Antimicrobial Potential of Chitosan Films Incorporated with Alcoholic Extract from Mimosa tenuiflora Leaves.

Antimicrobial films were prepared with chitosan containing the methanolic extract of M. tenuiflora leaves (FECT20%, FECT30%, and FECT40%), and their antimicrobial activities were evaluated by agar diffusion. The films were characterized by IR spectroscopy, scanning electron microscopy (SEM) and TG/DTG curves. TG/DTG curves showed thermal stability of chitosan-extract films up to 166 ºC. Micrographs of chitosan-extract films revealed an increase in porosity with the addition of extract. The FECT40% film showed inhibition zone diameters (IZ) against Micrococcus luteus, Staphylococcus aureus, Bacillus subtilis, and B. cereus, ranging from 1.0 ± 0.02 to 0.72 ± 0.09 cm. Only FECT30% and FECT40% inhibited the P. aeruginosa with IZs of 0.68 ± 0.02 and 0.77 ± 0.06 cm, respectively. In turn, the extract showed inhibition against B. subtilis and B. cereus, with IZs values of 0.92 ± 0.2 cm and 0.72 ± 0.05 cm, respectively. Additionally, the crude extract presented antioxidant potential with inhibition percentages of 32.74% ± 0.90 for ABTS and 27.04% ± 1.36 for DPPH. The antimicrobial and antioxidant activities of the crude extract, as well as the antimicrobial property of chitosan-extract films, suggests the potential of these biopolymers for the development of wound healing bandages and new food packaging alternatives.

Assessment of the photoprotective potential and structural characterization of secondary metabolites of Antarctic fungus Arthrinium sp.

Interest in Antarctic fungi has grown due to their resilience in harsh environments, suggesting the presence of valuable compounds from its organisms, such as those presenting photoprotective potential, since this environment suffers the most dangerous UV exposure in the world. Therefore, this research aimed to assess the photoprotective potential of compounds from sustainable marine sources, specifically seaweed-derived fungi from Antarctic continent. These studies led to discovery of photoprotective and antioxidant properties of metabolites from Arthrinium sp., an endophytic fungus from Antarctic brown algae Phaeurus antarcticus. From crude extract, fractions A-I were obtained and compounds 1-6 isolated from E and F fractions, namely 3-Hydroxybenzyl alcohol (1), (-)-orthosporin (2), norlichexanthone (3), anomalin B (4), anomalin A (5), and agonodepside B (6). Compounds 1, 2, and 6 were not previously reported in Arthrinium. Fraction F demonstrated excellent absorbance in both UVA and UVB regions, while compound 6 exhibited lower UVB absorbance, possibly due to synergistic effects. Fraction F and compound 6 displayed photostability and were non-phototoxic to HaCaT cells. They also exhibited antioxidant activity by reducing intracellular ROS production induced by UVA in keratinocyte monolayers and reconstructed human skin models (resulting in 34.6% and 30.2% fluorescence reduction) and did not show irritation potential in HET-CAM assay. Thus, both are promising candidates for use in sunscreens. It is noted that Fraction F does not require further purification, making it advantageous, although clinical studies are necessary to confirm its potential applicability for sunscreen formulations.

Annotation of GC-MS Data of Antimicrobial Constituents in the Antarctic Seaweed Phaeurus antarcticus by Molecular Networking.

Phaeurus antarcticus is a member of the Desmarestiaceae family endemic to the Antarctic Peninsula. Reports addressing its chemical composition and biological activities are scarce. Herein, bioactive non-polar compounds of P. antarcticus against pathogenic bacteria, Leishmania amazonensis and Neospora caninum parasites were targeted through GC-MS Molecular Networking and multivariate analysis (OPLS-DA). The effects on horseradish peroxidase (HRP) were also evaluated. P. antarcticus exhibited selective bacteriostatic and bactericidal activities against Staphylococcus aureus with MIC and MBC values from 6.25-100 µg mL-1 . Fractions HX-FC and HX-FD were the most active against L. amazonensis with EC50 ranging from 18.5-62.3 µg mL-1 . Additionally, fractions HX-FC and HX-FD showed potent inhibition of N. caninum at EC50 values of 2.8 and 6.3 µg mL-1 , respectively. All fractions inhibited HRP activity, indicating possible interactions with Heme proteins. It was possible to annotate compounds from tree mains clusters, containing terpenoids, steroids, fatty acids, and alcohols by correlating the spectral data of the GC-MS analysis with Molecular Networking and the OPLS-DA results.

