10-Day survival of Hyalella azteca as a function of water quality parameters.

Estuarine systems are among the most impacted ecosystems due to anthropogenic contaminants; however, they present unique challenges to toxicity testing with regard to varying water quality parameters. The euryhaline amphipod species, Hyalella azteca, is widely used in toxicity testing and well suited for testing estuarine water samples. Nevertheless, the influence of relevant water quality parameters on test endpoints must be quantified in order to efficiently use this species for routine monitoring. Here, we studied the influence of five water quality parameters: electrical conductivity, pH, un-ionized ammonia, dissolved oxygen and temperature, on H. azteca survival in a water column toxicity test. A model was developed to quantify and predict the independent and interacting effects of water quality variables on 10-day survival. The model allows simultaneous assessment of multiple potential predictors recorded during the tests. Data used for modeling came from 1089 tests performed on ambient water samples over a period of three years (2006-2008). The final model reflects significant effects of predictors and their two-way interactions. The effect of each level of all predictors on survival probability of H. azteca was examined by comparing levels of each predictor at a time, while holding all others at their lowest (reference) level. This study showed that predictors of survival in water column tests should not be evaluated in isolation in the interpretation of H. azteca water column tests. Our model provides a useful tool to predict expected control survival based on relevant water quality parameters, and thus enables the use of H. azteca tests for toxicity monitoring in estuaries with a wide range of water quality conditions.

Fish evacuation and emergency sheltering during wildfire disasters.

Wildfires are a serious and expanding threat in western North America, and wildfire encroachment on human populations leads to widespread evacuation and emergency housing operations for residents and their companion animals and livestock. Veterinarians are frequently part of wildfire response efforts and are called upon to assist in rescue, evacuation, and emergency housing operations as well as to provide medical care for evacuated animals. Although veterinarians are likely familiar with the principles of transporting and housing terrestrial animals, emergency response for aquatic companion animals presents unique logistic challenges. Veterinarians familiar with aquatic animal evacuation, housing, and care prior to a wildfire response can extend the scope of disaster recovery. This report offers general guidance for rescuing, evacuating, housing, and caring for aquatic animals in the wake of a wildfire.

Toxicity of a dissolved pyrethroid mixture to Hyalella azteca at environmentally relevant concentrations.

Use of pyrethroid pesticides, which are highly toxic to aquatic organisms, has increased substantially over the past decade. In 2006, the pyrethroid pesticides cyfluthrin and permethrin were measured in Sacramento-San Joaquin (SSJ) Delta (CA, USA) water at 5 and 24 ng/L (pptr), respectively. To elucidate any interactions between the two pyrethroids, a 10-d laboratory exposure was performed with 7- to 14-d-old amphipods (Hyalella azteca). Cyfluthrin and permethrin were tested singly and in combination at detected levels and also at half and twice the detected levels, both with and without the addition of 25 ppb of piperonyl butoxide (PBO). Mortality in all treatments was significantly higher than in controls, with the median lethal concentration (LC50) for permethrin with PBO (13.9 ng/L) and the LC50s with and without PBO for cyfluthrin (5.7 and 2.9 ng/L, respectively) at or below levels measured in SSJ Delta water samples. The LC50 for permethrin alone was estimated to be 48.9 ng/L. To evaluate combined toxicity, logistic regression models containing terms for concentrations of cyfluthrin, permethrin, and PBO, as well as models containing all possible combinations of these terms and interactions, were run and compared using Akaike's information criterion. The most parsimonious set of models indicated slight antagonism between cyfluthrin and permethrin. Results indicate that a dissolved mixture of cyfluthrin and permethrin is toxic at environmentally relevant concentrations in the water column.

Tracking pyrethroid toxicity in surface water samples: Exposure dynamics and toxicity identification tools for laboratory tests with Hyalella azteca (Amphipoda).

