Comparative susceptibility of two Neotropical predators, Eriopis connexa and Chrysoperla externa, to acetamiprid and pyriproxyfen: Short and long-term effects after egg exposure.

Compatibility assessments between selective insecticides and the natural enemies of pests are essential for integrated-pest-management programs. Chrysoperla externa and Eriopis connexa are two principal Neotropical predators of agricultural pests whose conservation in agroecosystems requires a toxicity evaluation of pesticides to minimize the impact on those beneficial insects on the environment. The objective of this work was to evaluate the toxicity of the insecticides pyriproxyfen and acetamiprid on C. externa and E. connexa eggs exposed to the maximum recommended field concentrations of each along with three successive dilutions. The survival and the immature developmental time were assessed daily until adulthood and the mean survival time calculated over a 10-day period. The cumulative survival of E. connexa was reduced at all concentrations of both insecticides, while that of C. externa was significantly decreased by ≥50 mg L-1 of acetamiprid and ≥37.6 mg L-1 of pyriproxyfen. In both species, the reductions occurred principally on the eggs and first larval instar. Survival curves, in general, differed from those of the controls, with the mean survival time of E. connexa being significantly shorter in insecticides treatments than that of the controls. Certain concentrations of each of the insecticide lengthened the egg and first-larval-instar developmental periods of E. connexa and C. externa, respectively. Also, pyriproxyfen reduced the first-larval-instar period and lengthened the fourth of E. connexa. Acetamiprid was more toxic to E. connexa than to C. externa at the two highest concentrations. Conversely, at those same concentrations of pyriproxyfen, the relative toxicity to the two species was reversed. The present work represents the first investigation on the comparative susceptibility of two relevant Neotropical biological control agents to acetamiprid and pyriproxyfen. Also, it highlights the necessity of assessing long-term effects in the compatibility studies between natural enemies of agricultural pests and insecticides.

Pesticide survey in water and suspended solids from the Uruguay River Basin, Argentina.

The Uruguay River is receptor of pollutants, such as pesticides, from agriculture activities along its course. The present study reports concentration levels of organochlorinate, organophosphorus, and other pesticides in water and suspended solids in nine sampling sites of the Uruguay River. Data analyses included principal component analysis (PCA) to assess differences between sampling sites contamination. Most of the tested pesticides were ubiquitous due to the widely use in the chemical control of pests implemented in the region. Detected concentrations of aldrin, chlordane, dieldrin, endrin, heptachlor epoxide, lindane, 4,4'-DDT, endosulfan, chlorpyrifos, diazinon, methyl-parathion, and malathion were found to be over regional and international concentration level guidelines, according to the European Union, the US Environmental Protection Agency, or the Argentinean Secretariat of Environment and Sustainable Development. For this reason, future studies in Uruguay River Basin are needed.

Effect on the growth and development and induction of abnormalities by a glyphosate commercial formulation and its active ingredient during two developmental stages of the South-American Creole frog, Leptodactylus latrans.

We evaluated the acute lethal and sublethal effects of technical-grade glyphosate (GLY) and the GLY-based commercial formulation Roundup ULTRA MAX® (RU) on two Gosner stages (Gss) 25 and 36 of the South-American Creole frog, Leptodactylus latrans. Bioassays were performed following standardized methods within a wide range of concentrations (0.0007-9.62 mg of acid equivalents per liter-a.e./L-of RU and 3-300 mg/L of GLY). The endpoints evaluated were mortality, swimming activity, growth, development, and the presence of morphologic abnormalities, especially in the mouthparts. No lethal effects were observed on larvae exposed to GLY during either Gs-25 or Gs-36. The concentrations inducing 50 % lethality in RU-exposed larvae at different exposure times and Gss ranged from 3.26 to 9.61 mg a.e./L. Swimming activity was affected by only RU. Effects on growth and development and the induction of morphologic abnormalities-like oral abnormalities and edema-were observed after exposure to either GLY or RU. Gs-25 was the most sensitive stage to both forms of the herbicide. The commercial formulation was much more toxic than the active ingredient on all the endpoints assessed. Effects on growth, development, and the induction of morphologic abnormalities observed in the range of environmental concentrations reported for agroecosystems of Argentina constitute an alert to the potential detrimental effects of the herbicide that could be affecting the fitness and survival of anurans in agroecosystems.

Toxicity assessment of four insecticides with different modes of action on pupae and adults of Eriopis connexa (Coleoptera: Coccinellidae), a relevant predator of the Neotropical Region.

