Feeding behavior of early life stages of the zebrafish Danio rerio is altered by exposure to glyphosate.

Glyphosate levels and the transfer of glyphosate across trophic levels have rarely been studied in zooplankton. The food preferences of zebrafish during the first-feeding stage (which is critical for the survival of organisms), were analyzed because of the requirement for live food. Larval survival begins to be affected when glyphosate intake exceeds 0.3666 µg/larvae/day, in the case that only the food is contaminated; if the medium is also contaminated, the effects on survival start from 0.2456 µg/larvae/day. It was shown that glyphosate was more likely to be incorporated through the medium than through the food (zooplankton), which supports the results of previous studies that have ruled out the potential for biomagnification. The bioconcentration factor (BCF) of glyphosate was determined using an ELISA tests specific to measure glyphosate in the fish D. rerio, the rotifers Brachionus calyciflorus and Lecane papuana, and the cladoceran Ceriodaphnia dubia. The experimental design consisted in exposing seven zebrafish adults per replica (four replicates) in three treatments 1, 5, and 10 mg/L of glyphosate for 96 h to obtain bioconcentration factors in the gills, liver, and muscle. These concentrations were selected as potential glyphosate concentrations right after application as double highest reported concentration. Glyphosate levels in zooplankton can represent up to 6.26% of the total weight of rotifers (BFC = 60.35) and in zebrafish adult organs were less than 8 µg/mg of tissue (BCF values < 6). Although glyphosate does not biomagnify, our results suggest that glyphosate affected the dynamics between zooplankton and zebrafish larvae, diminishing survival and feeding rates, given that zooplankton species bioconcentrate glyphosate in large quantities. The BCF values found in this contribution are higher than expected. Glyphosate exposure affected energy metabolism and feeding behavior of zebrafish larvae, which presented high mortality rates at environmentally relevant concentrations.

Synergistic, antagonistic, and additive effects of heavy metals (copper and cadmium) and polycyclic aromatic hydrocarbons (PAHs) under binary and tertiary combinations in key habitat-forming kelp species of Chile.

Heavy metals (HMs) and polycyclic aromatic hydrocarbons (PAHs) are persistent toxicants in coastal environments. Notably, in comparison to individual metal toxicity, knowledge about the effects of HMs and PAHs mixtures on kelps remains scarce. Accordingly, we performed in vitro experiments to determine the individual and combined effects of Cu, Cd, and PAHs on spore release, settlement, and germination on Macrocystis pyrifera and Lessonia spicata, two key-habitat forming kelp species of the coast of the Valparaíso Region in Chile. This region concentrates highly polluting industries, mainly due to unrestrained mining and fossil-fuel energy production. Single Cu, Cd, and PAHs treatments included concentrations in the ranges 5-200, 0.125-2000, and 0.05-100 µg/L, respectively, and a toxic-free treatment. Cu, Cd, and PAHs concentrations causing 20-50% (IC20, IC50) arrested spore release, settlement, and germination were determined, and the results shown in both species that single Cu, Cd, and PAHs IC20 values were generally lower on spore release than on spore settlement and germination, probably due to the absence of a cell wall in spores compared to later stages. Binary equitoxic IC20s mixture treatments changed from an antagonistic response to another with a greater inhibitory effect on spore release, from hour 1 to 7, whereas in IC50 treatments, the response was always antagonistic. The tertiary IC20 mixture of Cu+Cd+PAHs produced generally an antagonistic effect. Remarkably, all IC20 equitoxic mixture treatments showed a synergistic response on spore settlement in both kelps, suggesting that these toxicants are extremely harmful to kelp population persistence near highly polluted sites.

