic Elements (PTEs) in agricultural soils in Usangu Basin (Mbeya Region)-Tanzania were conducted. The study included three land-use types (paddy farming, maize farming, and conserved community forest areas). About 198 soil samples were collected from November to December 2019 across contrasting land management schemes (Group I dominated by agricultural areas versus Group II dominated by residential and agricultural areas). Total (aqua regia extracts) and bioavailable (Mehlich 3 extracts) PTEs concentrations were analyzed. For Group I and II areas, total and bioavailable concentrations (mg/kg dry weight, mean values) of some PTEs were: chromium 1662 ± 5.2 µg/kg for Group I and 1307 ± 3.9 µg/kg for Group II (Total), 55.1 ± 37.1 µg/kg for Group I and 19.2 ± 21.6 µg/kg for Group II (bioavailable); and lead 5272 ± 1650 µg/kg for Group I and 6656 ± 1994 µg/kg for Group II (Total), 1870 ± 800 µg/kg for Group I and 1730 ± 530 µg/kg for Group II (bioavailable). Soil total PTEs such as cadmium and lead were generally lower in Group I areas than in Group II areas. The reverse scenario was observed for copper. Farming areas had high PTEs concentration than non-farming areas because of anthropogenic activities. Overall, soil total concentrations of Fe (99.5%), As (87%), Se (66%), and Hg (12%) were above Tanzanian Maximum Allowable Limits. This study provides essential baseline information to support environmental risk assessment of PTEs in Tanzanian agro-ecosystem.

Toxic metals in East African agro-ecosystems: Key risks for sustainable food production.

The dramatic increase in world population underpins current escalating food demand, which requires increased productivity in the available arable land through agricultural intensification. Agricultural intensification involves increased agrochemicals use to increase land productivity. Increased uses of agrochemicals pose environmental and ecological risks such as contamination and water eutrophication. Consequently, toxic metals accumulate in plant products, thus entering the food chain leading to health concerns. To achieve this study, secondary data from peer-reviewed papers, universities, and government authorities were collected from a public database using Tanzania as a case study. Data from Science Direct, Web of Science, and other internet sources were gathered using specific keywords such as nutrient saturation and losses, water eutrophication, potentially toxic metal (PTEs), and impact of toxic metals on soils, water, and food safety. The reported toxic metal concentrations in agro-ecosystem worldwide are linked to agricultural intensification, mining, and urbanization. Statistical analysis of secondary data collected from East African agro-ecosystem had wide range of toxic metals concentration such as; mercury (0.001-11.0 mg Hg/kg), copper (0.14-312 mg Cu/kg), cadmium (0.02-13.8 mg Cd/kg), zinc (0.27-19.30 mg Zn/kg), lead (0.75-51.7 mg Pb/kg) and chromium (19.14-34.9 mg Cr/kg). In some cases, metal concentrations were above the FAO/WHO maximum permissible limits for soil health. To achieve high agricultural productivity and environmental safety, key research-informed policy needs are proposed: (i) development of regulatory guidelines for agrochemicals uses, (ii) establishment of agro-environmental quality indicators for soils and water assessment to monitor agro-ecosystem quality changes, and (iii) adoption of best farming practices such as split fertilization, cover cropping, reduced tillage, drip irrigation to ensure crop productivity and agro-ecosystem sustainability. Therefore, robust and representative evaluation of current soil contamination status, sources, and processes leading to pollution are paramount. To achieve safe and sustainable food production, management of potential toxic metal in agro-ecosystems is vital.

Assessment of arsenic status and distribution in Usangu agro-ecosystem-Tanzania.

This study was conducted to assess arsenic (As) status and distribution in Usangu agroecosystem-Tanzania, including three land use. About 198 soil samples were collected in ten irrigation schemes in three land uses. Total and bioavailable As were determined by acid digestion (Aqua regia (AQ)) and Mehlich 3 method (M3) to estimate status, distribution and bioavailability. Arsenic concentration were variable among land use and irrigation schemes where total arsenic ranged 567.74-2909.84 µg/kg and bioavailable As ranged 26.17-712.37 µg/kg. About 12-16% of total arsenic were available for plant uptake. Approximately 86.53% of studied agricultural soils had total As concentration above Tanzania maximum allowable limit. Bioavailable As were lower compared to total As and were within the acceptable threshold. Total arsenic concentration were variable among schemes and higher values were observed in schemes which are highly intensified and mechanized. Thus, this study provides essential site specific preliminary baseline information for As status and distribution in agricultural soils to initiate monitoring and management strategies for increased land productivity and environmental safety.

