Paula Jackman

Pharmaceutically active compounds in Atlantic Canadian sewage treatment plant effluents and receiving waters, and potential for environmental effects as measured by acute and chronic Aquatic Toxicity

Ten acidic and two neutral pharmaceuticals were detected in the effluents of eight sewage treatment plants (STPs) from across Atlantic Canada. Concentrations varied between nondetectable and 35 microg/L. The analgesic, nonsteroidal anti-inflammatory drugs ibuprofen and naproxen were predominant. Carbamazepine, a neutral compound used as an antiepileptic drug, was observed consistently at a median concentration of 79 ng/L. Acetaminophen was found in the effluents of the three largest mechanical STPs at a median concentration of 1.9 microg/L, but not in the lagoon treatment systems. The substantially longer hydraulic retention times may have contributed to more effective removal of acetaminophen in the lagoon treatment systems. Drugs generally were not detected at significant concentrations in the larger bodies of receiving water (Saint John River, Hillsborough River, and Bedford Bay, Canada). However, drug residues in the small receiving streams were 15 to 30% of the effluent median concentrations. Six compounds (caffeine, naproxen, salicylic acid, carbamazepine, metoprolol, and sotolol) were found to persist in a small stream for a distance of at least 17 km, suggesting that small stream exposure to pharmaceutically active residues may be relatively greater than that in large bodies of water. Bioassays assessing acute and chronic effects on four organisms were conducted on four high-use drugs: Acetaminophen, ibuprofen, naproxen, and salicylic acid (metabolite of acetyl salicylic acid). Results indicated no negative effects except for the chronic algal (Selanastrum capricornutum) growth test on ibuprofen (no-observed-effect concentration, 10 microg/L; lowest-observed-effect concentration, 32 microg/L). Effects of these four compounds on invertebrates and plants in the receiving environments are unlikely based on the concentrations measured.

Dispersion and Toxicity to Non-target Aquatic Organisms of Pesticides Used to Treat Sea Lice on Salmon in Net Pen Enclosures

Pesticides are used extensively in the finfish aquaculture industry to control sea lice infestations on farmed salmon. The most prevalent method of use is to enclose a net pen with an impervious tarpaulin and mix a pesticide solution within that enclosure. After treatment for short periods (1 h) the pesticide solution is released to the environment. Concerns have been raised that there is a potential risk to non-target aquatic organisms from those releases. The fate of dispersing pesticide solutions was measured after six simulated treatments in the Lower Bay of Fundy, New Brunswick. Three simulated treatments were done with azamethiphos and three with cypermethrin. Rhodamine dye was added to all pesticide solutions in order to facilitate tracking of the dispersing plume through real-time measurements of dye concentrations by a flow-through fluorometer coupled with a differential global positioning system (DGPS). Water samples were obtained from within the plumes at various times after release and analysed for pesticide content and toxicity to a benthic amphipod Eohaustorius estuaris. Dye concentrations were detectable for time periods after release which varied from 2 to 5.5 h. Distances travelled by the dye patches ranged from 900 to 3000 m and the dye concentrations at the final sampling period were generally 1/200-1/3000 the pre-release concentrations and cypermethrin concentrations were generally 1/1000-1/2000 the pre-release concentrations. Cypermethrin concentrations in water samples were closely correlated with dye concentrations, indicating that dye analyses were an accurate surrogate for cypermethrin concentrations. Most samples taken after the releases of azamethiphos were not toxic to test organisms in 48 h exposures and none were beyond 20 min post-release. By contrast, almost all samples taken after the release of cypermethrin, even up to 5-h post-release, were toxic. Data indicate the potential to cause toxic effects over areas of hectares from a single release of cypermethrin.

