Diclofenac eco-geno-toxicity in freshwater algae, rotifers and crustaceans.

This study assessed the eco-genotoxic impact of diclofenac (DCF) in sentinel species of the freshwater ecosystem. DCF residues are found in freshwater from few ng/L to tens of µg/L due to the inability of conventional wastewater treatment plants to ensure removal efficiency of the drug. An ample body of literature reports on the acute toxicity of DCF in non-target organisms without addressing potential chronic long-term effects on organisms at actual, environmental concentrations. Herein, assessment for acute and chronic toxicity was performed on organisms in vivo exposed to DCF, specifically on the green alga Raphidocelis subcapitata, the rotifer Brachionus calyciflorus and the crustacean Ceriodaphnia dubia. Furthermore, potential DNA damage and expression of antioxidant genes (MnSOD, Cu/ZnSOD and CAT) were evaluated in crustacean neonates. The toxicological risk of DCF was assessed as well as its. GENOTOXIC RISK: The acute toxicity was observed at concentrations far from those of environmental concern. Rotifers and crustaceans were much more chronically sensitive than the algae to DCF, observing besides, the median effect concentrations at tens of µg/L. In crustaceans, DNA damage was noted at units of µg/L, revealing concentrations of environmental concern. The dysregulated activity of SOD and CAT also showed the ability of DCF to provoke oxidative stress. On assessment of environmental risk, the chronic Risk Quotient (RQ) was above the threshold value of 1. Nevertheless, the genotoxic RQ was significantly greater than the chronic RQ, thus, the need of regulatory bodies to acknowledge the genotoxic impact as an environmental risk factor. To our knowledge, this study is the first investigation to perform environmental genotoxic risk assessment of DCF.

Characterization of Complexes between Imidacloprid and ß-Cyclodextrin: Evaluation of the Toxic Activity in Algae and Rotifers.

The development of new formulations can be driven by the knowledge of host-guest complexes using cyclodextrins which have the ability to include guest molecules within their hydrophobic cavities, improving the physicochemical properties of the guest. To rationally explore new pesticide formulations, the effects of cyclodextrins on the properties of such guest molecules need to be explored. Imidacloprid is a neonicotinoid systemic insecticide used worldwide. In this study, the inclusion complexes of Imidacloprid (IMI) with ß-cyclodextrin (ß-CD) were prepared in the solid state by co-precipitation and the physical mixing method, with a stoichiometry of 1:1 and 1:2 molar ratios. The obtained products, Imidacloprid:ß-cyclodextrin inclusion complex (IMI:ß-CD), were characterized in the solid state by Fourier transform-infrared (FT-IR) spectroscopy and X-ray powder diffractometry (XRD). In solution, the 1:1 stoichiometry for the inclusion complexes was established by the Job plot method, and the binding constant of IMI:ß-CD was determined by UV-vis titration. The toxicity was determined in producers and primary consumers of the freshwater trophic chain, the green alga Raphidocelis subcapitata and the rotifer Brachionus calyciflorus, respectively. The results indicated that Imidacloprid forms inclusion complexes with CDs showing improved physicochemical properties compared to free Imidacloprid. The formation of the inclusion complex reduced the chronic toxicity in rotifers when IMI concentrations were close to those of environmental concern (tenths/hundredths of micromoles/L). Therefore, CD inclusion complexes could provide important advantages to be considered for the future industrial production of new formulations.

Imidacloprid: Comparative toxicity, DNA damage, ROS production and risk assessment for aquatic non-target organisms.

Imidacloprid is a neonicotinoid systemic insecticide used worldwide. Despite its hazardous impact on non-target organisms, few studies have been conducted concerning the potential eco-genotoxic effects in invertebrates of surface waters where this pesticide is detected from units of ng/L to tens of µg/L. The aim of the present work was to determine the acute, the sub-chronic and the chronic toxicity of imidacloprid in producers and primary consumers of the freshwater trophic chain. The organisms under investigation were the green alga Raphidocelis subcapitata, the rotifer Brachionus calyciflorus, the cladoceran crustacean Ceriodaphnia dubia and the benthic ostracod Heterocypris incongruens. In addition, potential DNA damage and ROS production were evaluated in C. dubia. Furthermore, in accordance with European guidelines, toxicological risk assessment of imidacloprid was performed for all continents considering its global occurrence in surface waters. In addition, we assessed the genotoxicological risk and median inhibition of reproduction was observed at units of mg/L for rotifers and daphnids. Algae showed the lowest level of sensitivity to the pesticide with effective concentrations from units to hundreds of mg/L. DNA lesions were marked from 7 µg/L with a significant increase in damage as concentrations increased. Chronic toxicity risk quotient values were generally below to a threshold value of 1, with no consequential environmental concern other than for the Canadian areas. On the contrary, the genotoxicological risk quotient values were found higher than the threshold value in all continents.

