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The Itapemirim River is considered one of the most important water resources in the state of Espírito Santo, Brazil. However, environmental problems due to continuous anthropogenic contamination are threatening its potential use. This study assessed water quality by analyzing abiotic and toxicogenetic aspects of the water from four stations along the river. Samples were collected in both dry and rainy seasons. Most of the abiotic variables were below the threshold established by CONAMA Resolution No. 357/2005, and so were most of the metals. However, Al and Cu contents were above those allowed by legislation, ranging from 0.2 to 0.9 mg/L. Regarding toxicogenetic aspects, genotoxic effects were observed in meristematic cells of Allium cepa, in micronucleus test and comet assay of Oreochromis niloticus, and CHO-K1 cells. Mutagenic effects were significant at RI 02 (0.34), RI 03 (0.46), and RI 04 (0.12) stations on the first campaign in A. cepa F1 cells, compared to the negative control (0.0). The second campaign revealed the same results, but with the addition of samples from RI 01 (0.17) and RI 03 (0.18) showing mutagenicity in the micronucleus test with fish erythrocytes when compared to the negative control (0.3). Essentially, all the samples evaluated in both campaigns showed damage in A. cepa, O. niloticus, and CHO-K1 cells, thus demonstrating that the water quality of the Itapemirim River is compromised and requires action plans for its recovery.

Introdução: A agricultura é uma prática indispensável para o desenvolvimento das sociedades, através do fornecimento de alimentos e fomento da economia. Apesar de ser fundamental para a indústria alimentícia e ter grande importância econômica, é necessário analisar os prejuízos ambientais que essa atividade pode gerar quando não executada de forma consciente e sem as devidas responsabilidades em sua condução. Os defensivos agrícolas são muito utilizados em plantações no controle e combate ao crescimento de pragas, porém, em determinadas quantidades seu uso pode afetar os organismos existentes no solo como as minhocas, consideradas ótimos bioindicadores. Objetivos: Avaliar a resposta de minhocas (Eisenia fetida) expostas ao fungicida FOX e aos herbicidas Glifosato e Haloxyfop em concentrações de uso em soja. Material e métodos: Potes de plástico com capacidade de 2L foram divididos em 2 metade iguais e receberam em cada lado 500g de solo artificial. O solo, em uma metade foi umedecido com 40 ml de água (controle negativo-CN) e na outra uma solução contendo os defensivos agrícolas em concentrações usadas em plantações de soja. No centro do pote foram colocadas 10 minhocas e após 48h contabilizada sua distribuição entre as metade, sendo este resultado utilizado para calcular a porcentagem de fuga/atração. Resultados: Notou-se que no Glifosato a média de fuga das minhocas foi igual a 0%, resultado distinto dos defensivos FOX e Haloxyfop, cuja média foi 68% e 92%, respectivamente. Conclusão: De acordo com a ISO 17512-1, quando a fuga de organismos exposto à substância testada em direção ao CN está acima de 80%, considera-se a amostra tóxica, considerando a função do habitat limitada. Neste caso, o Haloxyfop mostrou alto índice de toxicidade, visto a alta porcentagem das minhocas em evitar a substância química. O fungicida FOX causou repelência e o herbicida Glifosato, por sua vez, mostrou-se indiferente nas concentrações testadas.

Sewage sludge has been used as a soil amendment because it is rich in plant essential nutrients and can serve as a substitute for chemical fertilizers. However, sewage sludge can contain a complex mixture of pollutants that are harmful to organisms. Considering that the management of the waste should not result in risks to environmental health, the toxic potential of sewage sludge to organisms at different trophic levels should be measured before it final disposal. Diplopod (Rhinocricus padbergi) and fish (Xiphophorus maculatus) were exposed to sewage sludge or sewage sludge treated with lime (biosolid) at different dilutions under laboratory conditions. Semiquantitative histopathological analyses were performed on target organs (midgut and fat body for the diplopod and gills for the fish). Toxins from sludge or biosolid caused tissue damage and activated detoxification processes in both the diplopod and fish. In comparison to other sludge concentrations, the highest sludge concentrations tested were more toxic, and millipedes likely reduced their food intake to avoid toxins, resulting in weight loss. Our data suggest that sewage sludge pathogens promoted specific histological alterations. Chemical analysis revealed that the concentrations of priority pollutants did not exceed the limits established by Brazilian law. Therefore, this outcome indicates that R. padbergi and X. maculatus are sensitive to low levels of toxins from sewage sludge.