Pesticides can be toxic to nontarget organisms including the natural enemies of agricultural pests, thus reducing the biodiversity of agroecosystems. The lethal and sublethal effects of four insecticides with different modes of action-pyriproxyfen, teflubenzuron, acetamiprid, and cypermethrin-were evaluated on pupae and adults of Eriopis connexa, an effective predator in horticultural crops. Pupal survival was reduced by pyriproxyfen (26 %) and cypermethrin (41 %). Malformations in adults emerged from treated pupae were observed after acetamiprid (82.7 and 100 % for 100 and 200 mg a.i./l, respectively), pyriproxyfen (48.6 %), and cypermethrin (13.3 %) treatments. A longer mean oviposition time was also observed in adults emerged from pupae treated with cypermethrin. Moreover, the latter insecticide as well as teflubenzuron did not reduce reproductive parameters, whereas females emerged from pyriproxyfen-treated pupae were not be able to lay eggs even when females showed large abdomens. Upon exposure of adults, survival was reduced to approximately 90 % by acetamiprid, but no reduction occurred with pyriproxyfen, teflubenzuron, or cypermethrin though the fecundity at fifth oviposition time of the female survivors was reduced. Pyriproxyfen decreased the hatching at all the oviposition times tested, whereas fertility was reduced in the fourth and fifth ovipositions by teflubenzuron and in the first and third ovipositions by cypermethrin. In conclusion, all four insecticides tested exhibited lethal or sublethal effects, or both, on E. connexa. The neurotoxic insecticides were more harmful than the insect-growth regulators, and pupae were more susceptible than adults. The toxicity of insecticides on the conservation of predators in agroecosystems of the Neotropical Region is discussed.

Toxicity Persistence of Chlorpyrifos in Runoff from Experimental Soybean Plots to the Non-target Amphipod Hyalella curvispina: Effect of Crop Management.

Toxicity persistence to the nontarget amphipod Hyalella curvispina in runoff events following chlorpyrifos applications to soy experimental plots was compared in conventional and no-till management. Two application scenarios were compared: an early-season application with the soil almost bare and a late-season application after the foliage had attained complete soil cover. H. curvispina was exposed to chlorpyrifos using two different test systems: a short-term (48 h) runoff water exposure and a long-term (10 days) soil exposure. Both commonly used crop management practices for soybean production resulted in runoff toxicity following pesticide applications and represent a toxicity risk for adjacent inland waters. Toxicity persistence was longer after the earlier than the late season application, likely because of higher volatilization and photodecomposition losses from the soy canopy than from the soil. For the early-season application, toxicity persisted longer in the no-till plots than in the conventional tillage plots. Suspended matter was higher in the conventional treatment. Chlorpyrifos sorption to suspended matter likely contributed to the shorter persistence. For the late-season application, toxicity persisted longer in the conventional treatment. The causes remain conjectural. The soil organic carbon content was higher in the no-till treatment. Sorption to organic matter might have contributed to the shorter chlorpyrifos toxicity persistence in no-till management. Late applications are more frequent and prevail longer throughout the soy growing season. Overall, the no-till management practice seems preferably because shorter toxicity persistence in runoff represents a lower environmental risk for the adjacent inland waters.

Differential uptake of endosulfan in the South American toad under sublethal exposure.

Agroecosystems are usually polluted with a wide variety of contaminants with pesticides being very frequently detected. Endosulfan, an organochlorine pesticide, has been shown to cause both lethal and sublethal effects on aquatic organisms such as amphibians and especially on its early developmental stages. In this context, the aim of this study was to evaluate the uptake of environmentally relevant concentrations of endosulfan and its correlation with differential sensitivity in the early development stages of the common South American toad, Rhinella arenarum. Embryos and larvae were exposed to sublethal concentrations of endosulfan for several periods of exposures. According to the developmental stage at which they were exposed, the uptake rate was different. Bioconcentration factors (BCFs) for embryos significantly decreased with exposure time and concentration (p < 0.05) reaching a BCF of ≤1679 for embryos at 96 h of exposure to 0.001 mg endosulfan L(-1). BCFs for larvae significantly increased with exposure time (p < 0.05) obtaining a maximum of 40 at 504 h. In our previous study, we reported that embryos were less sensitive to the effects of endosulfan than larvae, which is in line with the main tendency of embryos to bioconcentrate endosulfan as observed in this study.

