Short-term Toxicity Tests Research Articles

Bioassay methods for evaluating the toxicity of heavy metals, biocides and sewage effluent using microscopic stages of giant kelp Macrocystis pyrifera (Agardh): A preliminary report

Methods for using early life stages of giant kelp Macrocystis pyrifera in toxicity tests intended for evaluating the toxicity of municipal sewage effluents are given. Included are methods for both a short-term (48-h) toxicity test, and a long-term (16-day) toxicity test. Preliminary results of experiments with zinc sulfate, sodium pentachlorophenate, and a primary-treated sewage effluent showed that early life stages of Macrocystis have a variable sensitivity to these toxicants. No Observed Effect Concentrations (NOECs) for effects of zinc sulfate on germination of Macrocystis zoospores ranged from 1730 μg/liter to 5500 μg/liter in 3 separate 48-h experiments. The NOECs for zinc effects on the growth of Macrocystis zoospore germination tubes ranged from 550 to 1090 μg/liter in three 48-h experiments. The NOEC for zinc effects on sporophyte production by Macrocystis was 1070 μg/liter in a 16-day experiment. The fungicide and herbicide sodium pentachlorophenate significantly inhibited Macrocystis zoospore germination, and gametophyte reproduction at concentrations of 32 μg/liter and greater (NOEC < 32 μg/liter for both tests), and was more toxic to kelp than zinc. Primary-treated-sewage effluent significantly inhibited zoospore germination at concentrations greater than 1% effluent (NOEC = 0·56% effluent). The results of these preliminary experiments indicate that early life stages of Macrocystis pyrifera are amenable to evaluating the toxicity of a variety of toxicants, including sewage effluents, and that Macrocystis has a variable sensitivity to toxicants depending on the nature of the toxicant and the endpoint being tested.

Critical analysis of factors affecting the sensitivity of zooplankton and the reproducibility of toxicity test results

This paper concerns the variables that affect the sensitivity of test zooplankton and the reproducibility of results obtained from repeated toxicity tests. Among the variables considered are: (a) the nutrition of the test zooplankton, their health and diet; (b) culturing techniques, in particular, the effect of the ambient medium on demographic variables and testing results; (c) physical and chemical characteristics of the test compound; (d) purity of the test compound, and finally (e) the variation in results obtained with nominal and measured concentrations of the test compound. Critical factors addressed include the effects of inadequate cobalamine or selenium concentrations in the diet, adversely affecting fertility, which may result in misleading interpretation of short-term toxicity test results involving Ceriodaphnia or chronic studies using Daphnia. Other factors considered are, for example, the problems of poor health, as measured by fertility or by the reproducibility of static acute toxicity test results, and their effect on zooplankton sensitivity to toxicants. The observation that diets of several algae result in greater tolerance of zooplankton to toxicants than do diets based on synthetic food are discussed. It is also noted that organisms reared in clean, frequently replenished and renewed habitats and fed axenic cultures of algae, are healthier than animals maintained in static environments. The observation that the results of interlaboratory comparisons of zooplankton response to toxicants are more uniform and exhibit smaller standard deviations when the test animals are sustained on algal foods rather than on synthetic diets is emphasized. Variability due to test compound characteristics include the observation that test compounds of variable purity often are responsible for poor test results reproducibility, and that the reproducibility of the toxic response is often variable due to physical and chemical characteristics of the test chemical. Finally, poor interlaboratory comparisons may result when nominal concentrations are the only ones considered. Data are selected from the published literature to illustrate the above discussion.

Enzymatic methylation of sulfide, selenide, and organic thiols by Tetrahymena thermophila.

Cell extracts from the ciliate Tetrahymena thermophila catalyzed the S-adenosylmethionine-dependent methylation of sulfide. The product of the reaction, methanethiol, was detected by a radiometric assay and by a gas-chromatographic assay coupled to a sulfur-selective chemiluminescence detector. Extracts also catalyzed the methylation of selenide, and the product was shown by gas chromatography-mass spectrometry to be methaneselenol. The sulfide and selenide methyltransferase activities copurified with the aromatic thiol methyltransferase previously characterized from this organism (A.-M. Drotar and R. Fall, Pestic. Biochem. Physiol. 25:396-406, 1986), but heat inactivation experiments suggested the involvement of distinct sulfide and selenide methyltransferases. Short-term toxicity tests were carried out for sulfide, selenide, and their methylated derivatives; the monomethylated forms were somewhat more toxic than the nonmethylated or dimethylated compounds. Cell suspensions of T. thermophila exposed to sulfide, methanethiol, or their selenium analogs emitted methylated derivatives into the headspace. These results suggest that this freshwater protozoan is capable of the stepwise methylation of sulfide and selenide, leading to the release of volatile methylated sulfur or selenium gases.

