Multixenobiotic response of Collembola to soil contamination, the phisiological basis for bioindicative environmental monitoring.

Collembola are well-established models in ecotoxicological research, extensively employed to investigate the effects of various contaminants, including heavy metals. The Multixenobiotic Resistance Mechanism (MXR) is a physiological response based on transmembrane efflux proteins that play a pivotal role in pumping xenobiotics and conferring resistance. This mechanism is firmly established as a biomarker of aquatic contamination and has recently shown promise as a soil biomonitoring tool. In this study, we aimed to assess the feasibility of utilizing the MXR mechanism as a biomonitoring tool, specifically by investigating the response of two Collembola species exposed to soil contaminated in a real-life situation. Soil samples were obtained from the site of Brazil's largest mine disaster, a dam rupture in Brumadinho-MG. We explored MXR activity in the model species Folsomia candida and a tropical native species, Cyphoderus sp. Our findings reveal efflux activity in both species, confirmed by model MXR protein inhibitors. Moreover, we observed distinct MXR activity levels corresponding to the degree of heavy metal contamination in the soil samples. Consequently, our results underscore the potential of combining an established soil bioindicator, such as Collembola, with the physiological response of a molecular biomarker like MXR. This approach may represent a valuable strategy for biomonitoring terrestrial ecosystems.

First characterization of multixenobiotic activity in Collembola: An approach on cadmium-induced response.

ATP-binding cassette (ABC) efflux pumps mediate the activity of the Multixenobiotic Resistance (MXR) mechanism and have been proposed as a biomarker of environmental pollution mainly in aquatic invertebrates. MXR activity was never investigated in Collembola and represents a potential tool for soil biomonitoring. This study aimed to characterize for the first time the activity of ABC efflux pumps in the gut of collembolan species, and investigate its responsiveness to cadmium (Cd), a common stressor found in polluted soils. We performed in vitro rhodamine-B accumulation assays in the presence of model inhibitors of ABC efflux pumps: verapamil hydrochloride as P-gp (P-glycoprotein) inhibitor, and MK571, as MRPs (multidrug resistance-related proteins) inhibitor. We also performed rhodamine-B accumulation assays under Cd-exposure (209 µg/L;1 µM). Our results showed that all species presented basal (noninduced) level of MXR activity in their gut. Efflux pumps P-gp and/or MRPs activity were confirmed in Cyphoderus innominatus, Cyphoderus similis, and Folsomia candida, the standard species. The rhodamine-B accumulation assays performed with Cd, applied as soil pollutant, showed that the gut of non-standard species C. similis and Trogolaphysa sp. presented an increase of MXR activity for both P-gp and MRP transporters, indicating the potential of these species as test organisms for soil ecotoxicology studies in Neotropical region. Our findings suggest a functional role of ABC transporters in the collembolan gut and their cellular involvement in Cd defense response, corroborating that MXR phenotype in Collembola can be a promising tool for bioindication of soil contamination.

Estuarine fish assemblages present a species-specific difference in the multixenobiotics resistance activity.

We investigated the activity of the multixenobiotic resistance (MXR) phenotype, a biological defense system in aquatic organisms, in the fish assemblages of two tropical estuaries with different degrees of environmental impacts, the Paraiba River and Mamanguape River Estuaries. The aim of this work was to compare the activity of the MXR phenotype of different fishes to test the hypothesis that each species has an inherent activity level and to use this activity as a bioindicator of aquatic contamination. We assessed the MXR activity of the gills, using rhodamine B (RB) accumulation assay. The results demonstrated a species-specific difference in the MXR activity of fishes caught in the same estuarine zone. Also, the pelagic species Eucinostomus melanopterus, Eucinostomus argenteus, and Lutjanus jocu had higher RB accumulation, while the demersal species Sphoeroides testudineus and Sphoeroides greeleyi had the lowest RB accumulation, suggesting that the ecological characteristic of fish in the water column exerts an influence on MXR activity. Besides, we demonstrated the potential of using the gill MXR activity of the key estuarine species, the Brazilian silversides Atherinella brasiliensis, as a tool for biomonitoring estuaries.

Capacity of tissue water regulation is impaired in an osmoconformer living in impacted estuaries?

Estuarine osmoconformes rely on their ability to perform tissue and cell water regulation to cope with daily osmotic challenges that occur in the estuary. In addition, these animals currently must deal with pollutants present in the estuarine environment, which can disturb their capacity of water regulation. We collected the mangrove oyster Crassostrea rhizophorae in two tropical estuaries in the Northeast region of Brazil with different degrees of human interference: the Paraíba Estuary (impacted) and the Mamanguape Estuary (preserved). Tissue water content was analyzed after exposure to salinities 12, 24 and 36 for 24 h. Gill cell volume regulation was analyzed in vitro upon hypo- and hyper-osmotic conditions. We also analyzed gill MXR (multi-xenobiotic resistance) mechanism, as reference of environmental pollution. Gill and muscle of oysters from two sites of Paraíba Estuary, and from one site of Mamanguape Estuary were not able to maintain tissue water content upon hypo- and hyper-osmotic conditions. Gill cells of oyster from the same sites exhibited swelling followed by regulatory volume decrease upon hypo-osmotic condition. Gill MXR activity was increased in oysters from these sites. The best tissue and cell water regulation, and the lowest MXR activity, was found in oyster from downstream of Mamanguape Estuary, our reference site and the one most preserved. Tissue and cell water regulation proved to be a sensitive parameter to environmental pollution and could be considered as biomarker of aquatic contamination.

Hemolymph and gill carbonic anhydrase are more sensitive to aquatic contamination than mantle carbonic anhydrase in the mangrove oyster Crassostrea rhizophorae.

Carbonic anhydrase (CA) is a ubiquitous metalloenzyme of great importance in several physiological processes. Due to its physiological importance and sensitivity to various pollutants, CA activity has been used as biomarker of aquatic contamination. Considering that in bivalves the sensitivity of CA to pollutants seems to be tissue-specific, we proposed here to analyze CA activity of hemolymph, gill and mantle of Crassostrea rhizophorae collected in two tropical Brazilian estuaries with different levels of anthropogenic impact, in dry and rainy season. We found increased carbonic anhydrase activity in hemolymph, gill and mantle of oysters collected in the Paraíba Estuary (a site of high anthropogenic impact) when compared to oysters from Mamanguape Estuary (inserted in an area of environmental preservation), especially in the rainy season. CA of hemolymph and gill were more sensitive than mantle CA to aquatic contamination. This study enhances the suitability of carbonic anhydrase activity for field biomarker applications with bivalves and brings new and relevant information on hemolymph carbonic anhydrase activity as biomarker of aquatic contamination.