Quantum dots (QDs) have attracted broad attention due to their special optical properties and promising prospect in medical and biological applications. However, the process of QDs on cell membrane is worth further investigations because such process may lead to harmful effects on organisms and also important for QD application. In this study, adhesion of amino- and carboxyl-coated CdTe QDs (A-QDs and C-QDs) on cell membrane and the subsequent internalization are studied using a series of endocytosis-free model membranes, including giant and small unilamellar vesicles, supported lipid bilayers and giant plasma membrane vesicles (GPMVs). The adhered QD amounts on model membranes are quantified by a quartz crystal microbalance. The CdTe QD adhesion on model membranes is governed by electrostatic forces. Positively charged A-QDs adhere on GPMV surface and passively penetrate the plasma membrane via endocytosis-free mechanism, but negatively charged C-QDs cannot. Rat basophilic leukemia (RBL-2H3) cells are exposed to CdTe QDs to monitor the QD internalization process. Both A- and C-QDs are internalized by RBL-2H3 cells mainly via endocytosis. CdTe QDs do not accumulate on the plasma membrane of living cells due to the fast endocytosis and the weakened electrostatic attraction in biological medium, resulting in low chance of passive penetration. The suspended cells after trypsin digestion take more QDs than the adherent cells. A-QDs cause lower cell viability than C-QDs, probably because the approach of positively charged QDs to cells is favored and the smaller aggregates of A-QDs.

In the marine environment, microplastic contamination and acidification may occur simultaneously, this study evaluated the effects of ocean acidification and microplastics on oxidative stress responses and digestive enzymes in mussels. The thick shell mussels Mytilus coruscus were exposed to four concentrations of polystyrene microspheres (diameter 2 μm, 0, 10, 10⁴ and 10⁶ particles/L) under two pH levels (7.7 and 8.1) for 14 days followed by a 7-day recovery acclimation. Throughout the experiment, we found that microplastics and ocean acidification exerted little oxidative stress to the digestive gland. Only catalase (CAT) and glutathione (GSH) showed a significant increase along with increased microplastics during the experiment, but recovered to the control levels once these stressors were removed. No significant effects of pH and microplastics on glutathione peroxidase (GPx) and superoxide dismutase (SOD) were observed. The responses of digestive enzymes to both stressors were more pronounced than antioxidant enzymes. During the experiment, pepsin (PES), trypsin (TRS), alpha-amylase (AMS) and lipase (LPS) were significantly inhibited under microplastics exposure and this inhibition was aggravated by acidification conditions. Only PES and AMS tended to recover during the recovery period. Lysozyme (LZM) increased significantly under microplastic exposure conditions, but acidification did not exacerbate this effect. Therefore, combined stress of microplastics and ocean acidification slightly impacts oxidative responses but significantly inhibits digestive enzymes in mussels.

We describe a batch-extraction with simulated digestive fluid (salivary fluid, gastric fluid and intestinal fluid) to estimate the bioaccessibility of inhaled trace metals (TMs) in particulate matter less than 10 and 2.5 μm in aerodynamic diameter (PM₁₀ and PM₂.₅). Concentrations of the assayed TMs (As, Cd, Cr, Ni, Mn, Cu, Zn, Sb, Hg and Pb) were determined in PM₁₀ and PM₂.₅ samples by inductively coupled plasma-mass spectrometry. The TMs with the largest soluble fractions for airborne PM collected from winter and summer in saliva were Mn and Sb, respectively; in seasons this became Co in gastric fluid and Cu in intestinal fluid. Clearly, bioaccessibility is strongly dependent on particle size, the component of simulated digestive fluids (e.g., pH, digestive enzymes pepsin and trypsin), and the chemical properties of metal ions. The particle size and seasonal variation affected the inhaled bioaccessible fraction of PM-bound TMs during mucociliary clearance, which transported PM from the tracheal and the bronchial region to the digestive system. This study provides direct evidence for TMs in airborne PM being bioaccessible TMs are likely to possess an enhanced digestive toxic potential due to airborne PM pollution.

