Comparative toxicity of ten organic chemicals to ten common aquatic species

The susceptibilities of 10 aquatic organisms to 10 organic chemicals were determined using lethality tests. The species included six fishes, two crustaceans, a chironomid and an amphibian. The chemicals were selected to span the toxicity range from 26 g l⁻¹ to 1 μg l⁻¹ and include chemicals which were lethal by four modes of toxic action. There was no consistent relative susceptibility among the test species because the sensitivity to specific modes of toxic action varied among the chemicals. Nonetheless, the toxicities of the chemicals to any given species were highly correlated to the toxicities to other species, particularly among fishes. The 96-h median lethal concentration (LC₅₀) of the chemicals to rainbow trout (Salmo gairdneri) could be estimated from the 96-h LC₅₀ with fathead minnows (Pimephales promelas) with a correlation coefficient greater than 0.99. Equations for estimating the lethal concentration of chemicals with each species from the 96-h LC₅₀ for fathead minnows are presented.     

Comparison of chlorine and chlorine dioxide toxicity to fathead minnows and bluegill

The comparative toxicity of total residual chlorine (TRC) and chlorine dioxide (ClO₂) was evaluated by conducting 96 h flow-through bioassays with three types of fish. The fish were subjected to an intermittent exposure regime in which biocide residuals were present for approx. 2-h periods beginning at 0, 24, 48 and 72 h into the tests. These conditions simulated the antifouling procedure (1 h day⁻¹ biocide addition) used to control biofouling of nuclear reactor heat exchangers at the Savannah River Plant near Aiken, South Carolina. LC₅₀ values showed that ClO₂ was approx. 2–4 times more toxic than TRC to: (1) juvenile and 1-year-old fathead minnows (Pimphales promelas); and (2) young-of-the-year bluegill (Lepomis macrochirus).The TRC mean 96-h LC₅₀ values were: 0.08 mg l⁻¹ for juvenile fathead minnows, 0.35 mg l⁻¹ for adult fathead minnows and 0.44 mg l⁻¹ for young-of-the-year bluegills. The ClO₂ mean LC₅₀ values were: 0.02 mg l⁻¹ for juvenile fathead minnows, 0.17 mg l⁻¹ for adult fathead minnows and 0.15 mg l⁻¹ for young-of-the-year bluegills.     

A comparison of toxicity tests conducted in the laboratory and in experimental ponds using cadmium and the fathead minnow (Pimephales promelas)

Acute toxicity tests were conducted in the laboratory with fathead minnows (Pimephales promelas) to determine the 96-h LC₅₀ of cadmium under three conditions: (1) in laboratory water, (2) in water from experimental ponds, and (3) in pond water underlain by sediment. Cadmium was then applied at doses equivalent to the estimated LC₅₀ values to 0.07-ha ponds containing caged fathead minnows. A cadmium ion selective electrode, ultrafiltration, and equilibrium calculations were used to determine cadmium speciation, and several water quality characteristics were measured to correlate differences in mortality between test systems (laboratory and field) with observed differences in water quality. The LC₅₀ estimates (mg l⁻¹) for the bioassays were 4.39 for the laboratory water, 3.52 for the pond water with sediment, and 2.91 for the pond water. Concentrations of Cd²⁺ decreased and those of cadmium in the particulate (> 1.2 μm) and 300,000 mol. wt (0.018–1.2 μm) fractions increased over the 96-h; cadmium in these fractions was believed to consist of colloidal sized CdCO₃ precipitates. Concentrations of Cd²⁺ decreased at different rates between test systems, regulated by the degree of CdCO₃(s) supersaturation which in turn depended on pH and total metal concentrations. Differences in toxicity in the laboratory tests were attributed to differences in water hardness and Cd²⁺ concentrations. Mortality of fathead minnows was low (0–10%) during the 96-h test period in the ponds due to the higher pH, which produced supersaturated conditions resulting in the rapid formation of nontoxic CdCO₃ precipitates and a more rapid decrease in Cd²⁺ concentrations as compared to the laboratory bioassays.     