Polystyrene microplastic particles in combination with pesticides and antiviral drugs: Toxicity and genotoxicity in Ceriodaphnia dubia.

Freshwater ecosystems are recognized as non-negligible sources of plastic contamination for the marine environment that is the final acceptor of 53 thousand tons of plastic per year. In this context, microplastic particles are well known to directly pose a great threat to freshwater organisms, they also indirectly affect the aquatic ecosystem by adsorbing and acting as a vector for the transport of other pollutants ("Trojan horse effect"). Polystyrene is one of the most widely produced plastics on a global scale, and it is among the most abundant microplastic particles found in freshwaters. Nevertheless, to date few studies have focused on the eco-genotoxic effects on freshwater organisms caused by polystyrene microplastic particles (PS-MPs) in combination with other pollutants such as pharmaceuticals and pesticides. The aim of this study is to investigate chronic and sub-chronic effects of the microplastic polystyrene beads (PS-MP, 1.0 µm) both as individual xenobiotic and in combination (binary/ternary mixtures) with the acicloguanosine antiviral drug acyclovir (AC), and the neonicotinoid broad-spectrum insecticide imidacloprid (IMD) in one of the most sensitive non-target organisms of the freshwater food chain: the cladoceran crustacean Ceriodaphnia dubia. Considering that the individually selected xenobiotics have different modes of action and/or different biological sites, the Bliss independence was used as reference model for this research. Basically, when C. dubia neonates were exposed for 24 h to the mixtures during Comet assay, mostly an antagonistic genotoxic effect was observed. When neonates were exposed to the mixtures for 7 days, mostly an additive chronic toxic effect occurred at concentrations very close or even overlapping to the environmental ones ranging from units to tens of ng/L for PS-MPs, from tenths/hundredths to units of µg/L for AC and from units to hundreds of µg/L for IMD, revealing great environmental concern.

Toxic impact of polystyrene microplastic particles in freshwater organisms.

The ongoing COVID-19 pandemic is leading to an increase of the global production of plastics since the use of personal protective equipment (PPEs, i.e. gloves, gowns, masks, packaging items), has become mandatory to prevent the spread of the virus. Plastic breaks down into micro/nano particles due to physical or chemical or biological actions into environment. Due to small dimensions, ubiquitous and persistent nature, the plastic particles represent a significant threat to ecosystems and can entry into food chains. Among the plastic polymers used for PPEs, polystyrene is less studied regarding its eco-geno-toxicity. This study aims to investigate acute, chronic and subchronic effects of the microplastic polystyrene beads (PS-MP, size 1.0 µm) on three freshwater species, the alga Raphidocelis subcapitata, the rotifer Brachionus calyciflorus, the crustacean Ceriodaphnia dubia and the benthic ostracod Heterocypris incongruens. Furthermore, the potential genotoxicity and the ROS production due to the PS-MP were also determined in C. dubia. Results revealed that the acute effects occurred at concentrations of PS-MP in the order of dozens of mg/L in B. calyciflorus and C. dubia and hundreds of mg/L in H. incongruens. Regarding long-term toxicity, increasing chronic effects with EC50s in the order of units (C. dubia), hundreds (B. calyciflorus) and thousands (R. subcapitata) of µg/L were observed. Both for acute and chronic/sub chronic toxicity, daphnids were more sensitive to polystyrene than ostracods. Moreover, when C. dubia neonates were exposed to the PS-MP, alterations in genetic material as well as the production of ROS occurred, starting from concentrations in the order of units of µg/L, probably due to inflammatory responses. At last, the risk quotient (RQ) as a measure of risk posed by PS-MPs in freshwater environment, was calculated obtaining a value of 7.2, higher than the threshold value of 1.

Comparative assessment of antimicrobial, antiradical and cytotoxic activities of cannabidiol and its propyl analogue cannabidivarin.

Cannabidiol and cannabidivarin are phytocannabinoids produced by Cannabis indica and Cannabis sativa. Cannabidiol has been studied more extensively than its propyl analogue cannabidivarin. Therefore, we performed a battery of in vitro biological assays to compare the cytotoxic, antiradical and antibacterial activities of both cannabinoids. Potential mitochondrial metabolism alterations, DNA synthesis inhibition, and plasma membrane damage were studied by MTT assay, BrdU-ELISA and LDH assay of cancer and normal human cells exposed to cannabinoids. ABTS and DPPH assays were performed to observe the effects of the cannabinoids on free radicals. Microbial susceptibility tests were performed to study the activity of the cannabinoids in two bacterial species implicated in human infections, Escherichia coli and Staphylococcus aureus. The results showed that the cannabinoids induced medium levels of cytotoxicity in cancer and normal cells at concentrations ranging from 15.80 to 48.63 and from 31.89 to 151.70 µM, respectively, after 72 h of exposure. Cannabinoids did not exhibit a strong antioxidant capacity in scavenging ABTS or DPPH radicals. No evident differences were observed between the two cannabinoids in antimicrobial activity, except with respect to S. aureus, which showed greater susceptibility to cannabidiol than to cannabidivarin after 72 h of exposure.

