A realistic combined exposure scenario: effect of microplastics and atrazine on Piaractus mesopotamicus.

Microplastics, considered emerging environmental contaminants resulting from plastic degradation, are discovered in diverse aquatic ecosystems and can be unintentionally ingested by fish. Therefore, it is essential to characterize their interaction with other contaminants, such as agrochemicals, in aquatic environments. This study aimed to assess histological, enzymatic, and genotoxic biomarkers in juvenile pacú (Piaractus mesopotamicus) exposed to polyethylene (PE) microplastic particles and the herbicide atrazine, individually or combined, for 15 days. Four treatments were used: a negative control (CON), PE in the fish diet (0.1% w/w, FPE), atrazine through water (100 µg L-1, ATZ), and the mixture (ATZ+FPE). Results confirmed histological alterations in gills (edema and lamellar fusion) and liver (necrotic areas and congestion) of fish exposed to ATZ and ATZ+FPE. The number of goblet cells increased in the posterior intestine of fish under ATZ+FPE compared to CON and FPE. Enzyme activities (CAT, GST, AChE, and BChE) significantly increased in ATZ+FPE compared to CON. However, no genotoxic effect was demonstrated. These findings provide insights into the complex impacts of simultaneous exposure to atrazine and microplastics, emphasizing the need for continued research to guide effective environmental management strategies against these contaminants that represent a risk to aquatic organisms.

Anuran heart development and critical developmental periods: A comparative analysis of three neotropical anuran species.

The heart begins to form early during vertebrate development and is the first functional organ of the embryo. This study aimed to describe and compare the heart development in three Neotropical anuran species, Physalaemus albonotatus, Elachistocleis bicolor, and Scinax nasicus. Different Gosner Stages (GS) of embryos (GS 18-20) and premetamorphic (GS 21-25), prometamorphic (GS 26-41), and metamorphic (GS 42-46) tadpoles were analyzed using stereoscopic microscopy and Scanning Electronic Microscopy. Heart development was similar in the three analyzed species; however, some heterochronic events were identified between P. albonotatus and S. nasicus compared to E. bicolor. In addition, different patterns of melanophores arrangement were observed. During the embryonic and metamorphic periods, the main morphogenetic events occur: formation of the heart tube, regionalization of the heart compartments, development of spiral valve, onset of heartbeat, looping, and final displacement of the atrium and its complete septation. Both periods are critical for the normal morphogenesis and the correct functioning of the anuran heart. These results are useful to characterize the normal anuran heart morphology and to identify possible abnormalities caused by exposure to environmental contaminants.

Effects of the emulsifiable herbicide Dicamba on amphibian tadpoles: an underestimated toxicity risk?

The effects of exposure to the herbicide Dicamba (DIC) on tadpoles of two amphibian species, Scinax nasicus and Elachistocleis bicolor, were assessed. Mortality and biochemical sublethal effects were evaluated using acetylcholinesterase (AChE), glutathione S-transferase (GST), glutathione reductase (GR), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) activities and thyroid hormone (T4) levels. The LC50 value at 48h was 0.859 mg L-1 for S. nasicus and 0.221 mg L-1 for E. bicolor tadpoles. After exposure to sublethal DIC concentrations for 48 h, GST activity increased in S. nasicus but significantly decreased in E. bicolor with respect to controls. GR activity decreased only in S. nasicus at all the tested DIC concentrations. AChE activity was significantly inhibited in both S. nasicus and E. bicolor tadpoles at 48 h. DIC also caused significant changes in transamination, as evidenced by an increase in AST and ALT activities in both amphibian species. T4 levels were higher in DIC-treated tadpoles of both species than in controls. The DIC-induced biochemical alterations in glutathione system enzymes and transaminases indicate lesions in liver tissues and cellular function. Moreover, the observed AChE inhibition could lead to the accumulation of acetylcholine, excessively stimulating postsynaptic receptors, and the increase in T4 levels in both species may indicate an overactive thyroid. The commercial DIC formulation showed a high biotoxicity in the two amphibian native species after short-term exposure, controversially differing from the toxicity level indicated in the official fact sheet data. This fact highlights the need for an urgent re-categorization and reevaluation of DIC toxicity in native species.

Morphology and microchemistry of the otoliths of the inner ear of anuran larvae.

To navigate in space most vertebrates need precise positional cues provided by a variety of sensors, including structures in the inner ear, which are exquisitely sensitive to gravity and linear acceleration. Although these structures have been described in many vertebrates, no information is available for anuran larvae. The purpose of our study was to describe, for the first time, the size, complexity and microchemistry of the saccular otoliths of the larva of 13 anuran species from central Argentina, using electron microscopy and X-ray spectroscopy (N = 65). We concluded that a) these structures differ in area, perimeter, otolith relative size and fractal dimension, but are similar in terms of their microchemistry when compared by spatial guilds, b) that nektonic species have larger otoliths than nektonic-benthic and benthic species and c) that benthic species have larger otolith relative size than nektonic-benthic and nektonic species.

