Do frogs really eat cardamom? Understanding the myth of crop damage by amphibians in the Western Ghats,India

In the Western Ghats of India, amphibians are culled at cardamom plantations since they are perceived to consume cardamom. To better understand the relationship between amphibians and cardamom, a study was undertaken at these plantations, which harbor numerous threatened and range-restricted amphibians. We undertook questionnaire surveys with 298 respondents at 148 plantations across southern India. Time-activity budget and diet analysis surveys were undertaken to determine whether amphibians really consumed cardamom. The conception that amphibians eat cardamom was found to be widespread especially among small-sized plantations, leading to negative perceptions and a lack of interest in amphibian conservation. The plantation community perceives a substantial economic loss due to amphibians, even though this is non-existent as revealed by our field surveys. These perceptions would lead to a continued intolerance of amphibian presence in plantations. A suitable outreach initiative re-affirming facts and spreading awareness on the positive role of amphibians would need to be conducted to negate this age-old myth.     

The interactive effects of low pH, toxic metals, and DOC on a simulated temporary pond community

The effects of pH, potentially toxic metals, and dissolved organic carbon (DOC) on a freshwater, palustrine wetland assemblage (three amphibian species, one mosquito genus, and 18 genera of algae) were examined in 500 liter outdoor mesocosms. The design was fully factorial and exposure levels were set to represent concentrations that occur naturally in temporary ponds in central Pennsylvania, an acid-impacted region of the USA. Water chemistry was precisely regulated by use of an artificial pond water rather than using stream water or well water with partially unknown composition. Jefferson (Ambystoma jeffersonianum) and spotted salamanders (A. maculatum) were negatively impacted by low pH and high metal concentrations. Wood frogs (Rana sylvatica) were negatively impacted by high DOC and low pH. These negative effects included increased time to metamorphosis and decreased survival and wet body mass at metamorphosis. Mosquito emergence was unaffected by pH, metals, or DOC. All three abiotic factors (metals, pH, and DOC) interacted to determine algal taxonomic composition. Hence, abiotic conditions may determine amphibian reproductive success both directly and indirectly, via effects on algae, since a primary food source of salamanders, zooplankton, are affected by changes in algal composition. These complex interactions should be considered in evaluating the possible toxicity of pond waters to fauna and flora. Until now, pH alone has been considered the primary limiting factor for amphibians breeding in temporary ponds.     

Metal bioavailability and toxicity to fish in low-alkalinity lakes: A critical review

Fish in low-alkalinity lakes having pH of 6.0-6.5 or less often have higher body or tissue burdens of mercury, cadmium, and lead than do fish in nearby lakes with higher pH. The greater bioaccumulation of these metals in such waters seems to result partly from the greater aqueous abundances of biologically available forms (CH(3) Hg(+), Cd(2+), and Pb(2+)) at low pH. In addition, the low concentrations of aqueous calcium in low-alkalinity lakes increase the permeability of biological membranes to these metals, which in fish may cause greater uptake from both water and food. Fish exposed to aqueous inorganic aluminum in the laboratory and field accumulate the metal in and on the epithelial cells of the gills; however, there is little accumulation of aluminum in the blood or internal organs. In low-pH water, both sublethal and lethal toxicity of aluminum has been clearly demonstrated in both laboratory and field studies at environmental concentrations. In contrast, recently measured aqueous concentrations of total mercury, methylmercury, cadmium, and lead in low-alkalinity lakes are much lower than the aqueous concentrations known to cause acute or chronic toxicity in fish, although the vast majority of toxicological research has involved waters with much higher ionic strength than that in low-alkalinity lakes. Additional work with fish is needed to better assess (1) the toxicity of aqueous metals in low-alkalinity waters, and (2) the toxicological significance of dietary methylmercury and cadmium.     

Toxicity of road salt to Nova Scotia amphibians

The deposition of chemical pollutants into roadside wetlands from runoff is a current environmental concern. In northern latitudes, a major pollutant in runoff water is salt (NaCl), used as de-icing agents. In this study, 26 roadside ponds were surveyed for amphibian species richness and chloride concentration. Acute toxicity tests (LC₅₀) were performed on five locally common amphibian species using a range of environmentally significant NaCl concentrations. Field surveys indicated that spotted salamanders (Ambystoma maculatum) and wood frogs (Rana sylvatica) did not occupy high chloride ponds. American toads (Bufo americanus) showed no pond preference based on chloride concentration. Acute toxicity tests showed spotted salamanders and wood frogs were most sensitive to chloride, and American toads were the least. Spring peepers (Pseudacris crucifer) and green frogs (Rana clamitans) showed intermediate sensitivities. We concluded that chloride concentrations in ponds due to application of de-icing salts, influenced community structure by excluding salt intolerant species.     

