Estudo da produção de biopolímeros via enzimática através de inativação e lise celular e com células viáveis de Beijerinckia sp. 7070

Biopolímeros microbianos são polissacarídeos produzidos por microrganismos. A Beijerinckia sp. 7070 produz o biopolímero denominado clairana®. Biopolímeros microbianos podem ser produzidos utilizando enzimas purificadas (via enzimática). O objetivo desse trabalho foi avaliar a possibilidade de produzir o biopolímero clairana® via enzimática, verificar se a síntese e as enzimas envolvidas na síntese são intra e/ou extracelulares, se as enzimas permanecem ativas no meio e estudar a produção do biopolímero via enzimática, através de inativação e lise celular, e pelo processo convencional com células viáveis. As amostras foram cultivadas com a bactéria em meio líquido e submetidas à morte celular por inativação e lise celular após 30h, através de antibiótico e ultrasom, respectivamente. Amostras foram retiradas após 32, 46 e 54h de incubação, junto com amostras produzidas com células viáveis. O mesmo procedimento foi realizado em 46h e amostras foram coletadas após 48, 52 e 54h de incubação. Os polímeros foram recuperados, secos, pesados e analisados. Os resultados sugerem que é possível produzir o biopolímero clairanaâ via enzimática e que, provavelmente enzimas intra e extracelulares estão envolvidas, permanecendo ativas no meio e começando a atuar antes de 30h de fermentação. O processo via enzimática através de lise foi mais eficiente pois libera os polissacarídeos produzidos intracelularmente para o meio externo.

Caracterização de biopolímeros produzidos por Beijerinckia sp. 7070 em diferentes tempos de cultivo

Biopolímeros são polissacarídeos microbianos. O biopolímero produzido por Beijerinckia sp. 7070 possui comportamento pseudoplástico e apresenta alta viscosidade em baixas velocidades de deformação, conferindo ao polímero excelentes características de suspensão. O objetivo desse trabalho foi caracterizar o biopolímero produzido por Beijerinckia sp. 7070 em diferentes tempos de cultivo, quanto à produção total, produção de polímeros de fibra longa e curta, produtividade, viscosidade e composição química. Os polímeros produzidos em meio YM líquido foram recuperados em diferentes tempos de cultivo, secos e pesados para determinação da produção e produtividade. O tipo de fibra produzido durante o cultivo foi avaliado microscopicamente. Viscosidades aparentes de solução aquosa 1% foram determinadas a 6, 12, 30 e 60rpm, a 25º C, em um viscosímetro Brookfield. A composição do biopolímero foi determinada por cromatografia em camada delgada comparativa. As maiores produções totais encontradas foram em 30 e 72h, a maior produtividade em 48h e a maior viscosidade em 72h. Os polímeros de fibra longa apresentaram uma tendência de tornarem-se mais longos com o tempo. A viscosidade do polímero de fibra longa foi maior que a do de fibra curta. Todos os biopolímeros apresentaram os mesmos componentes (glucose, galactose, fucose e ácido glucurônico) mas em concentrações diferentes.

