Wagner Ezequiel Risso

Multiple biomarker responses in Prochilodus lineatus subjected to short-term in situ exposure to streams from agricultural areas in Southern Brazil.

In order to assess the quality of streams susceptible to contamination by pesticides we apply biochemical and genotoxic biomarkers in the Neotropical fish Prochilodus lineatus submitted to in situ tests. Fish were caged, for 96 h, in two streams located in areas with intensive use of pesticides, the Apertados (AP) and the Jacutinga (JC), and in a small stream (Godoy stream — GD) found inside a forest fragment adjacent to a State Park. Biochemical parameters, such as biotransformation enzymes 7-ethoxyresorufin-O-deethylase (EROD) and glutathione-S-transferase (GST), non-protein thiols (NPSH), lipoperoxidation (LPO), protein carbonylation (PCO) and acetylcholinesterase (AChE) were evaluated in various fish organs, as well as genotoxic biomarkers (damage to DNA and occurrence of micronuclei and erythrocyte nuclear abnormalities). Samples of water and sediment were collected for analysis of metals (Cu, Cr, Pb, Ni, Mn, Cd and Zn), organochloride pesticides, and triazine and glyphosate herbicides. We observed an increase in liver GST activity in fish at AP and gill GST activity in fish at JC. An increase in liver LPO was also observed in fish exposed to AP and JC. The same animals also exhibited increased DNA damage and erythrocyte nuclear abnormalities (ENAs) compared to the fish kept in GD. A number of compounds showed concentrations higher than the permitted levels, in particular, dichlorodiphenyltrichloroethane (DDT), its metabolites dichlorodiphenyldichloroethylene (DDE) and dichlorodiphenyldichloroethane (DDD), hexachlorocyclohexanes (HCH), heptachloride, diclofluanid and aldrins. These pesticides were detected at higher concentrations in water and sediment samples from AP, followed by JC and GD. The Integrated Biomarker Response Index (IBR) indicated that AP and JC (AP: 21.7 > JC: 18.5 > GD: 12.6) have the worst environmental quality. Integrated biomarker analysis revealed that the alterations observed related well with the levels of environmental contaminants, demonstrating the effectiveness of this biomonitoring approach.

Bioaccumulation of nickel and its biochemical and genotoxic effects on juveniles of the neotropical fish Prochilodus lineatus.

Juveniles of the freshwater fish Prochilodus lineatus were exposed to three concentrations of nickel (Ni): 25, 250 and 2500 µg L(-1) or water only for periods of 24 and 96 h to test for Ni bioaccumulation, its effects on antioxidant defenses and metallothioneins, and the occurrence of DNA damage. After exposure, the fish were sampled and tissue removed from the gills, liver, kidney and muscle to test for Ni accumulation and conduct biochemical (gills and liver) and genotoxic (blood cells and gills) analyses. The results showed that Ni accumulates in the organs in different proportions (kidney>liver>gills>muscle) and accumulation varied according to exposure time. Metallothionein (MT) levels increased in the liver and gills after exposure to Ni, implying that the presence of Ni in these tissues could induce MT synthesis. We also observed that Ni exposure affected antioxidant defenses, increasing lipid peroxidation in the liver of fish exposed to Ni for 96 h at the highest concentration tested. DNA damage increased in both blood cells and gills of fish exposed to all Ni concentrations, indicating the genotoxic potential of Ni on fish. We therefore concluded that Ni accumulates in various tissues and results in oxidative and DNA damage in P. lineatus, and that the maximum permitted Ni concentration set in Brazilian legislation (25 µg L(-1)) for freshwaters is not safe for this species.

Lead accumulation and metallothionein content in female rats of different ages and generations after daily intake of Pb-contaminated food.

Female Wistar rats of different ages (45, 90 and 140 days) and generations (mothers and offspring) were fed a feed containing 2.0mg of Pb kg-1 daily from weaning and the Pb accumulation was determined in different organs and in maternal milk, in addition metallothioneins (MTs) content was determined in the liver and kidneys. The results showed that Pb accumulation exhibited the following pattern: bone>liver>kidney>gut>blood cells>muscle>brain>ovary. Bones accumulated the most Pb in all animals, with its concentration increasing with age and prenatal exposure. Pb accumulation in the liver, kidney and blood cells, did not follow a consistent pattern with increasing age and our data did not indicate a relationship between the presence of MTs in liver and kidney and metal accumulation in these organs. However, in the offspring and with increasing age, Pb accumulated in more organs. Mothers fed with Pb produced contaminated milk, exposing their offspring to the metal via nursing Thus, increasing age and prenatal exposure increases susceptibility to Pb toxicity-induced damage.

