Levantamento e avaliação de critérios para a ampliação de escala da produção de biossurfactantes utilizando melaço como substrato

Biosurfactants are amphiphilic molecules synthesized by microorganisms such as bacteria, yeast or filamented fungi cultivated in various carbon sources among sucrose and hydrocarbons. These molecules are composed by a hydrophilic and hydrophobic part. They operate mostly at interfaces of fluids of different polarities. Because of this characteristic, they are potentially employed in numerous industries, such as the textile, medical, cosmetics, food and mainly in the petrochemical ones. Therefore industry has interest in developing new biosurfactant production processes in high scale, in order to become them economically competitive when compared to synthetic biosurfactants. This work aims to evaluate the biosurfactant production applying a non-conventional substrate sugar cane molasses proceeding from the sugar industry thus reducing the production costs. The strain identified as AP029/GLIIA, isolated from oil wells in Rio Grande do Norte state and used in these experiments belongs to the culture collection of Antibiotics Department of UFPE. The fermentation were carried out using different conditions according to a factorial planning 24 with duplicate at center point, in which the studied factors were molasse concentration, nitrate concentration, agitation and aeration ratio. The experiments were performed in a shaker at 38ºC of temperature. Samples were withdrawn in regular periods of time of up to 72 hours of fermentation in order to analyze substrate consumption, cellular concentration, superficial tension, critical micelle dilution (CMD-1 e CMD-2) as well as extracelullar protein production. The results showed a production of 3,480 g/L of biomass, a reduction of 41% on superficial tension, 67% of substrate consumption and 0,2805 g/L of extracellular protein

Recuperação e purificação de ramnolipídeos produzidos por pseudomonas aeruginosa P029-GVIIA utilizando melaço de cana como substrato

Biosurfactants are molecules produced by microorganisms mainly bacteria as Pseudomonas and Bacillus. Among the biosurfactants, rhamnolipids play an important role due to their tensoactive as well as emulsifying properties. Besides can be produced in a well consolidated way the production costs of biosurfactants are quite expansive mainly if downstream processing is goning to be considered. Actually, attention has been given to identification of biosurfactants as well as optimization of its fermentative processes including downstream ones. This work deals with the development of strategies to recovery and purification of rhamnolipids produced by Pseudomonas aeruginosa P029-GVIIA using sugar-cane molasses as substrate. Broth free of cells was used in order to investigate the best strategies to recovery and purification produced by this system. Between the studied acids (HCl and H2SO4) for the acid precipitation step, HCl was the best one as has been showed by the experimental design 24. Extraction has been carried out using petroleum ether and quantification has been done using the thioglycolic acid method. Adsorption studies were carried out with activated carbon in a batch mode using a 24 experimental design as well as combined with an hydrophobic resin Streamline Phenyl aiming to separate the produced biosurfactant. Biosurfactant partial identification was carried out using High Performance Liquid Chromatography (HPLC). Experiments in batch mode showed that adsorption has been controlled mainly by pH and temperature. It was observed a reduction of 41.4% for the liquid phase and the solid phase it was possible to adsorb up to 15 mg of rhamnolipd/g of activated carbon. The kinetics of adsorption has been well fitted to a pseudo-first order reaction with velocity constant (k1) of 1.93 x 10-2 min-1. Experiments in packed bed ranging concentration on eluent (acetone) has been shown the highest recovery factor of 98% when pure acetone has been used. The combined effect if using activated carbon with an hydrophobic resin Streamline Phenyl has been shown successful for the rhamnolipids purification. It has been possible to purify a fraction of the crude broth with 98% of purity when the eluted of activated carbon packed bed was used with pure acetone

