Univariate or multivariate approaches for histopathological biomarkers in the context of environmental quality assessments?

Although the simplification of multivariate histopathological data into univariate indices can be useful for the assessment of environmental quality, this implies a great loss of information. The objective of the present study was to evaluate the effectiveness, in the context of environmental quality assessment, of an approach that integrates individual histopathological responses in a discriminated manner with the results of contaminants by means of multivariate analyses. This analysis was compared to the diagnosis of environmental quality provided by the use of the univariate Bernet histopathological index. Contaminant loads (sediments and fish) and the liver histopathology of Cathorops spixii were integrated through multivariate analysis. Integrated individual histopathological responses allowed classifying environmental quality from more to less impacted sites, while the univariate index showed some inconsistencies with chemical loads and allowed identifying only the most impacted site.

The effects of anaerobic reactor post-treatments by rapid filtration systems and conventional techniques.

Although anaerobic reactors are an excellent alternative in the treatment of domestic effluents, they have the disadvantage of requiring post-treatment. Many technologies have been studied and, recently, rapid filtration systems have been presented as a viable alternative for post-treatment. This work compared post-treatment techniques for anaerobic upflow sludge blanket (UASB) reactors by rapid filtration systems (double filtration (DF); triple filtration with clinoptilolite (TFc); and triple filtration with activated carbon (TFac)) to conventional systems (facultative pond (FP); biological filter (BF); biological filter with recirculation and decantation (BFD)), verifying their potential for improvement of the final effluent quality. The UASB effluent post-treatments by FP, BF, BFD, DF, TFc, and TFac were evaluated. The removal of turbidity in both BFD and FP post-treatments was below 75%. The DF, TFc, and TFac treatments showed over 99% removal of the same parameters. COD removal in the FP, BF, and BFD post-treatments was over 10%, while in the DF, TFc, and TFac treatments, it was over 80%. The greatest total phosphorus removal was observed in TFc and TFac, whose values were over 99%. The best removal of ammoniacal nitrogen, 99% was observed in the TFc treatment. Regarding Al, Cd, Cr, Cu, Mn, Ni, Pb, and Zn removal, all rapid filtration systems showed better performance when compared to conventional systems. The DF, TFc, and TFac systems showed over 90% removal of most metals evaluated, while the FP and BF treatments presented values below 50% for most metals, and in the BFD system, the removal values were below 80% for most metals. The results indicate that rapid filtration systems were better at removing all evaluated parameters when compared to conventional systems.

Exposure to pollutants present in Iguaçu River Southern Brazil affect the health of Oreochromis niloticus (Linnaeus, 1758): Assessment histological, genotoxic and biochemical.

Urban sewage is a source of major contamination in aquatic systems and contributes to environmental and human health disturbances. This study investigates the effects of sewage-polluted waters from Iguaçu River on the health of juvenile Oreochromis niloticus. Two hundred four specimens were exposed to riverine water in four groups: no diluted, 25 and 50 % diluted water and a control group without tested water for 72 days. Biological samples were obtained for histopathological, neurotoxicity, antioxidant defenses, genotoxicity, metallothionines expression and polycyclic aromatic hydrocarbons (PAHs) metabolites. The results showed histopathological alterations in liver and gills, genotoxic alteration in erythrocytes, reduction of acetylcholinesterase activity in brain and muscle, activation of antioxidant defenses in the liver, recruitment of metals by metallothionein and the detection of PAHs metabolites in bile. These results demonstrate that juveniles of O. niloticus are susceptible to Iguaçu River exposure water and they can be used as indicator of water quality.

Bioaccumulation and bioconcentration of metals in Characidae from a Neotropical river basin under anthropic activities.