Pyrethroid insecticides are commonly used in pest control and are present at toxic concentrations in surface waters of agricultural and urban areas worldwide. Monitoring is challenging as a result of their high hydrophobicity and low toxicity thresholds, which often fall below the analytical methods detection limits (MDLs). Standard daphnid bioassays used in surface water monitoring are not sensitive enough to protect more susceptible invertebrate species such as the amphipod Hyalella azteca and chemical loss during toxicity testing is of concern. In the present study, we quantified toxicity loss during storage and testing, using both natural and synthetic water, and presented a tool to enhance toxic signal strength for improved sensitivity of H. azteca toxicity tests. The average half-life during storage in low-density polyethylene (LDPE) cubitainers (Fisher Scientific) at 4 °C of 5 pyrethroids (permethrin, bifenthrin, lambda-cyhalothrin, cyfluthrin, and esfenvalerate) and one organophosphate (chlorpyrifos; used as reference) was 1.4 d, and piperonyl butoxide (PBO) proved an effective tool to potentiate toxicity. We conclude that toxicity tests on ambient water samples containing these hydrophobic insecticides are likely to underestimate toxicity present in the field, and mimic short pulse rather than continuous exposures. Where these chemicals are of concern, the addition of PBO during testing can yield valuable information on their presence or absence. Environ Toxicol Chem 2018;37:462-472. © 2017 SETAC.

Toxicity of storm-water runoff after dormant spray application in a french prune orchard, Glenn County, California, USA: temporal patterns and the effect of ground covers.

Organophosphorous (OP) insecticides, especially diazinon, have been detected routinely in surface waters of the Sacramento and San Joaquin River watersheds, coincident with rainfall events following their application to dormant orchards during the winter months. Preventive best management practices (BMP) aim at reducing off-site movement of pesticides into surface waters. Two proposed BMPs are: The use of more hydrophobic pyrethroid insecticides believed to adsorb strongly to organic matter and soil and the use of various types of ground cover vegetation to increase the soil's capacity for water infiltration. To measure the effectiveness of these BMPs, storm water runoff was collected in a California prune orchard (Glenn County, CA, USA) during several rainstorms in the winter of 2001, after the organophosphate diazinon and the pyrethroid esfenvalerate were applied to different orchard sections. We tested and compared acute toxicity of orchard runoff from diazinon- and esfenvalerate-sprayed sections to two species of fish (Pimephales promelas, Onchorhynchus mykiss) and three aquatic invertebrates (Ceriodaphnia dubia, Simocephalus vetelus, Chironomus riparius), and determined the mitigating effect of three ground cover crops on toxicity and insecticide loading in diazinon-sprayed orchard rows. Runoff from the esfenvalerate-sprayed orchard section was less toxic to waterflea than runoff from the diazinon-sprayed section. However, runoff from the orchard section sprayed with esfenvalerate was highly toxic to fish larvae. Samples collected from both sections one month later were not toxic to fish, but remained highly toxic to invertebrates. The ground cover crops reduced total pesticide loading in runoff by approximately 50%. No differences were found between the types of vegetation used as ground covers.

Joint acute toxicity of esfenvalerate and diazinon to larval fathead minnows (Pimephales promelas).

California (USA) agriculture employs pyrethroid and organophosphate insecticides to control insects in orchards and other crops. Diazinon and esfenvalerate were selected for this study because of their application overlaps. Toxicological and biochemical responses of larval fathead minnows (Pimephales promelas) exposed singly and in combinations to esfenvalerate and diazinon were determined. Exposures were 96-h static renewal tests that used standard U.S. Environmental Protection Agency acute toxicity test methods. After pesticide exposures, larvae were evaluated for carboxylesterase and acetylcholinesterase activity, and histopathological effects. Carboxylesterase activity was examined because of its potential influence on the toxicity of both organophosphates and pyrethroids. In vivo studies demonstrated that diazinon significantly inhibited carboxylesterase activity at nominal water concentrations as low as 50 microg/L. However, esfenvalerate did not affect carboxylesterase activity at any concentration tested. Liver glycogen depletion was the only histopathological effect observed; this effect was demonstrated with the individual pesticides and pesticide combinations (i.e., mixtures). The combinations of diazinon and esfenvalerate causing acute toxicity to fathead minnow larvae appeared to be greater than additive (i.e., synergistic) in all three tests.

Transcriptomic profiling permits the identification of pollutant sources and effects in ambient water samples.

