Basal levels of biochemical biomarkers in the freshwater prawn Palaemon argentinus and their alterations due to the exposure of both insecticides cypermethrin and spirotetramat.

The aim of this study was to evaluate the sensitivity of the prawn Palaemon argentinus to the pyrethroid cypermethrin (CYP) and the tetramic acid spirotetramat (STM). These treatments were compared with prawns collected at a reference site to define their basal physiological state. Initially, physicochemical parameters and several pollutants at the selected site were analyzed. The LC50-96 h was determined in adult prawns. Then, prawns were exposed for 96 h to sublethal concentrations of CYP (0.0005 µg/l) and STM (0.44 mg/l) to evaluate the effects on some biochemical endpoints. A treatment combining both pesticides was also added at 5 % of these values. Controls with and without solvent (acetone) were included. The LC50-96 h values were 0.005 µg/l and 4.43 mg/l for CYP and STM, respectively. Moreover, some biomarkers linked to oxidative and energy metabolism were analyzed in the hepatopancreas and muscle of both essayed prawns and those at the basal state. The STM caused a significant decrease in total protein content (32 %) in contrast to the increase of protein carbonyl content (71 %) (p < 0.05). Also, glutathione S-transferase (52 %) and catalase (61 %) activities in the hepatopancreas of exposed prawns were higher compared to both the control and state basal groups (p < 0.05). In muscle, only a significant decrease in the lactate content (69 %) was caused by STM (p < 0.05). In addition, CYP caused a significant increase in the lactate dehydrogenase activity (110 %) in muscle and triacylglycerol content (73 %) in the hepatopancreas (p < 0.05). The integrated biomarker index (IBRv2) analysis showed that STM caused greater damage than CYP. Besides, the combined treatment showed an antagonistic interaction between both insecticides. The differential response of biomarkers to both CYP and STM exposure with respect to their basal levels shows a high sensitivity of P. argentinus demonstrating its potential role as a bioindicator organism.

Sensitivity of the Neotropical Solitary Bee Centris analis F. (Hymenoptera, Apidae) to the Reference Insecticide Dimethoate for Pesticide Risk Assessment.

Currently, only Apis mellifera is used in environmental regulation to evaluate the hazard of pesticides to pollinators. The low representativeness of pollinators and bee diversity in this approach may result in insufficient protection for the wild species. This scenario is intensified in tropical environments, where little is known about the effects of pesticides on solitary bees. We aimed to calculate the medium lethal dose (LD50) and medium lethal concentration (LC50) of the insecticide dimethoate in the Neotropical solitary bee Centris analis, a cavity-nesting, oil-collecting bee distributed from Brazil to Mexico. Males and females of C. analis were exposed orally to dimethoate for 48 h under laboratory conditions. Lethality was assessed every 24 h until 144 h after the beginning of the test. After the LD50 calculation, we compared the value with available LD50 values in the literature of other bee species using the species sensitivity distribution curve. In 48 h of exposure, males showed an LD50 value 1.33 times lower than females (32.78 and 43.84 ng active ingredient/bee, respectively). Centris analis was more sensitive to dimethoate than the model species A. mellifera and the solitary bee from temperate zones, Osmia lignaria. However, on a body weight basis, C. analis and A. mellifera had similar LD50 values. Ours is the first study that calculated an LD50 for a Neotropical solitary bee. Besides, the results are of crucial importance for a better understanding of the effects of pesticides on the tropical bee fauna and will help to improve the risk assessment of pesticides to bees under tropical conditions, giving attention to wild species, which are commonly neglected. Environ Toxicol Chem 2023;42:2758-2767. © 2023 SETAC.

Acute toxicity of three herbicide formulations of Astyanax altiparanae (Characiformes, Characidae), an emerging neotropical fish model species.

Herbicides are used in agriculture to control harmful crop weeds, prevent algae proliferation, and enhance macrophyte growth. Herbicide contamination of water bodies might exert toxic effects on fish in different development stages. Sperm, embryos, and adults of Astyanax altiparanae were used as a model to examine the detrimental effects of the following herbicide formulations: Roundup Transorb® (glyphosate), Arsenal® NA (imazapyr), and Reglone® (diquat). The lethal concentration 50 (LC50) values for adults using glyphosate and imazapyr were 3.14 mg/L and 4.59 mg/L, respectively, while the LC50 was higher than 28 mg/L for diquat. For the initial stages of embryo development, LC50 values were 16.52 mg/L glyphosate, 9.33 mg/L imazapyr, and 1084 mg/L diquat. Inhibition of sperm motility was noted at 252 mg/L glyphosate, 137 mg/L imazapyr, and 11,300 mg/L diquat, with an average sperm viability of 12.5%, 73.2%, and 89.3%, respectively, compared to 87.5% detected to control. A. altiparanae exhibited different sensitivities to the herbicide formulations investigated in the developmental stages evaluated. Roundup Transorb® exposure was more toxic for adults, while Arsenal® NA was most harmful for early embryonic development and inhibited sperm motility. Reglone® demonstrated low toxicity for A. altiparanae compared to Roundup Transorb® and Arsenal® NA. A. altiparanae may be considered an emerging fish model for toxicological studies for the neotropical region due to its wide distribution and biological characteristics.