The industrial treatment of dimension stones, such as marbles and granites, includes a stage of plate polishing, in which resins and abrasives are used, producing a fine grained waste with high moisture content. These wastes pass through decantation tanks in order to separate the solid and liquid phases. Until now, there is no knowledge about the mutagenic effects that this effluent can cause to organisms exposed to it. Thus, this study evaluated the mutagenic potential of dimension stone polishing wastes in onion root cells and fish erythrocytes. The onion seeds were germinated in Petri dishes with filter paper moistened in the liquid phase of the effluent. After germination, the onion roots were prepared for analysis of chromosomal aberrations in meristematic cells. The fishes were exposed during 72h to the solid phase of the effluent diluted in pure groundwater. Blood samples were used for counting of micronucleus and nuclear abnormalities. The onion seeds had similar germination and mitotic index in all treatments. However, it was observed in the seeds exposed to the polishing waste, numbers significantly higher of micronucleus, nuclear buds and other chromosomal aberrations when compared with the negative control. The fishes exposed to the waste showed numbers significantly higher of micronucleus when compared with the negative control. The fishes from all treatments showed significant increase in nuclear abnormalities when compared to the negative control. We concluded that the analysed wastes have mutagenic potential at the studied conditions; this effect can be related to the high content of phenolic compounds identified in the samples.

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.

A utilização de agrotóxicos se faz presente em todo o Brasil, alguns possuem efeitos mutagênicos em células vegetais já provados, e seu uso indiscriminado pode levar a contaminação do solo e da água. Dentre esses produtos químicos, destaca-se os herbicidas que são muito utilizados na agricultura, no qual encontra-se inserido o ácido diclorofenoxiacético ou 2,4-D. O uso deste herbicida é recomendado em culturas de milho, entretanto, ainda são escassos trabalhos na literatura que demonstram seu efeito nas células deste vegetal, o que levou este trabalho a avaliar os efeitos do herbicida 2,4-D, em diferentes concentrações, no ciclo celular de células da raiz de milho (Zea mays). Os resultados demonstraram que há redução no crescimento de raízes de milho conforme é aumentada a concentração do herbicida, demonstrando a sensibilidade de tal semente ao herbicida, que acaba por induzir sua taxa de crescimento.

This study presents the morphological description and histochemical characterization of gill filaments of the Brazilian endemic bivalve Diplodon expansus, aiming to broaden the morphological knowledge of this species and establish the structure of the gills that will serve as control in histopathological studies applied to biomonitoring. The gill filaments are divided into three zones: frontal, intermediate, and abfrontal. In the center of the filament, haemocytes circulate through the haemolymph vessel, which is internally lined by endothelium. The frontal surface of the filament is covered with cilia, the lateral surface exhibits aquifer ducts, and the abfrontal surface presents ciliated and nonciliated cells. The epithelium of the filaments is composed of ciliated cells, nonciliated absorptive cells, and mucocytes. The support of the filaments is made by two specialized structures called skeletal rod and skeletal loop. Based on the obtained information, the gill filaments of the studied species present some peculiar characteristics that are not yet reported in detail in the literature such as the simultaneous presence of skeletal rod and skeletal loop. On the other hand, the general constitution of the filament is similar to that described for both marine and limnic bivalves and seems to be suitable for ecotoxicological studies.