Contaminant exposure is one possible contributor to population declines of endangered fish species in the Sacramento-San Joaquin Estuary, California, including the endangered delta smelt (Hypomesus transpacificus). Herein we investigated transcriptional responses in larval delta smelt resulting from exposure to water samples collected at the Department of Water Resources Field Station at Hood, a site of concern, situated upstream of known delta smelt habitat and spawning sites and downstream of the Sacramento Regional Wastewater Treatment Plant (SRWTP). Microarray assessments indicate impacts on energy metabolism, DNA repair mechanisms and RNA processing, the immune system, development and muscle function. Transcription responses of fish exposed to water samples from Hood were compared with exposures to 9% effluent samples from SRWTP, water from the Sacramento River at Garcia Bend (SRGB), upstream of the effluent discharge, and SRGB water spiked with 2mg/L total ammonium (9% effluent equivalent). Results indicate that transcriptomic profiles from Hood are similar to 9% SRWTP effluent and ammonium spiked SRGB water, but significantly different from SRGB. SRGB samples however were also significantly different from laboratory controls, suggesting that SRWTP effluent is not solely responsible for the responses determined at Hood, that ammonium exposure likely enhances the effect of multiple-contaminant exposures, and that the observed mortality at Hood is due to the combination of both effluent discharge and contaminants arising from upstream of the tested sites.

Comparing the effectiveness of chronic water column tests with the crustaceans Hyalella azteca (order: Amphipoda) and Ceriodaphnia dubia (order: Cladocera) in detecting toxicity of current-use insecticides.

Standard U.S. Environmental Protection Agency laboratory tests are used to monitor water column toxicity in U.S. surface waters. The water flea Ceriodaphnia dubia is among the most sensitive test species for detecting insecticide toxicity in freshwater environments.Its usefulness is limited, however, when water conductivity exceeds 2,000 µS/cm (approximately 1 ppt salinity) and test effectiveness is insufficient. Water column toxicity tests using the euryhaline amphipod Hyalella azteca could complement C. dubia tests; however, standard chronic protocols do not exist. The present study compares the effectiveness of two water column toxicity tests in detecting the toxicity of two organophosphate (OP) and two pyrethroid insecticides: the short-term chronic C. dubia test, which measures mortality and fecundity, and a 10-d H. azteca test, which measures mortality and growth. Sensitivity was evaluated by comparing effect data, and end point variability was evaluated by comparing minimum significant differences. Tests were performed in synthetic water and filtered ambient water to quantify the influence of water matrix on effect concentrations. The H. azteca test detected pyrethroid toxicity far more effectively, while the C. dubia test was more sensitive to OPs. Among endpoints, H. azteca mortality was most robust. The results demonstrate that the H. azteca test is preferable when conductivity of water samples is 2,000 to 10,000 µS/cm or if contaminants of concern include pyrethroid insecticides.

Sublethal responses to ammonia exposure in the endangered delta smelt; Hypomesus transpacificus (Fam. Osmeridae).

The delta smelt (Hypomesus transpacificus) is an endangered pelagic fish species endemic to the Sacramento-San Joaquin Estuary in Northern California, which acts as an indicator of ecosystem health in its habitat range. Interrogative tools are required to successfully monitor effects of contaminants upon the delta smelt, and to research potential causes of population decline in this species. We used microarray technology to investigate genome-wide effects in fish exposed to ammonia; one of multiple contaminants arising from wastewater treatment plants and agricultural runoff. A 4-day exposure of 57-day old juveniles resulted in a total ammonium (NH(4)(+)-N) median lethal concentration (LC50) of 13 mg/L, and a corresponding un-ionized ammonia (NH(3)) LC50 of 147 µg/L. Using the previously designed delta smelt microarray we assessed altered gene transcription in juveniles exposed to 10mg/L NH(4)(+)-N from this 4-day exposure. Over half of the responding genes were associated with membrane integrity and function, however, neurological and muscular function was also affected. Amongst the notable pathways affected by ammonium exposure, directly associated with cellular membranes, are energy metabolism through oxidative phosphorylation, cellular responses to environmental stimuli, highlighted through signal transduction and molecular interactions, cellular processes encompassing transport and catabolism, along with cell motility, development, communication and cell death. To assess these impacts further, key genes were selected as potential biomarkers and investigated using quantitative PCR analysis on fish exposed to 2.5, 5, 10, 20 and 40 mg/L NH(4)(+)-N. Quantitative PCR results indicate biphasic responses, pivoting around the estimated no-observed effect concentration (NOEC; 5.0mg/L NH(4)(+)-N) and below. Genes significantly affected by ammonia exposure include claudin-10, Keratin-15, Septin-3, Transmembrane protein 4, superfamily 4 (membrane), Tropomyosin, Myosin light chain, Calmodulin (muscular), Tubulin cofactor beta (neurological), Sirtuin-6 (development), and Rhesus associated type C glycoprotein 1 (gill- and skin-specific ammonium transporter). The quantitation of the ammonium transporter may highlight the capacity of delta smelt to contend with elevated levels of ammonia, the peak response of which may be indicative of short-term thresholds of tolerance. Our study supports the notion that exposure to ammonia results in cell membrane destabilization, potentially affecting membrane permeability, enhancing uptake and thus synergistic effects of multiple-contaminant exposure.

