Cell-based assays as an alternative for the study of aquatic toxicity of pharmaceuticals

An increasing number and amount of pharmaceuticals for human and veterinary use currently reach the aquatic environment, and the determination of their effects on aquatic organisms becomes of major importance. The 96-h fish lethal test is one of the conventional assays required for environmental hazardous assessment, but it is extremely time-consuming and costly, and it raises ethical concerns. In a broad study, we compared the ability of cell-based assays to detect, in absolute terms, lethal toxicity in fish due to pharmaceuticals in order to select sensitive cell lines to be posteriorly used as an alternative to fish testing. This study also explored the sensitivity of the rat cardiomyoblast H9c2(2-1) cell line and the suitability of the sulforhodamine B colorimetric assay regarding 15 pharmaceuticals belonging to 9 different therapeutic classes. The relation between in vivo and in vitro data was expressed as LC_50,96h/EC₅₀ ratios, and 66% of concordant data were attained. Accordingly, it was possible to conclude that cell-based assays could be considered a suitable alternative to fish lethal testing for pharmaceuticals, which, after validation, may dramatically reduce the number of fish required for environmental hazardous assessment. Several cell lines were selected as promising alternatives, but H9c2(2-1), HepG2, PLHC-1, and RTG-2 could be considered suitable starting cell types for further studies, as relevant results were obtained with low exposure times.

     

Effects of insecticides used in strawberries on stingless bees Melipona quadrifasciata and Tetragonisca fiebrigi (Hymenoptera: Apidae)

Environmental Science and Pollution Research - The use of pesticides is considered one of the most important threats to pollinators, especially since they are widely used in agriculture for pest...     

Gastroenteric Viruses Detection in a Drinking Water Distribution-to-Consumption System in a Low-Income Community in Rio de Janeiro

Food and Environmental Virology - The availability of drinking water is one of the main determinants of quality of life, disease prevention and the promotion of health. Viruses are important agents...     

Hepatitis A Virus,Hepatitis E Virus,and Rotavirus in Foods of Animal Origin Traded at the Borders of Brazil,Argentina, and Uruguay

The aim of this study was to investigate hepatitis A virus (HAV), hepatitis E (HEV), and rotavirus (RV) in fresh and processed meat traded on the border of Brazil with Argentina and Uruguay. In total, 159 samples of raw and processed foods of animal origin were collected in Paso de los Libres, Argentina (n?=?53 raw meat, n?=?24 processed meat) and Rivera, Uruguay (n?=?55 raw meat, n?=?18 processed meat), or were seized by the Brazilian International Agricultural Surveillance System—VIGIAGRO (Brazil–Argentina border) (n?=?8 raw meat, n?=?1 bush meat). All samples were tested for the presence of HAV, HEV, and RV genomes. HAV genes were detected in 18.23% of samples and RV genes in 23.89%. No HEV-positive samples were detected. HAV was also detected in two of the VIGIAGRO samples. Processed meats from Argentina and Uruguay had a higher rate of HAV and RV than raw meat (P?>?0.05). The median HAV in the Argentinian and Uruguayan samples was 6.9?×?10⁴ and 3.5?×?10³ copies/g, respectively. The presence of RV viral genes in raw meats from Argentina was significant, and this was not observed in processed meats. The presence of HAV and RV genes in a significant portion of products from Argentina and Uruguay is a potential source of human infection. This also indicates precarious conditions of acquisition, processing, and manipulation, which could be improved by improved regulation of food across borders.     

AnSBBR applied to the treatment of wastewater from a personal care industry: Effect of organic load and fill time