Toxicity of sodium nitrilotriacetate (NTA) to the fathead minnow and an amphipod in soft water

Amphipods, Gammarus pseudolimnaeus Bousfield and fathead minnows, Pimephales promelas Rafinesque, were submitted to acute (96-h) and chronic (generation-cycle) bioassays with sodium nitrilotriacetic acid (NTA). All measurements are reported as Na₃NTA. The average 96-h TL50 values under flow-through conditions were 98 mg 1⁻¹NTA for the amphipod and 114 mg 1⁻¹ for the fathead minnow. The acute toxicity of NTA was caused in part by the high pH resulting from the addition of large amounts of NTA (> 100 mg 1⁻¹) to soft water. Controlling pH reduced the lethality of NTA by at least one-half to fathead minnow larvae. The chronic no-effect level of NTA to the amphipods was 19 mg 1⁻¹; in fathead minnows, it exceeded the highest exposure level (> 54 mg 1⁻¹).     

Reduction of aquatic toxicity of linear alkylbenzene sulfonate (LAS) by biodegradation

Partial biodegradation of LAS is shown to significantly reduce the specific toxicity (i.e. per unit weight) of the remaining LAS to Daphnia magna (water fleas) and Pimephales promelas (fathead minnows). This results from the fact that the longer homologs and more terminal isomers, which are the more toxic, are also the more rapidly degraded under bacterial action. The acute aquatic LC₅₀ of LAS may range from 0.5 to 50 mg/l depending mainly upon the chain length of the particular homolog. A high molecular weight commercial type LAS with LC₅₀ around 2 mg/l before biodegradation may show Daphnia LC₅₀'s of 30–40 mg/l. for the LAS remaining after 80–85% degradation.A further contribution to this toxicity reduction may occur if the methylene blue analytical method is used to determine the amount of LAS remaining, since some of the biodegradation intermediates show methylene blue activity but no significant toxicity. For example, sulfophenylundecanoate, a model of early intermediates, shows Daphnia and fathead lc₅₀'s 200 and 75 mg/l., respectively. Sulfophenylbutyrate, modeling somewhat later intermediates, gives lc₅₀ values around 5000–10,000 mg/l. Dialkyl tetralin/indane sulfonates (the major non-linear components in commercial LAS) exhibit 1/2–1/10 the toxicity of the corresponding LAS homologs.These results re-emphasize that analysis simply for methylene blue active substances (MBAS) gives no basis for predicting the aquatic toxicity of an environmental sample. And furthermore, that meaningful water quality criteria and standards cannot be established in terms of MBAS content while based on toxicity studies on intact, undegraded LAS.     

Effects of pH increases and sodium chloride additions on the acute toxicity of 2,4-dichlorophenol to the fathead minnow

The observable toxic effects produced by short-term exposure of fathead minnows (Pimephales promelas) to 2,4-dichlorophenol were reduced when the pH of the test water was increased by the addition of NaOH. After exposure for 192 h to 7.43 mg 2,4-dichlorophenol l^-1, the average survival of fathead minnows ranged from 28% at pH 7.57 to 100% at pH 9.08. Normal schooling behaviour was completely disrupted, and the equilibrium of most fish was affected after a 24-h exposure to 7.43 mg 2,4-dichlorophenol 1^-1 at pH 7.57, but neither schooling nor equilibrium were affected even after 192 h at pH 8.68 and 9.08. Schooling and swimming behaviour of fathead minnows exposed to 12.33 mg 2,4-dichlorophenol l^-1 were affected at all pH levels. Survival of these fish after 24 h ranged from 0% at pH 7.84–46% at pH 8.81. Sodium chloride in concentrations ranging from 0 to 13.9 mg l^-1 had no observable effects on the acute toxicity of 2,4-dichlorophenol to fathead minnows.     

