Cristiano V.M. Araújo

Sediment integrative assessment of the Bay of Cádiz (Spain): An ecotoxicological and chemical approach

This study consisted of the sediment toxicity assessment of the Bay of Cádiz based on two endpoints: growth inhibition for Cylindrotheca closterium (benthic microalgae) and fecundity inhibition for Tisbe battagliai (harpacticoid copepod). A new methodology to eliminate (but not as storage technique) the autochthonous biota present in the sediment samples by immersing them in liquid nitrogen (-196 degrees C) was also assessed. Sediment toxicity data showed different toxicity levels for both organisms. In general, T. battagliai was more sensitive; however a good correlation (r=0.75; p<0.05) between sediment toxicity results for both species was found. Data in pore water (pH, redox potential, and toxicity for microalgae and copepod) and sediment (pH, redox potential, organic carbon, and metal concentrations) demonstrated that ultra-freezing did not alter sample characteristics; thus, this technique can be adopted as a pre-treatment in whole-sediment toxicity tests in order to avoid misleading results due to presence of autochthonous biota. Multivariate statistical analysis such as cluster and principal component analysis using chemical and ecotoxicological data were employed. Silt and organic matter percentage and lead concentration were found to be the factors that explain about 77% of sediment toxicity in the Bay of Cádiz. Assay methodology determined in this study for both assayed species is considered adequate to be used in sediment toxicity monitoring programs. Results obtained using both species show that the Bay of Cádiz can be considered a moderately polluted zone.

Copper-driven avoidance and mortality in temperate and tropical tadpoles

Amphibians have experienced an accentuated population decline in the whole world due to many factors, one of them being anthropogenic contamination. The present study aimed to assess the potential effect of copper, as a worldwide and reference contaminant, on the immediate decline of exposed population due to avoidance and mortality responses in tadpoles of three species of amphibians across climatic zones: a South American species, Leptodactylus latrans, a North American species, Lithobates catesbeianus, and a European species, Pelophylax perezi. A non-forced exposure system with a copper gradient along seven compartments through which organisms could freely move was used to assess the ability of tadpoles to detect and avoid copper contamination. All species were able to avoid copper at a concentration as low as 100 μg L(-1). At the lowest (sublethal) concentrations (up to 200 μg L(-1)) avoidance played an exclusive role for the population decline, whereas at the highest concentrations (> 450 μg L(-1)) mortality was the response determining population decline. The median concentrations causing exposed population immediate decline were 93, 106 and 180 μg L(-1) for Le. latrans, Li. catesbeianus and P. perezi, respectively. Contaminants might, therefore, act as environmental disruptors both by generating low-quality habitats and by triggering avoidance of tadpoles, which could be an important response contributing to dispersion patterns, susceptibility to future stressors and decline of amphibian populations (together with mortality).

Short-term toxicity tests on the harpacticoid copepod Tisbe battagliai: Lethal and reproductive endpoints

Short-term bioassays based on lethal and reproductive responses of Tisbe battagliai were applied to determine responses of copepods to copper and LAS. Percentage of spawning females, fecundity (F), and total newborn production (N) for 48 and 72 h were calculated for both substances. It was observed percentage of spawning females was not affected by sublethal concentrations of both compounds. Following values were obtained: EC(50)(N)-48 h of 670+/-30 microgLASL(-1) and EC(50)(F)-48 h of 670+/-30 microgLASL(-1); and EC(50)(N)-72 h of 44.5+/-1.8 microgCuL(-1) and EC(50)(F)-72 h of 30.8+/-1.1 microgCuL(-1). Lethal effects of the two substance-types were also assessed, obtaining the LC(50)-24h of 1980+/-160 microgLASL(-1); and LC(50)-48 h of 83.1+/-10.5 microgCuL(-1) for nauplii; and LC(50)-72 h of 157+/-25 microgCuL(-1), and LC(50)-72 h of 2660+/-270 microgLASL(-1) for adults. Fecundity and total newborn production are sensitive endpoints for determining effects of toxicants.

