Abstract The toxic effects of venlafaxine (VLX) on aquatic organisms have already been verified and therefore are a proven matter of concern. Herein, we evaluated zebrafish embryos/adults after acute exposure to VLX. Embryos/larvae were exposed to different concentrations of VLX (100-1000 mg/L; 1.33 as a dilution factor), to evaluate mortality/developmental changos and to analyze biomarkers (0.002-100 mg/L). For adults, mortality, genotoxicity, and biomarkers were assessed in five different concentrations of VLX (1-100 mg/L). The median lethal concentration (LC50-168h) was 274.1 mg/L for embryos/larvae, and >100 mg/L for adults (LC50-96h). VLX decreased the heart rate frequency and caused premature hatching and lack of equilibrium in embryos/larvae exposed to different concentrations ranging from 100 to 562.5 mg/L. The activity of acetylcholinesterase (AChE) was inhibited in larvae exposed to 1, 25 and 100 mg/L. Glutathione-S-transferase (GST) activity was reduced in both larvae and adults after exposure to different concentrations, mainly at 25 mg/L. For both larvae and adults, lactate dehydrogenase (LDH) activity increased after 100 mg/L of VLX exposure. No DNA damage was observed in peripheral erythrocytes. Exposure to VLX may cause adverse effects on zebrafish in their early and adult life stages, interfering with embryo-larval development, and can induce physiological disturbances in adults.

Abstract The threats posed by insecticide resistance to Aedes aegypti in the context of controlling dengue have led to an urgent search for an environmentally safer alternative chemical with more effective larvicidal properties. Among many molecules tested, 2-methylanthraquinone showed the lowest LC50 for A. aegypti in a previous study and the highest LC50 for zebrafish embryos. Embryos were exposed at concentrations of 1.0, 2.19, 4.78, 10.46, 22.87, 50.0 and 100.0 mg/L, and malformations and mortality were significantly observed only at the highest exposures of 50 and 100 mg/L after 96 h. Micronucleus test and comet assay in zebrafish adults were both negative after exposures at 6.25, 12.5, 25.0, 50.0 and 100.0 mg/L for 96 h. Several biochemical biomarkers were analyzed in adults, and 2-methylanthraquinone did not interfere with acetylcholinesterase activity. The lactate dehydrogenase activity was higher at concentrations of 25 and 100 mg/L. Glutathione-S-Transferase (GST) activities were tested in the gill and body (muscle tail). The gill was more sensitive than body for GST activity after exposure to 2-methylanthraquinone, showing the highest activities, and 2-methylanthraquinone showed low toxicity to a non-target organism.

Resumo Objetivo: O carreamento decorrente das atividades de mineração atinge corpos d'água, causando contaminação da água e acidificação por metais tóxicos. Como conseqüência, a depleção de oxigênio ocorre, com o desaparecimento de certos organismos bênticos, perdas na biomassa do fitoplâncton e do zooplâncton, e mortalidade de peixes. Remediação da água contaminada da mineração envolve o uso de floculantes baseados em monômeros de acrilamida e poliacrilamida e de coagulantes à base de alumínio. Nosso objetivo foi avaliar a toxicidade aquática do floculante WF 424 e do coagulante WF 2000 PCA. Métodos Um polímero aniónico floculante (WF 424) e um complexo polinuclear coagulante de íons hidro-alumínio polimerizados (WF2000 PCA) foram testados quanto à embriotoxicidade para o peixe-zebra (Danio rerio, teste FET - OEDC, 236, 2013). Resultados Após 96 h de exposição ao WF424, não houve letalidade significativa, nem efeitos subletais como atraso na eclosão, alteração do batimento cardíaco e malformação no desenvolvimento embrionário, quando comparados com o grupo controle (p> 0,05). O composto coagulante WF2000 PCA não apresentou efeitos letais durante 96 h de exposição, e EC50 foi considerado acima do nível de concentração mais alto testado. Conclusões Para ambos os compostos, nenhuma malformação, atraso no desenvolvimento embrionário ou embriotoxicidade foram observados.