Differential Activities of Antioxidant Enzymes, Superoxide Dismutase, Peroxidase, and Catalase vis-à-vis Phosphine Resistance in Field Populations of Lesser Grain Borer (Rhyzopertha dominica) from India.

Susceptibility to phosphine was compared in 15 populations of lesser grain borer (Rhyzopertha dominica) collected from grain storage godowns across India. A high level of resistance to phosphine was noticed in R. dominica collected from northern India compared to those collected from northeastern regions of India. The median lethal concentration values varied from 0.024 mg/L to 1.991 mg/L, with 1.63 to 82.96-fold resistance compared to laboratory susceptible checks. Antioxidant enzymes have been reported to negate the reactive oxygen species generated upon encountering the fumigant phosphine. Distinct differences in the activity of antioxidant enzymes were noticed in the field populations exposed to phosphine. Peroxidase activity varied between 1.28 and 336.8 nmol H2O2 reduced/min/mg protein. The superoxide dismutase inhibition rate was between 81.29 and 99.66%, and catalase activity varied between 6.28 and 320.13 nmol H2O2 reduced/min/mg protein. The findings of our investigation show that the activities of peroxidase and superoxide dismutase are positively linked (p < 0.01) with an increase in resistance ratios, whereas catalase was found to have a negative association with resistance to phosphine. The reported results elucidate the differential activities of principal antioxidant enzymes in scavenging the oxyradicals (O2•-, H2O2,•OH) associated with tolerance to phosphine in R. dominica.

Influence of salinity on growth and survival of juvenile Sardinella brasiliensis / Influência da salinidade no crescimento e sobrevivência de juvenis de Sardinella brasiliensis

The aim of the study was to evaluate the influence of changes in seawater salinity on juvenile Brazilian sardine (Sardinella brasiliensis). Through two assays, the LC50 (96 h) and the zootechnical performance (42 days) were determined, respectively. In the first assay, six treatments of salinity 0, 7, 14, 21, 28 and 35 parts per thousand (ppt) with three replicates were established. For this, 100-L cylinder-conical tanks were used, with 30 individuals with 45-day after eclosion (DAE) per unit, without water renewal and feeding. In the groups of salinity 0 and 7 ppt, all fish died on the first day. The LC50 was estimated at salinity 11.13 ppt. The second trial was designed with five treatments (salinity 7, 14, 21, 28 and 35 ppt), with three replicates. Two thousand-L cylindrical-conical tanks were used, with 30 individuals (58 DAE) per unit, with water renewal and feeding until apparent satiation. In the treatment salinity 7 ppt, all fish died by the second day. At salinity 35 ppt, the highest growth rates (2.78 g) and survival (100%) were observed. Salinity 14 ppt had the lowest survival (83%) and growth (1.48 g). We concluded that the juvenile sardines can be adapted to environments with salinity from 14 ppt, with significant losses. However, salinity 35 ppt showed the highest survival and growth rates.(AU)

O objetivo do estudo foi avaliar a influência da variação da salinidade na água do mar em juvenis de sardinha brasileira (Sardinella brasiliensis). Por meio de dois ensaios, determinaram-se a CL50 (96 h) e o desempenho zootécnico (42 dias), respectivamente. No primeiro ensaio foram estabelecidos seis tratamentos de salinidade 0, 7, 14, 21, 28 e 35 parts per thousand (ppt) com três repetições. Para isso, foram utilizados tanques cilindro-cônicos de 100 L com 30 peixes (45 dias após eclosão ­ DAE) por unidade, sem renovação de água e sem alimentação. Nos grupos de salinidade 0 e 7 ppt todos os peixes morreram no primeiro dia. O LC50 foi estimado na salinidade 11,13 ppt. O segundo ensaio foi delineado com cinco tratamentos (salinidade 7, 14, 21, 28 e 35 ppt), com três repetições. Foram empregados tanques cilíndrico-cônicos de 200 L, com 30 indivíduos (58 DAE) por unidade, com renovação de água e alimentação até a saciedade aparente. No tratamento salinidade 7 ppt, todos os peixes morreram no segundo dia. Na salinidade 35 ppt foram observadas as maiores taxas de crescimento (2,78 g) e sobrevivência (100%). A salinidade 14 ppt teve a menor sobrevivência (83%) e crescimento (1,48 g). Concluímos que os juvenis de sardinha podem se adaptar a ambientes com salinidade de 14 ppt em diante, com perdas significativas, no entanto a salinidade 35 ppt apresentou as maiores taxas de sobrevivência e crescimento.(AU)

Toxicological effect and enzymatic disorder of non-studied emerging contaminants in Artemia salina model.

