FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / O aproveitamento de Ãguas residuÃrias à uma alternativa bastante difundida na atualidade, e uma das formas de reÃso à na atividade piscÃcola, considerando-se que os efluentes das EstaÃÃes de Tratamento de Esgotos sÃo fontes ricas em nutrientes que podem ser aproveitados pelos peixes. Entretanto, algumas restriÃÃes se apresentam com relaÃÃo à qualidade dos animais para aproveitamento na alimentaÃÃo humana. Dessa forma, essa pesquisa foi realizada utilizando esgotos domÃsticos tratados, na criaÃÃo de tilÃpias do Nilo, Oreochromis niloticus, observando-se os aspectos zootÃcnicos, toxicolÃgicos do ambiente de cultivo, bem como condicionantes qualitativos sanitÃrios, nutricional, e de aceitabilidade do pescado produzido. A unidade experimental foi implantada em Ãrea adjacente à EstaÃÃo de Tratamento de Esgotos (lagoas de estabilizaÃÃo), da zona leste de Teresina-Pi, com dezoito tanques de fibra de vidro, dispostos em trÃs (03) tratamentos com seis (06) repetiÃÃes (3X6). O tratamento T-01 foi abastecido somente com Ãgua tratada e forneceu-se raÃÃo industrializada; o tratamento T-02 recebeu esgoto tratado da lagoa de maturaÃÃo, sem suprimento adicional de raÃÃo ou oxigÃnio, e o tratamento T-03 foi abastecido com o mesmo efluente sem fornecimento de raÃÃo, mas, recebeu aeraÃÃo suplementar. Durante o ciclo experimental, foi realizada a renovaÃÃo diÃria de 15% do volume de Ãgua armazenado nos tanques experimentais, para reposiÃÃo de nutrientes e limpeza. Nos tanques foram estocados alevinos de tilÃpias, revertidos sexualmente para machos em lotes de 10 indivÃduos por tanque (aproximadamente 03 alevinos /mÂ). Foram analisados os principais parÃmetros de qualidade da Ãgua e do esgoto, que abastece os tanques, com especial destaque para as anÃlises destinadas ao monitoramento da piscicultura. Para verificaÃÃo da capacidade produtiva do ambiente de cultivo, foram avaliados os parÃmetros zootÃcnicos, tais como: crescimento, ganho de peso, produtividade e conversÃo alimentar aparente. TambÃm foram realizados os testes microbiolÃgicos, de genotoxicidade (micronÃcleos), de aceitabilidade e preferÃncia (anÃlise sensorial), alÃm do Ãndice do rigor mortis (IRM) e da Bioimpedancia. Este conjunto de testes foi utilizado para identificar o nÃvel de qualidade do pescado. Os indivÃduos submetidos ao tratamento T-01 (produtividade de 118,14 kg/ha.dia), tiveram o melhor desempenho zootÃcnico, seguido dos peixes do T-03 (produtividade de 48,61kg/ha.dia), e com o pior resultado ficou o grupo do tratamento T-02 (produtividade de 34,90 kg/ha.dia). Entretanto, nos testes que aferiram a qualidade sanitÃria do pescado e a palatabilidade nÃo foram registradas diferenÃas significativas. Conclui-se que a piscicultura com esgoto domÃstico tratado em lagoas de estabilizaÃÃo à viÃvel, mas, ainda sÃo necessÃrios novos estudos para o desenvolvimento desta prÃtica. / The use of wastewater is an alternative widespread nowadays. One way to reuse it in fish activity, considering that the effluent from Sewage Treatment Stations, are rich sources of nutrients that could be used by fish. However, some restrictions arise regarding the quality of animals for use in human diet. Thus, this research was conducted using treated sewage, to raise Nile tilapia (Oreochromis niloticus), observing zootechnical aspects, toxicology of the culture environment, as well as qualitative sanitary conditions, nutrition, and acceptability of fish produced. The experimental unit was deployed in an area adjacent to the Sewage Treatment Station (stabilization ponds), at the east side of Teresina-PI, with eighteen of fiberglass tanks, arranged in three treatments with six repetitions. Treatment T-01 was supplied only with treated water and industrial fish feed; treatment T-02 received treated sewage from the maturation pond, without additional supply of feed or oxygen, and treatment T-03 was stocked with the same sewage effluent without feed, but received supplemental aeration. During the experimental cycle, we performed a daily renewal of 15% of the volume of water stored in experimental tanks for nutrient replacement and cleaning. In ponds stocked fingerlings of tilapia, sexually reversed for males in batches of 10 individuals per tank (about 03 fingerlings / mÂ). We analyzed the main parameters of water quality and sewage, fueling the tanks, with special emphasis on tests for monitoring of fish farming. To verify the productive capacity of the culture environment, we assessed the growth parameters such as growth, weight gain, feed conversion and productivity. Were also conducted microbiological testing of genotoxicity (micronuclei), acceptability and preference (sensory analysis), and the level of rigor mortis (MRI) and bioimpedance. This set of tests was used to identify the level of fish quality. Individuals undergoing treatment T-01 (productivity 118,14 kg/ha.day), had the best performance, followed of treatment T-03 (productivity 48,61kg/ha.day), and T-02 (productivity 34,90 kg/ha.day). However in tests that assessed the quality, fish health and the flavor, there was not significant difference. It is concluded that the fish farming with treated domestic sewage in stabilization ponds, it is feasible. But new studies are still needed for development of the practice.