Bivalve mussels can concentrate pollutants which, in high amounts, can cause cellular, physiological and behavioral changes. The gill mucus is essential for many metabolic and behavioral procedures on marine mussels including the response to environmental pollution. The present work analyzed the mucous cell number in Mytella falcata gill filaments from three sites of Santos estuary (Brazil) with different levels of sediment contamination named as sites A, B, and C. Site A is located at a low impacted region of the estuary, and sites B and C are under influence of port activities, domestic sewage, and industries like a still mill located in front of site C (the most contaminated area with high amounts of polycyclic aromatic hydrocarbons). An increase in mucous cell number was detected in animals from sites B and C, when compared to site A; this increase occurred preferentially in the frontal zone and was related to sediment contamination. Mucous cell analysis can be used as an efficient biomarker. It is suggested that mucus produced in lateral and abfrontal zones is responsible for eliminating pollutants previously absorbed and mucus produced in frontal zone is responsible for pollutant arrest and elimination as pseudofeces.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T19:05:03Z No. of bitstreams: 1 S1415-47572009000100024.pdf: 81043 bytes, checksum: 04d9c31a7cba9d5aa733af9fb52b9ad1 (MD5) Made available in DSpace on 2013-08-22T19:05:03Z (GMT). No. of bitstreams: 1 S1415-47572009000100024.pdf: 81043 bytes, checksum: 04d9c31a7cba9d5aa733af9fb52b9ad1 (MD5) Previous issue date: 2009-01-01 Made available in DSpace on 2013-09-30T20:08:42Z (GMT). No. of bitstreams: 2 S1415-47572009000100024.pdf: 81043 bytes, checksum: 04d9c31a7cba9d5aa733af9fb52b9ad1 (MD5) S1415-47572009000100024.pdf.txt: 21127 bytes, checksum: 98afd83ff92a8a77363ba5b1fb75f38b (MD5) Previous issue date: 2009-01-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:54:50Z No. of bitstreams: 2 S1415-47572009000100024.pdf: 81043 bytes, checksum: 04d9c31a7cba9d5aa733af9fb52b9ad1 (MD5) S1415-47572009000100024.pdf.txt: 21127 bytes, checksum: 98afd83ff92a8a77363ba5b1fb75f38b (MD5) Made available in DSpace on 2014-05-20T13:54:50Z (GMT). No. of bitstreams: 2 S1415-47572009000100024.pdf: 81043 bytes, checksum: 04d9c31a7cba9d5aa733af9fb52b9ad1 (MD5) S1415-47572009000100024.pdf.txt: 21127 bytes, checksum: 98afd83ff92a8a77363ba5b1fb75f38b (MD5) Previous issue date: 2009-01-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação para o Desenvolvimento da UNESP (FUNDUNESP) The present study applied the comet assay to erythrocytes of Oreochromis niloticus with the aim of improving protocols to detect DNA damage in these cells, by using two distinct pHs (pH = 12.1 and pH > 13) and evaluating whether there is a correspondence between silver and ethidium bromide staining. Comets were visually examined and, the frequency of cells with and without damage was obtained, as well as the distribution of classes and scores. By using the Kruskal-Wallis test, our results revealed that pH 12.1 is more effective, although both pHs can be used. Our findings also suggest that silver staining can substitute ethidium bromide, an expensive and highly toxic stain that requires specific equipment for examination. Universidade Estadual Paulista Júlio de Mesquita Filho Instituto de Biociências Departamento de Biologia Universidade Federal do Pará (UFPA) Instituto de Biociências Departamento de Genética Universidade Estadual Paulista Júlio de Mesquita Filho Instituto de Biociências Departamento de Biologia

Chrysomya blowflies are originally from Africa and Australasia and were introduced in the American Continent, probably as a sheep parasite, in the 1970s. These flies are extremely important for medical-sanitary purposes and are useful to forensic entomology being used as an indicative of decomposition time in human corpses. The morphology of the larval and pupal stages of some species has been already studied by different authors demonstrating that it is possible to identify them in premature stages. In this study, Chrysomya megacephala egg ultramorphology was analyzed to describe its structure, generating data for further comparison between different species and genera. The blowflies were collected in Rio Claro city, SP, Brazil. Flies were attracted by fish carcasses and maintained in net cages; small portions of minced meat were placed in plastic pots for egg laying. Eggs were collected, fixed in alcoholic Bouin, prepared according to SEM routine and observed under Philips scanning electron microscope. C. megacephala eggs are oval with one flat face and another convex, measuring approximately 0.52 (±0.03) mm of length and 0.12 (±0.02) mm of width. The micropyle is circular in shape, with two perforations enclosed by countless small projections and is located in one of the extremities in the interface between the flat and convex faces. On the convex surface hexagonal imprints were observed, this surface is impermeable to water and gases. The flat surface has numerous round projections with different sizes, creating a permeable surface with a thinner chorion from which the larvae hatches. The larvae presents itself all wrinkled and with numerous cuticular projections with a thorn shape facing the posterior region. The cuticle projections with a thorn shape from C. megacephala larvae are probably a vestige of the thorns found in larvae of parasite Diptera.