Seaweeds and Corals from the Brazilian Coast: Review on Biotechnological Potential and Environmental Aspects.

Brazil has a megadiversity that includes marine species that are distributed along 800 km of shoreline. This biodiversity status holds promising biotechnological potential. Marine organisms are important sources of novel chemical species, with applications in the pharmaceutical, cosmetic, chemical, and nutraceutical fields. However, ecological pressures derived from anthropogenic actions, including the bioaccumulation of potentially toxic elements and microplastics, impact promising species. This review describes the current status of the biotechnological and environmental aspects of seaweeds and corals from the Brazilian coast, including publications from the last 5 years (from January 2018 to December 2022). The search was conducted in the main public databases (PubChem, PubMed, Science Direct, and Google Scholar) and in the Espacenet database (European Patent Office-EPO) and the Brazilian National Property Institute (INPI). Bioprospecting studies were reported for seventy-one seaweed species and fifteen corals, but few targeted the isolation of compounds. The antioxidant potential was the most investigated biological activity. Despite being potential sources of macro- and microelements, there is a literature gap regarding the presence of potentially toxic elements and other emergent contaminants, such as microplastics, in seaweeds and corals from the Brazilian coast.

Biological, biochemical and genotoxic effects of Sb in the midge Chironomus sancticaroli Strixino and Strixino, 1981 (Diptera: Chironomidae).

In aquatic systems, antimony (Sb) is found in the water column and associated with sediment particles being bioavailable to organisms. Consequently, toxic effects have been detected in benthic invertebrates, but the toxicity after Sb exposure in Chironomidae have not been investigated. Were investigated DNA damage, activities of cholinesterase (ChE), alpha and beta esterase (EST-α, EST-ß), glutathione S-transferase (GST), catalase (CAT), and superoxide dismutase (SOD) and lipid peroxidation after acute (48 h) and subchronic exposure (8 d). We also investigated the effects of subchronic (8 d) on development of larvae and chronic (25 d) Sb exposure on emergence and size of adults of Chironomus sancticaroli. Were analyze Sb nominal concentrations ranged from 0.5 to 800 µg.L-1. Genotoxic effects occurred at higher concentrations upon acute (50, 800 µg.L-1) and subchronic exposure (50 µg.L-1). Acute exposure increased ChE, EST-α, EST-ß, and GST activities. Subchronic Sb exposure increased EST-α activity at 0.2 µg.L-1 and GST activity at 5 µg.L-1. CAT activity increased at all concentrations while increasing lipid peroxidation levels were observed (1 µg.L-1, 5 µg.L-1 and 50 µg.L-1), indicating oxidative stress. All concentrations of Sb delayed larval development and decreased the number of emerging adults. At high concentrations (50, 500, 800 µg.L-1), the emerging adults were smaller. In conclusion, these varying genotoxic, biochemical and biological effects of Sb make a notable impact on the reproduction and population dynamics of C. sancticaroli.

Sublethal effects of microcystin-LR in the exposure and depuration time in a neotropical fish: Multibiomarker approach.