All-male fry are preferred to prevent uncontrolled reproduction before harvest in intensive Nile tilapia (Oreochromis niloticus) aquaculture. Males also grow faster than females. An alternative approach to direct hormonal masculinisation of tilapia fry is to produce fry that are genetically male. However, sex determination system in tilapia is fairly complex. Recent developments have resulted in a linkage map and genetic markers that can be used to analyse the sex determination system. To analyse the genetic sex determination mechanism and to develop marker-assisted selection in the Stirling Nile tilapia population, a fully inbred line of clonal females (XX) was verified using test crosses and DNA markers (mostly microsatellites) to use as a standard reference line in sex determination studies. A series of crosses were performed involving this line of females and a range of males. Three groups of crosses were selected (each group consisted of three families) from progeny sex ratio distributions, and designated as type ‘A’ (normal XY males x clonal XX females), type ‘B’ (putative YY males x clonal XX females) and type ‘C’ (unknown groups of males x clonal XX females), for sex linkage study. For type ‘A’, inheritance of DNA markers and phenotypic sex was investigated using screened markers from tilapia linkage group 1 (LG1) to confirm the LG1-associated pattern of inheritance of phenotypic sex and the structure of LG1. Screened markers from LG1, LG3 and LG23 were used to investigate the association of markers with sex in families of type ‘B’ and ‘C’. In addition, a genome-wide scan of markers from the other 21 LGs was performed to investigate any association between markers and sex, in only families of cross type ‘B’. LG1 associated pattern of inheritance of phenotypic sex was confirmed by genotype and QTL analyses in families of cross type ‘A’. Analyses of genotypes in families of type ‘B’ and ‘C’ showed strong association with LG1 markers but no association with LG3 and LG23 markers. Genome wide scan of markers from all other LGs did not show any significant association between any markers and the sex. The allelic inheritance of two tightly linked LG1 markers (UNH995 and UNH104) in families of type ‘B’ and ‘C’ identified polymorphism in the sex determining locus: one of the alleles was associated mostly with male offspring whereas another allele was associated with both progeny (mostly males in type ‘B’ families, and approximately equal numbers in type ‘C’ families). This knowledge was used to identify and separate supermales (‘YY’ males) that should sire higher proportions of male progeny, reared to become sexually mature for use as broodstock. Two of them were crossed with XX females (one clonal and one outbred) to observe the phenotypic expression of the strongest male-associated allele in progeny sex. The observations of 98% male (99 males out of 101 progeny) and 100% male (N=75) from these two crosses respectively, suggest that a marker-assisted selection (MAS) programme for genetically male Nile tilapia production could be practical. This study also suggests that the departures from the sex ratios predicted using a “simple” XX/XY model (i.e., YY x XX should give all-male progeny) were strongly associated with the XX/XY system, due to multiple alleles, rather than being associated with loci in other LGs (e.g., LG3, LG23). This study also tentatively names the allele(s) giving intermediate sex ratios as “ambivalent” and emphasizes that the presence and actions of such allele(s) at the same sex-determining locus could explain departures from predicted sex ratios observed in some earlier studies in Nile tilapia.