The effect of soil composition and hydration on the bioavailability and toxicity of cadmium to hibernating juvenile American toads (Bufo americanus)

The soil ecotoxicology literature has focused primarily on a few major taxa, to the neglect of other fossorial organisms such as amphibians. We selected cadmium (Cd) and the American toad (Bufo americanus) as a model contaminant and biological species to assess the impact of soil contamination on amphibian hibernation survival and post-hibernation condition. Soil sand composition (50, 70, 90%) and hydration (100, 150% water holding capacity (WHC)) were manipulated in addition to Cd concentration (0, 56, 165, 483 microg/g) to determine whether these soil properties affect toxicity. Soil Cd concentration significantly reduced survival and locomotor performance, and was correlated negatively with percent mass loss and positively with whole body Cd concentration. Higher sand content resulted in less mass loss and greater Cd uptake. Toads that were hibernated in 50% sand hydrated to 100% WHC had higher survival, less mass loss, and better sprint performance than those hibernated in 50% sand, 150% WHC. This study demonstrates that concentrations of Cd found in soil at highly contaminated sites can be bioaccumulated by hibernating amphibians and may reduce fitness. Differences in microhabitat use may cause species to vary in their exposure and susceptibility to soil contamination. The toxicity of Cd to amphibians could be greater in natural systems where there are multiple stressors and fluctuations in environmental variables.     

Assessing the cause of impacts on benthic organisms near Rouyn-Noranda, Quebec

Sediments from lakes near Rouyn-Noranda, Quebec, contain elevated concentrations of several metals, including Cd, Cu, Pb and Zn. Amphipods, fingernail clams, mayflies and tanytarsid midges were absent, and sediment toxicity was observed in chronic tests with Hyalella in sediments from Lac Dufault, the lake closest to Rouyn-Noranda. Bioaccumulation by Hyalella demonstrated elevated bioavailability of Cd, Co, Cr, Pb and Tl, but only Cd was accumulated to levels close to the toxic threshold. Copper, which is regulated by Hyalella, was not elevated in these amphipods, but it was elevated in overlying water in the toxicity tests. Toxic effects in Lac Dufault sediments are probably caused primarily by Cd, at least in amphipods, with a possible minor contribution from Cu. An integrated assessment, including sediment chemistry, benthic community composition, sediment toxicity, metal bioaccumulation in benthos, and comparison of bioaccumulation and/or overlying water concentrations with threshold effect concentrations, provides the best indication of effects and their cause.     

Species-specific responses to metals in organically enriched river water, with emphasis on effects of humic acids

Invertebrate communities in polluted rivers are often exposed to a wide variety of compounds. Due to complex interactions, 'pollution tolerant' species are not necessarily the most tolerant species to toxicants tested under standard laboratory conditions. It was hypothesized that the distribution of species in polluted rivers is not only dependent on the tolerance of species to toxicants, but also on species-specific capacities to modify or compensate for negative effects of toxicants. To test this hypothesis, species-specific responses to metals in organically enriched river water were studied under controlled conditions. The zebra mussel Dreissena polymorpha and the midge Chironomus riparius were exposed to metal-polluted water from the River Dommel. Additionally, the (interactive) effects of metals and humic acids (HA) on both species were evaluated. In spite of a lower tolerance of Chironomus riparius to metals in laboratory studies, the midge was the most tolerant of the two test species to metal-polluted site water. The results indicated that the sensitivities of the two test species determined in laboratory tests were inversely related to their sensitivities to polluted river water. In accordance with these results, midge larvae were protected from copper (Cu) toxicity by HA, while metal toxicity was not reduced (Cu) or even amplified (cadmium) by HA for the zebra mussel. Thus, the presence of (naturally occurring) HA in site water may partly account for discrepancies between responses of species to bioassays and toxicity tests. It is suggested that these differences in responses to metals in site water are strongly influenced by species-specific preferences for organic compounds (like HA). It is concluded that the response to organic compounds present in site water largely determines whether a species is classified as 'pollution tolerant' or 'pollution sensitive'.     