Embryonic and larval development is conditioned by water temperature and maternal origin of eggs in the sea urchin Arbacia dufresnii (Echinodermata: Echinoidea) / El desarrollo embrionario y larvario está condicionado por la temperatura del agua y el origen materno de los huevos en el erizo de mar Arbacia dufresnii (Echinodermata: Echinoidea)

Introduction: Embryonic and larval development in sea urchins is highly dependent on maternal nutritional status and on the environmental conditions of the seawater. Objective: To compare the development of Arbacia dufresnii in two different water temperatures and in progeny with varying maternal origins. Methods: We induced A. dufresnii females and males from Nuevo Gulf to spawn, collected the eggs of each female individually (progeny), separated them into two seawater temperatures (12 and 17 °C), and fertilized them. We recorded the percentage of fertilized eggs and embryos per developmental stage according to time, temperature and progeny. We measured larval growth by total length (TL) and midline body length (M) according to time post fecundation (DPF), temperature, and progeny. Results: Temperature did not affect fertilization, but embryo development was faster and more synchronized in the high temperature treatment. The generalized linear models indicate that embryo development depends on a quadruple interaction between the embryonic stage, time (h), seawater temperature and progeny. Larval growth was faster, producing larger larvae at the highest temperature. Larval growth depends on a triple interaction between time (DPF), seawater temperature and progeny. Conclusions: We found a temperature and progeny impact during embryonic and larval development and, in both cases, these factors generate a synergistic effect on developmental timing and larval size. This probably provides a survival advantage as a more rapid speed of development implies a decrease in the time spent in the water column, where the sea urchins are vulnerable to biotic and abiotic stressors.

Introducción: El desarrollo embrionario y larvario de los erizos de mar depende en gran medida del estado nutricional materno y de las condiciones ambientales del agua de mar. Objetivo: Comparar el desarrollo de Arbacia dufresnii en dos temperaturas de agua diferentes y en progenies con diferentes orígenes maternos. Métodos: Indujimos a las hembras y machos de A. dufresnii del Golfo Nuevo a desovar, recolectamos los huevos de cada hembra individualmente (progenie), los separamos en dos temperaturas de agua de mar (12 y 17 ° C) y los fertilizamos. Registramos el porcentaje de óvulos fecundados y el porcentaje de embriones por etapa de desarrollo según tiempo, temperatura y descendencia. Medimos el crecimiento larvario según la longitud total (TL) y la longitud corporal de la línea media (M) de acuerdo con el tiempo en días post fecundación, la temperatura y la progenie. Resultados: La temperatura no afectó la fertilización, pero el desarrollo del embrión fue más rápido y más sincronizado en el tratamiento de alta temperatura. Los modelos lineales generalizados indican que el desarrollo del embrión depende una interacción cuádruple entre el estadio embrionario, el tiempo (h), la temperatura del agua de mar y la progenie. El crecimiento larvario fue más rápido, produciendo larvas más grandes a la temperatura más alta. El crecimiento de las larvas depende de una triple interacción entre el tiempo (DPF), la temperatura del agua de mar y la progenie. Conclusiones: Encontramos un impacto en la temperatura y en la progenie durante el desarrollo embrionario y larvario y, en ambos casos, estos factores generaron un efecto sinérgico sobre el tiempo de desarrollo y el tamaño de las larvas. Esto probablemente proporciona una ventaja de supervivencia, ya que una velocidad de desarrollo más rápida implica una disminución en el tiempo que pasan en la columna de agua, donde los erizos de mar son vulnerables a los factores estresantes bióticos y abióticos.

Early Life Stage Assays in Zebrafish.

Fish embryo toxicity (FET) test using zebrafish (Danio rerio) has been established as an alternative assay to animal experimentation. The FET assay enables the assessment of multiple morphological endpoints during the development of zebrafish early life stages, showing high impact to the field of ecotoxicology on risk assessment of chemicals and pollutants. Moreover, it is also applied to screening drug-induced toxicity and human diseases, due to the high genetic and physiological orthology between zebrafish and humans. Here, we describe FET test, with all steps and several adaptations involved in the methodological procedures. To demonstrate the efficiency of this method, results using the reference substance 3,4-dichloroaniline (DCA) were included to demonstrate sublethal and teratogenic malformations on zebrafish embryos. Thus, there is a strong tendency for using FET tests as a replacement strategy of traditional tests in toxicology and ecotoxicology.

