John C. Chapman

The effects of three organic chemicals on the upper thermal tolerances of four freshwater fishes

The upper temperature tolerance limits of four freshwater fish species, silver perch Bidyanus bidyanus, eastern rainbowfish Melanotaenia duboulayi, western carp gudgeon Hypseleotris klunzingeri, and rainbow trout Oncorhynchus mykiss, were determined using the critical thermal maximum (CTMaximum) method. The CTMaximum tests were carried out with unexposed fish and fish exposed to sublethal concentrations of endosulfan, chlorpyrifos, and phenol to determine whether or not the CTMaximum was affected. The CTMaximum temperature of B. bidyanus decreased by 2.8, 3.8, and 0.3 degrees C on exposure to endosulfan, chlorpyrifos, and phenol, respectively. Similarly, in M. duboulayi, the CTMaximum was decreased by 4.1, 2.5, and 0 degrees C, while in H. klunzingeri it decreased by 3.1, 4.3, and 0.1degrees C, respectively, and in O. mykiss by 4.8, 5.9, and 0.7 degrees C, respectively. Exposure to sublethal test concentrations of endosulfan and chlorpyrifos caused significant (p < or = 0.0001) reductions in CTMaximum values for all fish species compared to that of unexposed fish. However, exposure to phenol did not cause any significant (p > or = 0.05) change of CTMaximum temperatures.

Response and recovery of acetylcholinesterase activity in freshwater shrimp, Paratya australiensis (Decapoda: Atyidae) exposed to selected anti-cholinesterase insecticides

The toxicity of carbaryl, chlorpyrifos, dimethoate and profenofos to the freshwater shrimp, Paratya australiensis was assessed by measuring acetylcholinesterase (AChE) inhibition after 96h exposures. Shrimp exposed to these pesticides exhibited significant AChE inhibition, with mortality in shrimp corresponding to 70-90% AChE inhibition. The sensitivity of P. australiensis to the four pesticides based on AChE inhibition can be given as chlorpyrifos > profenofos > carbaryl > dimethoate. Recovery of AChE activity was followed in shrimp after 96 h exposures to carbaryl, chlorpyrifos and dimethoate. Recovery after exposure to the carbamate pesticide carbaryl was more rapid than for the two organophosphorus pesticides, chlorpyrifos and dimethoate. The slow recovery of depressed AChE activity may mean that affected organisms in the natural system are unable to sustain physical activities such as searching for food or eluding predators. To investigate the ecological significance of AChE inhibition, chemotaxis behaviour was assessed in shrimp exposed to profenofos for 24h. Abnormal chemotaxis behaviour in the exposed shrimp was observed at concentrations representing 30-50% AChE inhibition. A clear relationship existed between the depression of AChE activity and observed chemotaxis responses, such as approaching and grasping the chemoattractant source. These results suggest that in vivo toxicity tests based on this specific biomarker are sensitive and present advantages over conventional acute tests based on mortality. Behavioural studies of test organisms conducted in conjunction with measurement of AChE inhibition will provide data to clarify the toxic effects caused by sublethal chemical concentrations of anti-cholinesterase compounds.

Acute and chronic toxicity of endosulfan to two Australian cladocerans and their applicability in deriving water quality criteria

The acute and chronic toxicity of endosulfan was determined in two Australian cladocerans, Ceriodaphnia dubia and Moinodaphnia macleayi. For C. dubia, the 48-h nominal EC50 (immobilization) was 490 μg/L and the chronic NOEC for reproductive impairment was 10 μg/L. For M. macleayi, the 48-h nominal EC50 (immobilization) was 215 μg/L, and the chronic NOEC was 20 μg/L. A water quality guideline for endosulfan based on cladoceran chronic toxicity may be estimated at 2 μg/L by using an application factor of 0.1 and the LOEC value for C. dubia of 20 μg/L. A much lower guideline of 4 ng/L may be obtained if the acute/chronic ratio calculated for C. dubia is applied to the lowest LC50 value determined for an Australian fish, 0.2 μg/L in the bony bream, Nematolosa erebi. This suggests that the concentration of endosulfan for Australian waters should be less than the level of 10 ng/L given in the Australian Water Quality guidelines for (fresh and marine waters) aquatic ecosystems. Factors which may modify these conclusions under site-specific conditions are discussed.

Disposal of hazardous wastes in Australia: Implications for marine pollution

Abstract Australian coastal waters have been contaminated by polychlorinated biphenyls (PCBs), dioxins and furans, and persistent organochlorine pesticides. Most PCB contamination is associated with large urban centres, while dioxins and furans have been of particular concern around former industrial sites in Sydney, in trade wastes in Melbourne, and in relation to a pulp mill proposed for Tasmania. Prolonged use of organochlorine pesticides has resulted in widespread contamination. Australian regulation of chemicals and hazardous wastes is complex due to the involvement of eight State and Territory Governments, the piecemeal introduction of controls, and their division into sectoral areas. National legislation to control agricultural, veterinary and industrial chemicals came into effect in 1989 and 1991. Strategies are being developed for the management of intractable wastes, while controls on sea dumping meet the requirements of the London Dumping Convention.