Bioremediation of PAHs - Limitations and soultions

Introduction 1.1 Occurrence of polycyclic aromatic hydrocarbons (PAH) in the environment Worldwide industrial and agricultural developments have released a large number of natural and synthetic hazardous compounds into the environment due to careless waste disposal, illegal waste dumping and accidental spills. As a result, there are numerous sites in the world that require cleanup of soils and groundwater. Polycyclic aromatic hydrocarbons (PAHs) are one of the major groups of these contaminants (Da Silva et al., 2003). PAHs constitute a diverse class of organic compounds consisting of two or more aromatic rings with various structural configurations (Prabhu and Phale, 2003). Being a derivative of benzene, PAHs are thermodynamically stable. In addition, these chemicals tend to adhere to particle surfaces, such as soils, because of their low water solubility and strong hydrophobicity, and this results in greater persistence under natural conditions. This persistence coupled with their potential carcinogenicity makes PAHs problematic environmental contaminants (Cerniglia, 1992; Sutherland, 1992). PAHs are widely found in high concentrations at many industrial sites, particularly those associated with petroleum, gas production and wood preserving industries (Wilson and Jones, 1993). 1.2 Remediation technologies Conventional techniques used for the remediation of soil polluted with organic contaminants include excavation of the contaminated soil and disposal to a landfill or capping - containment - of the contaminated areas of a site. These methods have some drawbacks. The first method simply moves the contamination elsewhere and may create significant risks in the excavation, handling and transport of hazardous material. Additionally, it is very difficult and increasingly expensive to find new landfill sites for the final disposal of the material. The cap and containment method is only an interim solution since the contamination remains on site, requiring monitoring and maintenance of the isolation barriers long into the future, with all the associated costs and potential liability. A better approach than these traditional methods is to completely destroy the pollutants, if possible, or transform them into harmless substances. Some technologies that have been used are high-temperature incineration and various types of chemical decomposition (for example, base-catalyzed dechlorination, UV oxidation). However, these methods have significant disadvantages, principally their technological complexity, high cost , and the lack of public acceptance. Bioremediation, on the contrast, is a promising option for the complete removal and destruction of contaminants. 1.3 Bioremediation of PAH contaminated soil & groundwater Bioremediation is the use of living organisms, primarily microorganisms, to degrade or detoxify hazardous wastes into harmless substances such as carbon dioxide, water and cell biomass Most PAHs are biodegradable unter natural conditions (Da Silva et al., 2003; Meysami and Baheri, 2003) and bioremediation for cleanup of PAH wastes has been extensively studied at both laboratory and commercial levels- It has been implemented at a number of contaminated sites, including the cleanup of the Exxon Valdez oil spill in Prince William Sound, Alaska in 1989, the Mega Borg spill off the Texas coast in 1990 and the Burgan Oil Field, Kuwait in 1994 (Purwaningsih, 2002). Different strategies for PAH bioremediation, such as in situ , ex situ or on site bioremediation were developed in recent years. In situ bioremediation is a technique that is applied to soil and groundwater at the site without removing the contaminated soil or groundwater, based on the provision of optimum conditions for microbiological contaminant breakdown.. Ex situ bioremediation of PAHs, on the other hand, is a technique applied to soil and groundwater which has been removed from the site via excavation (soil) or pumping (water). Hazardous contaminants are converted in controlled bioreactors into harmless compounds in an efficient manner. 1.4 Bioavailability of PAH in the subsurface Frequently, PAH contamination in the environment is occurs as contaminants that are sorbed onto soilparticles rather than in phase (NAPL, non aqueous phase liquids). It is known that the biodegradation rate of most PAHs sorbed onto soil is far lower than rates measured in solution cultures of microorganisms with pure solid pollutants (Alexander and Scow, 1989; Hamaker, 1972). It is generally believed that only that fraction of PAHs dissolved in the solution can be metabolized by microorganisms in soil. The amount of contaminant that can be readily taken up and degraded by microorganisms is defined as bioavailability (Bosma et al., 1997; Maier, 2000). Two phenomena have been suggested to cause the low bioavailability of PAHs in soil (Danielsson, 2000). The first one is strong adsorption of the contaminants to the soil constituents which then leads to very slow release rates of contaminants to the aqueous phase. Sorption is often well correlated with soil organic matter content (Means, 1980) and significantly reduces biodegradation (Manilal and Alexander, 1991). The second phenomenon is slow mass transfer of pollutants, such as pore diffusion in the soil aggregates or diffusion in the organic matter in the soil. The complex set of these physical, chemical and biological processes is schematically illustrated in Figure 1. As shown in Figure 1, biodegradation processes are taking place in the soil solution while diffusion processes occur in the narrow pores in and between soil aggregates (Danielsson, 2000). Seemingly contradictory studies can be found in the literature that indicate the rate and final extent of metabolism may be either lower or higher for sorbed PAHs by soil than those for pure PAHs (Van Loosdrecht et al., 1990). These contrasting results demonstrate that the bioavailability of organic contaminants sorbed onto soil is far from being well understood. Besides bioavailability, there are several other factors influencing the rate and extent of biodegradation of PAHs in soil including microbial population characteristics, physical and chemical properties of PAHs and environmental factors (temperature, moisture, pH, degree of contamination). Figure 1: Schematic diagram showing possible rate-limiting processes during bioremediation of hydrophobic organic contaminants in a contaminated soil-water system (not to scale) (Danielsson, 2000). 1.5 Increasing the bioavailability of PAH in soil Attempts to improve the biodegradation of PAHs in soil by increasing their bioavailability include the use of surfactants , solvents or solubility enhancers.. However, introduction of synthetic surfactant may result in the addition of one more pollutant. (Wang and Brusseau, 1993).A study conducted by Mulder et al. showed that the introduction of hydropropyl-ß-cyclodextrin (HPCD), a well-known PAH solubility enhancer, significantly increased the solubilization of PAHs although it did not improve the biodegradation rate of PAHs (Mulder et al., 1998), indicating that further research is required in order to develop a feasible and efficient remediation method. Enhancing the extent of PAHs mass transfer from the soil phase to the liquid might prove an efficient and environmentally low-risk alternative way of addressing the problem of slow PAH biodegradation in soil.