Site-specific water quality guidelines: 1. Derivation approaches based on physicochemical, ecotoxicological and ecological data

Generic water quality guidelines (WQGs) are developed by countries/regions as broad scale tools to assist with the protection of aquatic ecosystems from the impacts of toxicants. However, since generic WQGs cannot adequately account for the many environmental factors that may affect toxicity at a particular site, site-specific WQGs are often needed, especially for high environmental value ecosystems. The Australian and New Zealand Guidelines for Fresh and Marine Water Quality provide comprehensive guidance on methods for refining or deriving WQGs for site-specific purposes. This paper describes three such methods for deriving site-specific WQGs, namely: (1) using local reference water quality data, (2) using biological effects data from laboratory-based toxicity testing, and (3) using biological effects data from field surveys. Two case studies related to the assessment of impacts arising from mining operations in northern Australia are used to illustrate the application of these methods. Finally, the potential of several emerging methods designed to assess thresholds of ecological change from field data for deriving site-specific WQGs is discussed. Ideally, multiple lines of evidence approaches, integrating both laboratory and field data, are recommended for deriving site-specific WQGs.     

Reduction in labile copper in the 7-day Ceriodaphnia dubia toxicity test due to the interaction with zooplankton food

Due to the increased popularity of zooplankton toxicity tests, it is important to investigate potential confounding factors. Though zooplankton food has been studied extensively to meet the nutritional needs of the zooplankton, less research has been done on whether food addition reduces the toxicity of metals in the tested sample. This investigation combines toxicity tests and metal speciation analysis to determine whether the EPA recommended food of YCT (yeast, cerophyll, and trout chow) and Pseudokirchneriella subcapitata (formerly Selenastrum capricornutum) reduces copper toxicity by decreasing the concentration of labile copper. Toxicity tests were performed with Ceriodaphnia dubia on culture water spiked with 0, 787, and 1574 nM copper with five different food levels. A Chelex-100 cation exchange resin and a graphite furnace-atomic absorption spectrophotometer were used in conjunction with the toxicity tests to measure the amount of labile copper in the culture water. At the EPA recommended food dosage, the C. dubia food has a chelating capacity of approximately 500 nM Cu. For both concentrations of spiked culture water, the toxicity to C. dubia was reduced with increasing food level, which seemed to be both from a decrease in labile copper concentration and an increase in the nutritional condition of the zooplankton.     

Acute toxicity of copper,lead, cadmium,and zinc to early life stages of white sturgeon (Acipenser transmontanus) in laboratory and Columbia River water

Populations of white sturgeon (Acipenser transmontanus) are in decline in North America. This is attributed, primarily, to poor recruitment, and white sturgeon are listed as threatened or endangered in several parts of British Columbia, Canada, and the United States. In the Columbia River, effects of metals have been hypothesized as possible contributing factors. Previous work has demonstrated that early life stage white sturgeon are particularly sensitive to certain metals, and concerns over the level of protectiveness of water quality standards are justified. Here we report results from acute (96-h) toxicity tests for copper (Cu), cadmium (Cd), zinc (Zn), and lead (Pb) from parallel studies that were conducted in laboratory water and in the field with Columbia River water. Water effect ratios (WERs) and sensitivity parameters (i.e., median lethal accumulations, or LA50s) were calculated to assess relative bioavailability of these metals in Columbia River water compared to laboratory water, and to elucidate possible differences in sensitivity of early life stage white sturgeon to the same concentrations of metals when tested in the different water sources. For Cu and Pb, white sturgeon toxicity tests were initiated at two life stages, 8 and 40 days post-hatch (dph), and median lethal concentrations (LC50s) ranged between 9–25 μg Cu/L and 177–1,556 μg Pb/L. LC50s for 8 dph white sturgeon exposed to Cd in laboratory water and river water were 14.5 and 72 μg/L, respectively. Exposure of 8 dph white sturgeon to Zn in laboratory and river water resulted in LC50s of 150 and 625 μg/L, respectively. Threshold concentrations were consistently less in laboratory water compared with river water, and as a result, WERs were greater than 1 in all cases. In addition, LA50s were consistently greater in river water exposures compared with laboratory exposures in all paired tests. These results, in combination with results from the biotic ligand model, suggest that the observed differences in toxicity between river water exposures and laboratory water exposures were not entirely due to differences in water quality and metal bioavailability but rather in combination with differences in fish sensitivity. It is hypothesized that differences in concentrations of calcium in the different water sources might have resulted in differences in acquired sensitivity of sturgeon to metals. Canadian water quality guidelines, US national criteria for the protection of aquatic life, and water quality criteria for the state of Washington were less than LC50 values for all metals and life stages tested in laboratory and Columbia River water. With the exception, however, that 40 dph white sturgeon exposed to Cu in laboratory water resulted in threshold values that bordered US national criteria and criteria for the state of Washington.     