Decaimento bacteriano em lagoas de estabilização no Nordeste brasileiro

Stabilization pond system consisting in more sewage treatment used in Rio Grande do Norte (RN), Brazil, representing about 90% of all systems. Fecal bacteria are removed mainly facultative ponds and in maturation ponds. Many factors influence bacterial decay, such as the levels of pH and DO, temperature, light intensity, HDT and nutrient availability. The bacterial decay rate (Kb) is calculated considering many variables, but the hydraulic regime is a significant influence for microorganisms removal, and the dispersed flow which best characterizes a stabilization pond. However, some authors developed equations for the Kb accordant plug flow and complete mixing. This research study aimed to evaluate the bacterial decay of fecal coliform and Enterococcus sp. in stabilization ponds designed to treat domestic sewage, full-scale, in RN. All systems have assessed pretreatment, a facultative pond (LF) followed by two maturation (LM1 and LM2). The parameters availed were: temperature, pH, DO, BOD5, COD, fecal coliform, Enterococcus sp., Chlorophyll a, total suspended solids, fixed and volatile. In general, there were not significant differences for pH, DO and temperature in the ponds, except for the new systems, since they have low flow and hydraulic loads. The removal of organic matter in the ponds was low, about 70%, and nearly all are overloaded organic and operational problems. The bacterial removals were low, with average 96% for LF for fecal coliform, and 98% for Enterococcus sp.; LM1 were in itself a removal for fecal coliform about 71%, and 81% for Enterococcus sp.; LM2 have efficiency of 69% for fecal coliform, and 68% for Enterococcus sp. The equation proposed by Von Sperling (1999), according to the dispersed flow regime, generated empirical values of Kb more approximate to calculated values of Kb. On average, the calculated Kb to coliforms in the LF was 0.31 d-1, and for both maturation ponds were 0.35 d-1. For Enterococcus sp. the average was 0.40 d-1 for LF, 0.55 d-1 for LM1, and 0.58 d-1 for LM2. These results also showed that the Kb obtained in full-scale systems are smaller than those found in pilot-scale ponds. Moreover, one can say that the equation proposed by Marais (1974), according to the complete-mix regime, overestimates Kb. Actual results of Kb indicated that fecal coliforms are more resistant to adverse conditions present in stabilization ponds than Enterococcus sp., therefore, an indicator of microbiological safety and efficiency. The factors significant interventions in the rate of bacterial decay were concentrations of COD, the organic loading and HDT. The few Kb relationship between pH, DO and temperature were not significant. Finally, we conclude that it s essential to correct operation and maintenance, for not performing these activities is one of the main factors contributing to low rates of bacterial decay.

O que controla o crescimento do Fitoplâncton em lagoas costeiras tropicais? um modelo para a lagoa de Extremoz (RN)

The phytoplankton growth is dependent of several abiotic (nutrients, temperature) and biotic (predation by zooplankton) variables. In this work, a mathematical model was developed in Stella software to understand the planktonic dynamics of Extremoz Lagoon (RN) and to simulate scenarios of different environmental conditions. Data were collected monthly at two points of the lagoon. The state variables are phytoplankton and zooplankton and forcing variables are nitrogen, phosphorus and temperature. The results show that: a) the model are well coupled, especially when some constants assume different values; b) simulated nutrient concentrations reduction decreases phytoplankton biomass, but the increase of nutrients does not stimulate the growth; c) changes in the temperature does not change the phytoplankton biomass; d) changes in zooplankton biomass affect directly and reduces the phytoplankton, indicating a topdown control mechanism; e) changes in the nutrient concentration modified the biomass of zooplankton suggesting a rapid flow of energy between nutrients, phytoplankton and zooplankton and a ecosystem likely arranged in an inverted pyramid (higher concentration of zooplankton than phytoplankton)

Susceptibility of Saccharomyces cerevisiae and lactic acid bacteria from the alcohol industry to several antimicrobial compounds

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Caracterização molecular de estafilococos isolados de vacas com mastite subclínica e ordenhadores

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Período de infectividade de animais inoculados experimentalmente com Yersinia sp

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Exame microbiológico de sorvetes não pasteurizados

Realizou-se exame microbiológico em 24 amostras de sorvetes não pasteurizados, todos preparados de maneira não industrial e à base de leite (creme, nata, chocolate), fabricados por 12 sorveterias diferentes da cidade de Araraquara, SP. Colheram-se duas amostras de cada sorveteria com intervalo de 15 dias entre as colheitas. Realizaram-se as seguintes provas: contagem de bactérias aeróbicas ou facultativas mesófilas e psicrófilas e de Staphylococcus aureus; determinação do Número Mais Provável (NMP) de coliformes totais e fecais e da presença de Salmonella. As técnicas utilizadas foram aquelas convencionalmente usadas para tais determinações. Não foi encontrada Salmonella em nenhuma das amostras e de cerca de 16,6% delas isolou-se Staphylococcus aureus. em proporções variáveis verificou-se a presença de microrgarnismos deteriorantes e daqueles indicadores de poluição de origem fecal.