Heavy metals in yellowfin tuna (Thunnus albacares) and common dolphinfish (Coryphaena hippurus) landed on the Ecuadorian coast

Heavy metals are contaminants of great environmental concern due to their multiple origins (natural and anthropogenic), the ability to accumulate in organs and tissues, and the deleterious effects they can cause in organisms. Studies on the accumulation of metals in seafood, such as fish, have increased in importance due to the risk for human health when consuming fish contaminated by metals. The present work was aimed at verifying the concentrations of cadmium (Cd), mercury (Hg) and lead (Pb) in the muscular tissue and liver of yellowfin tuna (Thunnus albacares) and common dolphinfish (Coryphaena hippurus) from the Eastern Pacific Ocean landed in Manta city, Ecuador. Samples were analyzed by inductively coupled plasma mass spectroscopy (ICP-MS). Around half of the muscle samples of both species presented levels of Cd and Hg above the limits considered safe for human consumption established by the European Union. For Pb,most of the muscle samples were considered acceptable for consumption. Results indicate that both species should be consumed with some caution. Considering the tolerable weekly intake recommended for adults by the World Health Organization, results indicate that Hg is the main metal that limits the consumption of yellowfin tuna and common dolphinfish, with a recommended maximum ingestion, respectively, of 191 and 178 g per week for an adult.c

Ring test for whole-sediment toxicity assay with a benthic marine diatom

This work presents the results of an interlaboratory proficiency exercise for whole-sediment toxicity assays with the benthic marine diatom Cylindrotheca closterium. An assay protocol was established and followed by all participating laboratories. Cell growth after 72 h exposure was the endpoint used. Four sediment samples of unknown toxicity were assayed. The main problem encountered during this exercise was the differences in the cell growth of algae exposed to reference sediment. Those differences may be associated with changes in the physiological status of the initial culture due to temperature changes during transport to the other laboratories. In general, the method proposed presented good replicability (precision between replicates) and reproducibility (interlaboratory precision). Around 80% (17 out of 21) of results obtained were classified as satisfactory (Z-scores <2). The whole-sediment assay with C. closterium presented here can be considered sufficiently successful for possible use as a standard toxicity test. The assay is simple to perform, the proposed species is ecologically relevant as an integral component of microphytobenthos, and is widely distributed around the world. These positive factors suggest that the whole-sediment assay with the benthic marine diatom C. closterium can be used as a reliable tool in marine sediment quality assessment.

Active and passive spatial avoidance by aquatic organisms from environmental stressors: A complementary perspective and a critical review

Spatial avoidance is a mechanism by which many organisms prevent their exposure to environmental stressors, namely chemical contaminants. Numerous studies on active avoidance and drift by aquatic organisms, as well as the main approaches used to measure both responses, were reviewed. We put forward a particular recommendation regarding methodological approaches: active avoidance should preferably be evaluated under a dilution gradient in a multi-compartmented system instead of in a bi-compartmented system. Available data on spatial avoidance from contamination indicate that emigration can occur at even lower contaminant concentrations than sub-individual noxious effects (assessed with the traditional forced-exposure assays), challenging the widely accepted paradigm in ecotoxicology that contaminant-driven adverse consequences at the population level result from a time delayed cascade of sequentially linked biochemical, cellular, physiological, and finally whole organism deleterious effects. Therefore, contaminants should not be viewed solely as potential toxicants at the individual level, but also as potential disturbers of habitats, by making the latter, at least partially, unsuited to accommodate life. Also, exposure to contamination is needed to trigger avoidance, but uptake is not mandatory, which demands the concept of exposure to be expanded, to include also the mere perception of the stressor. Since emigration eventually leads to local population extinction, and thus to severe implications for ecosystem structure and functioning, we then recommend that avoidance data be incorporated in environmental risk assessment schemes.

Avoidance response of Danio rerio to a fungicide in a linear contamination gradient