Urban effluents affect the early development stages of Brazilian fish species with implications for their population dynamics.

The pollution from urban effluents discharged into natural waters is a major cause of aquatic biodiversity loss. Ecotoxicological testing contributes significantly to understand the risk of exposure to the biota and to establish conservation policies. The objective of the current study was to assess the toxicity of a river highly influenced by urban effluents (Atuba River, Curitiba city, Southern Brazil) to the early stages of development in four South American native fish species, investigating the consequences at the population level through mathematical modelling. The species chosen were Salminus brasiliensis, Prochilodus lineatus, Rhamdia quelen, and Pseudoplatystoma corruscans, ecologically important species encompassing different conservation statuses and vulnerability. The embryos were exposed from 8 to 96 h post fertilization to the Atuba River water, collected downstream of the largest wastewater treatment plant in the Metropolitan Region of Curitiba, and their survival rates and deformities were registered. The species S. brasiliensis and P. lineatus presented the highest mortality rates, showing high sensitivity to the pollutants present in the water. According to the individual-based mathematical model, these species showed high vulnerability and risk of extinction under the tested experimental conditions, even when different sensitivity scenarios of juveniles and adults were considered. The other two species, R. quelen and P. corruscans, showed a more resistant condition to mortality, but also presented high frequency and severity of deformities. These results emphasize the importance of testing the sensitivity of different Brazilian native species for the conservation of biodiversity and the application of models to predict the effects of pollutants at the population level.

Low concentration of 2,4,6-tribromophenol (TBP) represents a risk to South American silver catfish Ramdia quelen (Quoy and Gaimard, 1824) population.

The 2,4,6-tribromophenol (TBP) is an environmental persistent pollutant widely used as flame retardant, antimicrobial and insecticide agent in wood preservation and plastic production. Currently, TBP is found in environmental compartments such as soil, freshwater, groundwater, sewage sludge and domestic dust, but the effects to biota and the risk of exposure to aquatic vertebrates are still scarce. In the present study, Rhamdia quelen fish embryos (8 h post-fertilization - hpf) were exposed to 0.3 and 3.0 µg L-1 of TBP until 96 hpf. Biochemical biomarkers, hatching, survival and larvae/embryo malformations were evaluated after exposure. Additionally, a mathematical model was proposed to evaluate the effects along further generations. The results showed that TBP decreased the survival level but did not cause significant difference in the hatching rates. After 72 and 96 hpf, individuals from the highest tested concentration group showed more severe malformations than individuals from control and the lower concentrations groups. The deformities were concentrated on the embryos facial region where the sensorial structures related to fish behavior are present. The biochemical biomarkers revealed both oxidative stress and neurotoxicity signs after exposure to the contaminant, while the application of the mathematical model showed a decrease of population in both tested TBP concentrations. In conclusion, the current results demonstrated that TBP is toxic to R. quelen embryos and represents a risk to population after early life stage exposure.

Exposure to ayahuasca induces developmental and behavioral alterations on early life stages of zebrafish.

Ayahuasca is a psychoactive concoction prepared from the plants Banisteriopsis caapi and Psychotria viridis which are used ancestrally by Amazonian Indian populations and more recently, by Christian religious groups in Brazil and other countries. The aims of the present study were to identify the effects of ayahuasca on zebrafish embryo development and neurobehavior. Toxicity and developmental endpoints for zebrafish embryos were assessed from 0 to 1000 mg/L over 96 h of exposure. The effects on locomotor activity of zebrafish larvae were assessed using a video tracking system (ZebraBox) from 0 to 20 mg/L and after 120 and 144 h of exposure. The LC50 of ayahuasca in zebrafish was determined as 236.3 mg/L. Ayahuasca exposure caused significant developmental anomalies in zebrafish embryos, mainly at the highest concentration tested, including hatching delay, loss of equilibrium, edema and the accumulation of red blood cells. Embryo behavior was also significantly affected, with decreased locomotor activity at the highest concentration tested. These results are in accordance with data obtained in mammal studies highlighting the possible risks of uncontrolled use of ayahuasca. Further research employing more specific behavior analysis could provide additional data on both therapeutic benefits and possible toxicological risk of ayahuasca.