The objective of this work was to study the technological feasibility of treating wastewater from a personal care industry (PCI-WW) in a mechanically stirred anaerobic sequencing batch biofilm reactor (AnSBBR) containing immobilized biomass on polyurethane foam. An assessment was made on how system efficiency and stability would be affected by: increasing organic load; supplementation of nutrients and alkalinity; and different feed strategies. The AnSBBR operated with 8-h cycles, stirring speed of 400 rpm, temperature of 30 °C, and treated with 2.0 L wastewater per cycle. First the efficiency and stability of the AnSBBR were studied when submitted to an organic loading rate (OLR) of 3.1–9.4 gCOD/(L d), and when the PCI-WW was supplemented with nutrients (sucrose, urea, trace metals) and alkalinity. The AnSBBR was shown to be robust and presented stability and removal efficiency exceeding 90%. At an OLR of 12.0 gCOD/(L d) efficiency became difficult to maintain due to the presence of commercial cleansers and disinfectants in the wastewater lots. In a subsequent stage the AnSBBR treated the wastewater supplemented with alkalinity, but with no nutrients at varying feed strategies and maintaining an OLR of approximately 9.0 gCOD/(L d). The first strategy consists of feeding 2.0 L of the influent batchwise [OLR of 9.4 gCOD/(L d)]. In the second 1.0 L of influent was fed-batchwise and an additional 1.0 L was fed fed-batchwise [OLR of 9.2 gCOD/(L d)], i.e., in relation to the first strategy the feed volume was maintained but supplied in different periods. In the third strategy 1.0 L of treated effluent was maintained in the reactor and 1.0 L of influent was fed fed-batchwise [OLR of 9.0 gCOD/(L d)], i.e., in relation to the first strategy the feed volume was different but the feed period was the same and the OLR was maintained by increasing the influent concentration. Comparison of the first and second strategies revealed that organic matter removal efficiency was unaffected (exceeding 90%). The third strategy resulted in a reduction in average removal efficiency from 91 to 83% when compared to the first one. A kinetic study resulted in first order kinetic parameters ranges from 0.42 to 1.46 h⁻¹ at OLRs from 3.1 to 12.0 gCOD/(L d), respectively, and the second feed strategy [OLR of 9.2 gCOD/(L d)] was shown to be the most favorable.     

Effect of impeller type and mechanical agitation on the mass transfer and power consumption aspects of ASBR operation treating synthetic wastewater

The effect of flow type and rotor speed was investigated in a round-bottom reactor with 5 L useful volume containing 2.0 L of granular biomass. The reactor treated 2.0 L of synthetic wastewater with a concentration of 800 mgCOD/L in 8-h cycles at 30 degrees C. Five impellers, commonly used in biological processes, have been employed to this end, namely: a turbine and a paddle impeller with six-vertical-flat-blades, a turbine and a paddle impeller with six-45 degrees -inclined-flat-blades and a three-blade-helix impeller. Results showed that altering impeller type and rotor speed did not significantly affect system stability and performance. Average organic matter removal efficiency was about 84% for filtered samples, total volatile acids concentration was below 20 mgHAc/L and bicarbonate alkalinity a little less than 400 mgCaCO3/L for most of the investigated conditions. However, analysis of the first-order kinetic model constants showed that alteration in rotor speed resulted in an increase in the values of the kinetic constants (for instance, from 0.57 h(-1) at 50 rpm to 0.84 h(-1) at 75 rpm when the paddle impeller with six-45 degrees -inclined-flat-blades was used) and that axial flow in mechanically stirred reactors is preferable over radial-flow when the vertical-flat-blade impeller is compared to the inclined-flat-blade impeller (for instance at 75 rpm, from 0.52 h(-1) with the six-flat-blade-paddle impeller to 0.84 h(-1) with the six-45 degrees -inclined-flat-blade-paddle impeller), demonstrating that there is a rotor speed and an impeller type that maximize solid-liquid mass transfer in the reaction medium. Furthermore, power consumption studies in this reduced reactor volume showed that no high power transfer is required to improve mass transfer (less than 0.6 kW/10(3)m3).     

Prediction of the PCDD/F and dl-PCB 2005-WHO-TEQ content based on the contribution of six congeners: Toward a new screening approach for fish samples?

Current European Union regulation regarding polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) in food and feed is based on Toxic Equivalent Quotient (TEQ) concept. For confirmatory purpose, the isotope-dilution method associated to a measurement by gas chromatography coupled with high resolution mass spectrometry is usually the method of choice for precisely measuring the 29 target congeners in three separated fractions. Time and cost related to these analyses are very significant. Various kinds of screening concepts can be considered. In the present study, we elaborated and validated a prediction model for the 2005 World Health Organization TEQ in fish, based on the measurement of 4 PCDD/F and 2 non-ortho dl-PCB congeners, potentially analyzable in a single extracted fraction by gas chromatography coupled with mass spectrometry. Large independent datasets have been used for model elaboration (n = 108) and validation (n = 363, n = 357 and n = 6).     