Bivalve filter feeders are sessile animals that live in constant contact with water and its pollutants. Their gill is an organ highly exposed to these conditions due to its large surface and its involvement in gas exchanges and feeding. The bivalve Mytella falcata is found in estuaries of Latin America, on the Atlantic as well as the Pacific Coast. It is commonly consumed, and sometimes is the only source of protein of low-income communities. In this study, gill filaments of M. falcata were characterized using histology, histochemistry and transmission electron microscopy for future comparative studies among animals exposed to environmental pollutants. Gill filaments may be divided into abfrontal, intermediate and frontal zones. Filaments are interconnected by ciliary discs. In the center of filaments, haemocytes circulate through a haemolymph vessel internally lined by an endothelium and supported by an acellular connective tissue rich in polysaccharides and collagen. The abfrontal zone contains cuboidal cells, while the intermediate zone consists of a simple squamous epithelium. The frontal zone is composed of five columnar cell types: one absorptive, mainly characterized by the presence of pinocytic vesicles in the apical region of the cell; one secretory, rarely observed; and three ciliated with abundant mitochondria. All cells lining the filament exhibit numerous microvilli and seem to absorb substances from the environment. PAS staining was observed in mucous cells in the frontal and abfrontal zones. Bromophenol blue allowed the distinction of haemocytes and detection of a glycoprotein secretion in the secretory cells of the frontal region. The characteristics of M. falcata gill filaments observed in this study were very similar to those of other bivalves, especially other Mytilidae, and are suitable for histopathological studies on the effect of water-soluble pollutants.

Coffee wastewater (CWW) is an effluent produced through wet processing of coffee containing high concentration of organic matter, nutrients, salts and also agrochemicals. It is released directly into the argillaceous soil or into decantation tanks for later disposal into soils, by fertigation, subsurface infiltration or superficial draining. However, this practice is not followed by the monitoring the toxicity potential of this effluent. In this sense, the present work aimed to evaluate the phytotoxic, cytogenotoxic and mutagenic potential of CWW on seed germination, root elongation and cell cycle alterations in the plant model Lactuca sativa L. The effluent (CWW) collected was diluted in distilled water into six concentrations solutions (1.25%, 1.66%, 2.5%, 5.0%, 10%, 20%). A solution of raw CWW (100%) was also applied. Distilled water was used as negative control), and the DNA alkylating agent, metilmetano sulfonate (4×10(-4)M) as positive control. Physico-chemical parameters of the CWW was accessed and it was found that the effluent contained total phenols and inorganic matter in amounts within the limits established by the National Environment Council (CONAMA). Nevertheless, the biologicals assays performed demonstrated the phytotoxicity and cytogenotoxicty of CWW. Seed germination was totally inhibited after exposure of raw CWW. In addition, a decrease in seed germination speed as well as in root growth dose-dependently manner was noticed. Moreover, nuclear and chromosomal alterations were observed in the cell cycle, mostly arising from aneugenic action.

Fish gills are constituted by primary filaments and secondary lamellae, which are formed, basically, by three different cell types: pillar cells, respiratory cells and erythrocytes that circulate in the lamellae interior. This respiratory organ is very appreciated in environmental studies since it is constantly in contact with the water, accomplishing exchanges; besides this, it presents a high adaptation capacity. In this work, some fishes were reared in polluted waters treated by the flotation method (treated group) and compared with others that were reared in clean waters (control group). The ultramorphological, histological and histochemical analyses were used. The fishes from the treated group presented morphological alterations when compared with the control groups, such as lamellar fusion, aneurism and detachment of the respiratory epithelium. These alterations demonstrate the several adaptations, as morphologic as physiologic, that fishes can present when exposed to an inappropriate environment. The techniques employed are good tools to detect morphological biomarkers in gills of Oreochromis niloticus submitted to polluted waters.