Characterization of soil phosphate status, sorption and saturation in paddy wetlands in usangu basin-Tanzania.

Phosphorus (P) is a vital plant macronutrient required for plant growth which usually available in limited amount. P availability for plant uptake in highly weathered soil is controlled by soil erosion and high fixation. The availability of P applied from fertilizers depend on the soil pH, soil sorption capacity (PSC) and P saturation status (PSD), which determines P storage, losses, fixation, and additional P to be added with minimal loss to the environment. PSC and PSD are agro-environmental indicators used to estimate P availability and P loss to the environment. However, PSC and PSD of agricultural soils had been never studied in Tanzanian soils. This study was conducted to assess and estimate P availability, PSC and PSD and the risks of P losses in tropical soils from Usangu basin popular for paddy farming. In total, 198 soil samples from 10 paddy irrigation schemes were collected (November-December 2019) and analyzed for inherent P (PM3), metal oxides of Aluminium (Al M3), iron (Fe M3), and calcium (Ca M3) as main PSC and PSD determinant. The determined concentrations were in range of; P M3 014.9-974.69 mg/kg, Al M3 234.56-3789.36 mg/kg, Fe M3 456.78-2980.23 mg/kg, and Ca M3 234.67-973.34 mg/kg. Estimated PSCM3 ranged 5.62-34.85 mmol/kg with a mean value of 14.14 mmol/kg corresponding to high status, ensuring high P holding capacity for plant uptake. However, some soils had very low PSCM3 creating a risk of P loss to environment. Among soils, the estimated PSD M3 ranged from 0.01 to 17.57% and was below (<24%), indicating low P loss risks to surface and groundwater, however, some soils were observed to have PSDM3 above 15% which correspond to a critical degree of phosphate saturation of 25% in a watershed using oxalate extraction method. Therefore some sites were associated with high P loss to the environment, immediate and precautionary actions for sustainable P management to increase productivity, environmental safety and sustainability are needed to be in place.

Spatial distribution of sediment phosphorus in a Ramsar wetland.

Eutrophication is a significant threat to surface water biodiversity worldwide, with excessive phosphorus concentrations being among the most common causes. Wetland ditches under these conditions shift from primarily submerged aquatic vegetation to algae or duckweed dominance, leading to excessive shading and anoxic conditions. Phosphorus, from both point (e.g. wastewater treatment works) and diffuse (largely agricultural runoff) sources, is currently the central reason for failure in the majority of surface water bodies in England to meet required water quality guidelines. This study assesses phosphorus storage in the ditch systems at West Sedgemoor, a designated site of special scientific interest. Elevated phosphorus concentrations in sediment was observed across the Moor up to 4220 mg Kg-1, almost 10 times that which may be expected from background levels. The highest concentrations were generally observed at the more intensively farmed sites in the north of the moor, near key inlets and the outlet. Based upon their chemical and physical properties, clear distinction was observed between sites outside and within the Royal Society of the Protection of Birds nature reserve, using principal component analysis.

Environmental risks to freshwater organisms from the mycotoxins deoxynivalenol and zearalenone using Species Sensitivity Distributions.

In this study, laboratory experiments have addressed the acute toxicity of two common mycotoxins, deoxynivalenol (DON) and zearalenone (ZON), in a range of freshwater organisms (including rotifers Brachionus calyciflorus, insects Chironomus riparius (larvae), crustaceans Daphnia pulex and Thamnocephalus platyurus, cnidarians Hydra vulgaris, molluscs Lymnaea stagnalis (embryos) and Protozoa Tetrahymena thermophila). Acute EC50 values highlight crustaceans as the most sensitive organisms to DON, with T. platyurus having a 24 h EC50 of 0.14 and D. magna having a 48 h EC50 of 0.13 mg DON/L. During exposures to ZON, H. vulgaris and L. stagnalis embryos showed the highest sensitivity; mortality EC50 values were 1.1 (96 h) and 0.42 mg ZON/L (7 d), respectively. Combining these novel invertebrate toxicity results, along with recent published data for freshwater plant and fish toxicity for analysis of Species Sensitivity Distributions, provides freshwater HC5 values of 5.2 µg DON/L and 43 µg ZON/L, respectively. Using highest reported environmental concentrations and following REACH guidelines, risk ratios calculated here show the risk of ZON to freshwater organisms is low. In contrast, DON may periodically because for concern in streams subject to high agricultural run-off, likely during certain times of year where cereal crops are susceptible to higher fungal infections rates and may pose increased risks due to climate change.