Screening concentration of E1, E2 and EE2 in sewage effluents and surface waters of the "Pampas" region and the "Río de la Plata" estuary (Argentina).

Concentrations of estrone (E1), 17ß-estradiol (E2) and 17α-ethinylestradiol (EE2) were investigated for the first time in sewage effluents and receiving waters of the "Río de la Plata" estuary and neighboring areas by means of LC-MS/MS. E2 and EE2 were ubiquitous in the evaluated sewage effluent samples showing concentrations ranging between 122-631 and 65-187 ng/L, respectively. In surface waters, these estrogens were only detected in the "Girado" stream (Chascomús) at 369 and 43 ng/L, respectively. No significant relationship was found among the size of the served population and the concentration of the estrogens in the sewage effluent. The detection of these estrogens in receiving waters was dependent on the dilution capacity of the system. The studied estrogens were undetectable at the La Plata City water supply station. Conversely, concentrations found at the "Girado" stream indicate a potential ecotoxicological risk of these estrogens to the local aquatic biota.

The effect of cypermethrin, chlorpyrifos, and glyphosate active ingredients and formulations on Daphnia magna (Straus).

Acute effects of active ingredients (a.i.) and formulations (F) of widely used pesticides were assessed by means of the Daphnia magna toxicity test. Studied pesticides were the insecticides, cypermethrin and chlorpyrifos, and the herbicide, glyphosate. Results were analyzed and compared according to statistical endpoints (LCx) and log-probit regressions of toxicity data. The potency of acute toxicity followed the order: chlorpyrifos F > chlorpyrifos a.i. > cypermethrin F > cypermethrin a.i. ≫ glyphosate F > glyphosate a.i. Three to five orders of magnitude differences between the toxicity (µg/L to mg/L) of insecticides and the herbicide were observed. A pairwise comparison between a.i. and F indicated that all formulations were more potent. Additionally, for the case of glyphosate, evidence suggests that the adjuvant contributes to formulation toxicity rather than to the enhancement of a.i. potency.

Differential toxicity and uptake of Diazinon on embryo-larval development of Rhinella arenarum.

Diazinon, an anti-cholinesterase organophosphate, is an extensively used pesticide. The main objective of this work was to assess the lethal and sublethal effects of Diazinon and its comparison with the uptake by embryos and larvae of the common South American toad Rhinella arenarum by means of standardized bioassays during acute (96 h), short-term chronic (168 h) and chronic (504 h) exposures. Toxicity resulted time- and stage-dependent, thus the lethal concentration 50 for 96 h, 168 h and 504 h were 27.2; 20.1 and 6.8 mg Diazinon L(-1) for embryos and 8, 6.7 and 1.9 mg Diazinon L(-1) for larvae. It is noteworthy the remarkable differences found in the concentration which caused lethality with those causing adverse effects on development such as malformations (teratogenic effects). Therefore, the teratogenic index from 144 h was greater than two; the main adverse effects were axial flexures, irregular borders, wavy tail, microcephaly, malformed mouth and adhesive structures, gut miscoiling, underdeveloped gills, cloacal edema, desquamation and severe hydropsy. Moreover, the characteristic sublethal effect of Diazinon on larvae was abnormal behavior related to neurotoxicity with a NOEC-168 h of 4.5 mg Diazinon L(-1). Diazinon contents in R. arenarum were time-dependent and significantly related to exposure concentration for both embryos and larvae. Diazinon contents were also stage-dependent, as it was up to 27 times higher for organisms exposed from blastula stage onwards than early larvae. These facts and the Hazard Quotients, a numerical expression of ecological risk, of 2.73, which is above USEPA's Level of Concern, showed the threat that Diazinon represents for R. arenarum populations.

Integrated ecotoxicological assessment of bottom sediments from the Paraná basin, Argentina.

Paraná River, the six largest in the world, is receptor of pollution loads from tributaries traversing urban and industrialized areas, and extensive agriculture, particularly in its middle and low stretch along the Argentinean sector, where most of the productive activities of the country develop. Within the frame of monitoring surveys, the quality of bottom sediments from distal positions of twenty tributaries and three of the main course was evaluated. The assessment covered testing lethal and sublethal effects with the Hyalella curvispina based toxicity test, a benthic macrofauna survey and physicochemical variables of sediment matrix composition. A multivariate statistical analysis approach permitted integrating the obtained data from the different survey lines of evidence, explaining potential causes of the measured biological effects. The main perturbations detected were associated to tributaries in the middle sector of the basin, where anoxic conditions with high sulfide contents prevail mostly related to organic matter inputs of diverse combined activities, where sediments induce high lethality, and a consequent strong reduction of the benthic community population and diversity. The integrated survey approach proved being a robust tool in the assessment of causative-adverse effects relationships.