Abstract Aim: Leaching from mining activities reaches water bodies, causing water contamination and acidification by toxic metals. As a consequence, oxygen depletion occurs, with the disappearance of certain benthic organisms, losses in phytoplankton and zooplankton biomass, and fish mortality. Remediation of contaminated water from mining involves the use of flocculants based on acrylamide monomers and polyacrylamide and of coagulants based on aluminum. Our aim was to assess the aquatic toxicity of the flocculant WF 424 and the coagulant WF 2000 PCA. Methods A flocculant anionic polymer (WF 424) and a coagulant polynuclear complex of polymerized hydro-aluminum ions (WF2000 PCA) were tested for embryotoxicity to zebrafish (Danio rerio, FET test - OEDC, 236, 2013). Results After 96 h of exposure to WF424, there was no significant lethality, nor were there sublethal effects such as hatching delay, heartbeat alteration, and malformation in embryonic development, when compared with the control group (p > 0.05). The coagulant compound WF2000 PCA did not show lethal effects during 96 h of exposure, and EC50 was considered as above the highest concentration-level tested. Conclusions For both compounds, no malformation, embryonic development delay or embryotoxicity were observed.

Abstract Continued exposure to reactive oxygen species and inflammation are the rationale behind aging theories and associated diseases. Scientific evidence corroborates the ethnomedicinal use of the oil of pequi (Caryocar brasiliense Camb.), a typical Brazilian Cerrado fruit, against oxidative damage to biomolecules and inflammation. We aimed to investigate in vivo the antioxidant and anti-inflammatory effects of pequi oil on hemogram and DNA damage in healthy young adult and older middle-aged Swiss mice of both genders. Animals, aged 6-7 and 11-12 months, were orally treated for 15 days with pequi oil at 30 mg/day. Blood samples were used for hemogram and comet assay, and bone marrow for micronucleus test. Female controls of 11-12 months had significantly lower haemoglobin and hematocrit than those of 6-7 months. Treatment with pequi oil improved this state, removing the differences. Pequi oil had no genotoxic or clastogenic effects and significantly increased lymphocytes and decreased neutrophils+monocytes in females of 11-12 months, removing the significant differences observed between controls of 6-7 and 11-12 months. The results suggest that dietary supplementation with pequi oil could protect against anemia, inflammation and oxidative stress related to aging, helping to prevent aging-related chronic degenerative diseases, mainly for females.

Abstract Exercise is a double-edged sword: when practiced in moderation, it increases the expression of antioxidant enzymes, but when practiced strenuously it causes oxidative stress and cell damage. In this context, polymorphisms in the interleukin (IL)-6 gene should be investigated better because they can influence performance, at least in exercise that generates oxidative stress and leads to muscular injuries with consequent inflammation. In this work, we investigated the influence of IL-6 –174 G/C polymorphism on tissue damage and inflammation markers, lipid peroxidation, hemogram and lipid profile of runners before and after ingestion of 400 mg of pequi oil in capsules supplied daily for 14 consecutive days. The IL-6 genotypes were associated with significant differences in lipid peroxidation, with the CC mutant having lower values. There were also significant differences among these genotypes in the response to supplementation with pequi oil, exercise-induced damage and C-reactive protein (CRP) levels. The best protection against damage was observed with the heterozygous genotype. Although the CC genotype showed an increase in CRP levels after supplementation, the lack of a positive correlation between triglycerides and high-sensitivity CRP in this mutant genotype after supplementation indicated a protective effect of pequi. These findings deserve further investigation, particularly with regard to the quantification of circulating IL-6 concentrations.

Abstract Andirobeira is an Amazonian tree, the seeds of which produce a commercially valuable oil that is used in folk medicine and in the cosmetic industry. Andiroba oil contains components with anti-inflammatory, cicatrizing and insect-repellant actions. However, virtually nothing is known of the safety of this oil for humans. The aim of this work was therefore to investigate the hematotoxicity, genotoxicity and mutagenicity of andiroba oil using the comet and micronucleus assays, and to assess its antioxidant properties and lipidome as a means of addressing safety issues. For the experiments, andiroba oil was administered by gavage for 14 consecutive days in nulliparous female Swiss mice randomly distributed in four groups: negative control and three doses of oil (500, 1000 and 2000 mg/kg/day). These doses were chosen based on recommendations of the OECD guideline no. 474 (1997). GC/MS was used to investigate the free fatty acid, cholesterol and triterpene content of andiroba oil in a lipidomic analysis. No clinical or behavioral alterations were observed throughout the period of treatment, and exposure to andiroba oil at the doses and conditions used here did not result in hematotoxic, genotoxic or mutagenic effects. Tests in vitro showed that oil sample 3 from southwestern of Brazilian Amazon had a high antioxidant capacity that may protect biological systems from oxidative stress, although this activity remains to be demonstrated in vivo.

