Assessment of micronucleus frequency in the peripheral blood of female rats in persistent estrus treated with selective estrogen receptor modulators

The aim of this study was to evaluate micronucleus (MN) frequency in polychromatic erythrocytes (PCE) of female rats in persistent estrus (a model developed to mimic polycystic ovary syndrome) treated with selective estrogen receptor modulators (SERMs, tamoxifen, and raloxifene). Forty female Wistar-Hannover rats were divided into four groups of 10 animals each: Group I (normally cycling rats) and Group II (persistent estrus) both received only vehicle, while Group III (persistent estrus) was treated with tamoxifen (250 mu g/animal/day) and Group IV (persistent estrus) was treated with raloxifene (750 mu g/animal/day). Tamoxifen and raloxifene were given by oral gavage beginning on postnatal day 90 and continuing for 30 consecutive days. Peripheral blood samples were collected from tails 1 day following the last exposure. Blood smears were made on glass slides and stained with 10% Giemsa solution. ANOVA and a Tukey post-hoc test were used for data analysis. Mean percentages of MN were 1.82 +/- 0.13, 5.20 +/- 0.24, 3.32 +/- 0.13, and 3.04 +/- 0.12 in Groups I, II, III, and IV, respectively. The results indicate that tamoxifen and raloxifene similarly reduced the formation of MNPCE of female rats in persistent estrus (P < 0.0001 for Groups III and IV vs. Group II), using the dosages and time periods applied in the present study. The data suggest possibly antimutagenic effects of SERMs under high levels of estrogens. The findings also suggest that this is an interesting animal model for studying the genotoxicity of estrogens. Environ. Mol. Mutagen. 2012. (C) 2011 Wiley Periodicals, Inc.

Micronucleus frequency in children exposed to biomass burning in the Brazilian Legal Amazon region: a control case study

Background: The Amazon represents an area of 61% of Brazilian territory and is undergoing major changes resulting from disorderly economic development, especially the advance of agribusiness. Composition of the atmosphere is controlled by several natural and anthropogenic processes, and emission from biomass burning is one with the major impact on human health. The aim of this study was to evaluate genotoxic potential of air pollutants generated by biomass burning through micronucleus assay in exfoliated buccal cells of schoolchildren in the Brazilian Amazon region. Methods: The study was conducted during the dry seasons in two regions of the Brazilian Amazon. The assay was carried out on buccal epithelial cells of 574 schoolchildren between 6-16 years old. Results: The results show a significant difference between micronucleus frequencies in children exposed to biomass burning compared to those in a control area. Conclusions: The present study demonstrated that in situ biomonitoring using a sensitive and low cost assay (buccal micronucleus assay) may be an important tool for monitoring air quality in remote regions. It is difficult to attribute the increase in micronuclei frequency observed in our study to any specific toxic element integrated in the particulate matters. However, the contribution of the present study lies in the evidence that increased exposure to fine particulate matter generates an increased micronuclei frequency in oral epithelial cells of schoolchildren.

Biomonitoring study of seasonal anthropogenic influence at the Itamambuca beach (SP, Brazil)

The present work is focused on the study of a seasonal anthropogenic influence on the beach of Itamambuca (Ubatuba, SP, Brazil) carried out using Atherinella brasiliensis as biomonitor. In total 84 fish were caught between July 2004 and February 2005 in different locations at the beach and inside the Itamambuca river. The fish were pooled according to catch and their musculature was analyzed by Particle-Induced X-ray Emission (PIXE) and Rutherford Backscattering (RBS) techniques. While the concentration of light (matrix) elements like C and O were obtained using the RBS technique, major (Na, Mg, P, S, Cl, K and Ca) and trace (Si, Al, Ti, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Br and Sr) elements were measured by PIXE. The results show that differences were observed for several elemental concentrations of fish tissue between high season (spring-summer) and low season (winter-fall), indicating that increased human activity in the beach during high season may have some impact on the beach ecosystem. The role of the water salinity in the results is also discussed. (C) 2009 Elsevier B.V. All rights reserved.