Impact of the neonicotinoid acetamiprid on immature stages of the predator Eriopis connexa (Coleoptera: Coccinellidae).

Eriopis connexa is a native coccinelid predator in the Neotropical Region. In Argentina it is commonly found associated to sucking pests in several crops and among them aphids and whiteflies. These pests are usually controlled with newly developed systemic insecticides, such as the neonicotinoids. However, the compatibility between selective pesticides and natural enemies is required before incorporating them in integrated pest management (IPM) packages. Within this frame, the objective of this study was to evaluate the side effect of various concentrations/doses of one commonly used neonicotinoid in vegetal crops, acetamiprid, on immature stages of E. connexa by dipping or topical exposure for eggs and larvae, respectively. Acetamiprid reduced egg hatching from 34 to 100 %. Moreover, the embryogenesis was disrupted by insecticide at early embryo stage at all tested concentrations. Second larval instar was more susceptible to acetamiprid than the fourth one and this susceptibility was positively related with the tested concentrations. On the other hand, the survival reduction at larval stage reached 100 % from 20 mg a.i./L (10 % of maximum field concentration). Besides, the reproduction of the females developed from topical bioassays on fourth instar larvae was strongly affected, with reduction in fecundity and fertility from 22 to 44 % and from 37 to 45 %, respectively. Overall the results showed a high toxicity of acetamiprid on immature stages of E. connexa, demonstrating that this broadly used insecticide could reduce biocontrol services provided by this predator and could also likely disturb IPM programs.

Physicochemical and ecotoxicological based assessment of bottom sediments from the Luján River basin, Buenos Aires, Argentina.

The physicochemical analysis of bottom sediments of the Lujan River was done on samples from 14 sites situated along its course and covered grain size and organic matter, total N and P, sulfides, heavy metals, organochlorine, and pyrethroid pesticides. In addition, acute 10-day whole-sediment laboratory toxicity tests were carried with each sample, using the native amphipod Hyalella curvispina as test organism. In order to correlate both types of results, data were assessed by multivariate analysis, including principal component analysis (PCA). The physicochemical profile of samples resulted similar along the river course, though several anomalous data were registered in the middle course of the river, mainly in samples taken downstream a large industrial complex; with a few exceptions in upper basin sites characterized by the dominance of agricultural activities, the pesticides concentration were consistently below the analytical detection limits. Almost 50% of the samples induced adverse effects on the amphipod when testing sublethal and lethal end points. The toxicity of the samples in terms of survival rate was extremely high in two sites, in particular in samples taken downstream the Pilar industrial complex. The integration of a selection of physicochemical and toxicological parameters of the sediments by PCA allowed discriminating areas of the river basin according the type and intensity of their particular pollution condition.

Human pharmaceuticals in wastewaters from urbanized areas of Argentina.

The study contributes with a first survey of pharmaceuticals in municipal wastewaters discharging into fresh and estuarine waters from areas with varying degrees of urbanization of Argentina. Analyses were done on the soluble fraction by HPLC-MS after SPE extraction. In all of the samples were detected caffeine and ibuprofen within the range of 0.9-44.2 and 0.4-13.0 µg/L, and lower levels of carbamazepine, atenolol and diclofenac between 0.2-2.3, 0.2-1.7 and <0.03-1.2 µg/L, respectively. Profiles of compounds were similar in all studied locations.

Influence of existing site contamination on sensitivity of Rhinella fernandezae (Anura, Bufonidae) tadpoles to Lorsban®48E formulation of chlorpyrifos.