Os fundamentalismos surgiram no Ocidente a partir de questões religiosas e posteriormente difundiram-se para outras partes do mundo tomando outras conotações, principalmente políticas. As técnicas de manipulação genética difundiram-se pelas universidades, que formam mestres e doutores com os conhecimentos básicos sobre clonagem gênica, que se tornou de domínio público. Todos os insumos para clonagem gênica podem ser adquiridos por meio de catálogos via internet. Podem-se recrutar profissionais fanáticos e com a competência para a manipulação genética de organismos patogênicos, lado perverso da biotecnologia. Os conflitos étnicos, culturais e religiosos estão associados a um cenário de contrastes entre os países ricos e carentes de matéria-prima e aqueles pobres, mas detentores de insumos básicos e energia, e atingem a sua forma mais aguda nos fundamentalismos. Grupos de fanáticos têm pleno acesso a essa biotecnologia. Estariam assim as populações civis vulneráveis aos ataques do bioterrorismo com armas biológicas geneticamente modificadas?

Fundamentalism arose in the West based in religious matters and afterward diffused to other parts of the world with other connotations, especially political. Genetic manipulation techniques spread to universities, which has given masters and doctors the basic knowledge on gene cloning, which has become public domain. All inputs for gene cloning may be obtained through online catalogs. Fanatic professionals may be recruited, with qualification for genetic manipulation of pathogenic organisms, the negative side of biotechnology. Ethnic, cultural and religious conflicts are linked to a series of contrasts between countries that are rich but with a lack of raw materials and the poor countries that possess basic input and energy sources, when it reaches the highest fundamentalist form. Fanatic groups have complete access to this biotechnology. Are civilian populations in vulnerable to bioterrorist attacks involving genetically modified biological weapons?

Los fundamentalismos surgieron en el occidente basado en cuestiones religiosas y posteriormente se difundió para otras partes del mundo, tomando otras connotaciones, sobre todo políticas. Las técnicas de manipulación genética han propagado por las universidades, que forman másteres y doctores con los conocimientos básicos acerca de la clonación génica que se hizo dominio público. Todos los insumos para la clonación génica pueden ser adquiridos a través de catálogos a través de internet. Se pueden reclutar profesionales competentes para la manipulación genética de organismos patogénicos, lado perverso de la biotecnología. Los conflictos étnicos, culturales y religiosos están asociados a un escenario de contrastes entre los países ricos y carentes de materia prima y los países pobres poseedores de insumos básicos y energía, alcanzando su forma más aguda en el fundamentalismo. Grupos de fanáticos tienen total acceso a esa biotecnología. ¿Estarían, de esa forma las poblaciones civiles vulnerables a los ataques del bioterrorismo con armas biológicas genéticamente modificadas?

The frequency of spontaneous micronucleus (MN) formation in fish species needs to be determined to evaluate their usefulness for genotoxic biomonitoring. The definition of a good bioindicator takes into account the current knowledge of its metabolic traits as well as other factors including its feeding behavior and relationship to the environment. In this study, we compared the basal frequencies of micronucleated erythrocytes and nuclear abnormalities (NA) among different species of the fish Order Gymnotiformes (Rhamphichthys marmoratus, Steatogenys elegans, Sternopygus macrurus, Parapteronotus hasemani, Gymnotus mamiraua, Gymnotus arapaima, Brachyhypopomus beebei, Brachyhypopomus n. sp. BENN) sampled in several localities of the Eastern Amazon. A baseline of MN and NA frequency in these fish was determined, enabling the identification of potentially useful species as models for genotoxicity studies. Only one impacted sample collected at a site in the River Caripetuba showed a significant number of NAs, which may be due to the release of wastewater by neighbouring mining industries and by the burnt fuel released by the small boats used by a local community. Our results may provide support for further studies in areas of the Eastern Amazon affected by mining, deforestation and other anthropogenic activities.