High-throughput Analysis By Sp-ldi-ms For Fast Identification Of Adulterations In Commercial Balsamic Vinegars.

Balsamic vinegar (BV) is a typical and valuable Italian product, worldwide appreciated thanks to its characteristic flavors and potential health benefits. Several studies have been conducted to assess physicochemical and microbial compositions of BV, as well as its beneficial properties. Due to highly-disseminated claims of antioxidant, antihypertensive and antiglycemic properties, BV is a known target for frauds and adulterations. For that matter, product authentication, certifying its origin (region or country) and thus the processing conditions, is becoming a growing concern. Striving for fraud reduction as well as quality and safety assurance, reliable analytical strategies to rapidly evaluate BV quality are very interesting, also from an economical point of view. This work employs silica plate laser desorption/ionization mass spectrometry (SP-LDI-MS) for fast chemical profiling of commercial BV samples with protected geographical indication (PGI) and identification of its adulterated samples with low-priced vinegars, namely apple, alcohol and red/white wines.

Chromobacterium Amazonense Sp. Nov. Isolated From Water Samples From The Rio Negro, Amazon, Brazil.

The taxonomic position of a bacterium isolated from water samples from the Rio Negro, in Amazon, Brazil, was determined by using a polyphasic approach. The organism formed a distinct phyletic line in the Chromobacterium 16S rRNA gene tree and had chemotaxonomic and morphological properties consistent with its classification in this genus. It was found to be closely related to Chromobacterium vaccinii DSM 25150(T) (98.6 % 16S rRNA gene similarity) and shared 98.5 % 16S rRNA gene similarity with Chromobacterium piscinae LGM 3947(T). DNA-DNA relatedness studies showed that isolate CBMAI 310(T) belongs to distinct genomic species. The isolate was readily distinguished from the type strain of these species using a combination of phenotypic and chemotaxonomic properties. Thus, based on genotypic and phenotypic data, it is proposed that isolate CBMAI 310(T) (=DSM 26508(T)) be classified in the genus Chromobacterium as the type strain of a novel species, namely, Chromobacterium amazonense sp. nov.

Nitrogen Starvation For Lipid Accumulation In The Microalga Species Desmodesmus Sp.

Recently, to obtain lipids from microalgae has been the object of extensive research, since it is viewed as a promising feedstock for biodiesel production, especially when compared with crops such as soybean and sunflower, in terms of theoretical performance. The reduction of nutrient availability in culture media, especially nitrogen, stresses the microorganisms and affects cell growth, thus inducing lipid accumulation. This is an interesting step in biodiesel feedstock obtention from microalgae and should be better understood. In this study, four levels of nitrogen concentration in the BG-11 culture medium were evaluated in the growth of the chlorophycean microalga Desmodesmus sp. Both cell growth and lipid content were monitored over 7 days of cultivation, which yielded a final cell density of 33 × 10(6) cells mL(-1) with an initial NaNO3 concentration of 750 mg L(-1) in the medium and a maximum lipid content of 23 % with total nitrogen starvation. It was observed that the microalgae presented high lipid accumulation in the fourth day of cultivation with nitrogen starvation, although with moderate cell growth.

Variabilidade espacial e sazonal da concentração de elementos-traço em sedimentos do sistema estuarino de Santos-Cubatão (SP)

Multi-element analyses of sediment samples from the Santos-Cubatão Estuarine System were carried out to investigate the spatial and seasonal variability of trace-element concentrations. The study area contains a rich mangrove ecosystem that is a habitat for tens of thousands of resident and migratory birds, some of them endangered globally. Enrichments of metals in fine-grained surface sediments are, in decreasing order, Hg, Mn, La, Ca, Sr, Cd, Zn, Pb, Ba, Cu, Cr, Fe, Nb, Y, Ni and Ga, relative to pre-industrial background levels. The maximum enrichment ranged from 49 (Hg) to 3.1 (Ga). Mercury concentrations were greater in the Cubatão river than in other sites, while the other elements showed greater concentrations in the Morrão river. Concentrations of Mn were significantly greater in winter and autumn than in summer and spring. However, other elements (e.g. Cd and Pb) showed the opposite, with greater concentrations in summer and spring. This study suggests that seasonal changes in physical and chemical conditions may affect the degree of sediment enrichment and therefore make the assessment of contamination difficult. Consequently, these processes need to be considered when assessing water quality and the potential contamination of biota.