Emerging contaminants such as sunscreens, hair dyes and flame retardants have been found at important concentrations in surface water (river, lake, ocean), but their negative impact on different aquatic species is not fully known. This study evaluated the effect of benzophenone (BZ), 2,5-diaminotoluene sulfate (PTD), p-phenylenediamine (PPD) and tetrabromobisphenol A (TBPA) on survival (LC50) and the impact of sublethal concentrations (LC25) on the activity of enzymes linked to stress oxidative process in brine shrimp under two temperature conditions (22 °C and 28 °C) for 24 h and 48 h of exposure time. LC50 values obtained for each chemical substance and the activity of GST, AChE and LDH were significantly affected by the temperature conditions and exposure time. In contrast, GPx was only altered by the tested compound. TBBPA (LC50 from 17.05 up to 28.55 µg/L) and BZ (LC50 from 14.86 up to 24.49 mg/L) resulted in the most toxic substances for A. salina. The impact of dyes, such as PTD and PPD, on aquatic organisms is limited. These are the first results that show that not only dyes, but their respective by-products induce harmful effects in brine shrimp (LC50 for PTD and PPD were 23.6-396.3 and 52.0-164.9 mg/L respectively). Although this study model was very useful to evaluate the ecotoxicity of the different ECs, additional research is needed to increase available information related to the effects of dyes and other non-studied micropollutants on aquatic systems in general.

El mejillón Semimytilus patagonicus (Mytilida: Mytilidae) y el gusano Pseudonereis gallapagensis (Phyllodocida: Nereididae) como herramientas ecotoxicológica para la contaminación por detergentes en ambientes marinos / The mussel Semimytilus patagonicus (Mytilida: Mytilidae) and the worm Pseudonereis gallapagensis (Phyllodocida: Nereididae) as ecotoxicological tools for detergent pollution in marine environments

Introduction: The bivalve Semimytilus patagonicus is a potentially useful bioindicator because of its feeding mechanism, and the worm Pseudonereis gallapagensis is also interesting as a bioindicator because it is benthonic, abundant, and a food source for the squid Doryteuthis gahi. However, their sensitivity to contaminants has not been sufficiently studied. Objective: To test the usefulness of the mussel Semimytilus patagonicus and the polychaete Pseudonereis gallapagensis as ecotoxicological tools for detergents in the marine environment. Methods: We used 120 individuals of S. patagonicus from Miraflores and 120 of P. gallapagensis from Barranco (both near the city of Lima, Peru). For the bioassays, we used two anionic detergents (active ingredient, ai, Sodium Dodecylbenzene Sulfonate). For S. patagonicus, with an average valve length of 32.3 ± 6.4 mm, we tested "Double power Ariel®" (90 %) at concentrations of 17.5, 35, 70 and 140 mg ai l-1, evaluated after 48 and 72 h of exposure; and for P. gallapagensis, with a total body length of 20.4 ± 8.8 mm, we tested "Caricia®" at 62.5, 125, 250, 500 and 1 000 mg of ai l-1 at 24, 48 and 72 h of exposure. Results: The LC50 values (Mean Lethal Concentration) were 34.95 mg ia l-1 for S. patagonicus and 102.48 mg ia l-1 for P. gallapagensis at 72 h of exposure. The detergents were toxic for S. patagonicus and slightly toxic for P. gallapagensis. The risk classification for S. patagonicus is "harmful" and for P. gallapagensis "not classifiable". Conclusions: These two bioindicators allow evaluating the acute toxicity of SDBS-based commercial detergents in the marine aquatic environment.