Bioactivity of POPs and their effects in mosquitofish in Sydney Olympic Park, Australia

The site of the 2000 Olympic Games (Sydney Olympic Park (SOP), Sydney, Australia) was contaminated by persistent organic pollutants (POPs) prior to remediation in the 1990s. This study investigates the bioactivity of POPs in the sediment and water of wetlands across SOP by in vitro 2,3,7,8-TCDD equivalence (TCDDeq) measurement (H4IIE cell line bioassay). Further, it examines whether disturbance of these sediments is likely to mobilise ligands for this receptor into the water column. Exposure to aryl hydrocarbon receptor (AhR) ligands was measured in vivo using hepatic cytochrome P4501A (CYP1A) induction (EROD) in the mosquitofish (Gambusia holbrooki). Aqueous TCDDeq ranged from 0.013 to 0.057 pM in SOP wetlands which was significantly (p<0.05) less that in urban reference sites. These concentrations were not correlated to physical or chemical characteristics of the wetlands. In the sediments, TCDDeq ranged from 0.0016 to 7.06 microg/kg and these were not significantly (p>or=0.05) different to that measured in urban reference sites. Simulated disturbance of small quantities of sediment in water samples significantly (p<0.05) increased the levels of TCDDeq measured in the water. Sediment TCDDeq was correlated to sediment SigmaPAH concentration in 2006 and sediment SigmaPCB, SigmaDDT concentrations and fine sediment grain size in 2005. While fish at one SOP wetland had hepatic EROD activity elevated above the estimated basal level for this species, these were at the lower end of the range measured in urban impacted, non-remediated wetlands. EROD activity was positively correlated with both the sediment SigmaPCB load and aqueous TCDDeq. Increased catchment size was correlated with increased EROD activity suggesting an even spread of POPs throughout the residential areas of the Sydney metropolitan area. The concentration of bioactive POPs in the wetlands of SOP is therefore low relative to urban reference sites demonstrating the ongoing success of the remediation program.

Distribution of inorganic and organic contaminants in sediments from Sydney Olympic Park and the surrounding Sydney metropolitan area.

Organic and inorganic contamination was assessed for sediments from wetlands and water bodies within the Sydney Olympic Park (SOP, remediated sites) and its surroundings (unremediated sites) and urban reference sites in the Sydney Basin. Among the seven elements analysed (As, Cd, Cr, Cu, Pb, Ni and Zn), Zn concentrations were the highest, followed by Pb, Cu and Cr in the sediments of SOP. Significantly higher concentrations (p < 0.05) of DDTs were found in sediments of the Homebush Bay and Parramatta River sites compared with the urban reference sites, mainly due to past manufacturing of DDT in the Homebush Bay area. However, no significant differences (p > 0.05) in concentrations were found for polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) as well as DDTs between sediments from SOP and the urban reference sites. Source indicators suggest that PAHs in the sediments originated from combustion processes. Two distinct groups of dioxin profiles were observed within SOP and its surroundings. Levels of dioxins were more than 100 pg WHO-TEQ/g dry weight of sediment at five sites adjacent to the SOP boundaries. Based on the findings of the chemical profiles of the contaminants, the remediated sites in SOP can be regarded as similar to the urban reference sites within the Sydney Basin, while the adjacent unremediated sites have higher concentrations, especially of dioxins, that could still affect organisms in the aquatic environment.

Interactions between water temperature and contaminant toxicity to freshwater fish

Warming of freshwaters as a result of climate change is expected to have complex interactions with the toxicity of contaminants to aquatic organisms. The present study evaluated the effects of temperature on the acute toxicity of endosulfan, chlorpyrifos, and phenol to 3 warm water species of fish-silver perch, rainbowfish, and western carp gudgeon-and 1 cold water species, rainbow trout. Endosulfan was more toxic to silver perch at 30 °C and 35 °C than at 15 °C, 20 °C and 25 °C during short exposures of 24 h, but at 96 h, temperature had no effect on toxicity. Toxicity to rainbow trout increased with increasing temperature, whereas warm water species exhibited maximum toxicity at around 30 °C, decreasing again toward 35 °C. Chlorpyrifos became more toxic to all species with increasing temperature. Phenol toxicity to all species decreased at low to intermediate temperatures; but as temperatures increased further toward the upper thermal limit, phenol became more toxic. Increasing toxicity in the upper thermal range of cold water species may contribute to upstream range contraction in rivers with high toxicant loads. In contrast, warm water species may not exhibit a range shift within rivers as a result of interactions between temperature and toxicity. Catchment management to offset global warming at local scales may present opportunities to mitigate increased toxicity of contaminants to fish.