Five restriction site polymorphisms in the β-globin gene cluster (HincII-5'ε, HindIII-Gγ, HindIII-ªγ, HincII-'ψβ1 and HincII-3''ψβ1) were analyzed in three populations (n = 114) from Reconcavo Baiano, State of Bahia, Brazil. The groups included two urban populations from the towns of Cachoeira and Maragojipe and one rural Afro-descendant population, known as the "quilombo community", from Cachoeira municipality. The number of haplotypes found in the populations ranged from 10 to 13, which indicated higher diversity than in the parental populations. The haplotypes 2 (+----),3(----+),4(-+--+)and6(-++-+)onthe βA chromosomes were the most common, and two haplotypes, 9 (-++++)and 14 (++--+), were found exclusively in the Maragojipe population. The other haplotypes (1, 5, 9, 11, 12, 13, 14 and 16) had lower frequencies. Restriction site analysis and the derived haplotypes indicated homogeneity among the populations. Thirty-two individuals with hemoglobinopathies (17 sickle cell disease, 12 HbSC disease and 3 HbCC disease) were also analyzed. The haplotype frequencies of these patients differed significantly from those of the general population. In the sickle cell disease subgroup, the predominant haplotypes were BEN (Benin) and CAR (Central African Republic), with frequencies of 52.9% and 32.4%, respectively. The high frequency of the BEN haplotype agreed with the historical origin of the afro-descendant population in the state of Bahia. However, this frequency differed from that of Salvador, the state capital, where the CAR and BEN haplotypes have similar frequencies, probably as a consequence of domestic slave trade and subsequent internal migrations to other regions of Brazil.

Studies of genotoxicity in fish caused by cyanobacterial microcystins can be useful both in determining the sensitivity of native species, as well as comparing exposure routes. The genotoxicity caused by the microcystins LR and LA from a bloom collected in a eutrophic lake, was revealed in the fish Astyanax bimaculatus, a native species from South America. LC50 (72 h) was determined as 242.81 µg L-1 and LD50 (72 h) as 49.19 µg kg-1 bw. There was a significant increase of DNA damage in peripheral erythrocytes, following intraperitoneal injection (ip) with tested concentrations of 24.58 µg kg-1 bw and 36.88 µg kg-1 bw, as well as through body exposure to a concentration of 103.72 µg L-1. Micronucleus (MN) induction was observed after ip injections of 24.58 µg kg-1 bw and 36.88 µg kg-1 bw for 72 h, as well as following body exposure for 72 at 103.72 µg L-1. Thus, both exposure routes resulted in MN induction and DNA damage. Apoptosis-necrosis testing was carried out only by ip injection with concentrations of 24.58 µg -1 bw and 36.88 µg kg-1 bw. Exposure to microcystins at lower concentrations induced more apoptosis than necrosis in peripheral erythrocytes, whereas exposure at higher concentrations gave rise to both conditions. Thus, Astyanax bimaculatus can be considered as a species sensitive to the genotoxic effects caused by microcystins.

Pequi (Caryocar brasiliense Camb.), a typical fruit of Brazilian Cerrado, is well known in regional cookery and used in folk medicine to treat various illnesses. Mass spectrometry and chromatographic methods have identified the organic composition of pequi fruit pulp; however, NMR spectroscopy is used for the first time to characterize the nutritional components of organic and aqueous-ethanolic extracts. This spectroscopic technique determined the triacylglycerols in the pequi organic fraction, which is constituted mainly by oleate and palmitate esters, and detected the carbohydrate mixtures as the major components of aqueous and ethanolic fractions, respectively. In this study, presence of phenolic compounds was only evidenced in the ethanolic fraction.

The distribution of glutathione S-transferase (GST) GSTM1 and GSTT1 null phenotype frequencies in two Brazilian Amerindian tribes, the Munduruku tribe from Missão Cururu village (79 individuals) and the Kayabi tribe (41 individuals), was analyzed by polymerase chain reaction (PCR) amplification. The GST null phenotype frequencies for the Munduruku sample were 0% for GSTM1 and 27% for GSTT1 while for the Kayabi sample the null phenotype frequencies were 27% for GSTM1 and 29% for GSTT1. This is the first report of the absence of the GSTM1 null phenotype in any ethnic group.

An evaluation of the genotoxicity of domestic sewage was carried out in a municipal wastewater treatment plant. The study was carried out using Allium cepa root tip cells through cytological parameters such as aberrant cells in anaphase-telophase and the mitotic index, and the fish species Oreochromis niloticus and Tilapia rendalli, through the index of micronuclei in peripheral erythrocytes. In the Allium test, each of the four stages of the wasterwater treatment routine was analyzed, i.e., crude sewage, primary effluent, secondary effluent, and tertiary effluent. O. niloticus and T. rendalli specimens were maintained in an aquarium continuously receiving the final effluent, in an open system, before being discharged into Lake Paranoá. For micronuclei analysis, blood samples were drawn after 7, 21, 60, 102 and 142 days, respectively, in controlled conditions. The numbers of aberrant cells found in the Allium test did not differ among the four stages tested, nor when compared with the control. At all stages, the most concentrated sample was more toxic than the respective diluted samples, as demonstrated by the decreased mitotic index. Different sampling of the same group of fish revealed no increase in the micronuclei frequency, regardless of the period of exposure. A correlation between long-term exposure and micronuclei induction was not found either.