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / In this study, the microalgae Spirulina platensis was used to promote the treatment of the water from a cultivation of Nile tilapia (Oreochromis niloticus). Five treatments were carried (VD, CD, CS, SI and SE). In VD, fish were cultivated in fresh green water from a fish culture tank, containing several species of microalgae. In CD, tilapia fingerlings were cultivated in fresh clear water, without the presence of native microalgae. In CS, fingerlings, previously acclimated, were cultivated in clear water with salinity 10. In the treatment carried on the internal system (SI), the microalgae were cultured in a 1,500 L pool with tilapia effluent and artificial lighting. In the treatment SE, microalgae S. platensis were cultured with natural lighting in a 1,500 L boxe containing only the effluent of the fish cultivated in the clear water with salinity 10 (CS). The abiotic parameters analyzed during the cultures were: absorbance, pH, temperature, salinity, dissolved oxygen and total ammonia, nitrites, nitrates and total phosphates concentrations. The zootechnical development of fish was monitored and the production of microalgae biomass was determined. The results showed that fish growth in weight (W) and length (L) of treatment VD (W = 12.25  0.50 g and L = 7.1  0.1 cm) were higher than the treatments CD (W = 9.19  0.59 g and L = 6.3  0.1 cm) and CS (P = 9.90  0.69 g and L = 6.7  0.1 cm). The maximum cell density of S. platensis obtained in the treatment SE was 322,000 trichomes mL-1, whereas in SI were obtained 550,000 trichomes mL-1, which resulted in the greatest production of dry biomass of S. platensis (0.22 g L-1) and maximum yield (0.02 g L-1 day-1). SI was also the treatment which better reduced ammonia, nitrites, nitrates and phosphates concentrations ( 95%), leaving the effluent nutrients levels below to those established by n 357 CONAMA Resolution, making the water suitable for reuse in the fish culture or to be disposal in the natural environment. / Neste estudo, a microalga Spirulina platensis foi utilizada para promover o tratamento da Ãgua proveniente de cultivo da tilÃpia do Nilo (Oreochromis niloticus). Foram realizados 5 tratamentos (VD, CD, CS, SI e SE). Em VD os peixes foram cultivados em Ãgua verde e doce, proveniente de um tanque de piscicultura, contendo diversas espÃcies de microalgas. Em CD, os alevinos de tilÃpia foram cultivados em Ãguas claras e doce, sem a presenÃa de microalgas nativas. Em CS, os alevinos, previamente aclimatados, foram cultivados em Ãgua clara com salinidade 10. No tratamento realizado no sistema interno (SI), a microalga foi cultivada em uma piscina de 1.500 L contendo o efluente de tilÃpia com iluminaÃÃo artificial. No tratamento SE a microalga S. platensis foi cultivada em uma caixa de 1.500 L, contendo apenas o efluente do cultivo dos peixes em Ãguas claras com salinidade 10 (CS) com iluminaÃÃo natural. Os parÃmetros abiÃticos analisados durante os cultivos foram: absorbÃncia, pH, temperatura, salinidade, oxigÃnio dissolvido e as concentraÃÃes de amÃnia total, nitritos, nitratos e fosfatos totais. Foi realizado o acompanhamento do desenvolvimento zootÃcnico dos peixes e determinada a produÃÃo de biomassa da microalga. Os resultados revelaram que o crescimento dos peixes, em peso (P) e comprimento (L) finais do tratamento VD (P = 12,25  0,50 g e L = 7,1  0,1 cm) foram superiores aos dos tratamentos CD (P = 9,19  0,59 g e L = 6,3  0,1 cm) e CS (P = 9,90  0,69 g e L = 6,7  0,1 cm). A densidade celular mÃxima de S. platensis obtida no tratamento SE foi de 322 mil tricomas mL-1, enquanto que em SI foi de 550 mil tricomas mL-1, que resultou na maior produÃÃo de biomassa seca de S. platensis (0,22 g L-1) e produtividade mÃxima de 0,02 g L-1 dia-1. SI tambÃm foi o tratamento com a melhor reduÃÃo das concentraÃÃes de amÃnia, nitritos, nitratos e fosfatos, em mais 95%, o que deixou os nÃveis dos nutrientes no efluente abaixo dos estabelecidos como referÃncia pela resoluÃÃo n 357 do CONAMA, tornando essa Ãgua apta para o reuso no cultivo dos peixes ou descarte no ambiente natural.