Silver barb Barbodes gonionotus fry were exposed to polyvinyl chloride (PVC) fragments at increasing concentrations of 0.2, 0.5 and 1.0 mg/L for 96 h, following which whole body histological evaluation and analysis of the digestive enzymes trypsin and chymotrypsin were performed. Whole body trypsin and chymotrypsin activities increased significantly in fish exposed to 0.5 and 1.0 mg/L PVC as compared those exposed to zero or 0.2 mg/L PVC. In fish exposed to all tested concentrations, PVCs were observed in both the proximal and distal intestine, and fish exposed to 0.5–1.0 and 1.0 mg/L PVC, respectively, and these particles were associated with localized thickening of the mucosal epithelium. No tissue damage was evident in any other internal organs or gills. This lack of damage may be attributed to the absence of contaminants associated with the PVC fragments and their relatively smooth surface. The increased whole body trypsin and chymotrypsin activities may indicate an attempt to enhance digestion to compensate for epithelial thickening of the intestine and/or to digest the plastics.

[Departement_IRSTEA]Eaux [TR1_IRSTEA]BELCA | International audience | The feeding activity and subsequent assimilation of the products resulting from food digestion allow organisms to obtain energy for growth, maintenance and reproduction. Among these biological parameters, we studied digestive enzymes (amylase, cellulase and trypsin) in Gammarus fossarum to assess the impact of contaminants on their access to energy resources. However, to enable objective assessment of a toxic effect of decreased water quality on an organisms’ digestive capacity, it is necessary to establish reference values based on its natural variability as a function of changing biotic and abiotic factors. To limit the confounding influence of biotic factors, a caging approach with calibrated male organisms from the same population was used. This study applied an in situ deployment at 23 sites of the Rhone basin rivers, complemented by a laboratory experiment assessing the influence of two abiotic factors (temperature and conductivity). The results showed a small effect of conductivity on cellulase activity and a significant effect of temperature on digestive enzyme activity but only at the lowest temperature (7 °C). The experimental conditions allowed us to define an environmental reference value for digestive enzyme activities to select sites where the quality of the water impacted the digestive capacity of the organisms. In addition to the feeding rate, this study showed the relevance of digestive enzymes as biomarkers to be used as an early warning tool to reflect organisms’ health and the chemical quality of aquatic ecosystems.

A simple methodology for the determination of bioavailability of fourteen metals in coastal sediments has been developed by simulating the conditions of digestive process of marine fishes. With this aim, a representative sediment composite sample was treated with hydrochloric acid solutions at different pH values, temperatures and contact times, in the presence and absence of Pepsin and Trypsin. The addition of Pepsin and Trypsin did not affect the extraction of most elements. As a result of the present study, the digestion with a hydrochloric acid solution at pH 1, 40°C and 12h is proposed. Adjustments of the temperature and time reaction could be made according to the specific ecosystem under study. The amount of metal extracted by other methods based on acetic acid was lower than that extracted by HCl treatment proposed.

Methods standardisation in microplastics research is needed. Apart from reagent-dependent effects on microplastics, varying target particle sizes can hinder result comparison between studies. Human health concerns warrant recovery of small microplastics. We compared existing techniques using hydrogen peroxide, Proteinase-K, Trypsin and potassium hydroxide to digest bivalve tissue. Filterability, digestion efficacy, recoverability of microplastics and subsequent polymer identification using Raman spectroscopy and a matching software were assessed. Only KOH allowed filtration at ≤25 μm. When adding a neutralisation step prior to filtration, KOH digestates were filterable using 1.2-μm filters. Digestion efficacies were >95.0% for oysters, but lower for clams. KOH destroyed rayon at 60 °C but not at 40 °C. Acrylic fibre identification was affected due to changes in Raman spectra peaks. Despite those effects, we recommend KOH as the most viable extraction method for exposure risk studies, due to microplastics recovery from bivalve tissues of single-digit micrometre size.