The impact of pollutants in an organism can be observed by changes in functional complexity at different levels. Bivalve gills are suitable for histopathological analysis because of their structure and function. This study aimed at examining the morphology of Mytella falcata gill filaments from three sites in the Santos estuary (Sao Paulo, Brazil) with different levels of environmental degradation to identify possible changes in gill structure and discuss the significance of these alterations. For this purpose, histological, histochemical and ultrastructural techniques were used. The filaments of animals from site A (less impacted site) were intact, while in sites B and C, pathological changes were observed, such as: detachment of the epithelium in the intermediate zone, morphological changes of this epithelium, inflammatory process, increase in the number of mucous cells and cell turnover processes. These results suggest that the related changes are an attempt to prevent the entrance of pollutants through gill filaments into the entire organism and that cell turnover is the final way to compensate cell injury.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T18:41:55Z No. of bitstreams: 1 S1519-566X2003000100010.pdf: 156468 bytes, checksum: 73be7aae5057230b0120ffa416f8ed06 (MD5) Made available in DSpace on 2013-08-22T18:41:55Z (GMT). No. of bitstreams: 1 S1519-566X2003000100010.pdf: 156468 bytes, checksum: 73be7aae5057230b0120ffa416f8ed06 (MD5) Previous issue date: 2003-01-01 Made available in DSpace on 2013-09-30T19:28:50Z (GMT). No. of bitstreams: 2 S1519-566X2003000100010.pdf: 156468 bytes, checksum: 73be7aae5057230b0120ffa416f8ed06 (MD5) S1519-566X2003000100010.pdf.txt: 17723 bytes, checksum: 8009df87bed8a94367ba41007acc8165 (MD5) Previous issue date: 2003-01-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:07:52Z No. of bitstreams: 2 S1519-566X2003000100010.pdf: 156468 bytes, checksum: 73be7aae5057230b0120ffa416f8ed06 (MD5) S1519-566X2003000100010.pdf.txt: 17723 bytes, checksum: 8009df87bed8a94367ba41007acc8165 (MD5) Made available in DSpace on 2014-05-20T15:07:52Z (GMT). No. of bitstreams: 2 S1519-566X2003000100010.pdf: 156468 bytes, checksum: 73be7aae5057230b0120ffa416f8ed06 (MD5) S1519-566X2003000100010.pdf.txt: 17723 bytes, checksum: 8009df87bed8a94367ba41007acc8165 (MD5) Previous issue date: 2003-01-01 O gênero Gryllus destaca-se com um grande número de espécies descritas e representa um dos mais complexos gêneros na sistemática dos Orthoptera, caracterizado por um conjunto de espécies cosmopolitas e crípticas. Este trabalho compara os sons emitidos por espécimens de Gryllus coletados no campus da UNESP em Rio Claro (SP), bem como a morfologia e morfometria das pars stridens, com o objetivo de aplicar os resultados no reconhecimento de possíveis espécies crípticas e contribuir na discussão de processos de especiação. Três grupos de grilos foram discriminados por diferenças nas pars stridens e sons de chamado, caracterizados por diferentes ritmos, freqüências e composição das notas, indicando assim, a presença de três espécies no local analisado, morfologicamente pouco distintas. Diante dos resultados, sugere-se a utilização das características da pars stridens e do som de chamado como caracteres diagnósticos imprescindíveis na taxonomia dos Gryllus. The Gryllus genus represents one the most complex in the Orthoptera systematic, characterized by a set of cosmopolite and cryptic species, many of them already described. This study compared the songs emitted by Gryllus specimens collected on the UNESP campus in Rio Claro (SP) and the pars stridens morphology and morphometry, to use the results to recognize possible cryptic species and contribute to the discussion of speciation processes. Three groups of crickets were discriminated by differences in the pars stridens and calling songs, characterized by different rhythms, frequencies and note composition, that indicated the presence of three species in the location analyzed, that presented few morphological differences. It is suggested from the results that the use of the pars stridens characteristics and the calling song as diagnostic traits is essential in the Gryllus taxonomy. UNESP Depto. Biologia Faculdade União das Américas UNESP Depto. Biologia