Application of tracer techniques to continuous-flow toxicity testing

Frequent toxicant analyses are essential for good quality data in long-term continuous-flow tests. Due to the time consuming and costly chemical analyses, exposure levels are measured at best on a daily basis. These infrequent determinations may not detect variability in toxicant concentrations that could result in test failures. To minimize repetitive testing and improve data quality, a dye tracer method was evaluated. Rhodamine WT was selected as a toxicant tracer because of easy detectability, low toxicity to aquatic organisms, and negligible transformation in the aquatic environment.Results over a 24 h period showed that rhodamine reliably predicted the toxicant (diquat) concentrations with an r value of 0.99. Based upon these data, two replicate long-term tests with and without tracer were carried out exposing fathead minnows Pimephales promelas to diquat (1:1′-ethylene-2:2′-dipiridium dibromide). The test results indicated that the added rhodamine WT did not alter the diquat toxicity to fathead minnows using LC₅₀ and EC₅₀ values for comparisons. From these findings it is concluded that dye tracers are suitable toxicant surrogates. Their use in flow-through tests allow more frequent analyses which result in better data and minimizes experimental failure.     

Hepatic metallothionein level and mixed function oxidase activity in fingerling rainbow trout (Oncorhynchus mykiss) after acute exposure to pulp and paper mill effluents

Hepatic metallothionein (MT) levels and mixed function oxidase (MFO) activity (7-ethoxyresorufin-o-deethylase or EROD) were measured in fingerling rainbow trout (Oncorhynchus mykiss) exposed to sublethal concentrations of 12 pulp and paper effluents, after completion of 96 h static acute lethality assays. Barring one primary-treated effluent where MFO levels were significantly depressed and two secondary-treated effluents where no significant MFO induction were observed, all other effluents triggered significant induction of MT and EROD, regardless of mill process/treatment or of effluent lethality and chemical characteristics. MT and EROD inductions were significant, however, at higher concentrations for secondary-treated effluents than for primary-treated ones. Lethal (96 h LC₅₀s) to sublethal (MT and EROD lowest observable effect concentrations) ratios were variable and indicated that significant biochemical effects were present at effluent concentrations that were roughly 4–33 (MT) and 3–59 (EROD) times lower than the LC₃₀. Enzyme induction ranged from 1.3 to 2.5-fold for MT and from 1.3 to 9.4-fold for EROD compared to controls. Limited chemical data available suggest that there were indeed classes of compounds present capable of inducing MT or EROD. Observed patterns of MT/MFO responses also suggest that contaminant interactions may have interfered with induction for some of the effluents studied. Refinements of this combined (sub)lethal bioassay procedure are envisaged to determine whether it can provide an efficient means of detecting hazardous chemicals in industrial wastewaters.     

Acute toxicity tests with Daphnia magna straus and Pimephales promelas rafinesque in support of national pollutant discharge elimination permit requirements

The purpose of this study was to determine the static acute toxicity of aniline, p-chloro-m-cresol and 2(2,4,5-trichlorophenoxy) propionic acid (silvex) to daphnids (Daphnia magna) and bis(2-chloroethoxy)methane and 2-sec-butyl-4,6-dinitrophenol (dinoseb) to both daphnids and fathead minnows (Pimephales promelas). These data were needed to fulfill requirements established in the NPDES Permit (National Pollutant Discharge Elimination System) issued to the Michigan Division of The Dow Chemical Co. (Midland, Mich., U.S.A.) by the State of Michigan. Where appropriate, water quality-based effluent limitations could be recommended based on the acute toxicity data generated during this study. The results of the acute toxicity tests indicated that bis(2-chloroethoxy)methane was practically non-toxic to both daphnids and fathead minnows (LC₅₀ values of 201 and 184 mg l⁻¹, respectively); additionally, silvex was also found to be practically non-toxic to daphnids (LC₅₀ value > 140 mg l⁻¹). Aniline was highly toxic and p-chloro-m-cresol moderately toxic to daphnids, with calculated LC₅₀ values of 0.17 and 2.0 mg l⁻¹, respectively. Dinoseb was highly toxic both to daphnids and fathead minnows, with reported LC₅₀ values of 0.24 and 0.17 mg l⁻¹, respectively.     