Monitoring acute and chronic water column toxicity in the Northern Sacramento-San Joaquin Estuary, California, USA, using the euryhaline amphipod, Hyalella azteca: 2006 to 2007.

After the significant population decline of several pelagic fish species in the Northern Sacramento-San Joaquin (SSJ) Estuary (CA, USA) in 2002, a study was performed to monitor water column toxicity using the amphipod Hyalella azteca. From January 1, 2006 to December 31, 2007, water samples were collected biweekly from 15 to 16 sites located in large delta channels and main-stem rivers, selected based on prevalent distribution patterns of fish species of concern. Ten-day laboratory tests with H. azteca survival and relative growth as toxicity endpoints were conducted. The enzyme inhibitor piperonyl butoxide ([PBO], 25 µg/L) was added to synergize or antagonize pyrethroid or organophosphate (OP) insecticide toxicity, respectively. Significant amphipod mortality was observed in 5.6% of ambient samples. Addition of PBO significantly changed survival or growth in 1.1% and 10.1% of ambient samples, respectively. Sites in the Lower Sacramento River had the largest number of acutely toxic samples, high occurrence of PBO effects on amphipod growth (along with sites in the South Delta), and the highest total ammonia/ammonium concentrations (0.28 ± 0.15 mg/L). Ammonia/ammonium, or contaminants occurring in mixture with these, likely contributed to the observed toxicity. Pyrethroid insecticides were detected at potentially toxic concentrations. Overall, results of this study identified specific areas and contaminants of concern and showed that water in the Northern SSJ Estuary was at times acutely toxic to sensitive invertebrates.

Use of vegetated agricultural drainage ditches to decrease toxicity of irrigation runoff from tomato and alfalfa fields in California, USA.

The current study investigated the potential of vegetated drainage ditches for mitigating the impact of agricultural irrigation runoff on downstream aquatic ecosystems. Water column toxicity to larval fathead minnow (Pimephales promelas),and the amphipod Hyalella azteca was measured for 12 h or less at the ditch inflow and outflow, using custom-built in situ exposure systems. In addition, water and sediment samples were subject to standard toxicity tests with Ceriodaphnia dubia and H. azteca, respectively. No acute toxicity to larval fathead minnow was observed; however, runoff was highly toxic to invertebrates. Passage through a 389- to 402-m section of vegetated ditch had a mitigating effect and reduced toxicity to some degree. However, runoff from an alfalfa field treated with chlorpyrifos remained highly toxic to both invertebrate species, and runoff from a tomato field treated with permethrin remained highly toxic to H. azteca after passage through the ditch. Predicted toxic units calculated from insecticide concentrations in runoff and 96-h median lethal concentration (LC50) values generally agreed with C. dubia toxicity measured in the laboratory but significantly underestimated in situ toxicity to H. azteca. Sediments collected near the ditch outflow were toxic to H. azteca. Results from the current study demonstrate that experimental vegetated ditches were unable to eliminate the risk of irrigation runoff to aquatic ecosystems. In addition, protective measures based on chemical concentrations or laboratory toxicity tests with C. dubia do not ensure adequate protection of aquatic ecosystems from pyrethroid-associated toxicity.