Eutrophication is an ecological process that results in cyanobacterial blooms. Microcystin-LR is the most toxic variant of microcystins and may cause toxic effects in the organisms, mainly in hepatic tissues. The aims of this study were to use multiple biomarkers in order to evaluate the sublethal effects of a low concentration of MC-LR (1 µg/L) in fish Geophagus brasiliensis by waterborne exposure; and evaluate the depuration of this toxin during 15 days. A group of 30 fish was exposed to 1 µg/L of MC-LR solution for 96 h in a static bioassay. After this time, blood, brain, muscle, liver, gonad and gills were collected from half of the exposed fish group in order to evaluate chemical, biochemical, histological and genotoxic biomarkers. The rest of the fish group was submitted to the depuration experiment with free MC-LR water for 15 days. After this time the same tissues were collected and evaluated using biomarkers analysis. Toxic effects were found mostly in the fish liver from depuration time as alterations on the antioxidant system and histopathologies. The results showed that even low concentrations can cause sublethal effects to aquatic organisms, and cyanotoxins monitoring and regulation tools are required.

Progress in the environmental risk assessment of plant protection products in Brazil: An overview of birds and mammals and soil organisms proposals.

Since 2019, the Brazilian Institute of Environment and Renewable Natural Resources (IBAMA) has actively developed pesticide environmental risk assessment (ERA) frameworks adapted to Brazil's specific ecological contexts. This initiative, supported by funding from the Brazilian Ministry of Justice and in partnership with academic institutions, has led to a concerted effort to establish ERA protocols for various taxa, including birds and mammals, soil organisms, aquatic organisms, and reptiles and amphibians. The outcomes of this initiative were disseminated in two distinct workshops held in February and November of 2023, where the agency showcased its research to the technical-regulatory community. This article synthesizes the proposals for birds and mammals and soil organisms. First, we summarize the agency's proposals for both focal and generic species to be incorporated into the ERA and the methodologies for calculating exposure of these taxa to pesticides through agricultural practices, encompassing seed treatment and foliar applications. On this occasion, IBAMA also disclosed the risk assessment tool that the agency is developing for birds and mammals. IBAMA highlighted the knowledge gaps that must be bridged to progress from preliminary (lower-tier) to more comprehensive (higher-tier) assessments. Regarding soil organisms, during the workshop, the presenters shared findings on the most prevalent species of earthworms and enchytraeids in Brazil. They emphasized the need for additional data collection on a regional scale. The agency has also proposed methods for estimating soil organism exposure to pesticides at a screening level and identified specific data gaps that could be addressed to refine assessments at higher tiers. In summary, the workshop communicated the progress in establishing ERA guidelines, which we encapsulate here to benefit the technical-regulatory community. © 2024 The Author(s). Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Progress in the environmental risk assessment of plant protection products in Brazil: An overview of aquatic organism proposals.

Since 2019, the Brazilian Institute of Environment and Renewable Natural Resources (IBAMA) has actively developed pesticide environmental risk assessment (ERA) frameworks adapted to Brazil's specific ecological contexts. This endeavor, supported by funding from the Brazilian Ministry of Justice and in partnership with academic institutions, has led to a concerted effort to establish ERA protocols for various taxa, including birds and mammals, soil organisms, aquatic organisms, and reptiles and amphibians. The outcomes of this initiative were conveyed in two workshops held in February and November 2023, during which the agency communicated its findings to the technical-regulatory community. This article represents one of three articles that provide more detailed insights into the ERA propositions for all taxa. In this article, we summarize the proposals for aquatic organisms presented and discussed during the workshops, which can be used as an informational source by the technical-regulatory community. Integr Environ Assess Manag 2024;00:1-6. © 2024 The Author(s). Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Proposal for a tiered regulatory framework for the aquatic risk assessment of pesticides in Brazil.