Benthic macroinvertebrate assemblages in remediated wetlands around Sydney, Australia.

To investigate potential high organisational level impacts of persistent organic pollution in the wetlands in the Sydney Olympic Park (SOP) remediated site, the benthic macroinvertebrate assemblages of seven wetlands within SOP and two off-site reference wetlands were examined. Sediment cores were collected, stained and preserved from each study site and the macroinvertebrates identified to the appropriate taxonomic level (Class, Order, Family, Subfamily). Data were analysed for taxon richness and macroinvertebrate abundance and multivariate techniques were used to identify chemical/physical characteristics of the sediment, which were important influences on the differences in the assemblage between study sites. Macroinvertebrate abundance was highly variable between study sites and taxon richness was low across all sites. Oligochaetes, nematodes, ostracods and chironomids were the most common taxa found and were the most important in influencing differences between the macroinvertebrate assemblages among the study sites. Sediment grain size and chemical characteristics of the sediments (ΣPAH, ΣPCB, TCDDeq and heavy metal concentrations) were important in separating the study sites based on taxon richness and abundance. Canonical correspondence analysis separated the macroinvertebrate assemblages at newly two created wetlands from those at other study sites including the urban reference sites. Increased sediment POP contamination (particularly as measured TCDDeq and ΣDDT concentrations) is a likely contributor in excluding pollution sensitive taxa and, therefore, alterations to benthic macroinvertebrate assemblages. Further, the influence of TOC suggests the significance of catchment inputs in contributing to changes in macroinvertebrate assemblage. The SOP remediation led to the establishment of wetlands with benthic communities representative of those expected in urban wetlands.

Effects of temperature on ventilatory behavior of fish exposed to sublethal concentrations of endosulfan and chlorpyrifos

The ventilation amplitude and frequency of silver perch Bidyanus bidyanus, and the ventilation frequency of rainbow fish Melanotaenia duboulayi and rainbow trout Oncorhynchus mykiss, were determined at different temperatures upon exposure to endosulfan and chlorpyrifos, respectively. Silver perch and rainbow fish were tested at 15, 20, 25, 30, and 35 degrees C, while rainbow trout was tested at 10, 15, 20, and 25 degrees C. Although some trend of increasing amplitudes with increasing temperature was evident; there was no significant temperature response of ventilation frequency rates over time in silver perch pre-exposed to 10 microg L(-1) endosulfan for 18 h. The rate of ventilation frequency of rainbow fish pre-exposed to 200 microg L(-1) of chlorpyrifos for 96 h was lower in treatments than in the control at 15 degrees C. However, between 20 and 35 degrees C, rates were significantly higher in the treatments than those of the control. In rainbow trout pre-exposed to 100 microg L(-1) of chlorpyrifos, the rates of frequency were significantly lower than those of controls in temperatures between 10 and 20 degrees C but higher at 25 degrees C. The amplitude of silver perch seemed to increase with the increase in temperature; however, the corresponding temperature quotient values at various temperature regimes and over exposure time showed no significant differences. The ventilation frequency of rainbow fish and rainbow trout significantly increased at the higher test temperatures, and their corresponding temperature quotient values for both fish also increased at the elevated temperatures.

A DNA‐based assay for toxic chemicals in wastewater

Chemical toxicants, particularly metal ions, are a major contaminant in global waterways. Live-organism bioassays used to monitor chemical toxicants commonly involve measurements of activity or survival of a freshwater cladoceran (Ceriodaphnia dubia) or light emitted by the marine bacterium Vibrio fischeri, used in the commercial Microtox® bioassay. Here we describe a novel molecule-based assay system employing DNA as the chemical biosensor. Metals bind to DNA, causing structural changes that expel a bound (intercalated) fluorescent reporter dye. Analyses of test data using 48 wastewater samples potentially contaminated by metal ions show that the DNA-dye assay results correlate with those from C. dubia and Microtox bioassays. All three assays exhibit additive, antagonistic, and synergistic responses that cannot be predicted by knowing individual metal concentrations. Analyses of metals in these samples imply the presence of chemical toxicants other than metal ions. The DNA-dye assay is robust, has a 12-month shelf life, and is only slightly affected by sample pH in the range 4 to 9. The assay is completed in a matter of minutes, and its portability makes it well suited as a screening assay for use in the field. We conclude that the DNA-dye test is a surrogate bioassay suitable for screening chemical toxicity.