Lethal and sublethal toxicity of abamectin and difenoconazole (individually and in mixture) to early life stages of zebrafish.

In recent years, the need for the development of alternative test methods for the conventional acute fish toxicity test (AFT) with adult fish has often been discussed. In addition, concerns have been raised on the potential risks related with environmentally realistic pesticide mixtures since risk evaluations have traditionally been based on individual pesticides. The insecticide/acaricide abamectin and the fungicide difenoconazole are the main pesticides that are intensively used in Brazilian strawberry crop and are hence likely to occur simultaneously in edge-of-field waterbodies. The aim of the present study was therefore to evaluate the lethal and sublethal toxicity of single and mixture exposures of these pesticides to zebrafish early life stages (embryos and juveniles). By comparing the derived toxicity data of the individual compounds with that previously determined for zebrafish adults, the order of life stage sensitivity was juvenile > adult > embryo. The pesticide mixture revealed a dose-level dependent deviation of the independent action model, with antagonism at low dose levels and synergism at high dose levels. Sublethal parameters (especially those related with locomotion) were considerably more sensitive than lethality. Subsequently, the inclusion of sublethal parameters may greatly improve the sensitivity of FET tests and hence its suitability as a substitution of adult fish testing in risk assessment evaluations.

The effects of ocean acidification and a carbon dioxide capture and storage leak on the early life stages of the marine mussel Perna perna (Linneaus, 1758) and metal bioavailability.

The study assesses the effects of carbon dioxide capture and storage (CCS) leaks and ocean acidification (OA) on the metal bioavailability and reproduction of the mytilid Perna perna. In laboratory-scale experiments, CCS leakage scenarios (pH 7.0, 6.5, 6.0) and one OA (pH 7.6) scenario were tested using metal-contaminated sediment elutriates and seawater from Santos Bay. The OA treatment did not have an effect on fertilisation, while significant effects were observed in larval-development bioassays where only 16 to 27 % of larva developed normally. In treatments that simulated CO2 leaks, when compared with control, fertilisation success gradually decreased and no larva developed to the D-shaped stage. A fall in pH increased the bioavailability of metals to marine mussels. Larva shell size was significantly affected by both elutriates when compared with seawater; moreover, a significant difference occurred at pH 6.5 between elutriates in the fertilisation bioassay.

Characterization of the early development of Pseudoplatystoma reticulatum Eigenmann & Eigenmann, 1889 (Siluriformes: Pimelodidae) from the Paraguay River Basin

This study described the initial development of Pseudoplatystoma reticulatum, as well as changes in growth patterns. Morphometric and meristic variables were analyzed during embryonic, larval and juvenile periods, even as allometric growth coefficients for larvae and juveniles. Eggs showed an average diameter of 1.79 mm and broad perivitelline space (23.41%). The total length (TL) of the larvae ranged from 3.05 to 25.72 mm, and the total number of myomeres ranged from 45 to 50 (preanal = 13-16 and postanal = 30-35). Initial pigmentation is poor, concentrated at the extremities of the yolk sac. In the flexion stage, the pigmentation intensifies and forms a longitudinal stripe extending from the snout to the operculum and two longitudinal stripes on the body, one dorsal and the other ventral, that unite at the caudal peduncle. At first, there is a differential growth of the head and tail, and then there is the growth of the body, which shows changes in the ontogenetic priorities of feeding, swimming, and breathing capacities. Our results are very important for ecology, systematics, and hatchering, especially in terms of ontogenetic variation in morphology, growth, feeding, behavior, and mortality of P. reticulatum.