Aquatic risk assessment is essential to guarantee the sustainable use of pesticides and the conservation of water resources near agricultural fields. This article discusses a proposal for a tiered regulatory framework for the aquatic risk assessment of pesticides in Brazil. The first step is problem formulation, which includes establishing general and specific protection goals. In the exposure assessment, the Estimated Environmental Concentrations in water should be calculated based on realistic worst-case assumptions regarding application rate and frequency, the entry into the edge-of-field water body, and fate in the water body, using scenario-dependent models suggested by the Brazilian Environmental Agency. These calculations can be refined by including Efate studies with variable exposures to reflect realistic environmental conditions accurately and include mitigation measures that impact the modeling. In the hazard assessment, ecotoxicological data for toxicity to fish, aquatic invertebrates, algae, and aquatic plants should be required for all pesticides based on standardized protocols and species. Tier 2 has several refinement options, including incorporating toxicity data from additional test species and effect modeling. In Tier 3, population- and community-level effects are evaluated using semi-field studies. Considering the case study in Brazil, Tier 1 demonstrated that, from the 12 pesticides that were assessed, seven (58%) failed based on the value of the Risk Quotient. In Tier 2, when exposure refinement options and mitigation measures such as buffer zones are considered, all seven pesticides, for which Tier 1 indicated risk, still failed the assessment. The risk for four of these seven pesticides could be refined by considering toxicity information from additional species. Refinement options and mitigation measures that could be applied to the agricultural scenario in Brazil were discussed. In conclusion, the proposed tiered risk assessment is a feasible way to evaluate whether a pesticide will pose an unacceptable risk to aquatic organisms. Integr Environ Assess Manag 2024;00:1-15. © 2024 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Use of dry bean fields by birds and mammals in Brazil: Insights from a field study and its use in pesticide risk assessment.

The relationship between agriculture and wildlife can be both synergetic and challenging, as the increased surface of agricultural land makes it increasingly important for agriculture and wildlife to coexist. This study aims to describe the use of freshly drilled dry bean fields by birds and mammals in Brazilian Cerrado and Atlantic Forest sites and their diversity and abundance within in-crop and off-crop areas (with native permanent vegetation and other surrounding crop fields) at four different farms. A comprehensive survey was conducted, using various sampling methods, including point counts, foraging counts, trail cameras, and occasional encounters. In all, contacts for 12 518 birds across 306 species and 313 mammals across 34 species were registered. The off-crop areas exhibited greater species richness, abundance, and diversity than the in-crop areas on all farms. For birds, 47 species were recorded in-crop, of which 15 were classified as insectivores, 15 as granivores, seven as omnivores, seven as carnivores, and three as frugivores. The number of in-crop observations per species was small. The abundance off-crop was greater for 31 species observed in-crop, indicating that dry bean fields are probably not a preferred habitat for those species. Species classified as granivorous are most likely to feed on dry bean seeds. However, almost all granivorous species observed in-crop areas are too small to be able to feed on dry bean seeds. For mammals, nine species were recorded in-crop, of which four were classified as carnivores, three as omnivores, one as insectivore, and one as granivore. Additionally, despite the considerable effort in this study, no evidence was found that birds and mammals feed on dry bean seeds. The results highlight the importance of off-crop areas in dry bean fields, characterized by a more diverse and abundant bird community than in-crop. Integr Environ Assess Manag 2024;20:864-874. © 2023 The Authors. Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Trace metals in estuarine organisms from a port region in southern Brazil: consumption risk to the local population.

Metal contamination is a threat to estuarine environments. They can accumulate in the food chain and cause toxic effects on aquatic organisms and human health. This study evaluated the concentrations of metals (Cd, Cr, Cu, Fe, Ni, Pb, and Zn) in aquatic organisms of Antonina Bay (southern Brazil) to analyze whether the metal concentrations were in accordance with Brazilian food legislation and estimate the human risk of local seafood consumption. All analyzed organisms (Centropomus parallelus, Mugil curema, Genidens genidens, Crassostrea brasiliana, and Xiphopenaeus kroyeri) showed traces of metals in their tissues with different metal concentrations among species. Metal concentrations were generally higher in oyster C. brasiliana, and biomagnification was not observed. Cr and Zn concentrations were above the limits established by legislation for all species in at least one sample. The concentrations of the other metals were within permitted levels. However, concentrations of Cd, Cr, Fe, and Zn posed a human consumption risk. In general, the C. brasiliana oyster presented the highest risk for human consumption, probably due to its filtering habit. Thus, the results indicated that metal concentrations in the tissues of the Antonina Bay seafood can pose a risk to human health, and this chronic exposure to metals also can cause toxic effects on local aquatic biota.