NOGUEIRA, Sara Monaliza Sousa. Tratamento de efluentes de cultivos de tilápia do nilo (Oreochromis niloticus) com a microalga Spirulina platensis. 2012. 59 f. : Dissertação (mestrado) - Universidade Federal do Ceará, Centro de Ciências Agrárias, Departamento de Engenharia de Pesca, Fortaleza-CE. 2012 / Submitted by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-21T14:41:46Z No. of bitstreams: 1 2012_dis_smsnogueira.pdf: 611465 bytes, checksum: ea326d9588b86dea24e8d3c17335321a (MD5) / Approved for entry into archive by Nádja Goes (nmoraissoares@gmail.com) on 2016-07-21T14:41:59Z (GMT) No. of bitstreams: 1 2012_dis_smsnogueira.pdf: 611465 bytes, checksum: ea326d9588b86dea24e8d3c17335321a (MD5) / Made available in DSpace on 2016-07-21T14:41:59Z (GMT). No. of bitstreams: 1 2012_dis_smsnogueira.pdf: 611465 bytes, checksum: ea326d9588b86dea24e8d3c17335321a (MD5) Previous issue date: 2012 / In this study, the microalgae Spirulina platensis was used to promote the treatment of the water from a cultivation of Nile tilapia (Oreochromis niloticus). Five treatments were carried (VD, CD, CS, SI and SE). In VD, fish were cultivated in fresh green water from a fish culture tank, containing several species of microalgae. In CD, tilapia fingerlings were cultivated in fresh clear water, without the presence of native microalgae. In CS, fingerlings, previously acclimated, were cultivated in clear water with salinity 10. In the treatment carried on the internal system (SI), the microalgae were cultured in a 1,500 L pool with tilapia effluent and artificial lighting. In the treatment SE, microalgae S. platensis were cultured with natural lighting in a 1,500 L boxe containing only the effluent of the fish cultivated in the clear water with salinity 10 (CS). The abiotic parameters analyzed during the cultures were: absorbance, pH, temperature, salinity, dissolved oxygen and total ammonia, nitrites, nitrates and total phosphates concentrations. The zootechnical development of fish was monitored and the production of microalgae biomass was determined. The results showed that fish growth in weight (W) and length (L) of treatment VD (W = 12.25 ± 0.50 g and L = 7.1 ± 0.1 cm) were higher than the treatments CD (W = 9.19 ± 0.59 g and L = 6.3 ± 0.1 cm) and CS (P = 9.90 ± 0.69 g and L = 6.7 ± 0.1 cm). The maximum cell density of S. platensis obtained in the treatment SE was 322,000 trichomes mL-1, whereas in SI were obtained 550,000 trichomes mL-1, which resulted in the greatest production of dry biomass of S. platensis (0.22 g L-1) and maximum yield (0.02 g L-1 day-1). SI was also the treatment which better reduced ammonia, nitrites, nitrates and phosphates concentrations ( 95%), leaving the effluent nutrients levels below to those established by nº 357 CONAMA Resolution, making the water suitable for reuse in the fish culture or to be disposal in the natural environment. / Neste estudo, a microalga Spirulina platensis foi utilizada para promover o tratamento da água proveniente de cultivo da tilápia do Nilo (Oreochromis niloticus). Foram realizados 5 tratamentos (VD, CD, CS, SI e SE). Em VD os peixes foram cultivados em água verde e doce, proveniente de um tanque de piscicultura, contendo diversas espécies de microalgas. Em CD, os alevinos de tilápia foram cultivados em águas claras e doce, sem a presença de microalgas nativas. Em CS, os alevinos, previamente aclimatados, foram cultivados em água clara com salinidade 10. No tratamento realizado no sistema interno (SI), a microalga foi cultivada em uma piscina de 1.500 L contendo o efluente de tilápia com iluminação artificial. No tratamento SE a microalga S. platensis foi cultivada em uma caixa de 1.500 L, contendo apenas o efluente do cultivo dos peixes em águas claras com salinidade 10 (CS) com iluminação natural. Os parâmetros abióticos analisados durante os cultivos foram: absorbância, pH, temperatura, salinidade, oxigênio dissolvido e as concentrações de amônia total, nitritos, nitratos e fosfatos totais. Foi realizado o acompanhamento do desenvolvimento zootécnico dos peixes e