Applicability of Chronic Multiple Linear Regression Models for Predicting Zinc Toxicity in Australian and New Zealand Freshwaters.

Bioavailability models, for example, multiple linear regressions (MLRs) of water quality parameters, are increasingly being used to develop bioavailability-based water quality criteria for metals. However, models developed for the Northern Hemisphere cannot be adopted for Australia and New Zealand without first validating them against local species and local water chemistry characteristics. We investigated the applicability of zinc chronic bioavailability models to predict toxicity in a range of uncontaminated natural waters in Australia and New Zealand. Water chemistry data were compiled to guide a selection of waters with different zinc toxicity-modifying factors. Predicted toxicities using several bioavailability models were compared with observed chronic toxicities for the green alga Raphidocelis subcapitata and the native cladocerans Ceriodaphnia cf. dubia and Daphnia thomsoni. The most sensitive species to zinc in five New Zealand freshwaters was R. subcapitata (72-h growth rate), with toxicity ameliorated by high dissolved organic carbon (DOC) or low pH, and hardness having a minimal influence. Zinc toxicity to D. thomsoni (reproduction) was ameliorated by both high DOC and hardness in these same waters. No single trophic level-specific effect concentration, 10% (EC10) MLR was the best predictor of chronic toxicity to the cladocerans, and MLRs based on EC10 values both over- and under-predicted zinc toxicity. The EC50 MLRs better predicted toxicities to both the Australian and New Zealand cladocerans to within a factor of 2 of the observed toxicities in most waters. These findings suggest that existing MLRs may be useful for normalizing local ecotoxicity data to derive water quality criteria for Australia and New Zealand. The final choice of models will depend on their predictive ability, level of protection, and ease of use. Environ Toxicol Chem 2023;42:2614-2629. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Investigating the fate of copper in a laboratory-based toxicity test with embryos of Mytilus galloprovincialis: Copper mass balance of a closed bioassay.

The production of accurate and reliable data on metal toxicity during ecotoxicological bioassays is important for credible environmental risk assessments and management in aquatic environments. Actual measurements and reporting of contaminant concentrations in bioassays are, however, often disregarded; and potential contaminant loss attributable to adsorption processes (e.g., wall adsorption) in bioassays is widely omitted, which can have detrimental effects on calculated metal toxicity thresholds. In the present we assessed copper (Cu) mass balance during a standard 48-h bioassay test with blue mussel (Mytilus galloprovincialis) embryos to evaluate effects on calculated toxicity endpoints. We demonstrated that measured Cu concentrations at the test conclusion need to be used to quantify the risk of Cu toxicity because nominal Cu and initial Cu concentrations underestimate overall Cu toxicity by up to 1.5-fold, owing to Cu loss in solution attributable to adsorption and bioaccumulation processes. For the first time we provide evidence that extracellular adsorption to the biological surface of the embryos is the most important sink for total dissolved Cu in a bioassay. We also established that adsorbed extracellular Cu accumulation reduces Cu toxicity to embryos, potentially by inhibiting Cu from entering the cell of the mussel embryo. Environmental factors (e.g., salinity and dissolved organic carbon) did not influence the partitioning of Cu within the laboratory-based bioassay. The present results 1) demonstrate the importance of differentiating extra- and intracellular Cu pools to improve our understanding of Cu toxicity and associated processes, 2) reveal the potential for bias with respect to calculated Cu toxicity thresholds when results are based on nominal and initial Cu concentrations, and 3) point out the need to follow current guidelines for the testing of chemicals to standardize toxicity tests and data reporting. Environ Toxicol Chem 2019;38:561-574. © 2019 SETAC.

Copper toxicity to blue mussel embryos (Mytilus galloprovincialis): The effect of natural dissolved organic matter on copper toxicity in estuarine waters.

Copper (Cu) is a naturally occurring micronutrient of eco-toxicological concern in aquatic ecosystems. Current knowledge of Cu-speciation and bioavailability in natural saline environments is insufficient to adequately inform environmental protection policy for estuarine systems. We assessed the combined effect of two of the main drivers of metal bioavailability, salinity and natural dissolved organic carbon (DOC), on Cu-speciation and associated Cu-toxicity to blue mussel (Mytilus galloprovincialis) embryos in a standard 48-h bioassay. We placed special emphasis on measurement of Cu-speciation rather than modelling. Cu-toxicity was found to be a function of DOC and salinity. The varying protective effect of different DOC-types suggests that estuarine DOC is more protective against Cu-toxicity than oceanic DOC. Salinity was negatively correlated with [Cu48-h-EC50], indicating a salinity-induced alteration in the physiology of the exposed mussel embryos and/or Cu-DOC-reactivity. These two assumptions were supported by (1) the relative uniformity of bioavailable copper ([Cu']) across similar salinity treatments despite considerable variation in [Cu48-h-EC50] and DOC-concentrations, and (2) the fact that Cu-toxicity and [Cu'] were slightly higher in the 35 salinity treatment compared to the 25 salinity treatment. Stripping voltammetry studies determined the presence of only one strong Cu-binding ligand class (i.e., L1), either actively or passively released by the exposed embryos. [L1] was found to be proportional to the total dissolved Cu-concentration ([CuT]), suggesting a protective effect of Cu-binding-ligands, in addition to the protective effect of DOC. There was also a strong positive correlation between [L1] and [Cu48-h-EC50], implying that electrochemically defined ligand concentrations along with measurements of [Cu'], DOC-quality, and salinity can be used as proxies for 48-h-EC50 Cu-values in estuarine waters, which may result in a significant improvement to risk assessments of Cu in estuarine systems.