Estimating pesticide environmental concentrations in Latin America: The importance of developing local scenarios.

Data to assess pesticide exposure in soil and water are scarce and unevenly distributed in Latin America, especially due to the size of the region and the vast agricultural landscape. This makes it difficult to assess associated environmental risks. We suggest that the lack of pesticide exposure or monitoring data can be addressed by using validated models to provide estimated pesticide exposure concentrations in soil and water bodies. This exposure modeling approach has been used by regulatory agencies in other countries and regions such as the United States, Canada, and the European Union. In order to properly estimate pesticide exposure concentrations, we advocate for the development of local scenarios containing local weather, soil, and crop data to be used in the existing models. A sensitivity analysis of the models can be performed to determine parameters that are sensitive and therefore inputs to these parameters are derived locally. We believe the development of local scenarios in the region is attainable and can be a pragmatic approach for developing a more comprehensive picture of potential pesticide exposure in the region. Integr Environ Assess Manag 2021;17:901-904. © 2021 Syngenta Proteção de Cultivos Ltda.

In Vitro Evaluation of the Photoprotective Potential of Quinolinic Alkaloids Isolated from the Antarctic Marine Fungus Penicillium echinulatum for Topical Use.

Marine-derived fungi proved to be a rich source of biologically active compounds. The genus Penicillium has been extensively studied regarding their secondary metabolites and biological applications. However, the photoprotective effects of these metabolites remain underexplored. Herein, the photoprotective potential of Penicillium echinulatum, an Antarctic alga-associated fungus, was assessed by UV absorption, photostability study, and protection from UVA-induced ROS generation assay on human immortalized keratinocytes (HaCaT) and reconstructed human skin (RHS). The photosafety was evaluated by the photoreactivity (OECD TG 495) and phototoxicity assays, performed by 3T3 neutral red uptake (3T3 NRU PT, OECD TG 432) and by the RHS model. Through a bio-guided purification approach, four known alkaloids, (-)-cyclopenin (1), dehydrocyclopeptine (2), viridicatin (3), and viridicatol (4), were isolated. Compounds 3 and 4 presented absorption in UVB and UVA-II regions and were considered photostable after UVA irradiation. Despite compounds 3 and 4 showed phototoxic potential in 3T3 NRU PT, no phototoxicity was observed in the RHS model (reduction of cell viability < 30%), which indicates their very low acute photoirritation and high photosafety potential in humans. Viridicatin was considered weakly photoreactive, while viridicatol showed no photoreactivity; both compounds inhibited UVA-induced ROS generation in HaCaT cells, although viridicatol was not able to protect the RHS model against UVA-induced ROS production. Thus, the results highlighted the photoprotective and antioxidant potential of metabolites produced by P. echinulatum which can be considered a new class of molecules for photoprotection, since their photosafety and non-cytotoxicity were predicted using recommended in vitro methods for topical use.

Genotoxic, metabolic, and biological responses of Chironomus sancticaroli Strixino & Strixino, 1981 (Diptera: Chironomidae) after exposure to BBP.