Este estudo descreveu o desenvolvimento inicial de Pseudoplatystoma reticulatum, bem como as mudanças nos padrões de crescimento, testando a hipótese da ocorrência de crescimento diferencial durante a ontogenia inicial da espécie. Variáveis morfométricas e merísticas foram analisadas durante os períodos embrionário, larval e juvenil, assim como os coeficientes de crescimento alométrico para larvas e juvenis. Os ovos apresentaram diâmetro médio de 1,79 mm e espaço perivitelino amplo (23,41%). O comprimento total (CT) das larvas variou de 3,05 a 125,72 mm e o número total de miômeros de 45 a 50 (pré-anal=13-16 e pós-anal=30-35). A pigmentação inicial é escassa, concentrada nas extremidades do saco vitelino. A partir do estágio de flexão a pigmentação se intensifica, formando uma faixa longitudinal que se estende do focinho ao opérculo e duas no corpo, uma dorsal e outra ventral, que se unem no pedúnculo caudal. Inicialmente há um crescimento diferenciado da cabeça e da cauda, com posterior crescimento do tronco, que demonstra mudanças nas prioridades ontogenéticas para as capacidades de alimentação, natação e respiração. Nossos resultados são de grande importância para a ecologia, sistemática e larvicultura, especialmente em termos de variação ontogenética na morfologia, crescimento, alimentação, comportamento e mortalidade P. reticulatum.

Characterization of the early development of Pseudoplatystoma reticulatum Eigenmann & Eigenmann, 1889 (Siluriformes: Pimelodidae) from the Paraguay River Basin

This study described the initial development of Pseudoplatystoma reticulatum, as well as changes in growth patterns. Morphometric and meristic variables were analyzed during embryonic, larval and juvenile periods, even as allometric growth coefficients for larvae and juveniles. Eggs showed an average diameter of 1.79 mm and broad perivitelline space (23.41%). The total length (TL) of the larvae ranged from 3.05 to 25.72 mm, and the total number of myomeres ranged from 45 to 50 (preanal = 13-16 and postanal = 30-35). Initial pigmentation is poor, concentrated at the extremities of the yolk sac. In the flexion stage, the pigmentation intensifies and forms a longitudinal stripe extending from the snout to the operculum and two longitudinal stripes on the body, one dorsal and the other ventral, that unite at the caudal peduncle. At first, there is a differential growth of the head and tail, and then there is the growth of the body, which shows changes in the ontogenetic priorities of feeding, swimming, and breathing capacities. Our results are very important for ecology, systematics, and hatchering, especially in terms of ontogenetic variation in morphology, growth, feeding, behavior, and mortality of P. reticulatum.(AU)

Este estudo descreveu o desenvolvimento inicial de Pseudoplatystoma reticulatum, bem como as mudanças nos padrões de crescimento, testando a hipótese da ocorrência de crescimento diferencial durante a ontogenia inicial da espécie. Variáveis morfométricas e merísticas foram analisadas durante os períodos embrionário, larval e juvenil, assim como os coeficientes de crescimento alométrico para larvas e juvenis. Os ovos apresentaram diâmetro médio de 1,79 mm e espaço perivitelino amplo (23,41%). O comprimento total (CT) das larvas variou de 3,05 a 125,72 mm e o número total de miômeros de 45 a 50 (pré-anal=13-16 e pós-anal=30-35). A pigmentação inicial é escassa, concentrada nas extremidades do saco vitelino. A partir do estágio de flexão a pigmentação se intensifica, formando uma faixa longitudinal que se estende do focinho ao opérculo e duas no corpo, uma dorsal e outra ventral, que se unem no pedúnculo caudal. Inicialmente há um crescimento diferenciado da cabeça e da cauda, com posterior crescimento do tronco, que demonstra mudanças nas prioridades ontogenéticas para as capacidades de alimentação, natação e respiração. Nossos resultados são de grande importância para a ecologia, sistemática e larvicultura, especialmente em termos de variação ontogenética na morfologia, crescimento, alimentação, comportamento e mortalidade P. reticulatum.(AU)