Natural rates of metal mobilization and deposition in terrestrial and aquatic environments have been changed due to anthropic activities, exposing the native biota to dangerous effects related to bioconcentration and bioaccumulation of metals. This study assessed the concentrations of Cr, Ni, Cd, Pb, Cu, Mn, Co, and Zn in the water and riverbed sediment samples from the Verde River basin (VR), and in tissue samples from two native fishes, the Psalidodon paranae, a VR inhabitant, and the Psalidodon aff. fasciatus, a migratory species. Arithmetic mean values of metal concentrations recorded in waters were Cr: 46.16, Ni: 40.29, Cd 43.19, Pb: 57.74, Cu: 63.72, Mn: 98.36, Co: 64.53, Zn: 81.19, while for riverbed sediments were Cr: 11.84, Ni: 10.52, Cd: 7.14, Pb: 15.00, Cu: 22.16, Mn: 334.77, Co: 24.62, Zn: 434.44. For several analyzed samples, metal concentrations found were higher than Brazilian and international limits set for healthy aquatic life and human uses. Analyzed fish tissues also presented metal concentrations higher than Brazilian and international limits set, indicating a high ecological and health risk for the region. Psalidodon paranae showed affinity to bioconcentrate Pb, Zn, and Cd, while Psalidodon. aff. fasciatus tended to bioconcentrate Ni, Zn, and Cr. Multivariate analyses revealed spatial and temporal patterns in the metal contaminations in VR. These patterns were associated with rural and urban activities developed along VR, which practice inadequate soil handling, indiscriminate use of agrochemicals, and the dumping of domestic garbage and untreated and treated sewage into the river. The implementation of public policies for biomonitoring and pollution control by metals in VR is essential to safeguard regional water resources and their biota.

Micropollutants impair the survival of Oreochromis niloticus and threat local species from Iguaçu River, Southern of Brazil.

The wastewater contamination of urban rivers is a concern for biodiversity and a consequence from poor urban conservation policies. In the current study, the impact of urban and industrial activities was investigated in Iguaçu river (Southern Brazil) using juvenile Oreochromis niloticus, after trophic and chronic exposure (25, 50 and 100 %), over 81 days. After exposure liver, gills, gonads, brain, muscle, and blood were sampled for chemical, biochemical, histopathological, genotoxic and molecular analyses. Water levels of persistent organic pollutants such as polychlorinated biphenyls, organochlorine pesticides, polycyclic aromatics hydrocarbon (PAHs) and metals were investigated. The redox unbalance, histopathological and increase in vitellogenin expression in fish revealed both the bioavailability of micropollutants and their harmful effects. According to the results, the level of Iguaçu river pollution negatively impacts the health of O. niloticus revealing and highlighting the risk of this pollution exposure to biota and human populations.

Effects of different filtration techniques on quality and toxicology of post treatment effluent from an anaerobic reactor.

The early stages of the fish life cycle correspond to the phase that is most susceptible to deleterious effects caused by exposure to pollution. The aim of this study was to evaluate the quality of the anaerobic reactor effluent after three filtration-based treatments, namely, Double-Filtration in gravel and sand (DF), Triple-Filtration in gravel, sand and Activated Charcoal (TF-AC) and Triple-Filtration in gravel, sand and clinoptilolite (TF-C). The toxic effects on the population dynamics of larvae and embryos of catfish (Rhamdia quelen) to the final effluent were evaluated using an individual-based model (IBM). The results indicate that the three post-treatments produced effluents with significant improvement of the physicochemical parameters evaluated in relation to the anaerobic reactor effluent. In addition, all post-treatments improved the removal of metal ions. Experimental data showed high mortality rates for Rhamdia quelen embryos and larvae for most treatments, except for the effluent treated by TF-C. The results demonstrated that the concentration of ammoniacal nitrogen in relation to AR treatments (69.0); DF (44.0); TF-AC (46.6) and TF-C (0.33) in mg/L can be a limiting factor for the development of embryos and larvae during the ecotoxicity tests.

Availability of nutrients, removal of nicotine, heavy metals and pathogens in compounds obtained from smuggled cigarette tobacco compost associated with industrial sewage sludge.

This study evaluated the chemical and microbiological properties of the compost obtained from the tobacco of smuggled cigarettes (SCT) and industrial sewage sludge (ISS). The composting was carried out in three reactors from different combinations of residues. The compost was analyzed to verify the percentage of nicotine removal, heavy metals, nutrient content and the inactivation of pathogenic microorganisms. The concentration of heavy metals: Ni, Cd, Cr, Pb, Cu, Zn, Fe and Mn in the composts obtained from the three reactors was below the values set for their use in soils. The NPK content ranged between 8.31% and 12.43%, indicating that the compost produced can add nutritional benefits to the plants. The nicotine removal, 72.6% (R1), 96.4% (R2) and 99.6% (R3) indicated efficiency of the composting process in reactors in the degradation of this substance. The results of pathogenic microorganism analysis showed that the three composts obtained from reactors R1, R2 and R3 met the sanitation standards for agricultural use according to the normative of maximum limits of contaminants allowed in organic compounds. These results show that the treatment of SCT and ISS by the process of composting in reactors may be an ecologically viable alternative.