Benzyl butyl phthalate (BBP), which is widely used in industrial production, reaches the aquatic environment, mainly owing to improper disposal of plastic products. In the water, it remains adsorbed to sedimentary particles causing toxic effects in aquatic invertebrates such as Chironomidae, which are important in maintaining ecosystem dynamics and are an important link in the food chain. However, the effects of BBP on Chironomidae are still poorly known. Thus, the toxic effects of BBP on Chironomus sancticaroli at acute (48 h), subchronic (8 d), and chronic (25 d) exposures of concentrations between 0.1 and 2000 µg·L-1 were determined. Genotoxicity effects, changes in the oxidative stress pathway, and development and emergence of organisms were evaluated. Biochemical markers showed a reduction in cholinesterase (ChE) activity, indicating a neurotoxic effect on acute exposure (1-1000 µg·L-1). The antioxidant pathway, glutathione S-transferase (GST) activity showed reduction on acute (0.1; 1-2000 µg·L-1) and subchronic (1-2000 µg·L-1) exposures and reduction in superoxide dismutase (SOD) activity at all evaluated concentrations, suggesting oxidative stress. In contrast, lipid peroxidation was not observed. DNA damage occurred on acute (10 µg·L-1) and subchronic (10-2000 µg·L-1) exposures, indicating genotoxic effects. At concentrations above 10 µg·L-1, no emergence of adults occurred, while lower concentrations (0.1 and 1 µg·L-1) showed a reduction in the number of adults, mainly males. The observed effects indicate that BBP is genotoxic and causes biochemical alterations presenting high toxicity at the population level.

Multiple biomarkers response in a Neotropical fish exposed to paralytic shellfish toxins (PSTs).

The Alagados Reservoir (Southern Brazil) is used as water supply, and since 2002 there have been reports with a presence of cyanobacterial blooms and cyanotoxins. In order to assess the water quality and the ecological integrity of the reservoir, we evaluated biochemical, genotoxic and osmoregulatory biomarkers in the freshwater cichlid fish (Geophagus brasiliensis) that were exposed to PSTs. The fish were sampled in the Alagados Reservoir in February 2016 (Summer) and were divided in three groups: 1) Reservoir group (RES): fish were collected immediately after sampling; 2) Depuration group (DEP): fish were submitted to the depuration experiment for 90 days in the laboratory; and 3) Reproduction group (REP): fish were kept in the laboratory until the fertilization and the chemical analyses were performed on the offspring (F1 generation). In the RES and DEP the blood, brain, muscle, liver and gills were collected for biochemical, genotoxic and osmoregulatory biomarkers analysis. Our results showed that the fish from the Alagados Reservoir (RES) presented oxidative stress and DNA damage; and after 90 days (DEP), the antioxidant system and DNA damage were recovered. Although PSTs were considered a risk to the ecological integrity of this water body; PSTs concentrations were not found in the tissues of the F1 generation. In addition, the biomarkers used were useful tools to evaluate the effects of environment contamination. Therefore, it is necessary to develop new technologies and monitoring programs in order to reduce cyanobaterial blooms, cyanotoxins and human activities that cause the contamination in aquatic environments.

Antitumor Potential of Seaweed Derived-Endophytic Fungi.

The marine environment presents a high biodiversity and a valuable source of bioactive compounds with therapeutic and biotechnological potential. Among the organisms present in marine environment, the endophytic fungi isolated from seaweed stand out. These microorganisms have aroused interest in the scientific community regarding its various activities such as antiviral, antimicrobial, antioxidant, photoprotective, cytotoxic, genotoxic, anti-inflammatory, and anticancer, besides establishing important ecological relations with its hosts. Anticancer molecules derived from marine natural sources are a promising target against different types of cancer. The disease's high rates of morbidity and mortality affect millions of people world wild and the search for new therapeutic alternatives is needed. Thus, this review partially summarizes the methodologies for the isolation of seaweed-derived endophytic fungi, as well as describes the anticancer compounds isolated from such microorganisms, reported in the literature from 2009 to the present. In addition, it describes how some biotechnological processes can help in the discovery of bioactive compounds, especially with anticancer activity.

The accumulation dynamics, elimination and risk assessment of paralytic shellfish toxins in fish from a water supply reservoir.