LIVER DAMAGES AND NUCLEAR ABNORMALITIES IN ERYTHROCYTES OF Atherinella brasiliensis (ACTYNOPTERIGII, ATHERINOPSIDADE) FROM TWO BEACHES IN SOUTHEAST OF BRAZIL

There are few environmental studies using biomarkers for the species Atherinella brasiliensis in Brazil. In the present work, the presence of hepatic histopathological lesions and nuclear abnormalities in erythrocytes were investigated in A. brasiliensis from Lamberto, a beach under influence of domestic wastes and marine activities. For comparison, fish were also sampled in Puruba, a non-polluted beach, located in the northeastern of Sao Paulo State. The frequency of lesions found in liver was in higher numbers in individuals from Lamberto than Puruba beach. The most critical injuries observed in A. brasiliensis were the presence of necrotic areas, leucocytes infiltration and piknotic nucleus. A high occurrence of cells with vacuolization was also observed. The hepatic lesion index of the fish from Lamberto beach showed significant high values (I(org)=13) when compared with fish from Puruba beach (I(org)=7) suggesting the influence of the several human activities in the studied site. Notched and blebed nucleous were observed in this study, and significant differences were found between the studied sites. However, these differences did not reflect the total nuclear alterations.

Evaluation of genotoxicity and oxidative damage in painters exposed to low levels of toluene

Toluene is an organic solvent used in numerous processes and products, including industrial paints. Toluene neurotoxicity and reproductive toxicity are well recognized: however, its genotoxicity is still under discussion, and toluene is not classified as a carcinogenic solvent. Using the comet assay and the micronucleus test for detection of possible genotoxic effects of toluene, we monitored industrial painters from Rio Grande do Sul, Brazil. The putative involvement of oxidative stress in genetic damage and the influences of age, smoking, alcohol consumption, and exposure time were also assessed. Although all biomarkers of toluene exposure were below the biological exposure limits, painters presented significantly higher DNA damage (comet assay) than the control group; however, in the micronucleus assay, no significant difference was observed. Painters also showed alterations in hepatic enzymes and albumin levels, as well as oxidative damage, suggesting the involvement of oxidative stress. According to multiple linear regression analysis, blood toluene levels may account for the increased DNA damage in painters. In summary, this study showed that low levels of toluene exposure can cause genetic damage, and this is related to oxidative stress, age, and time of exposure. (C) 2012 Elsevier B.V. All rights reserved.

Nuclear abnormalities in erythrocytes and morphometric indexes in the catfish Cathorops spixii (Ariidae) from different sites on the southeastern Brazilian coast

Nuclear abnormalities in erythrocytes (NAE) were taken as biomarkers in the catfish Cathorops spixii (Ariidae) sampled in an estuary little affected by human activity (Cananeia) and in three regions (Santos Channel: SC, Santos Bay: SB and Sao Vicente Channel: SVC) of the Santos-Sao Vicente estuary impacted by various anthropogenic activities. Increases in NAE were observed in fish from SC and SVC sampled in the summer period as compared with specimens from the Cananeia estuary. These results suggest the presence of genotoxic compounds in these regions. However, the absence of significant differences in micronuclei frequency reflects slight mutagenic effects in these individuals. It is possible that the lower NAE frequency in specimens from SB might be associated with the greater remobilization and dilution of chemicals in this region. The low frequency of NAE in C. spixii from the Cananeia estuary is in accordance with the slight anthropogenic influence in this system, and may be suggestive of the absence of genotoxic and mutagenic effects in these organisms.

The protective effect of Canova homeopathic medicine in cyclophosphamide-treated non-human primates

Background: Canova activates macrophages and indirectly induces lymphocyte proliferation. Here we evaluated the effects of Canova in cyclophosphamide-treated non-human primates. Methods: Twelve Cebus apella were evaluated. Four animals were treated with Canova only. Eight animals were treated with two doses of cyclophosphamide (50 mg/kg) and four of these animals received Canova. Body weight, biochemistry and hematologic analyses were performed for 40 days. Micronucleus and comet assays were performed for the evaluation of DNA damage. Results: We observed that cyclophosphamide induced abnormal WBC count in all animals. However, the group treated with cyclophosphamide plus Canova presented a higher leukocyte count than that which received only cyclophosphamide. Cyclophosphamide induced micronucleus and DNA damage in all animals. The frequency of these alterations was significantly lower in the Canova group than in the group without this medicine. Conclusions: Our results demonstrated that Canova treatment minimizes cyclophosphamide myelotoxicity in C. apella. (C) 2012 Elsevier Ltd. All rights reserved.

An overview of biodiesel soil pollution: Data based on cytotoxicity and genotoxicity assessments

Biodiesel production has received considerable attention in the recent past as a nonpolluting fuel. However, this assertion has been based on its biodegradability and reduction in exhaust emissions. Assessments of water and soil biodiesel pollution are still limited. Spill simulation with biodiesel and their diesel blends in soils were carried out, aiming at analyzing their cytotoxic and genotoxic potentials. While the cytotoxicity observed may be related to diesel contaminants, the genotoxic and mutagenic effects can be ascribed to biodiesel pollutants. Thus, taking into account that our data stressed harmful effects on organisms exposed to biodiesel-polluted soils, the designation of this biofuel as an environmental-friendly fuel should be carefully reviewed to assure environmental quality. (C) 2011 Elsevier B.V. All rights reserved.