Geochemistry and toxicity of sediment porewater in a salt-impacted urban stormwater detention pond

A comprehensive study was carried out to investigate the impacts of road salts on the benthic compartment of a small urban detention facility, Rouge River Pond. Although the pond is an engineered water body, it is representative of many small urban lakes, ponds and wetlands, which receive road runoff and are probable high impact areas. Specific objectives of the study were to document the porewater chemistry of an aquatic system affected by elevated salt concentrations and to carry out a toxicological assessment of sediment porewater to determine what factors may cause porewater toxicity. The results indicate that the sediment porewater may itself attain high salt concentrations. The computations show that increased chloride levels have important implications on the Cd complexation, augmenting its concentration in porewater. The toxicity tests suggest that the toxicity in porewater is caused by metals or other toxic chemicals, rather than high levels of chloride.     

Analysis of copper,zinc, cadmium and chromium in fish tissues. A tool for detecting metal caused fish kills

In order to find the causative agent in frequently occuring fish kills in a Belgian river a series of toxicity tests has been conducted in which rudd (Scardinius erythrophtalmus) were exposed to acute lethal and subacute non lethal concentrations of copper, chromium, cadmium and zinc. The concentrations of these metals in gills, opercle, kidney, liver and muscle were measured. Metal levels in gills were the most valuable indicator of acute lethal exposure. This information was compared with levels found in rudd from a surface water storage reservoir and from the river Meuse. Fish collected after fish kills in the river Meuse were analysed. In one case copper could be identified as one of the toxicants concerned by fish tissue analysis. Metal levels in fish tissues can give valuable additional information concerning the cause of kills provided that background information is available about metal levels in water and normal tissue levels.     

Acute toxic hazard evaluations of glyphosate herbicide on terrestrial vertebrates of the oregon coast range

GOAL, SCOPE AND BACKGROUND: The degree to which dose responses of model organisms (lab rodents) can adequately predict dose responses of free-ranging wild mammals or amphibians is unknown, and the relative sensitivity of such species to body loading of a toxicant such as glyphosate is seldom reported. For relative effects of dosage, we compare sensitivity of nine wild vertebrate species to effects of high doses of glyphosate in Swiss-Webster laboratory mice both by gavage and by intraperitoneal injection. We also evaluate sublethal effects of herbicide exposure on behavior and reproductive success of one mammal and one amphibian species. METHODS: Comparisons of acute toxicity of glyphosate were made with intraperitoneal dosings of technical glyphosate isopropylamine salt to nine species of terrestrial vertebrates (five amphibians, four mammals) and compared with responses in Swiss-Webster laboratory mice. Animals collected from sites that had no recent herbicide application were allowed 7-14 days to equilibrate in captivity before treatment. RESULTS: Median lethal dose ranged from 800 to 1,340 mg kg(-1) in mammals, and 1,170 to >2,000 mg kg(-1) in amphibians, with Oregon vole being the most sensitive. White lab mice were in the middle of the mammalian range. Tailed frog, at >2,000 mg kg(-1) was the least sensitive. Calibration of IP sensitivity to oral administration by gavage indicated that roughly four times as much glyphosate must be administered to obtain a comparable estimate of lethality. Administration by gavage in highly concentrated solutions tended to cause physical injury, hence may prove less useful as a relative indicator of toxicity. When sublethal dosages were given to roughskin newts or chipmunks, mobility and use of cover appeared largely unaffected. DISCUSSION: Direct toxic effects of spraying glyphosate under normal forest management seem unlikely for the nine species examined. Nor could we detect significant indirect effects of exposure on behavior and use of cover features in two species. There may be effects on other aspects of the field biology of these animals, such as reproductive rates, which we did not investigate. Recent field data indicate that changes in habitat quality following herbicide application can result in high reproductive activity in species associated with the grasses and forbs that proliferate following field applications. CONCLUSIONS: When compared to field data on body burdens of wild mammals exposed after aerial application of glyphosate at maximum rates in forests, there seems to be a large margin of safety between dosages encountered and those causing either death or limitation of movement, foraging or shelter. RECOMMENDATIONS AND PERSPECTIVES: Margins of safety for small mammals and amphibians appear to be large under any probable exposure scenarios, however our results indicate high variability in responses among species. Uncertainty introduced into field studies from unknown sources of mortality (e.g, likely predation) must be considered when interpreting our results.     