Paralytic shellfish Toxins (PSTs) or saxitoxins are neurotoxins that block the neural transmission by binding to the voltage-gated sodium channels in the nerve cells. There are >50 analogues described, which could be biotransformed into a molecular form of greater or lesser toxicity. The Alagados Reservoir is used for water supply, and persistent cyanobacterial blooms as well as PSTs concentrations have been found in this water body since 2002. The aims of this study were to quantify the concentrations of PSTs in the water and fish samples from the Alagados Reservoir. In addition, we evaluated the elimination of PSTs for 90 days in fish and estimated the potential risk to human health. Water and fish samples were collected from the reservoir. For the water samples the phytoplankton and chemical analyses were carried out. Fish were divided into two sample times: Field Samples (FS) and Elimination Experiment Samples (EES), which were maintained for 90 days in filtered and dechlorinated water. For chemical analysis, the muscles of FS were collected on the fish sampling day and the muscles and feces of EES were collected at 7, 15, 30, 45, 60, 75 and 90 days. PSTs concentrations were present in water and fish samples, and they were estimated as a potential risk to humans; mainly for children. In addition, toxins were accumulated, biotransformed to other analogues and excreted by the fish. However, after 90 days, the toxins were still present in the water and fish muscle. Therefore, PSTs can remain for a long period in water, and fish can be a carrier of these neurotoxins. New approaches of monitoring and management are necessary in the actual global context of cyanobacteria and cyanotoxins.

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.

Depuration time and sublethal effects of microcystins in a freshwater fish from water supply reservoir.

Microcystins (MCs) are hepatotoxins that have been considered to be a worldwide problem due the effects that they can cause to environmental and human health systems. The Iraí Reservoir, located in the South of Brazil, is used as a water supply and MCs concentrations have been reported in this ecosystem. This study aimed to determine the MCs concentrations in the Iraí Reservoir and to evaluate the MCs depuration time and the health of Geophagus brasiliensis using biomarkers. Water and fish samples were collected in the Iraí Reservoir from August 2015 to May 2016. Phytoplankton and chemical analyses were conducted using water samples and the fish were divided into two groups; the Immediate Group (IMM) and the Depuration Group (DEP). In the IMM group, the blood, liver, muscle, brain and gills were collected, in order to evaluate the genotoxic, biochemical and chemical biomarkers. The DEP group was used in the depuration experiment for 90 days, and after this period the fish were submitted to the same procedure as the IMM group. Our results suggested that fish accumulated MCs and it may have caused oxidative stress, neurotoxicity and molecular damage. Furthermore, MCs concentrations increased during the depuration time and it resulted in molecular damage over the first 30 days. After 90 days, the recovery of the antioxidant system occurred. The depuration started on the 15th day, however, the toxins were still present in the samples. Therefore, the effects and the persistence of MCs are a risk to environmental systems and human health.

Neurotoxins in a water supply reservoir: An alert to environmental and human health.

Reservoirs are important source of power generation, recreation, and water supply. Nevertheless, human activities have favored the bloom of toxic cyanobacteria in many reservoirs, which has resulted in environmental, social, and economic problems. This study aims to evaluate the water quality of a reservoir in South Brazil through the analysis of cyanobacteria and cyanotoxins PSTs (Paralytic Shellfish Toxins) and biomarkers of environmental contamination in fish. For this purpose, water samples and fish (Geophagus brasiliensis) (Perciformes: Cichlidae) were collected from September 2013 to May 2014. The fish G. brasiliensis were separated in two groups. The first one "site group" was euthanized after the sampling and their weight and length were measured. The blood, brain, muscle and liver were collected for chemical, biochemical and genetics biomarkers analysis. The second group "depuration group" was submitted to depuration experiment for 40 days in clean water. After that, the same procedures as for the first group were carried out. Cylindrospermopsis raciborskii was the dominant cyanobacteria found in the reservoir, and it showed a density above the recommended limit by Brazilian legislation of 20,000 cells/mL. Results showed that the fish accumulate PSTs in the Reservoir and these were not eliminated after 40 days. The biochemical and genotoxic biomarkers showed a significant difference between "site groups" and "depuration groups", which suggests a recovery of the antioxidant system and a reduction of cellular damage after 40 days in clean water. In conjunction with results reported earlier by others, Alagados Reservoir, in South Brazil, appears to have a persistent contamination of cyanotoxins. Moreover, the mixture of contaminants which may be present in the water body can explain the seasonal differences in fish at the sampled points.