The present study examines the ability of juvenile Danio rerio to avoid pyrimethanil-contaminated water. An avoidance assay system was used with a contamination gradient formed by seven compartments, through which the fish could move and choose the preferred compartment(s). Additionally, the influence of fish movements in promoting the mixing between compartments and thus disruption of the gradient over time was also examined by testing sodium chloride (NaCl) at sublethal concentrations. Samples with pyrimethanil were obtained from the commercial formulation Mythos®, which was applied to mesocosm systems. Samples of the pyrimethanil-contaminated mesocosms water were collected and a series of seven concentrations (0.2 to 1.4mgL(-1) plus a control) diluted with reference (uncontaminated) mesocosm water were added to the system to form the gradient. After 4h exposure, fish avoidance in the three highest pyrimethanil concentrations ranged from 29 to 66%. The 4h-AC50 (concentration at which 50% of the fish avoided pyrimethanil after 4h exposure) was 1.10 (confidence interval: 1.07 to 1.12)mgL(-1). However, the avoidance pattern after 12h was strongly reduced and it was not possible to calculate the AC50. This is explained by the results of the NaCl experiment, which showed that the movement of fish in the system accelerates the mixing of the solutions between compartments. As pyrimethanil can trigger avoidance response in D. rerio, this fungicide, even at non-lethal concentrations, could be considered an environmental disturber.

Acute and chronic sensitivity, avoidance behavior and sensitive life stages of bullfrog tadpoles exposed to the biopesticide abamectin.

As compared to other aquatic organism groups, relatively few studies have been conducted so far evaluating the toxicity of pesticides to amphibians. This may at least partly be due to the fact that regulations for registering pesticides usually do not require testing amphibians. The sensitivity of amphibians is generally considered to be covered by that based on toxicity tests with other aquatic organisms (e.g. fish) although the impact of a pesticide on amphibians may be very different. In the present study, acute and chronic laboratory tests were conducted to evaluate the acute and chronic toxicity of abamectin (as Vertimec® 18EC) to bullfrog (Lithobates catesbeianus) tadpoles. Acute tests were conducted at two tadpole stages (Gosner stage 21G and 25G) and avoidance tests were also conducted with stage Gosner stage 21G tadpoles. Calculated acute toxicity values were greater than those reported for standard fish test species, hence supporting the use of fish toxicity data as surrogates for amphibians in acute risk assessments. Given the limited number and extent of available amphibian toxicity studies, however, research needs to increase our understanding of pesticide toxicity to amphibians are discussed.

Effects of cold-dark storage on growth of Cylindrotheca closterium and its sensitivity to copper.

Cylindrotheca closterium cells were maintained at low temperature (4+/-1 degrees C) and dark conditions up to 21 weeks to assess the effect on survival and physiological status. From a control culture under standard conditions, three densities were prepared: (A) 2 x 10(4), (B) 10 x 10(4), and (C) 25 x 10(4) cells ml(-1). Weekly, inoculums of each stored density were exposed to continuous light and at 20+/-1 degrees C. Sensitivity to copper for microalgal cultures was evaluated in order to assess possible changes in cells sensitivity due to storage. Concurrently, assays with a control culture were carried out in order to assess the sensitivity of C. closterium to copper and to be able to generate a standard sensitivity control chart with a mean value of EC50-72 h+/-2SD (standard deviation). Density-C presented higher cell yield values, between 40% and 80% relative to control culture. Cell density showed to be important feature that may be taken into account in cell storage experiments. There was an increase in sensitivity of cells submitted to storage; however results always kept in the range established as standard sensitivity with no statistically significant difference with regards to control culture. EC50-72 h mean value for the control culture was 29+/-10 mug Cul(-1), while for densities-A, B and C were 22+/-7; 23+/-9 and 23+/-8 microg Cul(-1), respectively. In spite of drastic changes in the environmental conditions due to storage, it is concluded that C. closterium cells stored during 5 months remained metabolically active and with no significant change in its sensitivity.

Microphytobenthos in ecotoxicology: A review of the use of marine benthic diatoms in bioassays

Contamination in coastal zones is an increasing problem that adversely affects biological diversity and the functioning of coastal ecosystems. Sediment is an important compartment of these zones since large quantities of diverse contaminants can accumulate there. Whole-sediment toxicity assays are of increasing importance, and several assay methods using mainly invertebrates have been developed. However, an important part of the benthic community, the microphytobenthos (represented principally by benthic diatoms and cyanobacteria), has surprisingly been neglected. Recently, comprehensive studies have been conducted using benthic marine microalgae with the object of establishing a toxicity assay method for sediment samples. The main results published to date in the literature and obtained by our own team have been compiled and are discussed in this review. The value and feasibility of using certain organisms of the microphytobenthos group in ecotoxicology studies are also discussed, and a sediment quality guideline based on multivariate procedure has been derived from data obtained in previous studies. Finally, future perspectives for research in this field are discussed.