Introducción: El bivalvo Semimytilus patagonicus es un bioindicador potencialmente útil por su mecanismo de alimentación, y el gusano Pseudonereis gallapagensis también es interesante como bioindicador por ser bentónico, abundante y fuente de alimento para el calamar Doryteuthis gahi. Sin embargo, su sensibilidad a los contaminantes no ha sido suficientemente estudiada. Objetivo: Probar la utilidad del mejillón S. patagonicus y el poliqueto P. gallapagensis como herramientas ecotoxicológicas para detergentes en el medio marino. Métodos: Se utilizaron 120 individuos de S. patagonicus de Miraflores y 120 de P. gallapagensis de Barranco (ambos cerca de la ciudad de Lima, Perú). Para los bioensayos se utilizaron dos detergentes aniónicos (ingrediente activo, ia, dodecilbenceno sulfonato de sodio). Para S. patagonicus, con una longitud valver promedio de 32.3 ± 6.4 mm, probamos Ariel Doble Poder® (90 %) a concentraciones de 17.5, 35, 70 y 140 mg·ia·l-1, evaluadas a las 48 y 72 h de exposición; y para P. gallapagensis, con una longitud corporal total de 20.4 ± 8.8 mm, probamos Caricia® a 62.5, 125, 250, 500 y 1 000 mg·ia·l-1 a las 24, 48 y 72 h de exposición. Resultados: Los valores de CL50 (Concentración Letal Media) fueron de 34.95 mg·ia·l-1 para S. patagonicus y 102.48 mg·ia·l-1 para P. gallapagensis a las 72 h de exposición. Los detergentes fueron tóxicos para S. patagonicus y levemente tóxicos para P. gallapagensis. La clasificación de riesgo para S. patagonicus es "nocivo" y para P. gallapagensis "no clasificable". Conclusiones: Estos dos bioindicadores permiten evaluar la toxicidad aguda del detergente comercial a base de SDBS en el ambiente acuático marino.

A systematic review of research conducted by pioneer groups in ecotoxicological studies with bees in Brazil: advances and perspectives.

Brazil presents the most threatened endemic or rare species among neotropical regions, with the Hymenoptera order, to which bees belong, classified as a high-risk category. In Brazil, the main cause of bee death is the indiscriminate use of pesticides. In this context, groups such as Bee Ecotoxicology and Conservation Laboratory (LECA in Portuguese) and Bees and Environmental Services (ASAs in Portuguese) have become a reference in studies evaluating the impacts of pesticides on bees since 1976. Thus, the objective of this review was to conduct a quantitative and qualitative review of the studies conducted by these groups to evaluate and compile the advances made over the years, identify potential knowledge gaps for future studies, and support the sensitivities of stingless bees when compared to the species Apis mellifera. The quantitative analyses showed that most studies were carried out in the genus Apis, under laboratory conditions. However, more recently (since 2003), studies have also focused on stingless bees and the neonicotinoid class of insecticides. The most relevant gaps identified were the lack of studies under field conditions and on bee biology. The qualitative analyses indicated that Brazilian stingless bees are more susceptible to pesticides than A. mellifera and require a much lower average dose, concentration, or lethal time to display morphological and behavioral damage or decreased lifespan. Thus, future studies should work towards establishing more representative protocols for stingless bees. Furthermore, public policies must be created for the protection and conservation of bees native to Brazil.

Virulence of native isolates of entomopathogenic fungi (Hypocreales) against the "sweetpotato whitefly" Bemisia tabaci (Hemiptera: Aleyrodidae), including the effects of temperature and fungicides.

Hypocrella, Moelleriella and related species in the Hypocreales (Ascomycota, Sordariomycetes) cause epizootics of whiteflies and scale insects in nature. However, studies on their host specificity, virulence, infection cycles, optimal development under laboratory conditions, and compatibility with other control methods, are unexplored for most species. Under laboratory conditions, the virulence of several isolates of field-collected hypocrealean fungi (Hypocrella, Moelleriella, Regiocrella, and Verticillium) was determined on Bemisia tabaci eggs and 4th instar nymphs. In addition to virulence, the effect of temperature and two commercial fungicides on growth rates and germination of the isolates was evaluated. None of the isolates infected the eggs, while M. libera, M. ochracea, and M. turbinata caused high nymphal mortality. Moelleriella libera was the most virulent isolate. At all temperatures, M. libera, Regiocrella sp. (P17H20), and Verticillium cf. pseudohemipterigenum had the highest germination and growth rates. The optimal growth temperature depended on the isolate, but at 23 °C and 25 °C, the probability of spore germination was higher for most isolates. Finally, the fungicides azoxystrobin and chlorothalonil inhibited growth rates and conidial germination at 24 and 48 h of exposure. This research produces vital knowledge on the virulence and infection cycles of poorly studied native species of entomopathogenic fungi. In addition, the results provide information on the optimal temperature for development in laboratory conditions and susceptibility to fungicides, which could contribute to future biological control strategies.