The toxic effects of trinitrotoluene (TNT) and its primary degradation products on two species of algae and the fathead minnow

The effects of alpha trinitrotoluene (alpha TNT) and its primary degradation product (TNTcc), commonly referred to as “pink water”, were determined on members of two trophic levels. The growth responses of the algae Selenastrum capricornutum and Microcystis aeruginosa were examined through static bioassays. Death and behavioral responses of the fathead minnow (Pimephales promelas) were determined using a proportional diluter. Alpha TNT and TNTcc were both more toxic to the fathead minnow than to either species of alga. Five and 15 mg l⁻¹ alpha TNT inhibited S. capricornutum and M. aeruginosa growth, respectively. TNTcc inhibited S. capricornutum growth at concentrations above 9 mg l⁻¹; it was lethal to M. aeruginosa at 50 mg l⁻¹, but stimulated growth at lower concentrations. The 96-h lc₅₀ values based on the death response of the fathead minnow to alpha TNT and TNTcc were 2.58 and 1.60 mg l⁻¹, respectively. The 96-h ec₅₀ values based on the behavioral responses were 0.46 and 0.64 mg l⁻¹, respectively. There was no response to concentrations of 0.05 mg l⁻¹ alpha TNT and 0.07 mg l⁻¹ TNTcc.     

Effect of size or age of goldfish and fathead minnows on use of pentachlorophenol as a reference toxicant

Acute bioassays of pentachlorophenol were conducted with fathead minnows and goldfish to determine the effect of testing different sized fish of the same age or testing different aged fish. With different sized goldfish the rate of mortality during the test was similar but the threshold lc₅₀ was higher for small fish. With different sized fathead minnows the rate of mortality during the test was significantly different but the threshold lc₅₀ was the same for large and small fish. With different aged fathead minnows the rate of mortality was different for 14-week-old fish and the same for 11-, 7-, and 4-week-old fish but there was no difference in threshold lc₅₀ due to age. Size selection of fatheads or goldfish within the ranges tested is unnecessary since differences are small. Since age of fatheads did not affect the 24-h, 96-h, or threshold lc₅₀'s, use of younger fish would allow smaller bioassay chambers or more fish per chamber.     

A simple, inexpensive in situ method for assessing acute toxicity of effluents to fish

Test chambers for conducting in situ fish bioassays were constructed from 81. polyethylene bottles. Yearling fathead minnows (Pimephales promelas) and young-of-the-year bluegill (Lepomis macrochirus) demonstrated greater than 50% survival in the chambers after 65 days of exposure in a reservoir, river and creek. Fathead minnow survival was substantially greater than that of bluegils. The chambers provide a simple, inexpensive, sensitive technique to screen effluents for toxicity.     

The effects of depressed pH on flagfish reproduction, growth and survival

Breeding communities of flagfish, Jordanella floridae, were exposed to northern Ontario lake water (hardness 28 mg l⁻¹ CaCo₃) adjusted to depressed pH levels of 6.0, 5.5, 5.0 and 4.5. Control water (pH 6.8) received no acid treatment. Egg production, egg fertility and fry growth was impaired (P < 0.05) at all exposure levels. Flagfish fry survival was reduced (P < 0.05) at pH 5.5 and 5.0 and no fry survived at pH 4.5. Variability of hatching in all treatments precluded any identifiable hatching response to depressed pH. Reduction in the reproductive processes monitored indicated the following order of sensitivity: egg production > fry survival > fry growth > egg fertility.Results of this study coincide with reproductive investigations on brook trout and fathead minnows indicating the “no effect” level of pH depression for successful reproduction to be pH 6.5.     