Pb2+ adsorption by a compost obtained from the treatment of tobacco from smuggled cigarettes and industrial sewage sludge.

The adsorption of Pb2+ by a compost obtained from the treatment of tobacco from smuggled cigarettes (SCT) and industrial sewage sludge (ISS) was investigated. The Pb2+ adsorption process was evaluated as function of different concentrations of adsorbent and adsorbate, pH variations, and contact time. Fourier transformed infrared spectroscopy (FTIR) and energy dispersive X-ray spectrometry (EDX) were adopted to obtain information regarding structural changes and a better understanding of the adsorption mechanism. The adsorbent maximum adsorption capacity for Pb2+, calculated using the Sips equation, was 21.454 mg/g with 3 g/L adsorbent at pH 5. The adsorption kinetics best adjustment was obtained using the pseudo-second-order model with a time of 240 min to reach the adsorption equilibrium. FTIR and EDX results suggest that Pb2+ might have bonded to phenolic, carboxylic, hydroxyl, and amine groups; they also show formation of organometallic complexes and cationic exchange between the compost and the solution. The study confirmed that the compost evaluated can be used as a potential adsorbent in environments contaminated with Pb2+.

Post-treatment of anaerobic reactor effluent for reuse using a triple filtration system.

This study evaluated the triple filtration technology efficiency as a post-treatment of anaerobic reactor effluent. This study was carried out employing different concentrations of ferric chloride as coagulant and peracetic acid or calcium hypochlorite as oxidant. The filtration rates used were 150 m3/m2 d and 120 m3/m2 d. The efficiency of the system was evaluated through physicochemical and microbiological parameters. The best conditions found were those using 20 mg/L ferric chloride, 120 m3/m2 d filtration rate and 0.8 and 1.6 mg/L free chlorine. These conditions resulted in turbidity <1.0 NTU, Total Organic Carbon <1.5 mg/L, Chemical Oxygen Demand <1.0 mg/L, Biochemical Oxygen Demand <1.0 mg/L, in addition 100% removal of Total Phosphorus and Linear Alkylate Sulfonate. The post-oxidation process promoted inactivation of 100% Total Coliforms and E. coli. The post-treatment was able to produce effluent with characteristics that enable its urban, damming, creation and maintenance of wetlands, industrial and agricultural reuse proposed by USEPA.

Exposure of male tilapia (Oreochromis niloticus) to copper by intraperitoneal injection: DNA damage and larval impairment.

The present work aimed to assess the genotoxic effects of intraperitoneally injected copper in males of O. niloticus, and to investigate its association with eventual changes in the quality of the gametes and in the generation of offspring abnormalities. Thus, to evaluate DNA damage, the alkaline comet assay was performed in sperm and blood; sperm parameters were analyzed to determine the quality of the gametes, and the incidence of morphological abnormalities of larvae, originated from eggs fertilized by males exposed to copper, was used as a measure of reproductive impairment. Male fish were exposed to three doses (1 mg/kg, 2 mg/kg, 4 mg/kg) of copper sulfate via intraperitoneal injection and observed after 96 h of exposure. We found that the quality of gametes was not affected by copper, as indicated by the lack of differences in sperm parameters between the exposed and the control groups. Nonetheless, copper induced an increase in sperm DNA damage at all tested doses. Taking into consideration the total number of abnormalities, larvae presented a higher proportion of morphological abnormalities at all the tested doses. Considering the morphological abnormalities separately, the 2 mg/kg and 4 mg/kg groups presented significantly higher frequencies of arrested development and spinal abnormalities than the control group. This work suggests that intraperitoneally injected copper is an inductor of DNA breakage in germ cells of O. niloticus males. Copper induces the formation of larval morphological abnormalities, causing reproductive impairment to this species. The fact that copper did not induce alterations on sperm parameters indicates that the parental genotoxic effects are inheritable to the offspring.