Indoor Air quality related to occupancy at an air-conditioned public building

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Análise farmacognóstica e atividade antibacteriana de extratos vegetais empregados em formulação para a higiene bucal

A utilização de extratos vegetais vem se tornando uma alternativa importante para a prevenção de doenças periodontais. Este trabalho objetivou desenvolver uma formulação de enxagüatório bucal, contendo, em associação, extratos hidroalcoólicos de Rosmarinus officinalis, Plantago major, Tabebuia impetiginosa, Achillea millefollium e Nasturtium officinale; avaliar sua composição farmacognóstica e sua atividade antibacteriana, como também da fórmula proposta. Foram realizados estudos de pré-formulação e análises farmacognósticas para as espécies vegetais. A atividade antibacteriana in vitro foi observada por meio dos métodos de difusão em disco de papel, por hole- plate e por template, frente a Staphylococcus aureus, Bacillus subtilis, Escherichia colik, Enterococcus faecalis e Pseudomonas aeruginosa. A concentração inibitória mínima (CIM) foi determinada por meio do método de macrodiluições sucessivas em caldo. Os resultados obtidos apresentaram-se de acordo com o histórico farmacognóstico das drogas estudadas. Todas as bactérias foram inibidas pelos extratos, observando-se que as espécies S. aureus e B. subtilis mostraram, aparentemente, maior sensibilidade. A CIM variou, em relação a sensibilidade de cada espécie bacteriana estudada, de 312,5 µL/mL a 1250 µL/mL para os extratos vegetais e de 625 µL/mL a 2500 µL/mL para o enxaguatório bucal. São necessários estudos complementares para a confirmação da eficácia deste produto e sua utilização na prevenção de doenças periodontais.

Detection of mixed microbial biofilms on central venous catheters removed from Intensive care Unit Patients

Cateteres venosos centrais inseridos em pacientes internados em unidade de terapia intensiva foram avaliados por métodos microbiológicos (cultura semi-quantitativa) e microscopia eletrônica de varredura a fim de detectar adesão microbiana e correlacionar com a cultura de sangue. Durante o período de estudo, foram avaliados 59 pacientes com cateter venoso central. A idade dos pacientes, sexo, sítio de inserção e tempo de permanência do cateter foram anotados. O cateter era de poliuretano não tunelizado e de único lúmen. O sangue para cultura foi coletado no momento da remoção do cateter. de 63 pontas de cateteres, 30 (47,6%) foram colonizadas e a infecção encontrada em 5 (23,8%) cateteres. A infecção foi mais prevalente em 26 pacientes (41,3%) com cateteres inseridos em veia subclávia do que nos 3 (3,2%) inseridos em veia jugular. A infecção foi observada com mais freqüência em cateteres com tempo de permanência maior do que sete dias. Os microrganismos isolados incluíram 32 estafilococos coagulase-negativa (29,7%), 61 bactérias Gram-negativas (52,9%), 9 estafilcocos coagulase-positiva (8,3%) e 3 leveduras (2,7%). Como agentes causais de infecções em unidade de terapia intensiva foram isolados E. aerogenes, P. aeruginosa, A. baumannii. Os antimicrobianos com maior atividade in vitro contra as bactérias Gram-negativas foram o imipenem e contra as Gram-positivas vancomicina, cefepime, penicilina, rifampicina e tetraciclina. As análises por microscopia eletrônica de varredura revelaram biofilmes sobre a superfície de todos os cateteres examinados.

Atividade antibacteriana do óleo essencial de Melampodium divaricatum (Rich.) DC., Asteraceae

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evolução do reparo por excisão de nucleotídeo

The phylogeny is one of the main activities of the modern taxonomists and a way to reconstruct the history of the life through comparative analysis of these sequences stored in their genomes aimed find any justification for the origin or evolution of them. Among the sequences with a high level of conservation are the genes of repair because it is important for the conservation and maintenance of genetic stability. Hence, variations in repair genes, as the genes of the nucleotide excision repair (NER), may indicate a possible gene transfer between species. This study aimed to examine the evolutionary history of the components of the NER. For this, sequences of UVRA, UVRB, UVRC and XPB were obtained from GenBank by Blast-p, considering 10-15 as cutoff to create a database. Phylogenetic studies were done using algorithms in PAUP programs, BAYES and PHYLIP package. Phylogenetic trees were build with protein sequences and with sequences of 16S ribosomal RNA for comparative analysis by the methods of parsimony, likelihood and Bayesian. The XPB tree shows that archaeal´s XPB helicases are similar to eukaryotic helicases. According to this data, we infer that the eukaryote nucleotide excision repair system had appeared in Archaea. At UVRA, UVRB and UVRC trees was found a monophyletic group formed by three species of epsilonproteobacterias class, three species of mollicutes class and archaeabacterias of Methanobacteria and Methanococci classes. This information is supported by a tree obtained with the proteins, UVRA, UVRB and UVRC concatenated. Thus, although there are arguments in the literature defending the horizontal transfer of the system uvrABC of bacteria to archaeabacterias, the analysis made in this study suggests that occurred a vertical transfer, from archaeabacteria, of both the NER genes: uvrABC and XPs. According the parsimony, this is the best way because of the occurrence of monophyletic groups, the time of divergence of classes and number of archaeabacterias species with uvrABC system