Hormonal induction of spawning in 4 species of frogs by coinjection with a gonadotropin-releasing hormone agonist and a dopamine antagonist

BackgroundIt is well known that many anurans do not reproduce easily in captivity. Some methods are based on administration of mammalian hormones such as human chorionic gonadotropin, which are not effective in many frogs. There is a need for simple, cost-effective alternative techniques to induce spawning.MethodsOur new method is based on the injection of a combination of a gonadotropin-releasing hormone (GnRH) agonist and a dopamine antagonist. We have named this formulation AMPHIPLEX, which is derived from the combination of the words amphibian and amplexus. This name refers to the specific reproductive behavior of frogs when the male mounts and clasps the female to induce ovulation and to fertilize the eggs as they are laid.ResultsWe describe the use of the method and demonstrate its applicability for captive breeding in 3 different anuran families. We tested several combinations of GnRH agonists with dopamine antagonists using Lithobates pipiens. The combination of des-Gly10, D-Ala6, Pro-LHRH (0.4 microrams/g body weight) and metoclopramide (10 micrograms/g BWt. MET) was most effective. It was used in-season, after short-term captivity and in frogs artificially hibernated under laboratory conditions. The AMPHIPLEX method was also effective in 3 Argentinian frogs, Ceratophrys ornata, Ceratophrys cranwelli and Odontophrynus americanus.ConclusionOur approach offers some advantages over other hormonally-based techniques. Both sexes are injected only once and at the same time, reducing handling stress. AMPHIPLEX is a new reproductive management tool for captive breeding in Anura.

Effects of glyphosate-based herbicides on survival, development, growth and sex ratios of wood frog (Lithobates sylvaticus) tadpoles. II: Agriculturally relevant exposures to Roundup WeatherMax® and Vision® under laboratory conditions

Glyphosate-based herbicides are currently the most commonly used herbicides in the world. They have been shown to affect survival, growth, development and sexual differentiation of tadpoles under chronic laboratory exposures but this has not been investigated under more environmentally realistic conditions. The purpose of this study is (1) to determine if an agriculturally relevant exposure to Roundup WeatherMax®, a relatively new and understudied formulation, influences the development of wood frog tadpoles (Lithobates sylvaticus) through effects on the mRNA levels of genes involved in the control of metamorphosis; (2) to compare results to the well-studied Vision® formulation (containing the isopropylamine salt of glyphosate [IPA] and polyethoxylated tallowamine [POEA] surfactant) and to determine which ingredient(s) in the formulations are responsible for potential effects on development; and (3) to compare results to recent field studies that used a similar experimental design. In the present laboratory study, wood frog tadpoles were exposed to an agriculturally relevant application (i.e., two pulses) of Roundup WeatherMax® and Vision® herbicides as well as the active ingredient (IPA) and the POEA surfactant of Vision®. Survival, development, growth, sex ratios and mRNA levels of genes involved in tadpole metamorphosis were measured. Results show that Roundup WeatherMax® (2.89 mg acid equivalent (a.e.)/L) caused 100% mortality after the first pulse. Tadpoles treated with a lower concentration of Roundup WeatherMax® (0.21 mg a.e./L) as well as Vision® (2.89 mg a.e./L), IPA and POEA had an increased condition factor (based on length and weight measures in the tadpoles) relative to controls at Gosner stage (Gs) 36/38. At Gs42, tadpoles treated with IPA and POEA had a decreased condition factor. Also at Gs42, the effect on condition factor was dependent on the sex of tadpoles and significant treatment effects were only detected in males. In most cases, treatment reduced the normal mRNA increase of key genes controlling development in tadpoles between Gs37 and Gs42, such as genes encoding thyroid hormone receptor beta in brain, glucocorticoid receptor in tail and deiodinase enzyme in brain and tail. We conclude that glyphosate-based herbicides have the potential to alter mRNA profiles during metamorphosis. However, studies in natural systems have yet to replicate these negative effects, which highlight the need for more ecologically relevant studies for risk assessment.

Analysis of gene expression in Homarus americanus larvae exposed to sublethal concentrations of endosulfan during metamorphosis.