Natural Methoxyphenol Compounds: Antimicrobial Activity against Foodborne Pathogens and Food Spoilage Bacteria, and Role in Antioxidant Processes.

The antibacterial and antioxidant activities of three methoxyphenol phytometabolites, eugenol, capsaicin, and vanillin, were determined. The in vitro antimicrobial potential was tested on three common foodborne pathogens (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus) and three food spoilage bacteria (Shewanella putrefaciens, Brochothrix thermosphacta, and Lactobacillus plantarum). The antioxidant assays were carried out for studying the free radical scavenging capacity and the anti-lipoperoxidant activity. The results showed that eugenol and capsaicin were the most active against both pathogens and spoilage bacteria. S. aureus was one of the most affected strains (median concentration of growth inhibition: IC50 eugenol = 0.75 mM; IC50 capsaicin = 0.68 mM; IC50 vanillin = 1.38 mM). All phytochemicals slightly inhibited the growth of L. plantarum. Eugenol was the most active molecule in the antioxidant assays. Only in the oxygen radical absorbing capacity (ORAC) test did vanillin show an antioxidant activity comparable to eugenol (eugenol ORAC value = 2.12 ± 0.08; vanillin ORAC value = 1.81 ± 0.19). This study, comparing the antimicrobial and antioxidant activities of three guaiacol derivatives, enhances their use in future applications as food additives for contrasting both common pathogens and spoilage bacteria and for improving the shelf life of preserved food.

Theobromacacao Criollo var. Beans: Biological Properties and Chemical Profile.

Theobroma cacao provides precious products such as polyphenol-rich beans that are useful for nutraceutical purposes. The geographical area may influence the chemical composition of raw cocoa beans in terms of the polyphenols and biological qualities of the products. This work aimed to investigate the biological properties and the chemical composition of two different samples of Criollo var. cocoa raw beans coming from two areas (Indonesia; Peru). Beans underwent biphasic extraction obtaining lipophilic and hydroalcoholic extracts. The extracts were tested for antiradical, antimutagenic, and antigenotoxic effects. Cell viability inhibition toward breast, gastric/esophageal colorectal adenocarcinoma, and hepatoblastoma human cell lines was evaluated. Extracts were chemically investigated through UV-Vis spectroscopy and ultra-high-pressure liquid chromatography electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-QqTOF MS/MS). Results showed that the Indonesian bean hydroalcoholic extracts were able to scavenge 2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) cation radical better than the Peruvian hydroalcoholic extracts (ECs50: 72.63 vs. 322.20 µg/mL). Extracts showed antimutagenic and antigenotoxic activity. The viability inhibitory effect on breast and hepatic cancer cells was reached only for the Indonesian hydroalcoholic extracts at hundreds of µg/mL. Phenylpropenoyl-L-amino acids, hydroxycinnamoyl aminoacids conjugates, and procyanidin compounds were found mainly in the hydroalcoholic extracts, whereas fatty acids and lyso-phospholipids were found mainly in lipophilic fractions. Fatty acid and (epi)catechins appeared to be affected by different environmental conditions of the geographical areas.

Capsaicin in Hot Chili Peppers: In Vitro Evaluation of Its Antiradical, Antiproliferative and Apoptotic Activities.

Capsaicin is a spicy capsaicinoid, produced as secondary metabolite by Capsicum fruits. This alkaloid has been used for years in folk medicine for its analgesic and antinflammatory properties although most data is referred to the raw fruit. In this study, the antiradical activity of the pure capsaicin has been studied using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays as well as its antiproliferative activity, using MTT assay, against two human tumour cell lines, the colorectal Caco-2 and the oesophageal OE19 cells. Furthermore, the antiproliferative activity observed on tumoral cells was compared with that of the human normal-like fibroblast cell line TelCOFS02MA. In addition, the apoptotic activity was evaluated using TUNEL assay. A higher radical scavenging activity was observed against ABTS radical cation than DPPH. Capsaicin showed also a higher cytotoxicity against cancer cells than normal-like cells with Selectivity index values greater than 2 at 72 h. Capsaicin induced apoptosis especially in OE19 cell line.