Short-term developmental toxicity testing: considerations in the validation process.

The development and validation of short-term tests that assess developmental toxicity have received increased attention due to the large number of agents that need to be tested and the desirability to reduce the overall use of animals in research and safety evaluation. The Short-Term In Vivo Reproductive Toxicity Test was developed to help meet this need and to provide a method for prioritizing agents for further testing. This report provides a brief overview of the risk assessment process and a focus on the aspects of test validation that should be considered when evaluating short-term tests. It is intended to give the reader an appreciation for many of the considerations that must go into developing and validating a short-term test, and to indicate areas within the risk assessment process where inclusion of such a test may be appropriate.

Evaluation of metabolic responses of Artemia salina to oil and oil dispersant as a potential indicator of toxicant stress.

Insight into the impact of a pollutant on marine organisms may be gained from four types of biological and chemical investigations: a) short-term acute toxicity tests,b)organism-environment transfer studies,c)physiological studies and d)field studies.In the case of oil pollution,field studies have limited value for the estimation of the impact of oil on natural communities since results are often confused with the effects of "detergents"used in cleanup processes. Both types of laboratory studies(acute toxicity and sublethal toxicity tests)provide valuable information for the toxicity of oils and oil dispersants.Since in the case of oil pollution treatment, oils and detergents are acting in combination,a realistic approach of laboratory studies must also include the combined action of these substances on marine organisms.Although acute toxicity studies which measure the mortality due to a toxicant are very useful for the determination of the range of animal tolerance,other effects causing physiological alterations may be detrimental to a population's survival. This paper concerns research on the effects of an oil,an oil dispersant and of the mixture of oil and dispersant on a physiological process;the respiration of the brine shrimp Artemia salina. MATERIAL AND METHODS Adults of Artemia hatched from commercially available cysts(New Technology Artemia salina Revolution)were used as test animals.The hatching of the cysts and rearing of Artemia was performed in constant temperature rooms at 22+0,5~ in synthetic sea water (Synthetica). The three toxic solutions tested were a crude oil(tunesian crude oil zarzaitine type),anoil dispersant(Finasol-OSR2)and their mixture. The type of oil/water mixture tested was oil-water dispersion(OWD).

Aquatic toxicological aspects of dithiocarbamates and related compounds. I. Short-term toxicity tests

Short-term toxicity of 26 dithiocarbamates (DCs) and related compounds was determined in tests with guppies (Poecilia reticulata), water fleas (Daphnia magna), green algae (Chlorella pyrenoidosa) and bacteria (Photobacterium phosphoreum). A screening test for the inhibition of nitrification was also conducted. It was found that both ethylenebisdithiocarbamates and dialkyldithiocarbamates were toxic to the organisms tested. Experiments were carried out to determine the n-octanol/water partition coefficient of a number of substances. In the case of DCs and their degradation products (metal derivatives were not considered) it was possible to draw up quantitative structure-activity relationships for the various test species. Various aspects of tests conducted on these fungicides and their degradation products (hydrolysis, biodegradation, polymerization and volatilization) are discussed, as are a number of biochemical mechanisms of toxicity. It is concluded that DCs are cytotoxic substances and therefore must be regarded as broad-spectrum biocides.

A new fathead minnow (Pimephales promelas) subchronic toxicity test

AbstractThe most commonly used test for evaluating the toxicity of effluents to fish and invertebrates has been the acute lethality test. However, effluents frequently are not acutely toxic, and so assessing the sublethal effects on fish and invertebrates is important. Described herein is a rapid method to estimate the chronic toxicity of effluents to the fathead minnow (Pimephales promelas). Tests are initiated using newly hatched minnow larvae and are run for 7 d under static‐renewal conditions. It is a cost‐effective, simple, short‐term toxicity test, and survival and growth are the measures of effects. Approximately 70 tests using this technique have been run with various types of effluents. Included in this article are results from representative effluent tests and also from tests conducted using Dursban, zinc and copper. The results indicate that this subchronic toxicity test gives a good estimate of chronic toxicity.

Microtox and Spirillum volutans tests for assessing toxicity of environmental samples.

Two short-term bacterial toxicity tests that have received a great deal of attention during the last few years are the Microtox and Spirillum volutans assays. The former involves monitoring changes in the light output from luminescent bacteria (Photobacterium phosphoreum) in a temperature controlled photometer when exposed to various concentrations of toxic substances. In the S. volutans assay, motility patterns of this large aquatic bacterium are used as the test endpoints. As part of the authors' program in assembling and establishing a battery of short-term bacterial toxicity tests, they examined the use of Microtox and S. volutans for determining the toxicity of a wide variety of environmental samples. Samples examined included potable and surface waters, industrial effluents, soil/sludge extracts and landfill leachates. Some of the results are presented in this paper.