Metals bioaccumulation and biomarkers responses in the Neotropical freshwater clam Anodontites trapesialis: Implications for monitoring coal mining areas.

As one of the most impactful industries, coal mining can promote several alterations at surrounding environment. In surface water, elevated concentrations of metals like Mn, Zn, Fe and Al are often observed. Thus, the aim of this study was to investigate the bioaccumulation and the sub-lethal effects of these metals on various organs of the Neotropical bivalve Anodontites trapesialis confined along a stream located near a coal mine, in order to assess a set of biomarkers that could be used for effectively monitoring coal mining areas. Clams were caged, for 96h, at two sites located upstream (Up1 and Up2) and two sites downstream (Dw1 and Dw2) from the mine. Metals bioaccumulation was determined in gills, mantle, digestive gland, muscle and hemolymph and the following biomarkers were measured in A. trapesialis tissues: total antioxidant capacity against peroxyl radicals, metallothionein content, lipid peroxidation (LPO), proteins carbonylation, glutathione S-transferase activity, superoxide dismutase activity and acetylcholinesterase (AChE) activity. The results showed that Al and Fe bioaccumulation in the gills and hemolymph, Al bioaccumulation in the mantle and muscle, increased LPO in the gills (Dw1 and Dw2) and mantle (Dw1), as well as reduced AChE activity in the muscle (Dw1 and Dw2) should be considered effective biomarkers for monitoring coal mining areas. A. trapesialis proved to be an efficient biological model, considering that biomarkers responses were observed in the clams after only 96h of confinement at Dw sites, accordingly this species could be a good candidate for monitoring Neotropical freshwaters.

Triple‐mixture of Zn, Mn, and Fe increases bioaccumulation and causes oxidative stress in freshwater neotropical fish

Metal bioaccumulation and oxidative stress biomarkers were determined in Prochilodus lineatus to understand the effects of short-term exposure to a triple-mixture of Zn, Mn, and Fe. Three independent tests were carried out, in which fish were exposed to 3 concentrations of Zn (0.18, 1.0, and 5.0 mg L-1 ), Mn (0.1, 0.5, and 5.0 mg L-1 ), and in the mix test to Fe (5.0 mg L-1 ) and a mixture of Zn (1.0 mg L-1 ) + Mn (0.5 mg L-1 ), with and without Fe. After exposure for 96 h, tissues were removed for metal bioaccumulation analysis and oxidative stress biomarkers were determined in liver, along with DNA damage in blood cells. Our results revealed that Zn and Mn were bioaccumulated in fish tissues after exposure to 5.0 mg L-1 , whereas Fe only bioaccumulated in muscle and gills after mixture exposure. Results indicated that 1 metal interfered with the others bioaccumulation. In P. lineatus, 5 mg L-1 of both Mn and Fe were toxic, because damage was observed (lipid peroxidation [LPO] in liver and DNA damage in blood cells), whereas Zn induced liver responses (metallothionein [MT] and reduced glutathione [GSH] increases) to prevent damage. In terms of bioaccumulation and alterations of oxidative stress biomarkers, we showed that Zn, Mn, and Fe triple-mixture enhances individual metal toxicity in Neotropical fish P. lineatus. Environ Toxicol Chem 2018;37:1749-1756.

Chromium accumulation and biomarker responses in the Neotropical fish Prochilodus lineatus caged in a river under the influence of tannery activities.

In order to assess the effects of tannery effluents (TE) in organism health, juveniles of Prochilodus lineatus were submitted to in situ tests at four different river locations: site A - upstream of the tannery; site B - next to the tannery; and sites C and D - downstream of the tannery. After 96 h exposure in the river, samples of fish tissue, river water and sediment were collected in order to quantify chromium (Cr) concentrations. Tissue samples were used to assess the activity of ethoxyresorufin-O-deethylase (EROD) and glutathione S-transferase (GST), the content of glutathione (GSH) and metallothionein (MT) and the occurrence of lipid peroxidation (LPO) and DNA damage. Higher Cr concentrations were detected in the water and sediments from site B and in the liver of fish confined at site B, compared to the other sites. Fish caged at site B demonstrated higher levels of liver MT and hepatic EROD activity in relation to fish caged at the other sites. Moreover, fish from site B presented increased liver and branchial GST activities, as well as more GSH in the liver, than fish from site A. There were no significant variations in the occurrence of LPO and DNA damage among fish caged at the different sites. Thus, TE increased Cr levels in the water, sediments, and fish livers and stimulated the synthesis of MT and GSH and the activities of EROD and GST. In conclusion, TE affect the quality of the river and promote changes in biochemical biomarkers and Cr accumulation in P. lineatus.