Um novo índice de qualidade das águas para proteção da vida aquática aplicado ao Rio Atibaia, região de Campinas/Paulínia-SP

In this paper a water quality index is developed to subsidize management actions in the Atibaia River for upon protection of aquatic organisms. This index is composed of two measurable environmental parameters normaly, ammonia and dissolved oxygen, the latter representing the contribution of organic matter. Concentrations of these two variables were normalized on a scale from 0 to 100 and translated into statements of quality (excellent, good, regular, bad and very bad). The index was applied to three monitoring points in the Atibaia River and compared to other indices used by the State of São Paulo Environmental Agency (CETESB). The results showed that the degradation in this watershed follows the urban population density. The developed index is more restricted than the other ones routinely used to infer water quality.

Levantamento fitossociológico e caráter sucessional de um trecho de floresta secundária ciliar em Juquitiba, SP, Brasil

The objective of this study was to evaluate the horizontal and vertical structures of tree community in regeneration in a fragment of a secondary riparian forest at approximately 30 years of age and to identify the most abundant species in each fragment of the forest to determine the sucessional stage. An area of 800 m² was subdivided into 16 samples of 10 x 5 m and all individuals with DBH ≥ 1 cm were sampled and identified for the following analyzes: horizontal parameters (DR, FR, DoR, IVC and IVI), vertical parameters (PSR and RNR) and mixed parameters, from of value of increased importance index (IVIa). The survey measured 689 individuals, belonging to 38 families, 74 genus and 109 species. The total density was 8,614 individuals/ha. The index of Shannon´s diversity was 3.99 and the index of Pielou´s equability was 0.85. Tibouchina pulchra, Psychotria suterella and Endlicheria paniculata obtained high values of IVIa. Guarea macrophylla, Gomidesia anacardiaefolia, Xylopia langsdorffiana and Endlicheria paniculata achieved high values of RNT, indicating adequate natural regeneration in the plot. The initial secondary and umbrophylous species showed the highest ecological importance in this fragment of the forest, with the highest values of sociologic position and importance index. Furthermore, the presence of late secondary species in all layers suggest that the studied fragment is in intermediate succession degree.

Classificação e padronização dos tomates cv. Carmem e Débora dentro da CEAGESP - SP

The creation of the Brazilian Program for the Modernization of the Horticulture by the Secretariat of Agriculture and Supplying of the State of São Paulo at CEAGESP, determined the standardization of fruit and vegetables in the follow aspects: degree of coloration, format, calibers, defects and packing. Therefore, the main goal of this research is to correlate the classification given by the Brazilian Program with the one used by the wholesalers at CEAGESP, verifying if the established norms are being fulfilled for cultivar Carmen and Debora (SAKATA SEED). The results showed, that for cultivar Carmem, for the averages of the observed values it does not move away from the norms created by the Program for sizes small and medium. However, for the case of cultivar Debora, the results showed differences between the adopted classifications. The tomatoes were devaluated, because had been commercialized below of the standardization indicated for the Brazilian Program.

Fundações de construções submetidas a esforços de tração em solo de alta porosidade da região de Campinas - SP

This paper presents the behavior of three bored piles conducted in diabasic soil submitted to uplift forces. The piles were built at the site for Experimental Studies in Soil Mechanics and Foundations of UNICAMP, located in the city of Campinas, Brazil. Field tests have already been conducted at the site (SPT, CPT, DMT and PMT), as well as laboratory tests by using sample soils taken from a well up to 17 m deep. The water table is not checked until a depth of 17 m. In order to check the behavior of the piles when submitted to uplift forces, slow static load tests were carried out as the recommendations of NBR 12131. The carrying capacity of these piles was also provided by means of theoretical methods, appropriate for uplift forces, and through semi-empirical methods appropriate for compression forces, considering only the portion of lateral resistance. The values estimated by using the considered methods were compared to those obtained by means of load tests. One of the tested piles was extracted from the soil to be the subject of a study on its geometry.