Comparative toxicity of copper and acridine to fish, Daphnia and algae

AbstractA comparison was made of the sensitivity of fish, Daphnia and algae to the toxic effects of copper and acridine. A series of toxicity tests was conducted with these organisms, and the following biological endpoints determined: LC50s for fish, LC50s and effects on reproduction of Daphnia and 50% inhibition of the growth rate of algae. The 96‐h LC50s for bluegill (Lepomis macrochirus) and Daphnia magna exposed to copper were 2.2 and 0.13 mg/L, respectively. A chronic exposure to 0.03 mg/L of copper for 14 d significantly decreased reproduction in Daphnia. Exposure to 0.4 and 0.2 mg/L copper inhibited the growth rate of Selenastrum capricornutum and Chlorella vulgaris, respectively, by 50%. The 96‐h LC50s for fathead minnows (Pimephales promelas) and D. magna exposed to acridine were 2.3 and 3.1 mg/L, respectively. A chronic exposure to 1.25 mg/L acridine for 14 d significantly inhibited reproduction in Daphnia, and an exposure to 0.9 mg/L inhibited the growth rate of S. capricornutum by 50%. Based on the biological endpoints determined in these tests, Daphnia were more sensitive to copper than were fish or algae. In contrast, the most sensitive biological endpoint in tests with acridine was the inhibition of algal growth. Comparison of these test results indicates that short‐term toxicity tests used for screening toxicants for possible environmental effects should include both plant and animal species.

Lethal and sublethal effects of marine sediment extracts on fish cells and chromosomes

The cost of conducting conventional chronic bioassays with every potentially toxic compound found in marine ecosystems is prohibitive; therefore short-term toxicity tests which can be used for rapid screening were developed. The tests employ cultured fish cells to measure lethal, sublethal or genotoxic effects of pure compounds and complex mixtures. The sensitivity of these tests has been proven under laboratory conditions; the following study used two of these tests, the anaphase aberration test and a cytotoxicity assay, under field conditions. Sediment was collected from 97 stations within Puget Sound, Washington. Serial washings of the sediment in methanol and dichloromethane yielded an organic extract which was dried, dissolved in DMSO and incubated as a series of dilutions with rainbow trout gonad (RTG-2) cells. The toxic effects of the extract were measured by examining the rate of cell proliferation and the percentage of damaged anaphase figures. Anaphase figures were considered to be abnormal if they exhibited non-disjunctions, chromosome fragments, or chromosome bridges. A second cell line (bluegill fry, BF-2) was also tested for cell proliferation and was included because, unlike the RTG-2 cell line, it contains little or no mixed function oxygenase activity. Of 97 stations tested, 35 showed no genotoxic activity, 42 showed high genotoxic activity (P≤.01) and the remainder were intermediate. Among the toxic sites were several deep water stations adjacent to municipal sewage outfalls and four urban waterways contaminated by industrial and municipal effluents. Extracts from areas that showed genotoxic effects also inhibited cell proliferation and were cytotoxic to RTG-2 cells. Few effects were noted in the MFO deficient BF-2 cells. Short term in vitro tests provide aquatic toxicologists with a versatile and cost effective tool for screening complex environments. Through these tests one can identify compounds or geographic regions that exhibit high cytotoxic or genotoxic potential.

Improved methods for harvesting and counting synchronous populations of Artemia nauplii for use in developmental toxicology

Artemia nauplli have, within recent years, gained popularity as a test organism for short-term toxicity testing. Because nauplii exhibit rapid development and growth within 48 hr after hatch, their potential as a model organism for teratology screening has been considered. To do this, synchronous populations of nauplii at different developmental intervals must be available. A dual-chamber hatching vessel which enables multiple harvesting of synchronous populations of nauplii from one sample of prepared cysts has been developed. An accumulation period of 2 hr defines the synchronous population and produces around 1000 nauplii among five hatching vessels. The body lengths of nauplii harvested 2 to 9 hr after the initial harvest were equivalent to and lower than that of animals incubated at the hatching temperature (31 ± 1°C); e.g., the body length of freshly harvested nauplii at 9 hr was 473 ± 6.2 μm (±SE) and for nauplii incubated for 8 to 10 hr, the body lengths were 643 ± 10.7 and 702 ± 10.9 μm, respectively. A counting method was developed that enabled precise counting up to 400 nauplii in suspension and distinguishing dead from live nauplii. Measurements of body length, body water volume, and whole animal DNA and protein of nauplii harvested sequentially from the same of sample prepared cysts indicated that in addition to temporal differences in hatching, the nauplii may differ both physically and chemically.