Comparative toxicity of fipronil, malathion, and thiamethoxam on the stingless bee Tetragonisca fiebrigi (Schwarz, 1938)

Stingless bees are important pollinators for various plant crops. We investigated the susceptibility of Tetragonisca fiebrigito sublethal concentrations of insecticides fipronil, malathion, and thiamethoxam (administered through contact and ingestion) by determining the LC50values after 24hoursof exposure and analyzing changes in the activity of esterase isoenzymes and the chromatin in brain cells. The LC50values showed that all three insecticides were highly toxic through contact and ingestion. Electrophoretic analysis revealed that the relative EST-4 (carboxylesterase) activity in T. fiebrigi was partially inhibited by malathion and fipronil ingestion. Moreover, the EST-4 band intensity was increased following high-concentration thiamethoxam (contact) exposure, indicating the increased relative activity of this isoenzyme to detoxify the compound. In the cytochemical analysis of brain cells, the critical electrolyte concentration (CEC) points for the control stingless bees and malathion ingestion-exposed and thiamethoxam-exposed (contact and ingestion) stingless bees were in the range of 0.20-0.30 M MgCl2, whereas that for malathion contact-exposed bees was 0.15 M MgCl2, indicating chromatin relaxation and suggesting an increase in gene expression. In conclusion, T. fiebrigistingless bees are susceptible to the insecticides tested, and the parameters analyzed may be used as biomarkers to detect the presence of these compounds.(AU)

Impact of copper sulfate on survival, behavior, midgut morphology, and antioxidant activity of Partamona helleri (Apidae: Meliponini).

Copper sulfate (CuSO4) is widely used in agriculture as a pesticide and foliar fertilizer. However, the possible environmental risks associated with CuSO4 use, particularly related to pollinating insects, have been poorly studied. In this study, we evaluated both lethal and sublethal effects of CuSO4 on the stingless bee Partamona helleri. Foragers were orally exposed to five concentrations of CuSO4 (5000, 1666.7, 554.2, 183.4, 58.4 µg mL-1), and the concentration killing 50% (LC50) was estimated. This concentration (142.95 µg mL-1) was subsequently used in behavioral, midgut morphology, and antioxidant activity analyses. Bee mortality increased with the ingestion of increasing concentrations of CuSO4. Ingestion at the estimated LC50 resulted in altered walking behavior and damage to the midgut epithelium and peritrophic matrix of bees. Furthermore, the LC50 increased the catalase or superoxide dismutase activities and levels of the lipid peroxidation biomarker malondialdehyde. Furthermore, the in situ detection of caspase-3 and LC3, proteins related to apoptosis and autophagy, respectively, revealed that these processes are intensified in the midgut of treated bees. These data show that the ingestion of CuSO4 can have considerable sublethal effects on the walking behavior and midgut of stingless bees, and therefore could pose potential risks to pollinators including native bees. Graphical abstract.

Mercury chloride toxicity in juveniles Prochilodus argenteus a species from southeastern Brazil.

The aim of this study was to determine the 96-h LC50 (lethal concentration for 50% of the test population in 96 h of observation) of mercury chloride (HgCl2) and evaluated its absorption in the gills, liver, and muscle of juvenile "curimatã-pacu," Prochilodus argenteus. The fish were exposed for 4 days to different concentrations of HgCl2: 0.000, 0.0375, 00.075, 0.150, 0.225, 0.300, and 0.400 mg L-1 to get the 96-h LC50, and the concentration of Hg in the tissues. The fish were exposed to different levels of HgCl2 (0.000, 0.0375, 00.075, 0.150, 0.225, and 0.300 mg L-1), for 4 and 7 days, when different tissues (gills, liver, and muscle) were sampled. The mercury concentration values obtained were compared between tissues, days, and concentration of mercury in the water. The 96-h LC50 for P. argenteus was 0.339 mg L-1 of HgCl2, within the range found for other neotropical and tropical species, showing a medium sensitivity. The concentration of Hg in the tissues increased from the muscle to the gills and liver, and according to the increasing concentration of HgCl2 in the water. The fish liver is the most suitable tissue for environmental monitoring and for the evaluation of the consumed fished. Despite being the tissue that least accumulated mercury, the muscle must be evaluated because it is the part consumed by the population.

Expression of ion transport proteins and routine metabolism in juveniles of tropical gar (Atractosteus tropicus) exposed to ammonia.

Tropical gar (Atractosteus tropicus) thrives in aquatic habitats with high levels of total nitrogen (TAN) and unionized ammonia (NH3). However, the tolerance of TAN and NH3, the excretion mechanisms involved, and the effects of these chemicals on routine metabolism are still unknown. Therefore, our objectives were to assess the acute toxicity of TAN and NH3 in A. tropicus juveniles after a 96-h exposure (LC50-96 h) to NH4Cl and after chronic exposure to two concentrations (15% and 30% of LC50-96 h TAN) for 12 days, as well as to evaluate the transcriptional effects associated with Rhesus proteins (rhag, rhbg, rhcg) and ion transporters (NHE, NKA, NKCC, and CFTR) in gills and skin; and to determine the effects of TAN and NH3 on routine metabolism through oxygen consumption (µM g-1 h-1) and gill ventilation frequency (beats min-1). LC50-96 h values were 100.20 ± 11.21 mg/L for TAN and 3.756 ± 0.259 mg/L for NH3. The genes encoding Rhesus proteins and ion transporters in gills and skin showed a differential expression according to TAN concentrations and exposure time. Oxygen consumption on day 12 showed significant differences between treatments with 15% and 30% TAN. Gill ventilation frequency on day 12 was higher in fish exposed to 30% TAN. In conclusion, A. tropicus juveniles are highly tolerant to TAN, showing upregulation of the genes involved in TAN excretion through gills and skin, which affects routine oxygen consumption and energetic cost. These findings are relevant for understanding adaptations in the physiological response of a tropical ancestral air-breathing fish.