Composição florística de uma floresta ripária na Reserva Estadual de Porto Ferreira, SP

The point-centred quarter method (63 points) was applied in Porto Ferreira State Reserve (21º49'S and 47º25'W) in an area (1.08ha) on the right margin of Moji Guaçu river, including two woody individuals per quarter - one with DBH < 10cm and at least 130cm high, the other with DBH > 10cm. The results obtained were compared with those published by other authors for a riparian forest (Mata da Figueira) at Moji Guaçu Ecological Station (about 100 km upstream on the same river). At Porto Ferreira 107 species were found, of which 80 were exclusive, compared with the Mata da Figueira where of the 59 species listed, 31 were exclusive. The two area shared 27 common species, thus accounting for a low Sørensen similarity of 48.6%. The great environmental heterogeneity of the floodplains, as well as the degree of anthropic disturbance, could account for this floristic variation. The greatest numbers of species were shown by Leguminosae (20), Myrtaceae (17), Rutaceae (9), Euphorbiaceae (7), and Lauraceae, Meliaceae, Moraceae and Rubiaceae (6 species each). There appears to be little difference at the family level among the periodically flooded and non-flooded forests of the State of São Paulo, but the species show different degreees of preference for habitat. The floristic composition of the two areas presented a mixture of typical species with others of non-flooded forests. The latter would occur on the floodplain probably by a) adaptation of the root system to relatively short flooding periods; b) shorter periods of flooding on the higher points of the microrelief of the floodplain, and c) greater aeration due to running water.

Flora arbustivo-arbórea do fragmento de floresta estacional semidecidual do Ribeirão Cachoeira, município de Campinas, SP

An inventory of the woody flora (trees and shrubs), was carried out in the Ribeirão Cachoeria forest (233.7ha, 650m high, 46°55'58''W, 22°50'13''S), the second largest and best conserved fragment of semideciduous tropical forest in the municipality of Campinas, São Paulo state, Southeastern Brazil. The soil is a red-yellow podsol and the climate is of Köppen's Cwag type. Collections were made from August/1996 to September/1997. Only fertile individuals with a perimeter at breast height of 9cm or greater were included in the survey. One hyndred and seventhy five species were identified, belonging to 119 genera and 49 families. The most important families were Myrtaceae (14 species), Rutaceae and Fabaceae (13), Caesalpiniaceae (11), Solanaceae (9), and Rubiaceae (8). Some species were found for the first time in the region: Tachigali multijuga Benth. and Schoepfia brasiliensis A.DC. The flowering peak for most species was from August to October. Maximum fruit production was from August to November. Most species are zoochoric (58%), but 23% were anemochoric and 19% autochoric. The floristic composition of this forest and another 20 forests from São Paulo state were compared. The results obtained indicate the existence of distinct groups of forests. The most homogeneus group contains forests from the municipality of Campinas with similarity of 40%. This suggests that these forests are possibly fragments of a original continuous forest in the Campinas region.

Lianas da estação ecológica do Noroeste Paulista, São José do Rio Preto/Mirassol, SP, Brasil

Lianas are characteristic, abundant and ecologically important members of tropical forest but they have been neglected in floristics and phytossociological studies. This work presents a floristic survey of the lianas species at Estação Ecológica do Noroeste Paulista (EENP), and a comparison of the list of species recorded in this work with those reported for other fragments of São Paulo state. The EENP (20º48'36'' S and 49º22'50'' W) is at 468 m of altitude and comprises an area of 168,43 ha, divided into three fragments of vegetation. Samples of lianas were collected in the interior and along the edges of the forest fragments. It was identified 105 species: 99 Magnoliopsida (60 genera and 22 families); six Liliopsida (three genera and three families). The richest families in species comprised 59% of the total of lianas sampled. The dendrogram of similarity showed a low similarity between the forest situated in the littoral (Atlantic Forest) and those located in the interior of the state of São Paulo. Some other authors, also analysing the similarity of forest of the interior and Atlantic Forest of São Paulo state, but considering only the trees reported similar result.

Produção de serapilheira e decomposição do material foliar em um cerradão na Estação Ecológica de Jataí, município de Luiz Antônio, SP, Brasil

We estimate litter production and leaf decomposition rate in a cerradão area, physiognomy little studied and very threatened in São Paulo State. During the period of study, litter production was 5646.9 kg.ha-1.year-1, which the 'leaf' fraction corresponded to 4081.2 kg.ha¹.year¹; the 'branch' fraction, to 1066.1 kg.ha-1.year-1; the 'reproductive structures' fraction, to 434.1 kg.ha-1.year-1; and the 'miscellaneous' fraction to 65.5 kg.ha-1.year-1. Litter production was highly seasonal and negatively correlated with relative humidity and air temperature. Leaf production was negatively correlated with relative humidity, rainfall, and air temperature. There was no significant difference between litter production found in this study and those in two other sites with cerradão and semideciduous forest, but these physiognomies differed significantly from the cerrado sensu stricto. Leaf decomposition rate (K) was 0.56. Half-life of the decomposing material was 1.8 years and turnover time was 2.3 years.