Redescription of Licnophora chattoni Villeneuve-Brachon, 1939 (Ciliophora, Spirotrichea), associated with Zyzzyzus warreni Calder, 1988 (Cnidaria, Hydrozoa)

Licnophora chattoni, found in association with Zyzzyzus warreni, a tubulariid hydroid epizoic in sponges from Sao Sebastiao (SP, Brazil), is redescribed and illustrated using light and electron microscopy. The ciliate has a flexible, transparent body formed by an oval anterior region linked to the posterior basal disc via a flexible neck region. Numerous cortical granules are observed scattered throughout the body and densely packed along the neck. The adoral zone is formed by about 81 external and 24 infundibular paramembranelles. The paroral membrane, formed by a row of long cilia arranged in monolcinetids, extends through a groove in the body to the adhesive disc. Two dorsal kinetids are present along the right body margin and around the neck. The adhesive disc (18 mu m in diameter) lacks cilia in the area above the velum. The velum covers a row of dikinetids bearing long cilia and four dikineties, two or three of which are interrupted on the ventral surface. Nine to twelve macronuclear nodules connected by isthmuses are distributed in the cytoplasm, plus two nodules located in the adhesive disc and between those there is an ovate micronucleus. (C) 2011 Published by Elsevier GmbH.

Sewage sludge does not induce genotoxicity and carcinogenesis

Through a series of experiments, the genotoxic/mutagenic and carcinogenic potential of sewage sludge was assessed. Male Wistar rats were randomly assigned to four groups: Group 1 - negative control; Group 2 - liver carcinogenesis initiated by diethylnitrosamine (DEN; 200 mg/kg i.p.); Group 3 and G4-liver carcinogenesis initiated by DEN and fed 10,000 ppm or 50,000 ppm of sewage sludge. The animals were submitted to a 70% partial hepatectomy at the 3rd week. Livers were processed for routine histological analysis and immunohistochemistry, in order to detect glutathione S-transferase positive altered hepatocyte foci (GST-P+ AHF). Peripheral blood samples for the comet assay were obtained from the periorbital plexus immediately prior to sacrificing. Polychromatic erythrocytes (PCEs) were analyzed in femoral bone-marrow smears, and the frequencies of those micronucleated (MNPCEs) registered. There was no sewage-sludge-induced increase in frequency of either DNA damage in peripheral blood leucocytes, or MNPCEs in the femoral bone marrow. Also, there was no increase in the levels of DNA damage, in the frequency of MNPCEs, and in the development of GST-P AHF when compared with the respective control group.

Biomonitoring of Microcystin and Aflatoxin Co-Occurrence in Aquaculture Using Immunohistochemistry and Genotoxicity Assays

This work investigated the effects of co-occurring aflatoxin B-1 (AFB(1)) and microcystin (MC) in aquaculture, using immunohistochemistry and genotoxicity methods. Tilapia (Oreochromis niloticus) were exposed to AFB(1) by intraperitoneal and MC (cell extract of Microcystis aeruginosa) by intraperitoneal and immersion routes. The interaction of MC-AFB(1) was evaluated co-exposing the intraperitoneal doses. Blood samples were collected after 8, 24, and 48h to analyze the micronucleus frequency and comet score. The interaction of MC-AFB(1) showed a synergic mutagenic response by higher micronucleus frequency of co-exposed group. A slight genotoxic synergism was also observed in the comet score. Immunohistochemistry detected MC in al lthe fish liver tissues exposed to MC by intraperitoneal route, and only the immersed group with the highest dose of MC showed a positive response. Although MC was non-detectable in the edible muscle, the combination of immunohistochemistry with genotoxicity assay was an attractive biomonitoring tool in aquaculture, where the animals were frequently exposed to co-occurring synergic hazards.

Genotoxic and mutagenic effects of erythrosine B, a xanthene food dye, on HepG2 cells

Erythrosine (ErB) is a xanthene and an US Food and Drug Administration approved dye used in foods, drugs and cosmetics. Although its utilization is permitted, ErB is described as inhibitor of enzymes and protein-protein interactions and is toxic to pituitary and spermatogenesis processes. However, the genotoxicity and mutagenicity of ErB is inconclusive in the literature. This study aimed to analyze the genotoxicity of this dye using the alkaline comet assay and is the first investigation to evaluate ErB mutagenicity using the cytokinesis block micronucleus cytome (CBMN-Cyt) assay in HepG2 cells. These cells were chosen because they produce phase I and phase II enzymes that can mimic in vivo metabolism. The cells were treated with seven concentrations (0.1-70.0 mu g mL(-1)) of ErB, and the results showed genotoxicity at the two highest concentrations and mutagenicity at six concentrations. Furthermore, as micronuclei result from clastogenic and aneugenic processes, while comet assay is often considered more sensitive and detects DNA single strain breaks, we suggest that an aneugenic is responsible for the observed damage. Although ErB is approved for use in the food, cosmetic and pharmaceutical industries, it must be used carefully because it damages the DNA structure. (C) 2012 Elsevier Ltd. All rights reserved.