TOXICIDAD AGUDA DEL HERBICIDA PARAQUAT EN Oreochromis niloticus (CICHLIDAE) Y Macrobrachium olfersii (PALAEMONIDAE) / Paraquat's herbicide acute toxicity in Oreochromis niloticus (Cichlidae) and Macrobrachium olfersii (Palaemonidae)

RESUMEN El Paraquat es un herbicida utilizado en la actividad agropecuaria para controlar hierbas, su modo de acción es por medio de contacto y no selectivo. Debido a su alta solubilidad en agua y baja volatilidad representa un riesgo potencial para organismos acuáticos, principalmente los que son cultivados con aguas superficiales que reciben impacto de la actividad agrícola. La tilapia Oreochromis niloticus y el langostino Macrobrachium olfersii son organismos de importancia comercial para la industria acuícola del estado de Veracruz, México. El objetivo de este estudio fue determinar la Concentración Letal Media (CL50) del herbicida Dasurquat® (ingrediente activo Paraquat) a través de un bioensayo de toxicidad aguda (96 horas). Se utilizaron como especímenes de prueba a juveniles de tilapia O. niloticus (peso promedio = 10 mg, longitud total= 8,75 mm) y otro el ensayo fue con poslarvas de langostino M. olfersii (peso promedio = 5 mg, longitud total= 5,72 mm). Se emplearon cinco concentraciones (5, 10, 20, 40 y 80 μL-1 para el bioensayo con juveniles de tilapia; para el bioensayo con poslarvas de langostino las concentraciones fueron 0,1, 0,2, 0,5, 0,7 y 1 μL-1). El diseño experimental consideró un control negativo, con dos repeticiones y dos réplicas por cada tratamiento. El análisis de datos se realizó con el método Probit para determinar la CL50 a 96 horas, se obtuvo un valor para juveniles de O. niloticus de 17,49 μL-1 con intervalo de confianza (95 %) con límite inferior de 13,75 μL-1 y límite superior 22,25 μL-1, para las larvas de M. olfersii se obtuvo un valor de 0,31 μL-1 con intervalo de confianza (95 %) con límite inferior de 0,26 μL-1 y límite superior 0,35 μL-1. El análisis de varianza demostró que no existió diferencia estadística significativa (p > 0.05) entre las réplicas de los tratamientos. Se concluye que es necesario continuar con estudios para evaluar su toxicidad en organismos acuáticos debido al amplio uso de este herbicida en la actividad agropecuaria, y determinar su riesgo para otras actividades productivas además de la acuicola.

ABSTRACT Paraquat is an herbicide used in the agricultural industry for weed and undergrowth control, its mode of action is by contact and nonselective. Due to its high-water solubility and low volatility, it represents a potential risk for non-target organisms. Oreochromis niloticus tilapia and the prawn Macrobrachium olfersii are important commercial species for Veracruz's aquaculture industry. The objective of this study was to determine the Mean Lethal Concentration (LC50) of the herbicide Dasurquat® (active ingredient Paraquat) through an acute toxicity bioassay (96 hours). Juvenile O. niloticus (average weight = 10 mg, total length = 8.75 mm) and juvenile M. olfersii postlarvae (average weight = 5 mg, total length = 5.72mm) were used as test specimens. Five herbicide concentrations (5, 10, 20, 40 and 80 μL-1 for tilapia and 0.1, 0.2, 0.5, 0.7 y 1 μL-1 for prawn) were applied. The experimental design considered a negative control, with two replications and two replicates for each treatment. Probit analysis determined that the LC50 at 96 hours for O. niloticus was 17.49 μL-1, with a lower 95 % confidence limit of 13.75 μL-1 and an upper limit of 22.25 μL-1, whereas for M. olfersii the LC50 at 96 hours was 0.31 μL-1, with a lower 95 % confidence limit of 0.26 μL-1 and an upper limit of 0.35 μL-1. The analysis of variance showed that there was no significant statistical difference (p > 0.05) between the replicates of the treatments. It is concluded that it is necessary to continue evaluating its toxicity in aquatic organisms due to the wide use of this herbicide in Veracruz's agricultural activity, to determine its potential risk to other activities.