determinada a produção de biomassa da microalga. Os resultados revelaram que o crescimento dos peixes, em peso (P) e comprimento (L) finais do tratamento VD (P = 12,25 ± 0,50 g e L = 7,1 ± 0,1 cm) foram superiores aos dos tratamentos CD (P = 9,19 ± 0,59 g e L = 6,3 ± 0,1 cm) e CS (P = 9,90 ± 0,69 g e L = 6,7 ± 0,1 cm). A densidade celular máxima de S. platensis obtida no tratamento SE foi de 322 mil tricomas mL-1, enquanto que em SI foi de 550 mil tricomas mL-1, que resultou na maior produção de biomassa seca de S. platensis (0,22 g L-1) e produtividade máxima de 0,02 g L-1 dia-1. SI também foi o tratamento com a melhor redução das concentrações de amônia, nitritos, nitratos e fosfatos, em mais 95%, o que deixou os níveis dos nutrientes no efluente abaixo dos estabelecidos como referência pela resolução nº 357 do CONAMA, tornando essa água apta para o reuso no cultivo dos peixes ou descarte no ambiente natural.

<p>As the human population increases and the demand for aquaculture increases, aquaculturists are coming up with new ways to mitigate stress in fish to increase their production. Cannabidiol (CBD) is an up and coming nutraceutical that may have potential to reduce stress in not only humans but also other vertebrates such as fish. In this project the effect of CBD on the stress physiology and immunology of Nile tilapia was evaluated both <i>in-vitro</i> and <i>in-vivo</i>. In the <i>in-vitro</i> study, spleen cell proliferation was conducted to observe the effect of CBD on fish T and B cells and were compared to mouse T and B cell proliferation. In the <i>in-vivo</i> study, the fish were reared in a recirculating aquaculture system. The effect of CBD on the stress physiology of the fish in short term and long term were evaluated. Based on the short-term acute study, a longer chronic study was designed where tilapia were fed with and without CBD (0.001% of feed weight) and with and without hydrocortisone stress hormone (0.01% of body weight) every day for four weeks. This experiment compared the various growth and feed utilization parameters as well as physiological and immunological parameters such as, plasma cortisol, blood glucose and protein levels, liver and spleen somatic indices (HSI and SSI, respectively), and lysozyme activity of the fish. From our current research, CBD shows potential in stress modulation and in immune modulation. It may have different effects based on the species, whether they need to enhance their immune response or reduce inflammation to be healthy. It also seems to have had different effect on different parts of the immune system. Hematological parameters were not significantly affected by acute stress. CBD did not make any substantial difference in growth. However, in the presence of stress, CBD was able to lower lysozyme activity down to the normal control levels. By administering the proper dosage of CBD on a case by case basis, health benefits can be achieved. Further investigation into the matter may not just be useful in stress mediation in aquatic organisms but may also have implications in human medicine as well.</p>

Made available in DSpace on 2014-12-17T14:01:59Z (GMT). No. of bitstreams: 1 EmersonESM.pdf: 931852 bytes, checksum: f768665fe7e8aa27e4023b4def16f98d (MD5) Previous issue date: 2009-03-27 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Considering their commercial importance, as these are the species of freshwater fish more commercialized in Brazil, their occurence in different kinds of aquatic environments (lakes, rivers and dams) and for being tolerant to a wide range of variation of various physical parameters and chemical water, the fish species Oreochromis niloticus, Cyprinus carpio and Colossoma macropomum were chosen for this study, furthermore, to test the toxicity we used the herbicide Roundup. The fingerlings of tilapia (Oreochromis niloticus), commun carp (Cyprinus carpio) and tambaqui (Colossoma macropomum) were submitted to the herbicide roundup in the following concentrations: 0.0 (control); 18,06; 19,10; 20,14; 21,18 and 22,22 mg.L-1, 0.0 (control); 13,89; 14,86; 15,83; 16,81 and 17,78 mg.L-1, and 0.0 (control); 18,06; 19,10; 20,14; 21,18 and 22,22 mg.L-1, respectively, three for 96 hours. The LC50 - 96h for O. niloticus, C. carpio and C. macropomum was 21,63, 15,33 and 20,06 mg.L-1 of the herbicide roundup, respectively. The results show that this herbicide is classified as slightly toxic to the three species. The values of dissolved oxygen, pH and temperature recorded in the aquarium control and aquarium experimental of the three fish species have remained without significant variations during the tests, which reduces the possibility of death caused by sudden variations of these parameters during the 96 hours the experiment. The values of LC50 between different species of fish were observed, noting that the species O.niloticus, C. carpio and C. macropomum showed no expressive differences. The values of environmental risk of Roundup were calculated to obtain more stringent parameters in assessing the dangerousness of those on nontargets. The risk of environmental contamination by Roundup for the Nile tilapia, common carp, and tambaqui are low for the lowest application rate (1 L.ha-1) and depths (1.5 and 2.0 m). The dilution of 100%, the highest recommended dose (5 L.ha-1) and depths (1.5 and 2.0 m) the risk is moderate for the three species. The values of the Risk Ratio (QR) were greater than 0,1, indicating that the values of the CAE and LC50 are above acceptable levels and there is a need, this study, a refinement in ecotoxicological tests / Considerando a import?ncia comercial, por se tratarem das esp?cies de ?gua-doce mais comercializadas no Brasil, por ocorrerem em diversos tipos de ambientes aqu?ticos (lagos, rios e a?udes) e ainda por serem tolerantes a um amplo espectro de varia??o de diversos par?metros f?sicos e qu?micos da ?gua, as esp?cies de peixes Oreochromis niloticus, Cyprinus carpio e Colossoma macropomum foram escolhidas para este estudo com a finalidade de testar a toxicidade do herbicida roundup. Diante do exposto, o presente trabalho teve como objetivo determinar a toxicidade aguda e avaliar o risco de contamina??o ambiental dessas tr?s esp?cies de peixes pelo herbicida Roundup. Os alevinos de Til?pia do Nilo (Oreochromis niloticus), Carpa-comum (Cyprinus carpio) e Tambaqui (Colossoma macropomum) foram submetidos ao herbicida roundup nas seguintes concentra??es: 0,0 (controle); 18,06; 19,10; 20,14; 21,18 e 22,22 mg.L-1, 0,0 (controle); 13,89; 14,86; 15,83; 16,81 e 17,78 mg.L-1 e 0,0 (controle); 18,06; 19,10; 20,14; 21,18 e 22,22 mg.L-1, respectivamente, os tr?s por 96 horas. A CL50 96h para O. niloticus, C. carpio e C. macropomum foi de 21,63, 15,33 e 20,06 mg.L-1 do herbicida roundup, respectivamente. Os resultados obtidos demonstram que esse herbicida est? classificado como levemente t?xico para as tr?s esp?cies. Os valores de oxig?nio dissolvido, pH e temperatura registrados no aqu?rio controle e nos aqu?rios experimentais das tr?s esp?cies estudadas mantiveram-se sem varia??es significativas ao longo dos testes, o que reduz a possibilidade de mortalidade causada por varia??es bruscas desses par?metros, durante as 96 horas de experimento. Os valores das CL50 entre as diferentes esp?cies de peixes foram observadas, constatando que as esp?cie Oreochromis niloticus, Colossoma macropomum e Cyprinus carpio n?o apresentaram diferen?as expressivas. Os valores de risco ambiental do Roundup foram calculados para a obten??o de par?metros mais rigorosos na avalia??o da periculosidade desses sobre organismos n?o-alvos. O risco de contamina??o ambiental por roundup para a til?pia do Nilo, carpa-comum e tambaqui s?o baixos para a menor dose de aplica??o (1 L.ha-1) e nas profundidades (1,5 e 2,0 m). Na dilui??o de 100%, na maior dose recomendada (5 L.ha-1) e nas profundidades (1,5 e 2,0 m) o risco ? moderado para as tr?s esp?cies. Os valores de Quociente de Risco (QR) foram maiores que 0,1, indicando que os valores da rela??o CAE e CL50 est?o acima dos n?veis aceit?veis, havendo a necessidade de um refinamento nas an?lises ecotoxicol?gicas