Biological endpoints, enzyme activities, and blood cell parameters in two anuran tadpole species in rice agroecosystems of mid-eastern Argentina.

Different biological variables of tadpoles, including survival, development and growth rates, and biomarkers [cholinesterases, glutathione-S-transferases (GST), and blood cell morphology] were evaluated in two anuran species, Scinax squalirostris (Hylidae) and Leptodactylus mystacinus (Leptodactylidae), using in situ experimental chambers in a rice field (RF) sprayed with insecticide Lambda-cyhalothrin (LTC) by aircraft in Santa Fe Province, Argentina. We found a significant decrease in body weight (0.62 ± 0.04 g) of L. mystacinus and an increased development rate of S. squalirostris in individuals from RF (41 ± 1; Gosner) with respect to individuals from the reference site (RS: 0.93 ± 0.04 g and 37 ± 0; respectively). In S. squalirostris, individuals from RF mean values of butyrylcholinesterase activities decreased at 48 (4.09 ± 0.32 nmol min-1 mg-1 of TP) and 96 h (3.74 ± 0.20 nmol min-1 mg-1 of TP), whereas inhibition of acetylcholinesterase was observed at 96 h (47.44 ± 2.78 nmol min-1 mg-1 of TP). In L. mystacinus from RF, an induction of acetylcholinesterase activity was observed at 96 h (36.01 ± 1.09 nmol min-1 mg-1 of TP). Glutathione-S-transferase levels varied between species, being higher in L. mystacinus individuals but lower in S. squalirostris from RF at 48 (272.29 ±11.78 and 71.87 ± 1.70 nmol min-1 mg-1 of TP; respectively) and 96 h (279.25 ± 13.06 and 57.62 ± 4.58 nmol min-1 mg-1 of TP, respectively). Blood cell parameters revealed a lower number of mitotic cells (MC: 0.36 ± 0.31%o for S. squalirostris and 0.08 ± 0.05 %o for L. mystacinus) and higher number of eosinophils (E: 3.45 ± 1.75 %o for S. squalirostris and 7.64 ± 0.98 %o for L. mystacinus) in individuals from the RF than in individuals from the RS (MC: 2.55 ± 0.74 %o for S. squalirostris and 1.87 ± 0.72%o for L. mystacinus; and E: 0.13 ± 0.09 for S. squalirostris and 3.20 ± 0.80 for L. mystacinus). Overall, our results demonstrate the existence of apparent differences in sensitivity between species in a series of sublethal responses to short-term exposure in RF after the application of Lambda-cyhalothrin. We suggest that the integral use of biological endpoints (development and growth) together with biomarkers (cholinesterase, GST, and blood cell parameters) may be a promising integral procedure for investigating pesticide exposure in wild frog populations.

Dinámica de un ensamble de anuros en un humedal del tramo inferior del Rio Paraná Argentina / Dynamics of anuran assemblage in a wetland from the southern section of Paran River Argentina

Se describe la diversidad, composición y estructura de tamaños de un ensamble de anuros del Parque Nacional Pre-Delta (PNPD) durante un período anual. Se colocaron trampas de caída desde dic 2004 hasta dic 2005. Se analizó la relación de la riqueza con la temperatura, humedad relativa, número de días con lluvia, precipitación y nivel del río mediante coeficientes de correlación simple y parcial. Se estudió la fenología de las especies más abundantes realizando un análisis de contingencia, contrastando para cada especie las frecuencias mensuales con la frecuencia media de captura en el año. Asimismo, se comparó la frecuencia de cada especie con las frecuencias unidas de las restantes, y se analizó la variación mensual de la longitud hocico-cloaca (LHC). Se capturó un total de 16 especies pertenecientes a 5 familias. Los coeficientes de correlación simple mostraron que la riqueza estuvo asociada con la temperatura y el nivel del río, no obteniéndose los mismos resultados para los coeficientes de correlación parcial. La mayor actividad se observó en ene-abr y en nov-dic 2005. Se hallaron diferencias significativas en la fenología de cada especie. En los meses cálidos se observó disminución en la LHC. Se sugiere que los ciclos anuales en las estructuras de tamaños estarían relacionados con las épocas de reclutamiento de juveniles. Por último, se concluye que los patrones de actividad de las distintas especies y sus ciclos reproductivos, influyen en la estructura temporal del ensamble de anuros estudiados.