Agricultural pesticide runoff has been suspected as the cause of numerous fish kills in rivers throughout Prince Edward Island but the impact on the surrounding marine environment is unknown. Endosulfan, an organochlorine pesticide, is a potent neurotoxin and molt inhibitor used to combat the Colorado potato beetle however it has the potential to affect non-target organisms including the American lobster (Homarus americanus). Metamorphosis is a critical stage of development and the effects of contaminant exposure during this time are largely unknown in lobster. A 14day endosulfan exposure was performed to identify the effects on survival, development and gene expression in lobster larvae during metamorphosis; all of which were predicted to be negatively impacted. The higher endosulfan concentrations resulted in greater mortality and a significant increase in the number of days required to reach metamorphosis in surviving animals. A custom made H. americanus microarray was used for monitoring the changes in expression of 14,592 genes at the termination of the exposure. Genes with >1.5 fold change and identified as being significant at p<0.05 using one-way ANOVA were selected for further analysis. A total of 707 genes were identified as being significantly differentiated, however with only ~40% annotation of the array, the majority of these genes were unknown. Annotated genes of interest were involved in many processes: development, metabolism, immunity and oxidative stress response and gene regulation. Nine genes of interest (histone H1, farnesoic acid O-methyltransferase, cuticle protein, glutathione S-transferase, thioredoxin, NADH dehydrogenase, ecdysone nuclear receptor Fushi tarazu F1 (FTZ-F1), ferritin and ecdysone inducible protein E75 (EIP-E75)) were selected for RT-qPCR validation of the microarray results. The RT-qPCR method was more sensitive than the microarray yet detected similar expression patterns. The two highest endosulfan concentrations resulted in increased mortalities, developmental delays in reaching metamorphosis and significant changes in gene expression. This research provides a foundation for using microarray gene expression profiles as screening tools for exploring the impact of environmental contaminants on lobster.

Ecotoxicogenomics: Microarray interlaboratory comparability.

Transcriptomic analysis can complement traditional ecotoxicology data by providing mechanistic insight, and by identifying sub-lethal organismal responses and contaminant classes underlying observed toxicity. Before transcriptomic information can be used in monitoring and risk assessment, it is necessary to determine its reproducibility and detect key steps impacting the reliable identification of differentially expressed genes. A custom 15K-probe microarray was used to conduct transcriptomics analyses across six laboratories with estuarine amphipods exposed to cyfluthrin-spiked or control sediments (10 days). Two sample types were generated, one consisted of total RNA extracts (Ex) from exposed and control samples (extracted by one laboratory) and the other consisted of exposed and control whole body amphipods (WB) from which each laboratory extracted RNA. Our findings indicate that gene expression microarray results are repeatable. Differentially expressed data had a higher degree of repeatability across all laboratories in samples with similar RNA quality (Ex) when compared to WB samples with more variable RNA quality. Despite such variability a subset of genes were consistently identified as differentially expressed across all laboratories and sample types. We found that the differences among the individual laboratory results can be attributed to several factors including RNA quality and technical expertise, but the overall results can be improved by following consistent protocols and with appropriate training.

Natural Recovery Reduces Impact of the 1970 Arrow Oil Spill

ABSTRACT In 1970 the tanker Arrow ran aground releasing 2,000 m3 of Bunker C crude oil along 300 km of Nova Scotias coastline, of which only 10% was subjected to cleanup, the rest was left to degr...

Toxicity testing of freshwater sediment collected near freshwater aquaculture facilities in the Maritimes, Canada

In the Atlantic region of Canada, there are close to 50 land-based freshwater aquaculture facilities, most of which discharge wastewater to freshwater receiving environments. This study was designed to assess the chemical and toxicological characteristics of sediments in those receiving environments. Thirty sediment samples were collected from 3 locations (upstream, outfall and downstream) at seven freshwater aquaculture facilities. Toxicity tests conducted included amphipod growth, amphipod survival and Microtox™. Sediments were also analysed for ammonia/porewater ammonia, redox and sulphide. Porewater ammonia concentration for the amphipod survival test ranged from 0.01 to 42 mg/L at the conclusion of the 14-day survival test. Ammonia concentration in sediment ranged from 0.3–202 μg/g, sulphide concentration ranged from 0.15 to 17.4 μg/g, yet redox ranged from 32 to 594 mV. Microtox™ IC50 values ranged from 108,00 to >164,000 mg/L, yet amphipod survival ranged from 0 to 100%, depending on sampling locations. Amphipod survival was significantly related (P < 0.05) to porewater ammonia (at time = 0 and 14 days) and Microtox™ IC50 was significantly related (P < 0.05) to ammonia, sulphide and redox levels. These results indicate that discharges from some the land-based aquaculture facilities are impacting sediment dwelling benthic invertebrates at the outfall but that impact largely disappears by 100 m downstream. Furthermore those impacts were rarely detected during the early winter season, when biomass production was at the lowest.