Sublethal effects of propiconazole on the metabolism of lambari Deuterodon iguape (Eigenmann 1907), a native species from Brazil.

The objective of this study was to analyze the sublethal effects of propiconazole on Deuterodon iguape, a native fish common in Brazil, which has potential for aquaculture and use as a bioindicator. The hypothesis was to test whether D. iguape has a metabolism similar to Danio rerio so that its use in bioassays may be validated. Lethal concentration (LC50) and metabolic rates were studied in fish exposed to propiconazole. Specific oxygen consumption and ammonia excretion for D. iguape and D. rerio increased by 0.01 µg L-1 and then decreased as the propiconazole concentration increased. The decrease in the averages of specific oxygen consumption at the concentration of 0.1 µg L-1 represented a reduction in the metabolic rate compared to the control of 71% for D. iguape and 40% D. rerio. For the ammonia excretion, at the same concentration, there was a reduction of 68.7% and 45.4% for D. iguape and D. rerio, respectively. When comparing ammonia excretion of the two species for each concentration of propiconazole, there was a significant difference (p < 0.05) in relation to the control and for the highest concentration (0.1 µg L-1). As for specific oxygen consumption, there was a statistically significant difference only for the concentration of 0.1 µg L-1. D. iguape proved to be a good and useful bioindicator for ichthyologists or ecologists in studies of moderate pesticide contamination in freshwater aquatic environments, as its metabolic response was similar to D. rerio.

Determination of chemical structure and anti-Trypanosoma cruzi activity of extracts from the roots of Lonchocarpus cultratus (Vell.) A.M.G. Azevedo & H.C. Lima.

Trypanosoma cruzi is the agent of Chagas disease, an infection that affects around 8 million people worldwide. The search for new anti-T. cruzi drugs are relevant, mainly because the treatment of this disease is limited to two drugs. The objective of this study was to investigate the trypanocidal and cytotoxic activity and elucidate the chemical profile of extracts from the roots of the Lonchocarpus cultratus. Roots from L. cultratus were submitted to successive extractions with hexane, dichloromethane, and methanol, resulting in LCH, LCD, and LCM extracts, respectively. Characterization of extracts was done using 1H-RMN, 13C-RMN, CC and TLC. Treatment of T. cruzi forms (epimastigotes, trypomastigotes, and amastigotes) with crescent concentrations of LCH, LCD, and LCM was done for 72, 48, and 48 h, respectively. After this, the percentage of inhibition and IC50/LC50 were calculated. Benznidazole was used as a positive control. Murine macrophages were treated with different concentrations of both extracts for 48 h, and after, the cellular viability was determined by the MTT method and CC50 was calculated. The chalcones derricin and lonchocarpine were identified in the hexane extract, and for the first time in the genus Lonchocarpus, the presence of a dihydrolonchocarpine derivative was observed. Other chalcones such as isocordoin and erioschalcone B were detected in the dichloromethane extract. The dichloromethane extract showed higher activity against all tested forms of T. cruzi than the other two extracts, with IC50 values of 10.98, 2.42, and 0.83 µg/mL, respectively; these values are very close to those of benznidazole. Although the dichloromethane extract presented a cytotoxic effect against mammalian cells, it showed selectivity against amastigotes. The methanolic extract showed the lowest anti-T. cruzi activity but was non-toxic to peritoneal murine macrophages. Thus, the genus Lonchocarpus had demonstrated in the past action against epimastigotes forms of T. cruzi but is the first time that the activity against infective forms is showed, which leading to further studies with in vivo tests.

Reproductive toxicity of Roundup WG® herbicide: impairments in ovarian follicles of model organism Danio rerio.

Glyphosate-based herbicides are widely used in global agriculture, and their effects on different non-target animal organisms have been the focus of many toxicological studies. Regarding the potential role of glyphosate-based herbicides as an endocrine disruptor, the present study aims to investigate the effects of the herbicide Roundup WG® (RWG) on female reproduction, specifically on the ovarian maturation of Danio rerio. Adult females were exposed to low concentrations of RWG (0.065, 0.65, and 6.5 mg L-1) for 15 days, and then the ovaries were submitted to structural and morphometric procedures, accompanied by analysis of the vitellin protein content. Our results showed an increase of initial ovarian follicle numbers, decrease of late ovarian follicles, and smaller diameter of ovarian follicles in fish exposed to 0.065 and 6.5 mg L-1. The thickness of vitelline envelope was reduced, and the vitellin protein content was increased in the ovarian follicle in the two highest concentrations. Ultrastructural changes in the ovarian follicular component were evident and expressed by the cell index; vacuolization in follicular cells, increase of perivitelline space, and impaired mitochondria in oocytes were observed. Therefore, RWG adversely affects the ovarian maturation in D. rerio, and these changes can lead to reproductive toxicity, compromising population dynamics.