Composting of smuggled cigarettes tobacco and industrial sewage sludge in reactors: Physicochemical, phytotoxic and spectroscopic study.

The maturity of smuggled cigarette tobacco (SCT) and industrial sewage sludge (ISS) compost during composting in reactors was evaluated through physicochemical, phytotoxic and spectroscopic parameters. The temperatures reached peaks above 52 °C in the three reactors and were enough to achieve the stability of the compounds. The electric conductivity was in the optimal interval for farming uses and the pH alkaline band was favorable to produce inorganic nitrogen. The reduction in the C∕N ratio and the increase in cation exchange capacity (CEC) indicated an increase in the compost humification. After 120 days, the seed germination index (SGI) reached 95 % in reactor 3. In the three treatments, the reduction in E2/E6 and E4/E6 ratios (UV/Vis), the increase in humification indices, obtained through FTIR and the aromatic carbon resonance (13C NMR) indicated a high degree of aromaticity. The composting process in reactors was efficient to degrade different proportions of SCT and ISS, resulting in mature composts.

Time does matter! Acute copper exposure abolishes rhythmicity of clock gene in Danio rerio.

The circadian clock is a key cellular timing system that coordinates physiology and behavior. Light is a key regulator of the clock mechanism via its activation of Per and Cry clock gene expression. Evidence points to a key role of reactive oxygen species (ROS) in resetting this process. In this context, the aim of the present study was to explore copper as a ROS generator, using an innovative approach investigating its effects on circadian timing. Liver and brain from Danio rerio specimens exposed to 0, 5, 25 and 45 µg/L copper concentrations were obtained. Daily oscillations of superoxide dismutase (SOD) and catalase (CAT) enzymatic activity and their correlations both with clock genes (per1, per2, and cry1a) and with organism energy cost were determined. CAT expression correlates with per2 and cry1a and, thus, provides data to support the hypothesis of hydrogen peroxide production by a phototransducing flavin-containing oxidase. Higher SOD activity is correlated with higher intracellular ATP levels. Copper disturbed the daily oscillation of antioxidant enzymes and clock genes, with disturbed per1 rhythmicity in both the brain and liver, while cry1a rhythmicity was abolished in the liver at 25 µg/L copper. Coordination between the SOD and the CAT enzymes was lost when copper concentrations exceeded the limits established by international laws. These results indicate that organism synchronization with the environment may be impaired due to acute copper exposure.

Metals and arsenic in fish from a Ramsar site under past and present human pressures: Consumption risk factors to the local population.

The risk of metals and As in seafood for traditional populations living in a Marine Protected Areas (MPA) is seldom assessed, although the risk of human exposure to contaminants is one of the indicators associated with the socioeconomic goals of MPAs. The current study aimed to estimate the potential risk of some metals (Cd, Pb, and Zn) and arsenic (As) for human health through the ingestion of fish locally harvested in a Ramsar site, the Cananéia-Iguape-Peruíbe Environmental Protected Area (APA-CIP). Previous studies showed environmental impacts in this area due to former mining activities and urbanization. Cathorops spixii, a catfish largely consumed by the local population, was collected along the estuary in three seasons with different rain regimes. Metals and As loads in muscle tissue were quantified and it was estimated (i) the target hazard quotient (THQ) and (ii) the daily intake (EDI) for metals and As, (iii) the cancer risk (CRisk) only for As, and (iv) the number of eligible meals per month. Cd, Pb, and As were found at concentrations above action levels for human consumption. Depending on the level of exposure of the local population, the consumption of C. spixii may pose risk to human health. Highest THQs were estimated for fish collected in sites closer to the main contamination sources in the APA-CIP, i.e. the mouth of Ribeira de Iguape River (P1) and the city of Cananéia (P4, P5, and P6). Arsenic showed high levels of cancer risk, although restricted to the area close to the city. The exposure of the local population to metal and As contaminated seafood cannot be disregarded in environmental studies and management of the APA-CIP.

How and where to perform biomonitoring studies: different levels of toxic metal pollution are detected in the Alagados Reservoir in Southern Brazil.

