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.

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.

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.

Assessing biochar impact on earthworms: Implications for soil quality promotion.

Potential harmful effects of spent coffee grounds (SCGs)-derived biochar on earthworms (Lumbricus terrestris) were investigated through two complementary experiments, which assessed the avoidance response of earthworms to biochar-amended soils (experiment 1), and the response of oxidative stress biomarkers and digestive enzymes (experiment 2). The main results were: 1) the highest dose of biochar (5% w/w) caused a significant avoidance response of earthworms (75% individuals avoided these treated soils after 48 h); 2) signs of oxidative stress were early detected in earthworms exposed to biochar (1 and 5% w/w) as indicated by the integrated biological response index; 3) earthworms exposed to biochar-amended soils for 30 d experienced a significant increase of digestive enzyme activities measured in both the gastrointestinal tissue and the luminal content; 4) interaction between earthworms and biochar led to a higher soil extracellular enzyme activities in the 1% biochar treatment than that of control and 5% biochar treatments. These findings suggest that the joint application of SCG-biochar and L. terrestris is a workable approach for improving soil quality in terms of soil biochemical promotion, although earthworms may develop some physiological mechanisms of biochar tolerance (antioxidant defenses).

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.

Neuropeptide glutamic-isoleucine (NEI) specifically stimulates the secretory activity of gonadotrophs in primary cultures of female rat pituitary cells.

The neuropeptide EI (NEI) is derived from proMCH. It activates GnRH neurons, and has been shown to stimulate the LH release following intracerebroventricular administration in several experimental models. The aim of the present paper was to evaluate NEI actions on pituitary hormone secretion and cell morphology in vitro. Pituitary cells from female rats were treated with NEI for a wide range of concentrations (1-400x10(-8)M) and time periods (1-5h). The media were collected and LH, FSH, PRL, and GH measured by RIA. The interaction between NEI (1, 10 and 100x10(-8)M) and GnRH (0.1 and 1x10(-9)M) was also tested. Pituitary cells were harvested for electron microscopy, and the immunogold immunocytochemistry of LH was assayed after 2 and 4h of NEI incubation. NEI (100x10(-8)M) induced a significant LH secretion after 2h of stimulus, reaching a maximum response 4h later. A rapid and remarkable LH release was induced by NEI (400x10(-8)M) 1h after stimulus, attaining its highest level at 2h. However, PRL, GH and FSH were not affected. NEI provoked ultrastructural changes in the gonadotrophs, which showed accumulations of LH-immunoreactive granules near the plasma membrane and exocytotic images, while the other populations exhibited no changes. Although NEI (10x10(-8)M), caused no action when used alone, its co-incubation with GnRH (1x10(-9)M), promoted a slight but significant increase in LH. These results demonstrate that NEI acts at the pituitary level through a direct action on gonadotrophs, as well as through interaction with GnRH.

Plasma B-esterase and glutathione S-transferase activity in the toad Chaunus schneideri (Amphibia, Anura) inhabiting rice agroecosystems of Argentina.

B-esterase (BChE: butyrylcholinesterase and CbE: carboxylesterase) and glutathione S-transferase (GST) activity were measured in the plasma of Chaunus schneideri collected in rice fields and surrounding environments and in a reference pristine forest. The chemical criterion based on in-vitro reactivation of BChE activity using pyridine-2-aldoxime methochloride (2-PAM) was also determined. Mean values of plasma BchE, CbE, and GST activity for samples from agricultural areas were different from those for samples from pristine forest. Plasma samples from the two agricultural areas showed positive reactivation of BChE activity after incubation with 2-PAM. Based on our experimental evidence we suggest B-esterases and gluthatione S-transferases can be used in field monitoring as biomarkers of exposure of wildlife to pesticides, because the analysis in non-destructive and is sensitive to anti-ChE agrochemicals. Chemical reactivation of BChE is also a complementary method for assessing the effects of pesticides on toads inhabiting rice fields. Further studies are urgently needed to investigate adverse effects of massive exposure to pesticides experienced by native populations of anurans.