The residual oxygen bioassay: A rapid procedure to predict effluent toxicity to rainbow trout

A rapid bioassay procedure is described which for the toxic effluents tested, is sensitive to concentrations in the 96-h lc₅₀ range. The procedure requires less than eight hours to complete and is based on the consumption of available oxygen by fish in sealed containers.The procedure was evaluated using sodium pentachlorophenate, 3 kraft pulp mill bleach plant effluents and a chloralkali plant waste. The threshold concentrations obtained from the rapid procedure were directly comparable to 96-h lc₅₀ values obtained by standard methods. For replicate tests with sodium pentachlorophenate, no significant difference (p < 0.05) between means of data obtained by the two bioassay procedures could be demonstrated, using the Student “t”-test.     

Copper lethality to rainbow trout in waters of various hardness and pH

The 96 h median lethal concentration (LC₅₀) of total dissolved copper varied from 20 μg 1⁻¹ in soft acid water to 520 μg l⁻¹ in hard alkaline water, in tests with hardness ranging from 30 to 360 mg l⁻¹ as CaCO₃ and pH from 5 to 9. The 3-dimensional response surface was complex, although an increase in hardness usually made copper less toxic. A good prediction of copper LC₅₀ at usual combinations of hardness and pH was given by the equation: LC₅₀ = antilog (1.933 + 0.0592 P_T + 0.4912 H_T + 0.4035 P_TH_T + 0.4813 P²_T + 0.1403 H²_TThe transformed variables are and A somewhat less accurate equation is provided for extreme combinations of hardness and pH.Trout of 10 g weight were 2.5 times more resistant than 0.7 g trout. Effect of size was apparently the same at different combinations of hardness and pH, and was predictable by an equation of the form LC₅₀ = Constant × Weight ^0.348.Ionic copper (Cu²⁺) and two ionized hydroxides (CuOH⁺ and Cu₂OH²⁺₂) seemed to be the toxic species of copper, since they yielded the smoothest response surface with the best fit to measured LC₅₀'s. The sum of these ions produced LC₅₀'s ranging from 0.09 μg l⁻¹ copper in soft alkaline water to 230 μg l⁻¹ in hard acid water. The ions were different in relative toxicity, or became more toxic at high pH, or both.     

Factors influencing acute toxicity estimates of hydrogen sulfide to freshwater invertebrates

Acute bioassay tests of hydrogen sulfide were run on Assellus militaris Hay, Crangonyx richmondensis laurentianus Bousfield. Gammarus pseudolimnacus Bousfield. Bactis vagans McDonough. Ephemera simulans Walker and Hexagenia limbata (Serville). Size and type of test chamber, type of substrate for barrowing forms or those seeking shelter in gravel, oxygen concentration, pH, and season of collection influenced the sensitivity of organisms. Hydrogen sulfide exposure at sublethal levels reduced feeding activity of Gammarus. Data indicate that test conditions should approximate natural habitat conditions as closely as practical. The most acceptable 96-h LC₅₀ hydrogen sulfide concentrations for the various species are: Assellus 1.07 mg 1⁻¹Crangonyx 0.84 mg 1⁻¹. Gammarus 0.059 mg 1⁻¹. Baetis 0.020 mg 1⁻¹. Ephemera 0.316 mg 1⁻¹, and Hexagenia, 0.111 mg 1⁻¹. Chronic exposure tests now in progress suggest that the no-effect levels are 8–12 per cent of the 96-h LC₅₀.     

Disruption of precopulatory behavior in the amphipod Anisogammarus pugettensis upon exposure to bleached kraft pulpmill effluent

Marine amphipods in precopula, Anisogammarus pugettensis (Dana), were used in static 117 h bioassays with neutralized, filtered, bleached kraft pulp mill effluent (BKME) and daily solution replacement. The effluent interfered with precopula behavior; high concentrations (40% of full strength BKME) resulted in rapid release of paired amphipods. A concentration-related time to 50% release of paired individuals was observed. For one batch of BKME, the behavioral threshold concentration (EC₅₀) was estimated at 10–15% of full strength BKME. The estimated 96 h LC₅₀ for underyearling coho salmon (Oncorhynchus kisutch) for the same effluent tested in freshwater was 36% BKME—illustrating a greater sensitivity of the amphipod behavioral bioassay in comparison to the salmonid acute lethal toxicity test. The amphipod precopula bioassay for toxicity testing has some merit in that it is moderately quantitative under given test conditions and does not require elaborate equipment.     