Physico-chemical factors controlling the speciation of phosphorus in English and Welsh rivers.

Phosphorus is a finite resource essential for global food production. However, excessive loss to river systems from diffuse sources (typically agricultural) and point sources (e.g. waste water treatment works and industrial effluent) can lead to negative environmental impacts, including changes to diatom and invertebrate community structure. Current environmental quality standards for phosphorus in the UK have been based on reactive phosphorus, which is poorly defined and comprises an unknown proportion of soluble reactive phosphorus and chemically extractable particulate phosphorus. This research assesses the influencing factors that may control soluble reactive phosphorus concentrations in rivers, including dissolved iron, as well as partitioning processes associated with the presence of total suspended solids, and questions the reliability of the assumptions used when setting environmental quality standards. The extensive phosphorus speciation monitoring carried out across a wide geographic area of England and Wales shows that not all phosphorus as measured by the molybdenum blue method is either soluble or necessarily bioavailable, particularly at concentrations in the range in which the Environmental Quality Standard for 'Good' status (typically less than 100 µg P L-1) has been set. Phosphorus speciation can change due to physico-chemical processes which vary spatially and/or temporally, including precipitation with iron and partitioning with suspended solids.

Key Opportunities to Replace, Reduce, and Refine Regulatory Fish Acute Toxicity Tests.

Fish acute toxicity tests are conducted as part of regulatory hazard identification and risk-assessment packages for industrial chemicals and plant protection products. The aim of these tests is to determine the concentration which would be lethal to 50% of the animals treated. These tests are therefore associated with suffering in the test animals, and Organisation for Economic Co-operation and Development test guideline 203 (fish, acute toxicity) studies are the most widely conducted regulatory vertebrate ecotoxicology tests for prospective chemical safety assessment. There is great scope to apply the 3Rs principles-the reduction, refinement, and replacement of animals-in this area of testing. An expert ecotoxicology working group, led by the UK National Centre for the Replacement, Refinement and Reduction of Animals in Research, including members from government, academia, and industry, reviewed global fish acute test data requirements for the major chemical sectors. The present study highlights ongoing initiatives and provides an overview of the key challenges and opportunities associated with replacing, reducing, and/or refining fish acute toxicity studies-without compromising environmental protection. Environ Toxicol Chem 2020;39:2076-2089. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Expression dynamics of genes in the hypothalamic-pituitary-thyroid (HPT) cascade and their responses to 3,3',5-triiodo-l-thyronine (T3) highlights potential vulnerability to thyroid-disrupting chemicals in zebrafish (Danio rerio) embryo-larvae.

Some chemicals in the environment disrupt thyroid hormone (TH) systems leading to alterations in organism development, but their effect mechanisms are poorly understood. In fish, this has been limited by a lack of fundamental knowledge on thyroid gene ontogeny and tissue expression in early life stages. Here we established detailed expression profiles for a suite of genes in the hypothalamic-pituitary-thyroid (HPT) axis of zebrafish (Danio rerio) between 24-120 h post fertilisation (hpf) and quantified their responses following exposure to 3,3',5-triiodo-L-thyronine (T3) using whole mount in situ hybridisation (WISH) and qRT-PCR (using whole-body extracts). All of the selected genes in the HPT axis demonstrated dynamic transcript expression profiles across the developmental stages examined. The expression of thyroid receptor alpha (thraa) was observed in the brain, gastrointestinal tract, craniofacial tissues and pectoral fins, while thyroid receptor beta (thrb) expression occurred in the brain, otic vesicles, liver and lower jaw. The TH deiodinases (dio1, dio2 and dio3b) were expressed in the liver, pronephric ducts and brain and the patterns differed depending on life stage. Both dio1 and dio2 were also expressed in the intestinal bulb (96-120 hpf), and dio2 expression occurred also in the pituitary (48-120 hpf). Exposure of zebrafish embryo-larvae to T3 (30 and 100 µg L-1) for periods of 48, 96 or 120 hpf resulted in the up-regulation of thraa, thrb, dio3b, thyroid follicle synthesis proteins (pax8) and corticotropin-releasing hormone (crhb) and down-regulation of dio1, dio2, glucuronidation enzymes (ugt1ab) and thyroid stimulating hormone (tshb) (assessed via qRT-PCR) and responses differed across life stage and tissues. T3 induced thraa expression in the pineal gland, pectoral fins, brain, somites, gastrointestinal tract, craniofacial tissues, liver and pronephric ducts. T3 enhanced thrb expression in the brain, jaw cartilage and intestine, while thrb expression was suppressed in the liver. T3 exposure suppressed the transcript levels of dio1 and dio2 in the liver, brain, gastrointestinal tract and craniofacial tissues, while dio2 signalling was also suppressed in the pituitary gland. Dio3b expression was induced by T3 exposure in the jaw cartilage, pectoral fins and brain. The involvement of THs in the development of numerous body tissues and the responsiveness of these tissues to T3 in zebrafish highlights their potential vulnerability to exposure to environmental thyroid-disrupting chemicals.