Single and combined effects of Zn, Mn and Fe on the Neotropical freshwater bivalve Anodontites trapesialis: Bioaccumulation and biochemical biomarkers

Important concentrations of Zn, Mn and Fe were detected in a stream near a coal mining area and promoted, in field, biomarkers alterations in the bivalve Anodontites trapesialis. In order to understand the isolated and mixed effects of these metals on these Neotropical bivalves, we run short-term experiments under laboratory controlled conditions. After 96 h-exposure, tissues (gills, mantle, digestive gland, muscle, hemolymph) were removed for metal bioaccumulation analysis, oxidative stress biomarkers (reactive oxygen species (ROS), total antioxidant capacity, lipoperoxidation (LPO), proteins carbonylation (PC), metallothionein (MT), activity of superoxide dismutase and glutathione S-transferase and hemocytes DNA damage) and cholinesterase (ChE versus ASCh activity) activity evaluation. We run three independent tests. In Zn test, clams were exposed to three concentrations of Zn (0.18 mg L-1, 1.0 mg L-1, 5.0 mg L-1); in Mn test, clams were exposed to three concentrations of Mn (0.1 mg L-1, 0.5 mg L-1, 5.0 mg L-1) and in Mix test, clams were exposed to the mixture Zn (1 mg L-1) + Mn (0.5 mg L-1), with and without Fe (5.0 mg L-1). After single exposure to 5.0 mg L-1, Zn bioaccumulated in all tissues, but only in mantle and hemolymph after exposure to 1.0 mg L-1. The increased MT in gills of A. trapesialis exposed to Zn appears to be sufficient to avoid damage, since LPO occurred only in digestive glands from animals exposed to 5.0 mg L-1. We suggested that A. trapesialis had a metabolic suppression in consequence of Mn presence, based on the following results: the decrease of ROS in gills, the decrease of the Zn and Mn concentrations in tissues and the decrease of ChE versus ASCh activity in muscle. Despite this, animals exposed to Mn suffer oxidative damages (LPO and PC) in the mantle and digestive gland and MT increased in the mantle. These results showed A. trapesialis responded differently to each metal and Mn caused more damage. When exposed to Fe, gills level of ROS was increased, despite no changes in metal accumulation occurred. On the other hand, after exposure to the mixtures, tissues bioaccumulated Zn and previously observed damages caused by Mn and Fe disappeared. Consequently, biomarkers were less affected under mixture treatments, demonstrating mixtures effects or responses were not simply a combination of single exposures to Zn, Mn and Fe, but depend on metals toxicokinetics.

Influence of plant densities and fertilization on maize grains by near-infrared spectroscopy

ABSTRACT Near-infrared spectroscopy is a rapid process analytical technology for measuring a wide range of constituents of biological materials. The objective of the current study was to investigate maize grains grown under different levels of fertilization and plant densities using near-infrared spectroscopy. Maize grains from the same plant density had their protein content increased with doses of nitrogen. In addition, it was observed the decrease in protein content with the increase in plant densities. Except for ash content, models for the assessed parameters indicated that near-infrared spectra has the potential for fast assessment of maize grains.

Safety and Quality in the Agricultural Product Chain in Brazil

An agriculture-intensive country should be aware of natural toxins, including both mycotoxins and cyanotoxins, which are closely associated with the quality of raw materials, for food safety and industry. The major production chains – corn, wheat, beef, and broiler chicken – are the top components of agribusiness, and they should be tracked by reliable and practical tools. The corn chain is of particular concern in food production; intensive controls, multi-year mycotoxin monitoring, and improved harmless/sustainable management methods for uninterrupted farming in the tropicsubtropics are needed to achieve a long-lasting trend. The rapid control of natural toxins (mycotoxin and cyanotoxin) has focused on immunochemical methods developed with highly specific monoclonal antibodies (mAb) matched with chroma‐ tographic methods. In parallel, the promising widespread application of nondestructive analytical methods based on NIR (Near Infrared Reflectance)