Effects of the widely employed insecticide Lorsban(®)48E formulation of chlorpyrifos (CPF) was studied on Rhinella fernandezae tadpoles, a native species of Argentina, Brazil, Paraguay and Uruguay, under the hypothesis of a differential response of organisms from ponds of two sites with different degree of anthropogenic disturbance: S1 an unpolluted area, and S2 area with high degree of antropogenic disturbance. To collect a representative sample of the genotypic variability of each population, small portions from six clutches were taken randomly from each site when the period of clutching was finished. Embryos and tadpoles were maintained under controlled laboratory conditions. Toxicity tests were conducted under standardized conditions to study acute and chronic lethal (mortality) and sublethal effects (behavior, growth, and abnormalities), within the range of concentrations of 0.010 to 5 mg/L. Chronic effects were assessed with organisms from one of the demes (S1). CPF showed high toxicity on the tadpoles, inducing lethal and sublethal effects at 96 h exposure within a narrow range of concentrations from 0.066 to 0.887 mg/L. Results indicate that R. fernandezae tadpoles are below the 30th percentile in the species sensitivity distribution of existing data. The acute LC50, NOEC, and LOEC values were 0.151, 0.066, and 0.133 mg/L for S1, and 0.293, 0.177, and 0.266 mg/L for S2, respectively. Considering all acute end-points evaluated, the effects of CPF showed no significant differences (p = 0.3484) between the studied populations. CPF has more severe effects at higher concentrations than at higher times of exposure. Contaminants in S2 do not seem to induce local adaptation. Sublethal effects data and measured environmental concentrations indicate potential risk for populations inhabiting agroecosystems.

Toxicity of the effluent from an anaerobic bioreactor treating cereal residues on Lactuca sativa.

Effluents generated during the process of anaerobic digestion should be treated before their disposal into the environment. The aim of this study was evaluating the effectiveness of the effluent treatment system from an anaerobic bioreactor, assessing the toxicity reduction with the Lactuca sativa seed germination and root elongation inhibition test. Three sampling points were selected along the effluent treatment system: inflow into the first treatment pond, outflow from the third pond and recirculated flow to the bioreactor. Effluent dilutions tested for each sampling point were 25% and 50% (v/v), undiluted sample and controls. The pH, conductivity, temperature, dissolved oxygen, BOD5 and COD were measured. The decrease in the organic and inorganic loads was correlated with a reduction in the phytotoxicity. The use of the seed toxicity test allows evaluating the quality and effectiveness of the studied effluent treatment system.

Global metabolic response in the bile of pejerrey (Odontesthes bonariensis, Pisces) sublethally exposed to the pyrethroid cypermethrin.

The metabolic profile of Odontesthes bonariensis and its global response to the insecticide cypermethrin were studied using HPLC-MS-based metabolomics. Three experiments using either juveniles or adults of O. bonariensis were performed by exposing fish (6, 24, or 96 h) to sublethal concentrations of cypermethrin (5 or 10 µg/L). Metabolic profiling was performed on either whole bile or aqueous and organic extracts. Chromatography was performed using a C18 column and an ACN/H2O mobile phase. Electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) interfaces were used in positive and negative modes. Full scan MS data were processed using the XCMS software, log-transformed, and analyzed using either regression analysis or principal component analysis (PCA). The highest amount of information (1163 peaks) was yielded by analyzing the whole bile with the ESI⁻ interface. Complementary information, useful for metabolite confirmation, was obtained from the aqueous and organic extracts and using the ESI⁺ and APCI interfaces. The bile metabolic profile of O. bonariensis was characterized by some abundant metabolite ions corresponding with taurine conjugated bile acids, which were useful as reference peaks. A characteristic global metabolic response to cypermethrin was identified in the bile of O. bonariensis. A ten-fold or higher variation in abundance was observed in the whole bile of exposed fish for a small group of peaks (32), and these peaks corresponded to an even smaller number of metabolites (nineteen). Both regression analysis and PCA were useful in identifying those peaks, better explaining differences between exposed and control groups, but slight differences were suggested by each of those methods. Using unsupervised PCA scores, we were able to distinguish organisms from each treatment on the basis of the metabolic changes induced by the cypermethrin, this variability being explained mainly by only one principal component (PC3, 17.7 percent total variance). Two cypermethrin metabolites were identified as major contributors within the augmented peaks: the known glucuronide of 4'-hydroxy-cypermethrin and the sulfate of 4'-hydroxycypermethrin, not previously reported in fish bile. The HPLC-MS-based metabolomic approach demonstrated to be a powerful ecotoxicological tool for identifying biological responses to pollutants, discovering new metabolic pathways and proposing specific biomarkers using non model organisms.

Short and long-term effects of endosulfan, cypermethrin, spinosad, and methoxyfenozide on adults of Chrysoperla externa (Neuroptera: Chrysopidae).