Studies on the toxicity of tribromoethene, cyclohexene and bromocyclohexane to different freshwater organisms

In order to get an impression about the possible environmental effects of tribromethene. cyclohexene and bromocyclohexane, short-term toxicity tests were carried out with green algae ( Chlorella pyrenoidosa), crustaceans ( Daphnia magna) and fishes ( Poecilia reticulata) and long-term toxicity tests with Daphnia magna. The LC(EC) 50 values from the short-term tests with tribromoethene, cyclohexene and bromocyclohexane were 3.5–7.2, 3.8–5.3 and 2.2–2.5 mg 1 − respectively, whereas the lowest NOEC (No Observed Effect Concentration)-values from these tests were 0.52, 0.22 and 1.1 mg 1 −1 respectively; based on the criterion reproduction in the long-term tests the NOEC-values were 3.1, 2.4 and 0.5 mg 1 −1 respectively; All these values were corrected for the actual concentrations during the experiments and the tests were carried out in completely closed systems: in open systems the concentration of the test compounds decreased more than 90% within 48 h. Based on the log p (octanol-water partition coefficient) the calculated bio-accumulation factor value for tribromoethene and bromocyclohexane was 160, and for cyclohexene was 16. As for water quality criteria, values of 0.2 mg 1 −1 for tribromoethene, 0.01 mg 1 −1 for cyclohexene and 0.4 mg 1 −1 for tribromoethene 0.01 mg 1 −1 for cyclohexene and 0.4 mg 1 −1 for bromocyclohexane are proposed.

In vitro antifungal activity of eugenol and vanillin against Candida albicans and Cryptococcus neoformans.

Eugenol and vanillin were examined for in vitro antifungal activity against the medically important yeasts, Candida albicans and Cryptococcus neoformans. Minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) were determined for each compound against 31 strains of C. albicans and 33 strains of C. neoformans. With eugenol, the mean MIC's for C. albicans and C. neoformans were 625 and 293 micrograms/mL, respectively, while the mean MFC's were 1209 and 521 micrograms/mL, respectively. With vanillin, the mean MIC's for C. albicans and C. neoformans were 1250 and 738 micrograms/mL, respectively, while the mean MFC's were 5000 and 1761 micrograms/mL, respectively. With C. albicans, inhibition and retardation of growth were similar for yeast and mycelial forms, but germ tube formation was inhibited at concentrations lower than those which inhibited growth. Short-term toxicity tests with mice using the intraperitoneal route gave maximum tolerated doses of 62.5 mg/kg for eugenol and 125 mg/kg for vanillin and excluded their use as therapeutic agents for systemic mycoses.

Effects of hexavalent chromium in rainbow trout ( Salmo gairdneri) after prolonged exposure at two different pH levels

Toxic effects were studied in rainbow trout ( Salmo gairdneri) exposed to Na 2CrO 4 solutions of different concentrations (0.02, 0.2, and 2.0 mg/liter Cr) and pH (7.8 and 6.5). The study involved embryo-through-juvenile and alevin-through-juvenile exposure for 32 weeks and exposure of yearling trout for 1, 3, 6, and 12 weeks. The experiments were started at the same time of the year and were carried out concurrently. When survival was used as the criterion, Cr was more toxic at pH 6.5 than at pH 7.8 in all life stages studied. For trout in the embryo-through-juvenile exposure study the lowest concentrations inducing an increase in mortality were 0.2 mg/liter Cr at pH 6.5 and 2.0 mg/liter Cr at pH 7.8. Embryo hatchability was only affected at the highest exposure concentration at a pH of 6.5. No growth retardation was detected at the termination of the experiments. In addition to survival and growth, biochemical, hematological, and histological changes were studied as indicators of toxicity in yearling trout. These parameters also showed that fish were more susceptible to Cr at the lower pH. The observed effects are discussed in relation to previously reported findings in short-term toxicity tests.