Análise comparativa dos genes de reparo do DNA em Gluconacetobacter diazotrophicus e Gluconobacter oxydans

Gluconacetobacter diazotrophicus é uma alfa-proteobactéria Gram-negativa, tolerante a meios ácidos, fixadora de nitrogênio atmosférico e foi a primeira bactéria diazotrófica endofítica isolada da cana-de-açúcar. Por sua vez, Gluconobacter oxydans, também alfa-proteobactéria Gram-negativa, possui a capacidade de oxidar incompletamente alcoóis e carboidratos. Ambas de interesse biotecnológico e industrial, essas bactérias tiveram seus genomas seqüenciados completamente em 2007. Desta forma, foi de interesse desse trabalho analisar e comparar os genes de reparo do DNA devido sua importância na manutenção da integridade genômica. Sendo assim, as vias de reparo presentes nos dois organismos foram identificadas, utilizando como base uma terceira alfa-proteobactéria, a Caulobacter crescentus, cujos genes de reparo foram descritos por um trabalho anterior e também os genes bem estabelecidos para o reparo do DNA em Escherichia coli. Para esse estudo, um banco de dados contendo ortólogos para os genes de reparo de DNA encontrados nos organismos foi criado e análises comparativas por similaridade usando o pacote Blast e o software Clustal foram feitas. Este estudo demonstrou que as principais vias de reparo ao DNA reparos por excisão, reparo direto, reparo recombinacional e reparo pelo sistema SOS estão presentes nos organismos analisados, demonstrando, na maioria das vezes, boa similaridade com E. coli. Interessantemente, foram encontradas duplicações gênicas nos quais uma das cópias estava presente no cromossomo e a outra, no plasmídeo, como no caso de UvrD, DnaE e Ssb, possivelmente caracterizando eventos de transferência lateral. Por fim, uma grande novidade foi a identificação de ortólogos para RecB em G. diazotrophicus e G. oxydans e de ortólogos duplicados de RecD em G. diazotrophicus. Até o momento, não havia sido relatada a presença de membros da via de iniciação RecBCD do reparo recombinacional em alfaproteobactérias

Estudos ecogenômicos e bioprospectivos de Shewanella ssp

Bacteria trom Shewanella and Geobacter ganera are the most studied iron-reducing microorganisms particularly due to their electron transport systems and contribution to some industrial and environmental problems, including steel corrosion, bioenergy and bioremediation of petroleum-impacted sites. The present study was focused in two ways: the first is an in silico comparative ecogenomic study of Shewanella spp. with sequenced genomes, and the second is an experimental metagenomic work to detect iron-reducing Shewanella through PCR-DGGE of a metabolic gene. The in silico study resulted in positive correIation between copy number of 16S rDNA and genome size in Shewanella spp., with clusters of rrn near lhe origin of replication. This way, the genus is inferred as opportunist. There are no compact genomes and their sequences length varied, ranging from 4306142 nt in S. amazonensis SB2B to 5935403 nt in S. woodyi ATCC 51908, without correIation to temperature range characteristic of each specie. Intragenomic 16S rDNA sequences possess little divergence, but reasonable to resuIt in different phyIogenetic trees, depending on the sequence that is chosen to compare. For moIecuIar detection of iron-reducing Shewanella, it is proposed the mtrB gene as new biomarker. because it codes to a fundamental protein at Fe (III)-reduction. The specific primers were designed and evaluated in silico and resulted in a fragment of 360 pb. In the second study, these primers were tested in a genomic sample from S. oneidensis MR-1, amplifying the expected region. After this successfuI resuIt, the primer set was used as a tool to assess the iron-reducing communities of ShewaneIla genus under an environmental stress, i.e. crude oil contamination in mangrove sediment in Rio Grande do Norte State (Brazil). The primers presented high specificity and the reactions performed resulted in one single band of ampIification in the metagenomic samples. The fingerprinting obtained at DGGE reveaIed temporal variation of Shewanella spp. in analyzed samples. The resuIts presented show the detection of a biotechnological important group of microorganisms, the iron-reducing Shewanella spp. using a metabolic gane as target. It is concluded there are eight or more 16S rDNA sequences in Shewanella genus, with little divergence among them that affects the phylogeny; the pair of primers designed to ampIify mtrB sequences is a viable alternative to detect iron-reducing ShewanelIa in metagenomic approaches; such bacteria are present in the mangrove sediment anaIyzed, with temporal variations in the samples. This is the first experimental study that screened the iron-reducing Shewanella genus in a metagenomic experiment of mangrove sediments subjected to oil contamination through a key metabolic gene