The susceptibility assessment of natural enemies to pesticides is relevant before the use of selective pesticides and biological control agents within the framework of integrated pest management programs. Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae) is a predator considered a potential biocontrol agent of agricultural pests in the Neothropical Region. The aim of this study was to evaluate the short and long-term effects of two broad spectrum insecticides (cypermethrin and endosulfan) and two biorational (spinosad and methoxyfenozide) registered in Argentina on young C. externa adults under laboratory conditions by ingestion through drinking water. The assessed end-points were: survivorship during preoviposition period, preoviposition time, fecundity and fertility of females, and survivorship of the progeny. Total survivorship of adults was adversely affected only by cypermethrin that reduced the survivorship of adults irrespective of sex. Although endosulfan did not induce significant total mortality, it reduced the survivorship of females. All tested insecticides delayed the reproductive maturity of adults. Cypermethrin, endosulfan, and spinosad reduced the fecundity of females. Fertility was affected only by spinosad. Methoxyfenozide was the insecticide inducing the lowest effects on reproductive parameters. No effects were observed on the survival of progeny with any of the tested compounds. In accordance with the International Organization for Biological Control and Noxious Animals and Plants guidelines the insecticides were classified according to its lethal effects in the following toxicity classes: methoxyfenozide, spinosad, and endosulfan class 1 (innocuous): cypermethrin class 2 (moderately toxic). However if sublethal effects are taken into account, spinosad and endosulfan should not be considered innocuous.

Effects of pesticide formulations and active ingredients on the coelenterate Hydra attenuata (Pallas, 1766).

Lethal effects of active ingredients and formulations of widely used soybean pesticides were assessed with the Hydra attenuata toxicity test. Studied pesticides were insecticides chlorpyrifos and cypermethrin, and herbicide glyphosate. Results indicate the following toxicity trend: chlorpyrifos > cypermethrin > glyphosate. Tested active ingredients of insecticides and respective formulations did not significantly differ between them. Glyphosate formulation exhibited higher toxicity at low concentrations (LC(1-10)) respect to active ingredient, reversing this behavior at higher concentrations (LC(50-90)). Comparing H. attenuata sensitivity with existent toxicity data for aquatic organisms indicates that this species is poorly sensitive to tested insecticides and highly sensitive to the herbicide.

Reduced body condition and enzymatic alterations in frogs inhabiting intensive crop production areas.

Parameters indicative of general condition and health were compared amongst adult frogs inhabiting uncultivated lands and fields subjected to agricultural practices typical of the humid pampas of Argentina. Whereas no significant differences existed in the parasite load and external malformations prevalence rate of adult frogs from either environments, a reduced condition factor was clearly demonstrated in frogs from agricultural lands. This conclusion was reached for four frog species of different life habits: the terrestrial fossorial Rhinella fernandezae, the terrestrial Leptodactylus latinasus, the semi-aquatic Leptodactylus ocellatus, and the arborescent Hypsiboas pulchellus. A distinct pattern of enzymatic modifications was furthermore observed in L. ocellatus and H. pulchellus from agricultural lands, including elevated hepatic activities of catalase and cholinesterase, and an inhibition of liver GST activity. Further studies should investigate the causes and consequences of the systematically low condition factor observed in frogs from agricultural fields of the humid pampas of Argentina.

Lethal and sublethal effects of cypermethrin to Hypsiboas pulchellus tadpoles.

The study of the effects of the insecticide cypermethrin (CY) technical grade and its Sherpa® commercial formulation on Hypsiboas pulchellus tadpoles assessing lethality, behavior, growth, and abnormalities under standardized laboratory conditions is reported. Observed behaviors were identified and categorized by means of a ranking system according to the loss of mobility. Results of acute lethal effects indicate higher potency for Sherpa® at this level of assessment. All effects on behavior showed an increasing degree of injury as insecticide concentration increased. Organisms exposed to technical grade CY showed lower body length with respect to controls from 3.44 µg CY/L to higher concentrations, whereas those exposed to Sherpa® exhibited lower growth from 0.83 µg CY/L. Both forms of the tested insecticide caused abnormalities between 0.34 and 4.18 µg CY/L, but 100% of malformed individuals was detected from 34.4 µg CY/L for those exposed to the technical grade CY, and from 8.36 µg CY/L for those exposed to Sherpa®. This study proposes the use of easily identifiable and distinguishable sublethal end-points. The high input loads of CY in natural environments, the detected concentrations in the field, in addition to the low levels of this insecticide required to induce sublethal effects (which could eventually lead to death), allow for the conclusion that the insecticide is a risk factor for amphibians inhabiting agroecosystems.