The acute toxicity of agricultural surfactants to the tadpoles of four Australian and two exotic frogs

Nonionic surfactants are frequently incorporated into pesticide formulations, and are therefore a group of chemicals to which amphibians may be exposed in agricultural or urban landscapes. However, little is known about the effects of surfactant exposure in amphibians. Feeding stage tadpoles of Bufo marinus, Xenopus laevis and four species of Australian frogs (Crinia insignifera, Heleioporus eyrei, Limnodynastes dorsalis and Litoria moorei) were exposed to nonylphenol ethoxylate (NPE) and alcohol alkoxylate in static-renewal acute toxicity tests. All species exhibited nonspecific narcosis following exposure to both these surfactants. The 48-h EC50 values for NPE ranged between 1.1 mg/l (mild narcosis) and 12.1 mg/l (full narcosis). The 48-h EC50 values for alcohol alkoxylate ranged between 5.3 mg/l (mild narcosis) and 25.4 mg/l (full narcosis). Replicate acute toxicity tests with B. narinus exposed to NPE at 30 degrees C over 96 h indicated that the narcotic effects were not particularly time dependant. The mean 24, 48, 72, and 96-h EC50 (mild narcosis) values were 3.6, 3.7, 3.5 and 3.5 mg/l, respectively. The mean 24, 48, 72 and 96-h EC50 (full narcosis) were 4.0, 4.1, 4.2 and 4.0, respectively. Acute toxicity tests with B. marinus exposed to NPE at 30 degrees C under conditions of low dissolved oxygen (0.8-2.3 mg/l) produced a two to threefold increase in toxicity.     

The effect of acidification on the accumulation and toxicity of metals to freshwater invertebrates

This paper reviews the literature to determine if lowered water pH (a) affects metal bioaccumulation in freshwater invertebrates, (b) enhances the toxicity of a given metal, and (c) increases waterborne metal concentrations to levels toxic to invertebrates. The elements considered are mercury, lead, cadmium and aluminum. The available evidence suggests that of these elements only mercury is biomagnified in aquatic foodchains. The bioaccumulation of all these elements is influenced by water pH, but data concerning invertebrates is meagre for mercury and lead. The effect of pH on mercury and lead toxicity to invertebrates is unclear and may be largely species specific. Cadmium toxicity is reduced by lower pH, while aluminum toxicity to invertebrates is markedly higher due to changes in aluminum speciation at low pH.     

Interaction between components of electroplating industry wastes. Influence of the receiving water on the toxicity of the effluent

A copper-cadmium-nickel-zinc mixture was assessed in seven different river waters to study metal toxicity to the ciliate protozoan Colpidium campylum, the interactions occurring between metals, and the influence of the receiving water on toxicity. In the range of concentrations tested, which are representative of electroplating industry wastes, the main part of the toxicity can be attributed to copper and to cadmium-copper synergy. A classification of waters, based on a principal component analysis (PCA), was used to examine the main parameters of the water, which can affect the toxicity of metal mixtures. It appears that the mineralization of the water, more than the total organic carbon (TOC), is an important parameter for the expression of toxicity. A strategy for the estimation of ecotoxicological hazard assessment, based on a simplified factorial experiment is proposed. It enables one to study, in a two-step bioassay, the toxicity of an effluent, the influence of river water on its toxicity, and the effects of contact time and dilution. By applying PCA to data from very different waters, it may be possible to estimate the ecotoxicological risk associated with the discharge of an effluent, on the basis of the chemistry of the receiving water.     