Consequences of surface coatings and soil ageing on the toxicity of cadmium telluride quantum dots to the earthworm Eisenia fetida.

The bioaccumulation potential and toxic effects of engineered nanomaterials (ENMs) to earthworms are poorly understood. Two studies were conducted following OECD TG 222 on Eisenia fetida to assess the effects of CdTe QDs with different coatings and soil ageing respectively. Earthworms were exposed to carboxylate (COOH), ammonium (NH4+), or polyethylene glycol (PEG) coated CdTe QDs, or a micron scale (bulk) CdTe material, at nominal concentrations of 50, 500 and 2000 mg CdTe QD kg-1 dry weight (dw) for 28 days in Lufa 2.2 soil. In the fresh soil study, earthworms accumulated similar amounts of Cd and Te in the CdTe-bulk exposures, while the accumulation of Cd was higher than Te during the exposures to CdTe QDs. However, neither the total Cd, nor Te concentrations in the earthworms, were easily explained by the extractable metal fractions in the soil or particle dissolution. There were no effects on survival, but some retardation of growth was observed at the higher doses. Inhibition of Na+/K+-ATPase activity with disturbances to tissue electrolytes, as well as tissue Cu and Mn were observed, but without depletion of total glutathione in the fresh soil experiment. Additionally, juvenile production was the most sensitive endpoint, with estimated nominal EC50 of values >2000, 108, 65, 96 mg CdTe kg-1 for bulk, PEG-, COOH- and NH4+-coated CdTe QDs, respectively. In the aged soil study, the accumulation of Cd and Te was higher than in the fresh soil study in all CdTe QD exposures. Survival of the adult worms was reduced in the top CdTe-COOH and -NH4+ QD exposures by 55 ±â€¯5 and 60 ±â€¯25%, respectively; and with decreases in growth. The nominal EC50 values for juvenile production in the aged soil were 165, 88, 78 and 63 mg CdTe kg-1 for bulk, PEG-, COOH- and NH4+-coated CdTe QDs, respectively. In conclusion, exposure to nanoscale CdTe QDs, regardless of coating, caused more severe toxic effects that the CdTe bulk material and the toxicity increased after soil ageing. There were some coating-mediated effects, likely due to differences in the metal content and behaviour of the materials.

Interpretation of sexual secondary characteristics (SSCs) in regulatory testing for endocrine activity in fish.

Secondary sexual characteristics (SSCs) are important features that have evolved in many fish species because of inter-individual competition for mates. SSCs are crucial not only for sexual selection, but also for other components of the reproductive process and parental care. Externally, they are especially clear in males (for instance, tubercles, fatpad, anal finnage, colouration) but are also externally present in the females (for instance, ovipositor). These characters are under hormonal control and as such there has been much interest in incorporating them as measures in fish test methods to assess the potential endocrine activity of chemicals. Here we describe the external SSCs in typical laboratory test species for endocrine testing - fathead minnow (Pimephales promelas), Japanese medaka (Oryzias latipes), zebrafish (Danio rerio) and the three-spined stickleback (Gasterosteus aculeatus L.). We also provide some examples and discuss the utility of SSC responses to the endocrine activity of chemicals in the field and the laboratory. This paper is not aimed to provide a comprehensive review of SSCs in fish but presents a view on the assessment of SSCs in regulatory testing. Due to the current regulatory importance of establishing an endocrine mode-of-action for chemicals, we also consider other, non-endocrine factors that may lead to SSC responses in fish. We conclude with recommendations for how the assessment of SSCs in fish could be usefully incorporated into the endocrine hazard and risk assessment of chemicals.