Inactivation of LPS and RNase A on photocatalytically active surfaces

TiO₂ coated surfaces are able to generate highly reactive oxidizing species under mild UV-A light exposure in the presence of water and oxygen. We have demonstrated that these radicals are sufficient to eliminate different pathogenic bacteria, by breaking their cell walls. The photocatalytic activity of surfaces coated with titanium dioxide offers therefore an alternative possibility of disinfection. However, restriction of bacterial growth does not protect surfaces from bacterial derived contaminations, such as endotoxins. Lipopolysaccharides (LPS) and Ribonuclease A (RNAse A) represent the two most abundant contaminations, causing severe problems in biomedical and immunological research as well as in the pharmaceutical industry. Due to their high stability, complete removal of these contaminants is technically challenging. Using irradiated TiO₂ coated glass plates, RNAse A and LPS containing contaminations could be completely inactivated. By establishing highly sensitive immuno-based assays, destruction of the contaminants was quantified and shown to be independent of the initial concentrations, following a zero-order reaction. Exposure for 96 h resulted in a reduction of 11 ng of LPS and 7 units of RNase A cm⁻² surface. These amounts are comparable to contamination levels found under standard working conditions. Titanium dioxide coatings provide therefore a powerful tool for auto-disinfection and self-cleaning of surfaces.     

Study of environmental burden of lead in children using teeth as bioindicator

The south region of Sao Paulo city hosts the Guarapiranga dam, responsible for water supply to 25% of the city population. Their surroundings have been subject to intense and irregular occupation by people from very low socioeconomics classes. Measurements undertaken on sediment and particulate materials in the dam revealed concentrations of lead, copper, zinc and cadmium above internationally accepted limits. Epidemiological and toxicological studies undertaken by the World Health Organization in individuals exhibiting lead concentrations in blood, near or below the maximum recommended (10 µg dl^? 1), surprisingly revealed that toxic effects are more intense in individuals belonging to low socioeconomics classes. Motivated by these facts, we aimed at the investigation of chronic incorporation of lead, as well as the use of our BIOKINETICS code, which is based on an accepted ICRP biokinetics model for lead, in order to extrapolate the results from teeth to other organs. The focus of our data taking was children from poor families, living in a small, restrict and allegedly contaminated area in São Paulo city. Thus, a total of 74 human teeth were collected. The average concentration of lead in teeth of children 5 to 10 years old was determined by means of a high-resolution inductively coupled plasma mass spectrometer (ICP-MS). For standardization of the measurements, an animal bone certified material (H-Animal Bone), from the International Atomic Energy Agency, was analyzed. The amount of lead in children living in the surroundings of the dam, was approximately 40% higher than those from the control region, and the average lead concentration was equal to 1.3 μg g^? 1 approximately. Grouping the results in terms of gender, tooth type and condition, it was concluded that a carious molar of boys is a much more efficient contamination pathway for lead, resulting in concentrations 70% higher than in the control region. We also inferred the average concentrations of lead in other organs of these children, by making use of our BIOKINETIC code.     

Growth and Histopathological Effects of Chronic Exposition of Marine Pejerrey Odontesthes argentinensis Larvae to Petroleum Water-Soluble Fraction (WSF)

The water-soluble fraction (WSF) of petroleum contains a mixture of polycyclic aromatic hydrocarbons, volatile hydrocarbons, phenols, and heterocyclic compounds, considered deleterious to aquatic biota. Marine “pejerrey” Odontesthes argentinensis (Teleostei: Atherinopsidae) has a great commercial importance in local fisheries and a high potential for aquaculture. The aim of this study was to evaluate the histopathological effects in “pejerrey” larvae exposed to different concentrations of petroleum WSF. The chronic toxicity test was conducted with newly hatched larvae exposed for 21 days to sublethal concentrations of WSF (2.5, 5, 10, and 20 % of WSF), plus one control. Survival and growth were significantly lower in the highest concentration. Several histopathological changes were found in the gills (e.g., hyperplasia, aneurisms, edema, and necrosis), kidney (e.g., nuclear alterations, decrease in the hematopoietic cells), and liver (e.g., hypertrophy, karyorrhexis, and karyopyknosis). An index of branchial lesion was proposed to standardize gill lesions to different pollutants.