Comparative study of beta-carotene and microencapsulated beta-carotene: Evaluation of their genotoxic and antigenotoxic effects

beta-Carotene (BC) is one of the natural pigments that is most commonly added to food; however, the utilization of BC is limited due to its instability. Microencapsulation techniques are commonly used because they can protect the microencapsulated material from oxidization. Nevertheless, the properties of the encapsulated compounds must be studied. We compared the antigenotoxic potential of pure and microencapsulated beta-carotene (mBC) in Wistar rats. Two doses of BC or mBC (2.5 or 5.0 mg/kg) were administered by gavage over a period of 14 days. The final gavage was followed by an injection of doxorubicin (DXR). After 24 h the animals were euthanized. The micronucleus test results showed that when both mBC and DXR were given, only the higher dose was antigenotoxic. The results of the comet assay show that when given in association with DXR, mBC had protective effects in the liver. The differences between the results obtained with BC and mBC suggest that possibly the carotenoid biodisponibility was modified by the process of microencapsulation. In conclusion, mBC does not lose its protective properties, but higher doses must be used to observe antigenotoxic effects. This is the first time that the genotoxicity and antigenotoxicity of a microencapsulated compound was evaluated in vivo. (C) 2012 Elsevier Ltd. All rights reserved.

Protective effect of bixin on cisplatin-induced genotoxicity in PC12 cells

Bixin is the main carotenoid found in annatto seeds (Bixa orellana L.) and is responsible for their reddish-orange color. The antioxidant properties of this compound are associated with its ability to scavenge free radicals, which may reduce damage and protect tissues against toxicity caused by anticancer drugs such as cisplatin. In this study, the genotoxicity and antigenotoxicity of bixin on cisplatin-induced toxicity in PC12 cells was assessed. Cytotoxicity was evaluated using the mu assay, mutagenicity, genotoxicity, and protective effect of bixin were evaluated using the micronucleus test and comet assay. PC12 cells were treated with bixin (0.05, 0.08, and 0.10 mu g/mL), cisplatin (0.1 mu g/mL) or a combination of both bixin and cisplatin. Bixin was neither cytotoxic nor genotoxic compared to the controls. In the combined treatment bixin significantly reduced the percentage of DNA in tail and the frequency of micronuclei induced by cisplatin. This result suggests that bixin can function as a protective agent, reducing cisplatin-induced DNA damage in PC12 cells, and it is possible that this protection could also extend to neuronal cells. Further studies are being conducted to better understand the mechanisms involved in the activity of this protective agent prior to using it therapeutically. (C) 2011 Elsevier Ltd. All rights reserved.

Antigenotoxic Effects of Piquia (Caryocar villosum) in Multiple Rat Organs

This study investigated the in vivo genotoxicity of piquia pulp (Caryocar villosum) and its potential antigenotoxicity on doxorubicin (DXR)-induced DNA damage by comet assay and micronucleus test. In addition, the phytochemicals present in piquia pulp were determined. Piquia fruit pulp (75, 150 or 300 mg/kg b.w.) was administered by gavage to Wistar rats for 14 days, and the animals received an injection of saline or DXR (15 mg/kg b.w., i.p.) 24 h before they were euthanized. The phytochemical analysis revealed the presence of carotenoids; phenolic compounds, including flavonoids; tannins and alpha-tocopherol in piquia pulp. No statistically significant differences were observed in the evaluated parameters, demonstrating the absence of cytotoxic and genotoxic effects of piquia pulp at all tested doses. In liver, kidney, cardiac and bone marrow cells, piquia significantly reduced the DNA damage induced by DXR. Our results showed that the lowest piquia dose caused the largest decrease in DNA damage and the highest dose caused the smallest decrease, demonstrating an inverse dose-response of piquia pulp. Furthermore, we observed a difference in the potential antigenotoxic effects in several tissues. In conclusion, our results demonstrated that piquia pulp was not genotoxic and inhibited the genotoxicity induced by DXR, but some of the protective effects that were observed depended on the doses and experimental conditions. Therefore, further investigations are needed to clarify how piquia pulp positively affects human health.