Citalopram (CTP) and mirtazapine (MTP) are two typical psychoactive drugs used for the depression treatment. As emerging pollutants, CTP and MTP have raised concern because of their harmful effect on aquatic organisms. Therefore, the ecotoxicological risk of these two pollutants to aquatic organisms should be given more attention. In this study, the effects of CTP and MTP on the feeding rate, heartbeat, nutritional enzymes, and their related gene expression of D. magna were investigated under single and binary mixture pollutant exposure. Subsequently, the recovery of exposed D. magna was studied to assess the toxic persistence of those pollutants. After 24-h exposure, the ingestion rate decreased by 34.2% and 21.5%, in the group of 1.45 mg/L CTP (C-H) and binary mixture with high concentration (Mix-H), respectively. After 24-h recovery, the feeding rate of D. magna was stimulated by a compensatory response. Over the exposure period, the heartbeat rate of D. magna increased significantly in the groups of CTP, MTP, and their binary mixture with low concentration (Mix-L), and then, their heartbeat rate was recovered during the recovery period. The activity of α-amylase (AMS) and trypsin were significantly changed in most of the exposed daphnia, both during the exposure and recovery period. CTP/MTP exposure stimulated the expression of the AMS gene. MTP and Mix-H exposure inhibited the expression of the trypsin gene and the other groups stimulated its expression. After 24-h recovery, the stimulating or inhibitory effects were alleviated. There were different responses between gene expression and enzyme activity. In conclusion, our results highlighted the toxic effects at high concentrations of single and mixed pollution of CTP and MTP on the feeding rate, heartbeat, AMS and trypsin enzyme activity, and expression of related genes of D. magna to assess the environment risk of them.

Mangrove crabs are ecosystem engineers through their bioturbation activity. On Mayotte Island, the abundance of Neosarmatium africanum decreased in wastewater-impacted areas. Previous analyses showed that global crab metabolism is impacted by wastewater, with a burst in O₂ consumption that may be caused by osmo-respiratory trade-offs since gill functioning was impacted. As the hepatopancreas is a key metabolic organ, the purpose of this study was to investigate the physiological effects of wastewater and ammonia-N 5-h exposure on crabs to better understand the potential trade-offs underlying the global metabolic state. Catalase, superoxide dismutase, glutathione S-transferase, total digestive protease, and serine protease (trypsin and chymotrypsin) activities were assessed. Histological analyses were performed to determine structural modifications. No effect of short-term wastewater and ammonia-N exposure was found in antioxidant defenses or digestive enzyme activity. However, histological changes of B-cells indicate an increase in intracellular digestive activity through higher vacuolization processes and tubule dilation in wastewater-exposed crabs.

This paper examined the gut digestive enzymes, trypsin and leucine aminopeptidase (LAP), in gypsy moth (Lymantria dispar, L.) larvae exposed to cadmium. We analyzed the 3-day acute effect, chronic effects from hatching until sacrifice, and recovery from long-term dietary treatment with cadmium concentrations of 10 and 30 μg Cd/g dry food. The activities of both examined enzymes declined at the higher level of cadmium after both acute and chronic treatments and did not recover within 3 days of feeding a diet with no added cadmium. Leucine aminopeptidase was more sensitive because its activity was inhibited after both short-term treatments. Three trypsin and one leucine aminopeptidase isoform were detected by electrophoresis. Egg hatches (full-sib families) differed in enzyme activities, index of phenotypic plasticity, and isozyme expression after different treatments. Statistically significant positive correlations between these enzymes pointed to common genetic regulation. Moreover, variances were higher for the control group than for cadmium treatment groups implying that these proteolytic enzymes did not participate directly in detoxification. These results suggest that, with additional research to discover the mechanisms of enzyme inhibition, trypsin and leucine aminopeptidase might be used as biomarkers to indicate the severity of gastrointestinal disease due to cadmium intake.