Substitutes for phosphate containing washing products: Their toxicity and biodegradability in the aquatic environment

The toxicity of nitrolotriacetic acid (NTA), citrex-S-5, sodium alluminium silicate (NAS), carboxymethyloxysuccinate (CMOS), linear alkylbenzene sulfonate (LAS) and soap was examined using bacteria ( Pseudomonas fluorescens ), algae ( Chlorella vulgaris and Microcystis aeruginosa ), crustaceans ( Daphnia magna ), insects ( Aedes aegypti ), fishes ( Poecilia reticulata and Oryzias latipes ) and amphibians ( Xenopus laevis ). The biodegradability of NTA, citrex-S-5, CMOS and LAS was studied in the OECD-test. The acute toxicity of the compounds differed largely with the organisms tested (up to a factor 1,000). Based on the results of the short-term toxicity tests (with all test organisms) and the long-term toxicity tests (with Daphnia magna and in case of LAS also with Poecilia reticulata ) the following no-observed-effect concentrations (NOEC) could be derived: LAS 3.2 mg/1; NAS and soap 10 mg/1; CMOS 32 mg/1; NTA 100 mg/1 and citrex-S-5 320 mg/1. However, testing different samples of one test compound may result in marked differences in toxicity (e.g. NAS: a factor 32). All compounds tested showed to be biodegradable as the percentage DOC decreased more than 80% within four weeks. CMOS biodegraded quickly, whereas citrex-S-5 and LAS showed a somewhat slower rate of degradation. In contrast to the other compounds NTA required an adaptation period of 1 week.

Proposal for a short-term toxicity test with Artemia nauplii

Although standardization of toxicity tests on aquatic organisms is an urgent necessity, very little has yet been achieved for the marine environment. As a first step in this direction, a simple, inexpensive and reliable short-term routine test with Artemia larvae is proposed. This test is the result of an extensive study in our laboratory, taking into account the advantages and disadvantages of Artemia bioassays published by various authors. The major advantage of the brine shrimp as a test species is its continuous availability under the form of dry cysts which can be hatched very easily; this eliminates all biological, technological, and financial problems of stock recruitment and/or culturing. The acute test presented is based on the determination of the LC 50-24 hr of instar II–III nauplii of a specific Artemia strain. Presently this test is the subject of an intercalibration exercise in North America; an analogous exercise is now in progress in the European Economic Community countries.

CAN A BATTERY OF SHORT‐TERM TOXICITY TESTS PREDICT CHRONIC TOXICOLOGIC EFFECTS? WORKSHOP PAPER

Annals of the New York Academy of SciencesVolume 329, Issue 1 p. 131-136 CAN A BATTERY OF SHORT-TERM TOXICITY TESTS PREDICT CHRONIC TOXICOLOGIC EFFECTS? WORKSHOP PAPER Mary Henry, Mary Henry United Slates Army Medical Bioengineering Research and Development Laboratory Fort Detrick Frederick, Maryland 21701Search for more papers by this author Mary Henry, Mary Henry United Slates Army Medical Bioengineering Research and Development Laboratory Fort Detrick Frederick, Maryland 21701Search for more papers by this author First published: October 1979 https://doi.org/10.1111/j.1749-6632.1979.tb15340.xCitations: 1AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume329, Issue1Public Control of Environmenal Health HazardsOctober 1979Pages 131-136 RelatedInformation

Reproducibility of short-term and reproduction toxicity experiments with Daphnia magna and comparison of the sensitivity of Daphnia magna with Daphnia pulex and Daphnia cucullata in short-term experiments

Short-term toxicity tests with Daphnia magna are reasonably reproducible. The sensitivity of Daphnia pulex, Daphnia cucullata and Daphnia magna proved to be about the same in short-term tests with 15 different chemical compounds.

A strategy for the prediction and detection of effects of pollution on natural communities

The author describes experimental and observational approaches to establishing standards for pollutants and stresses the need for the use of more realistic experimental models than, for example, the simple short-term toxicity test and for more sensitive procedures to detect and analyse environmental change and damage than the simple classification systems and indices so widely used in the past.

Acute and chronic toxicity of furylfuramide in rats and mice.

As a model experiment on the study of toxicity of a compound, acute, short-term and long-term toxicity tests on Furylfuramide (2-(2-Furyl)-3-(5-nitro-2-8- furyl) acryl amide) were performed using Nitrofurazone (5-nitro-2-furaldehyde semicarbazone) and sorbic acid as control compounds. As the most conspicuous changes, hypertrophy of liver cells with clear cytoplasm was noted within a week of feeding. These cells were first located around the central vein and increased in number with time toward the peripheral portion of the lobule. In spite of the long-term experiments up to two years in rats and mice, no progressive lesion of the liver followed by the hypertrophy. No cumulative effect of Furylfuramide was noted in any organ. The hypertrophy of liver cells was reversible, when animals were switched to basal diet and kept on it for a certain length of time. A concentration of 0.125% in the diet was considered to be the tolerable safe dose of Furylfuramide. High incidence of spontaneous tumors developed in mice fed on Furylfuramide and control diet for two years was described. There was no difference in incidences between groups.