Molecular mechanisms and tissue targets of brominated flame retardants, BDE-47 and TBBPA, in embryo-larval life stages of zebrafish (Danio rerio).

Brominated flame retardants are known to disrupt thyroid hormone (TH) homeostasis in several vertebrate species, but the molecular mechanisms underlying this process and their effects on TH-sensitive tissues during the stages of early development are not well characterised. In this study, we exposed zebrafish (Danio rerio) embryo-larvae to 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) and tetrabromobisphenol A (TBBPA) via the water for 96 h from fertilisation and assessed for lethality, effects on development and on the expression of a suite of genes in the hypothalamic-pituitary-thyroid (HPT) axis via both real time quantitative PCR (qRT-PCR) on whole body extracts and whole mount in situ hybridisation (WISH) to identify tissue targets. The 96-h lethal median concentration (96h-LC50) for TBBPA was 0.9 µM and mortality was preceded by retardation of development (smaller animals) and morphological deformities including, oedemas in the pericardial region and tail, small heads, swollen yolk sac extension. Exposure to BDE-47 did not affect zebrafish embryo-larvae survival at any of the concentrations tested (1-100 µM) but caused yolk sac and craniofacial deformities, a curved spine and shorter tail at the highest exposure concentration. TBBPA exposure resulted in higher levels of mRNAs for genes encoding deiodinases (dio1), transport proteins (ttr), the thyroid follicle synthesis protein paired box 8 (pax8) and glucuronidation enzymes (ugt1ab) and lower levels of dio3b mRNAs in whole body extracts, with responses varying with developmental stage. BDE-47 exposure resulted in higher levels of thrb, dio1, dio2, pax8 and ugt1ab mRNAs and lower levels of ttr mRNAs in whole body extracts. TBBPA and BDE-47 therefore appear to disrupt the TH system at multiple levels, increasing TH conjugation and clearance, disrupting thyroid follicle development and altering TH transport. Compensatory responses in TH production/ metabolism by deiodinases were also evident. WISH analyses further revealed that both TBBPA and BDE-47 caused tissue-specific changes in thyroid receptor and deiodinase enzyme expression, with the brain, liver, pronephric ducts and craniofacial tissues appearing particularly responsive to altered TH signalling. Given the important role of TRs in mediating the actions of THs during key developmental processes and deiodinases in the control of peripheral TH levels, these transcriptional alterations may have implications for TH sensitive target genes involved in brain and skeletal development. These findings further highlight the potential vulnerability of the thyroid system to disruption by BFRs during early developmental windows.

Impacts of the mycotoxin zearalenone on growth and photosynthetic responses in laboratory populations of freshwater macrophytes (Lemna minor) and microalgae (Pseudokirchneriella subcapitata).

Mycotoxins are an important class of chemicals of emerging concern, recently detected in aquatic environments, potentially reflecting the influence of fungicide resistance and climatic factors on fungal diseases in agricultural crops. Zearalenone (ZON) is a mycotoxin formed by Fusarium spp. and is known for its biological activity in animal tissues; both in vitro and in vivo. ZON has been reported in US and Polish surface waters at 0.7 - 96 ng/L, with agricultural run-off and wastewater treatment plants being the likely sources of mycotoxins. As some mycotoxins can induce phytotoxicity, laboratory studies were conducted to evaluate the toxicity of ZON (as measured concentrations) to freshwater algae (Pseudokirchneriella subcapitata) and macrophytes (Lemna minor) following OECD test guidelines 201 and 221, respectively. Zinc sulphate was used as a positive control. In the OECD 201 algal static study (72 h at 24 ±â€¯1 °C), exposure to ZON gave average specific growth rate (cell density) EC50 and yield (cell density) EC50 values of > 3.1 and 0.92 (0.74 - 1.8) mg/L, respectively. ZON was less toxic in the OECD 221 static study and after 7 d at 24 ±â€¯1 °C. L. minor growth was significantly reduced based on frond number and frond area at 11.4 mg ZON/L, showing a higher tolerance than reported for other mycotoxins with Lemna spp. Chlorophyll fluorescence parameters were used as biomarkers of impacts on photosystem II efficiency, with no effect seen in algae but, with responses being observed in L. minor between 5.2 - 14.4 mg ZON/L. ZON toxicity seen here is not of immediate concern in context with environmental levels, but this study highlights that other freshwater organisms including algae are more sensitive to mycotoxins than Lemna sp., the only current source of toxicity data for freshwater plants.