The Alagados Reservoir located in the Brazilian city of Ponta Grossa is used to supply water for human consumption but is impacted by toxic metals. The current study combined chemical, biochemical, and multivariate analyses to determine the bioavailability of toxic metals at three sites (AL-A, AL-B, and AL-C) within the Alagados Reservoir. Metal bioaccumulation was analyzed in the liver, gills, and muscle tissue of a native fish species (Geophagus brasiliensis), and neurotoxicity, xenobiotic metabolism, and oxidative stress were evaluated using biochemical biomarkers. Additionally, histopathological studies were performed on the gills and the liver using scanning electron microscopy (SEM) and conventional light microscopy (LM), respectively. Overall, the bioaccumulation of metals, biomarkers of oxidative stress, and melanomacrophage counts indicate that the AL-C and the AL-A sites are the most and least affected by metals, respectively. The AL-B site presented the lowest acetylcholinesterase enzyme activity, a finding which was probably associated with the agricultural activities around this area of the reservoir. The biomarkers clearly revealed that toxic metals negatively affect all three sites studied herein and that human activity is the major source of pollutants. Despite the existence of different pollution levels within the Alagados Reservoir, it is still used as a human water supply.

Translocation of metal ions from soil to tobacco roots and their concentration in the plant parts.

This paper presents a study on the translocation factors (TFs) and bioconcentration factors (BCFs) of copper (Cu), manganese (Mn), zinc (Zn), cobalt (Co), chromium (Cr), cadmium (Cd), lead (Pb), iron (Fe), nickel (Ni), and arsenic (As) ions in roots, stems, and leaves of tobacco. The results revealed that during the tobacco growth, the roots are able to increase the sensitiveness of the physiological control, reducing the translocation of the metals Ni (0.38) and Pb (0.48) to the leaves. Cd and Zn presented factors TF and BCF >1 in the three tissues under analysis, which indicates the high potential for transportation and accumulation of these metals in all plant tissues. The TF values for Cr (0.65) and As (0.63) revealed low translocation of these ions to the aerial parts, indicating low mobility of ions from the roots. Therefore, tobacco can be considered an efficient accumulator of Ni, Cr, As and Pb in roots and Cd and Zn in all plant parts.

Brain effects of manganese exposure in mice pups during prenatal and breastfeeding periods.

Manganese (Mn) is a trace element essential for brain development and functioning of the central nervous system. However, there is a lack of information concerning the neurotoxicity of Mn under realistic doses in early stages of development, though excess of Mn results in a progressive disorder of the nervous system called manganism. In the current study, adult mice were exposed to three doses of Mn for 60 days through daily gavages, while mice pups were exposed to the same Mn doses during developmental period (gestational and breast-feeding). From the latter group of mice, a group was exposed for more 60 days to the same Mn doses. Chemical analysis revealed a dose-dependent bioaccumulation of Mn in mice's brain. Biochemical parameters revealed that (1) Mn affects non-protein thiol levels, glutathione S-tranferase and acetylcholinesterase activities, as well as the levels of oxidized lipids and proteins in mice brain, though lipids and proteins alterations were found only after exposure to high and unrealistic doses; (2) Realistic doses of Mn affects the activity of brain AChE and finally; (3) Pups' brain were affected by Mn even whether only the parental females had been previously exposed. The current study shows evidences of chemical stress in mice exposed to Mn during the early period of development and an efficient mechanism of Mn elimination under higher doses. These findings open new lines of investigation regarding manganese toxicity in vertebrates mainly in the early stages of development.

Toxicological interactions of silver nanoparticles and organochlorine pesticides in mouse peritoneal macrophages.

Nanotechnology occupies a prominent space in economy and science due to the beneficial properties of nanomaterials. However, nanoparticles may pose risks to living organisms due to their adsorption and pro-oxidative properties. The aim of the current study was to investigate the effects of polymer-coated silver nanoparticles (AgNPs) and organochlorine pesticides (OCPs), as well as their combined effects on mouse peritoneal macrophages. Macrophages were isolated and exposed to three concentrations of AgNPs (groups: N1 = 30, N2 = 300 and N3 = 3000 ng.ml(-1)), two concentrations of OCPs (groups: P1 = 30 and P2 = 300 ng.ml(-1)) and the six possible combinations of these two contaminants for 24 h. AgNPs had irregular shape, Feret diameter of 8.7 ± 7.5 nm and zeta potential of -28.7 ± 3.9 mV in water and -10.7 ± 1.04 mV in culture medium. OCP mixtures and the lower concentrations of AgNPs had no detectable effects on cell parameters, but the highest AgNPs concentration showed high toxicity (trypan blue and MTT assays) resulting in morphological changes, increase of nitric oxide levels and phagocytic index. Foremost, the association of N3 and P2 led to distinct effects from those observed under single exposure.