Acute toxicity and behavioral responses of coho salmon (Oncorhynchus kisutch) and shiner perch (Cymatogaster aggregata) to chlorine in heated sea-water

The acute toxicity and behavioral response to chlorinated and heated sea-water was determined for coho salmon smolts and 1–3 month old shiner perch. LC₅₀'s were determined for 7.5, 15, 30 and 60 min exposure times; 13, 16 and 20°C (Δt = 0, 3, 7°C) temperatures and total residual oxidant (TRO) concentrations ranging from 0.077 to 1.035 mg l⁻¹. The mean 60 min LC₅₀ for shiner perch was significantly reduced (P ≤ 0.05) from 308 μg l⁻¹ TRO at 13°C to 230 μg l⁻¹ TRO at 20°C. The 60 min LC₅₀ for coho salmon decreased from 208 μg l⁻¹ TRO at 13°C to 130 μg l⁻¹ at 20°C. The LC₅₀'s for coho salmon in chlorinated sea-water averaged 55% of those for shiner perch. The relationship between TRO concentration, exposure time, and percent survival in chlorinated sea-water at 13°C is presented for both species.A significant (P ≤ 0.01) avoidance threshold for coho salmon occurred at 2 μg l⁻¹ TRO and was reinforced with increasing temperature. A significant (P ≤ 0.01) avoidance threshold for shiner perch occurred at 175 μg l⁻¹ TRO, while a significant preference (P ≤ 0.05 or 0.01) response at 16°C and 20°C occurred at 10, 25, 50 and 100 μg l⁻¹ TRO. The ecological implications of the toxicity tests and the behavioral responses are discussed.     

Toxicity, uptake and survey studies of boron in the marine environment

Little is known about the toxic and bioaccumulative dangers to aquatic life posed by borate discharge recently initiated by coastal British Columbia groundwood pulp mills. Bioassays with sodium metaborate and underyearling coho salmon (Onchorhynchus kisutch) in fresh water yielded a 283-h lc₅₀ of 113 μg ml⁻¹ (104, 123 = 95% confidence limits). Toxicity to underyearling coho in sea water appeared considerably greater with a 283 h lc₅₀ of 12.2 μg ml⁻¹ (10.89, 14.56 = 95% confidence limits). The disparity between fresh and saltwater boron toxicity to coho is not understood at this time. In salmonids, boron enters the tissues slowly, necessitating prolonged bioassay tests. Sockeye salmon (O. nerka) and juvenile oysters (Crassostrea gigas) exposed to sublethal doses of boron take up boron roughly in relation to its availability. Oysters show no bioaccumulative potential or prolonged retention of boron following cessation of dosage. Field surveys conducted before and after industrial borate emission confirm the lack of evidence for tissue bioaccumulation. Results of a survey of boron levels in receiving waters are reported. No hazard to salmonids of oysters at the present level of industrial discharge of boron (≤1 μg B ml⁻¹) is apparent from this work.     

The acute toxicity of vanadium and copper to eyed eggs of rainbow trout (Salmo gairdneri)

The effects of vanadium (25–595 mg l⁻¹) and of copper (0.03–4.78 mg l⁻¹) on embryonic survival and hatching of eyed eggs of rainbow trout, Salmo gairdneri, were investigated. Copper was approx. 300-fold more toxic than vanadium (96-h LC₅₀ = 0.4 and 118 mg l⁻¹, respectively) but had little effect on the timing of hatch. Vanadium induced premature hatching of eyed eggs at concentrations from 44 to 595 mg l⁻¹. Concentrations of copper required to produce lethality in eyed eggs were similar to concentrations required to produce mortality in juveniles. Vanadium concentrations approx. 15 times higher were required to produce mortality in eyed eggs than in juveniles. Therefore, acute exposure of eyed rainbow trout eggs to vanadium is not a sensitive toxicity test for use in establishing water quality criteria or maximum acceptable toxicant concentrations.