The frequencies of mitomycin C and cyclophosphamide-induced kidney and gill erythrocyte micronuclei were compared in Tilapia rendalli and Oreochromis niloticus. Cyclophosphamide (CP) and mitomycin C (MMC) were previously used to investigate fish sensitivity to micronuclei induction in kidney erythrocytes. Fish were treated with a single dose of cyclophosphamide (40 mg/kg) or mitomycin C (2 mg/kg). Peripheral blood samples and kidney blood samples were obtained, 1, 2, 3, 5, 7, and 15 days after the intra-abdominal injection of MMC. T. rendalli showed to be more sensitive than O. niloticus to micronuclei production. Both compounds were used at the maximum tolerated dose (MTD), and mitomycin C proved to be more toxic than cyclophosphamide. The frequency of micronucleated erythrocytes observed varied significantly among the treated individuals. The difference between the kidney and gill micronuclei frequencies was not significant. Higher levels of micronucleated erythrocytes were observed in Tilapia rendalli than in O. niloticus. T. rendalli produced erythrocytes with deformed nuclei, which O. niloticus did not.

Este estudo fez uma avaliação da indução de micronúcleos em eritrócitos de sangue periférico de peixes Tilapia rendalli, Oreochromis niloticus e Cyprinus carpio após o tratamento com mitomicina C, ciclofosfamida, 5-fluorouracil e bleomicina. Foram colhidas amostras periódicas de sangue com 2, 7, 14 e 30 dias após o tratamento único. Os tratamentos com citocalasina B tiveram como objetivo analisar as proporções entre células binucleadas/mononucleadas nos diferentes períodos de tratamento, para a comparação com os picos de micronúcleos entre as diferentes espécies, uma vez que o orgão hematopoiético nos peixes é o rim cefálico e o sangue analisado é de origem periférica, o que nos dá uma noção do tempo do ciclo celular dos eritrócitos. A análise das taxas de aumento das freqüências de micronúcleos ao longo do tempo demonstrou que do 2° ao 7°dias elas foram crescentes, decresceram ao 14° dia e mantiveram-se estáveis até ao 30°. Os 4 compostos químicos aumentaram significativamente as freqüências de micronúcleos ao longo de todo o período de tratamento, sendo que a ciclofosfamida e a mitomicina C foram as mais efetivas. A T. rendalli foi a espécie mais sensível a todos os tratamentos, enquanto que C. carpio foi a mais resistente. Entre 2 e 7 dias pós-tratamento observaram-se as maiores induções de micronúcleos. A partir do 14° dia de tratamento as freqüências de micronúcleos tenderam a diminuir e as diferenças entre as espécies não foram significativas.

Fish are often used for screening genotoxicity of water. For such programs, a knowledge of the sensitivity to clastogens, spontaneous micronucleus frequency and cell cycle kinetics of the target tissue is necessary. To investigate the pattern of inter-specific sensitivity to micronucleus induction three species of fish, Tilapia rendalli, Oreochromis niloticus and Cyprinus carpio, were exposed to the clastogens bleomycin (BLM), cyclophosphamide (CP), 5-fluorouracil (5-FU), and mitomycin C (MMC). The binucleate/mononucleate ratio in peripheral erythrocytes exposed to cytochalasin B was also used to evaluate the time-dependent response of micronucleus formation during hematopoesis in the kidney and the micronucleus peak in peripheral erythrocytes. Micronucleus frequencies induced by CP were significantly greater than their respective controls for the three fish species throughout all treatment periods. During the whole evaluation period (30 days) CP was also the most effective clastogen. In general, until the 14th day of evaluation period T. rendalii was the most sensitive species to clastogens. No difference in micronucleus frequencies among species was observed in the 4th evaluation (at the 30th day). A micronucleus peak was observed at the 7th day after treatment. After the 14th day the frequencies were stabilized. The cytochalasin B experiment was carried out to demonstrate that micronuclei induced in the young kidney erythrocyte cells were detected in the circulating blood 2-4 days later.