The applied indicators of water quality may underestimate the risk of chemical exposure to human population in reservoirs utilized for human supply-Southern Brazil.

The knowledge concerning associations between chronic chemical exposure and many disorders with complex etiology involving gene-environment interactions is increasing, and new methods must be developed to improve water quality monitoring. The complexity of chemical mixtures in polluted aquatic environments makes the evaluation of toxic potential in those sites difficult, but the use of biomarkers and bioindicators has been recognized as a reliable tool to assess risk of exposure to biota and also the human population. In order to evaluate the use of fish and biomarkers to assess toxic potential and bioavailability of chemicals in human-related hydric resources, an in situ experiment was accomplished in two water reservoirs designated for human supply, which were previously evaluated by the local environmental regulatory agency through a set of physical, chemical, and classical biological parameters. Molecular, biochemical, and morphological biomarkers were performed in caged Oreochromis niloticus kept for 6 months in the studied reservoirs to assess potentially useful biomarkers to evaluate the quality of water for human supply. Chemical analysis of toxic metals in liver and muscle and polycyclic aromatic hydrocarbons (PAHs) in bile was considered to assess the bioavailability of pollutants and highlight human activity impact. The reservoir previously classified by a governmental agency as less impacted presented more risk of exposure to biota. These results were supported by chemical analysis, vitellogenin expression, histopathological findings (gonads, liver, and gills), as well as indicators of neurotoxic effects and oxidative stress in liver. The inclusion of some biomarkers as parameters in regulatory monitoring programs in reservoirs designated for human supply is strongly suggested to evaluate the risks of exposure to the human population. Thus, a revision of the traditional biological and physicochemical analysis utilized to establish the conditions of water quality is necessary.

Assessing genotoxic effects in fish from a marine protected area influenced by former mining activities and other stressors.

The goal of the current study was to evaluate different genotoxicity tools in order to assess a marine protected area (MPA) affected by former mining activities and urban settlements. A catfish (Cathorops spixii) was analyzed for genotoxic effects at the (i) molecular and at the (ii) chromosomal levels. Through factor analysis, genotoxicity was found to be linked to levels of metals bioaccumulated and PAH metabolites in the bile. Micronucleus and nuclear alteration were less vulnerable to the effects of confounding factors in mildly contaminated areas since they were more frequently associated with bioaccumulated metals than the DNA analysis. The different genotoxicity responses allowed for the identification of sources of pollution in the MPA. This approach was important for detecting environmental risks related to genotoxic contaminants in a mildly contaminated MPA.

Post-treatment of anaerobic reactor effluent using coagulation/oxidation followed by double filtration.

This study evaluates the efficacy of a sanitary sewage treatment system, proposing post-treatment of the effluent generated by the upflow anaerobic sludge blanket UASB reactor, through a Fenton coagulation/oxidation ((ferric chloride (FC) or ferrous sulfate (FS) and peracetic acid (PAA)), followed by a double filtration system, composed of a gravel ascending drainage filter and a sand descending filter. Following the assessment of treatability, the system efficiency was evaluated using physicochemical and microbiological parameters. In all treatments performed in the pilot unit, total suspended solids (TSS) were completely removed, leading to a decrease in turbidity greater than 90% and close to 100% removal of total phosphorous. In the FC and PAA combination, the effluent was oxygenated prior to filtration, enabling a more significant removal of biochemical oxygen demand (BOD), which characterizes aerobic degradation even in a quick sand filter. The treatments carried out in the presence of the PAA oxidizing agent showed a more significant bleaching of the effluent. Concerning the microbiological parameters, the simultaneous use of PAA and FC contributed to the partial inactivation of the assessed microorganisms. A 65% recovery of the effluent was obtained with the proposed treatment system, considering the volume employed in filter backwashing.