Susceptibility of Fall Armyworms (Spodoptera frugiperda J.E.) from Mexico and Puerto Rico to Bt Proteins.

Fall armyworm is one of the main pests of conventional and Bacillus thuringiensis (Bt) corn in many countries in the Americas, Africa, Asia and in Australia. We conducted diet-overlay bioassays to determine the status of susceptibility to four Bt proteins (Cry1A.105, Cry2Ab2, Cry1F and Cry1Ac) in three different populations of fall armyworm from Mexico, and one population from Puerto Rico. Bioassays showed that fall armyworms from Puerto Rico were resistant to Cry1F with a resistance ratio 50 (RR50) higher than 10,000 ng/cm2 and to Cry1Ac with a RR50 = 12.2 ng/cm2, displaying the highest median lethal concentration (LC50) values to all Bt proteins tested. The effective concentration 50 (EC50) values further confirmed the loss of susceptibility to Cry1F and Cry1Ac in this population. However, LC50 and EC50 results with Cry1A.105 and Cry2Ab2 revealed that fall armyworm from Puerto Rico remained largely susceptible to these two proteins. The Mexican populations were highly susceptible to all the Bt proteins tested and displayed the lowest LC50 and EC50 values to all Bt proteins. Our results suggest that Cry1F and Cry1Ac resistance is stable in fall armyworm from Puerto Rico. However, this population remains susceptible to Cry1A.105 and Cry2Ab2. Results with Mexican fall armyworms suggest that possible deployment of Bt corn in Mexico will not be immediately challenged by Bt-resistant genes in those regions.

Efficacy of carvacrol on Rhipicephalus (Boophilus) microplus engorged female ticks (Canestrini, 1887) (Acari: Ixodidae): effects on mortality and reproduction.

The activity of carvacrol was evaluated in R. microplus female ticks by estimating the mean lethal concentration (LC50) and the acaricidal efficacy. Ticks were subjected to the Adult Immersion Test (AIT) to calculate LC50. From this value, the AIT was performed again at the concentrations of 20%, 40%, 60%, 80% and 100% of the LC50. Two control groups were established: one treated with distilled water and one with the solvent (ethanol 50%). The following parameters were evaluated: female weight before oviposition, egg mass weight, pre-oviposition and incubation period, hatching percentage, egg production index, fecundity rate, estimated reproduction, reduction in oviposition and hatching, and product efficacy. The obtained LC50 was 20.11 mg/mL. Egg mass weight, egg production index, pre-oviposition and fecundity rates of groups treated with 80% and 100% of LC50 were statistically different when compared to control groups. The product efficacy in groups treated with 100% of LC50 was 61.10%.

Low concentrations of acetylcarvacrol induce drastic morphological damages in ovaries of surviving Rhipicephalus sanguineus sensu lato ticks (Acari: Ixodidae).

Rhipicephalus sanguineus sensu lato (s. l.) ticks are targets of acaricidal treatments due to their great veterinary and medical importance. Acetylcarvacrol stands out as a promising acaricidal substance for its increased biocidal activity and stability. Additionally, its toxicity to mammals is reduced when compared to the parent molecule, carvacrol. The present study aimed to evaluate the effects of acetylcarvacrol on the morphology of ovaries of engorged R. sanguineus s. l. ticks. The animals were subjected to the Adult Immersion Test to calculate the lethal concentrations (LC50 and LC90) of acetylcarvacrol. Subsequently, the surviving ticks were dissected for collection of the ovaries. The samples were processed through routine histological techniques. The histological sections were stained with hematoxylin and eosin, bromophenol blue (protein detection) and PAS (polysaccharide detection). The alterations found in the ovaries were evaluated by morphometric and semiquantitative analysis. The LC50 and LC90 were 17.805 and 26.164 µL/mL, respectively. The most severe morphological alterations were disappearance of the female germ cells (oocytes), nucleolus vacuolization, thicker and irregular chorion, and decrease in size of the oocytes. Also, the content of proteins and carbohydrates in the oocytes were heavily affected by the chemical, as evidenced by a non-homogeneous staining pattern. The group exposed to the highest concentration of acetylcarvacrol (20 µL/mL) exhibited a statistically greater score in the semiquantitative analysis when compared to the other groups. The morphological changes in the ovaries may reduce the ticks' offspring production or generate descendants that will struggle to carry out essential biochemical processes during their lives. Thus, acetylcarvacrol may be a promising alternative to control tick infestation by impairing the reproduction of this parasite.