A comprehensive toolkit of plant cell wall glycan-directed monoclonal antibodies.

A collection of 130 new plant cell wall glycan-directed monoclonal antibodies (mAbs) was generated with the aim of facilitating in-depth analysis of cell wall glycans. An enzyme-linked immunosorbent assay-based screen against a diverse panel of 54 plant polysaccharides was used to characterize the binding patterns of these new mAbs, together with 50 other previously generated mAbs, against plant cell wall glycans. Hierarchical clustering analysis was used to group these mAbs based on the polysaccharide recognition patterns observed. The mAb groupings in the resulting cladogram were further verified by immunolocalization studies in Arabidopsis (Arabidopsis thaliana) stems. The mAbs could be resolved into 19 clades of antibodies that recognize distinct epitopes present on all major classes of plant cell wall glycans, including arabinogalactans (both protein- and polysaccharide-linked), pectins (homogalacturonan, rhamnogalacturonan I), xyloglucans, xylans, mannans, and glucans. In most cases, multiple subclades of antibodies were observed to bind to each glycan class, suggesting that the mAbs in these subgroups recognize distinct epitopes present on the cell wall glycans. The epitopes recognized by many of the mAbs in the toolkit, particularly those recognizing arabinose- and/or galactose-containing structures, are present on more than one glycan class, consistent with the known structural diversity and complexity of plant cell wall glycans. Thus, these cell wall glycan-directed mAbs should be viewed and utilized as epitope-specific, rather than polymer-specific, probes. The current world-wide toolkit of approximately 180 glycan-directed antibodies from various laboratories provides a large and diverse set of probes for studies of plant cell wall structure, function, dynamics, and biosynthesis.

Synthesis and immunological properties of a tetrasaccharide portion of the B side chain of rhamnogalacturonan II (RG-II).

A highly convergent strategy was used for the synthesis of a tetrasaccharide [3-aminopropyl beta-L-arabinofuranosyl-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-[alpha-L-rhamnopyranosyl-(1-->3)]-alpha-L-arabinopyranoside] portion of the B side chain of the plant cell-wall pectic polysaccharide rhamnogalacturonan II (RG-II). The terminal nonreducing beta-L-arabinofuranosyl residue of the target compound was installed by using an arabinofuranosyl donor that was protected with a 3,5-O-(di-tert-butylsilane) group to facilitate nucleophilic attack from the beta-face. The synthetic strategy also employed a chemoselective glycosylation of a trichloroacetimidate donor with a thioglycosyl acceptor; this gave a product that could be used immediately in a subsequent glycosylation. The reducing end of the tetrasaccharide contained an aminopropyl group to facilitate conjugation to keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA). Mice that were immunized with a KLH-tetrasaccharide conjugate produced antibodies that recognized RG-II isolated from Arabidopsis thaliana cell walls, but did not recognize RG-II obtained from red wine. Our data suggest that the arabinopyranosyl residue exists in the (4)C(1) conformation in the tetrasaccharide and in A. thaliana RG-II, whereas it has the (1)C(4) conformation in wine RG-II. It is proposed that differences in the conformation of side chain B might account for the ability of antibodies to discriminate between RG-II that was isolated from Arabidopsis and wine.

Dietary exposure to low pesticide doses causes long-term immunosuppression in the leopard frog (Rana pipiens).

This study examines the relationship between dietary exposure of pesticides, DDT, and dieldrin and immunosuppression in the northern leopard frog (Rana pipiens). Immune function was measured before, during, and after a 10-week exposure period with the use of both adaptive and innate immunity responses. Exposure to low doses (75 ng/g body wt DDT or 2.1 ng/g dieldrin total dose over the 10 weeks) resulted in significant suppressive effects on antibody production and secondary delayed-type hypersensitivity (DTH). The high doses (750 ng/g DDT and 21 ng/g dieldrin), however, did not affect antibody production, DTH, or oxidative burst in a predictable dose-response manner. The differences in magnitude and direction of the effects of the two dosing regimes were likely due to differences in chemical exposure on the basis of feeding and effectiveness of chemical uptake. The low dose results demonstrated that moderate concentrations of pesticides, frequently observed in the environment, are able to weaken the immune response of R. pipiens.

Immunosuppression in the northern leopard frog (Rana pipiens) induced by pesticide exposure.

An injection study and a field study were used to investigate the hypothesis that environmental xenobiotics have the potential to alter the immune function of northern leopard frogs (Rana pipiens). Three assays, IgM-specific antibody response to keyhole limpet hemocyanin linked to dinitrophenyl (KLH-DNP), zymozan induced chemiluminescence (CL) of whole blood and the delayed-type hypersensitivity (DTH), were used to assay humoral, innate and cell-mediated immune endpoints. Sublethal doses of DDT (923 ng/g wet wt), malathion (990 ng/g wet wt), and dieldrin (50 ng/g wet wt) were used in the injection study. In all pesticide-injected groups, antibody response was dramatically suppressed, DTH reactions were enhanced, and respiratory burst was lower. When the order of administration of pesticides and antigens was reversed, no differences in immune function between the control and dosed groups were apparent, indicating that frogs exposed to pathogens prior to pesticide exposure can still respond. A field study found significant differences in immune function between frog populations in pesticide-exposed and pesticide-free locations. The antibody response and CL were suppressed and the DTH enhanced in frogs from Essex County (ON, Canada). Overall, the results suggest that exposure to these pesticides can cause both stimulatory and suppressive immune changes in adult frogs and is doing so in wild populations.