Seasonality of contamination, toxicity, and quality values in sediments from littoral ecosystems in the Gulf of Cádiz (SW Spain)

To seasonally evaluate littoral contamination, toxicity and quality values of sediments from the Gulf of Cádiz, we measured chemical concentrations and conducted toxicity tests in winter and summer and linked these results by means of multivariate analysis. Sediment samples were subjected to two separate, replicated sediment toxicity tests (Microdeutopus gryllotalpa amphipod survival, and Ruditapes philippinarum clam reburial), and to comprehensive sediment chemistry analyses (grain size, organic carbon, 14 heavy metals, and the surfactant linear alkylbenzenesulfonate (LAS)). Only sediments associated with an untreated urban discharge were toxic and related to high levels of surfactant LAS, Ag, and Pb. Multivariate analysis indicated that variables and chemicals associated with geochemical matrix and background levels (specific surface, Fe, Zn, Cu, V, Ni, and Co), chemicals associated with untreated urban discharge sources, and toxicity effects showed no seasonal variability. Only copper concentrations showed seasonal differences, being toxic during the winter and not toxic during summer. Multivariate analysis permits us to derive sediment quality values (SQVs); in terms of concentrations at or below which biological effects were not measured (mg kg(-1) dry sediment), are: LAS, 2.6; lead, 66.8; silver, 0.78; copper, 69.6.     

Bioaccumulation of trace elements in omnivorous amphibian larvae: implications for amphibian health and contaminant transport

Despite the influence that amphibians have on the flow of energy and nutrients in ecological systems, the role that amphibians play in transporting contaminants through food webs has received very little attention. This study was undertaken to investigate bioaccumulation of trace elements in amphibians relative to other small aquatic organisms in a contaminated wetland. We collected bullfrog larvae (Rana catesbeiana) along with three other species of small vertebrates and four species of invertebrates from a site contaminated with a wide array of trace elements and analyzed them for trace element concentrations and stable nitrogen and carbon isotope composition. We found that amphibian larvae accumulated the highest concentrations of most trace elements, possibly due to their feeding ecology. These results suggest that omnivorous amphibian larvae can serve as a critical link for trace element trophic transfer. Their propensity to accumulate trace elements may have important implications for amphibian health in contaminated environments and should be further investigated.     

Toxic substances in rivers and streams

Many of the toxic substances entering freshwaters today are those which were present several decades ago, but others have become significant recently. The effects of toxicants in flowing waters are modified by unidirectional transport and dispersion which afford the potential for a degree of 'self-purification'. The chemical quality of the receiving water also affects toxicity. Biological factors also contribute to the ultimate effect of pollutants. The potential for accumulation of toxic substances within tissues increases the significance of certain pollutants which may be present in water even though ambient concentrations are very low. The biota of flowing waters may be restored, following catastrophic entry of pollutants, by drift from unaffected regions upstream. The range of potential toxic substances is very extensive and includes inorganic poisons, organic poisons, heavy metals, pesticides and PCBs. Metals, pesticides and PCBs have the greatest potential for bioaccumulation. Few generalisations can be made regarding the effects of toxic substances on the biota. Each species tends to respond to different toxicants in different ways and even at different stages in its life-history. Toxicity tests conducted under controlled laboratory conditions sometimes produce conflicting results: it is not then to be unexpected that field observations should sometimes vary widely. Determinations of toxicity in laboratory tests must be applied with caution to field conditions and it is not wise to extrapolate findings to other species or environments.     

Influence of organic solvents on the sensitivity of a bioluminescence toxicity test with Vibrio harveyi

The possibility of using organic solvents (OSs) to increase the susceptibility of bioluminescent microorganisms in a bioassay for assessing the toxicity of chemicals dissolved in water was investigated. To conduct the tests acetonitrile, dimethyl sulfoxide, ethanol, methanol, and isopropanol were used as OSs and Cd, Hg, and Zn as reference toxicants. The addition of OSs modified the toxicity of the three metals to Vibrio harveyi, according on the bioluminescence assay used. The sensitivity of the luminescence bioassay for Hg increased in the presence of the five OSs, thus indicating a greater toxic effect. However, the sensitivity of the assay for the other two metals, Cd and Zn, increased or decreased (lesser toxic effect) depending on the concentration at which the OSs were used. No correlation was observed between the concentration of the five OSs and the toxicity of the three reference toxicants. From this it can be deduced that none of these OSs could be recommended for increasing generically the sensitivity of toxicity biotests using V. harveyi.