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This study assessed the adhesion of Pseudomonas aeruginosa UFPEDA 416 (ATCC 27853) exposed to curcumin-mediated Photodynamic Antimicrobial Chemotherapy (PACT). Initially, the Minimum Inhibitory Concentration of curcumin was determined (0.63 μg/mL). After, P. aeruginosa UFPEDA 416 was exposed to the MIC solution of curcumin and then irradiated with a high-power blue LED light at 480 nm for 20, 40, 60 and 120 minutes (28.03 mW/cm2). At each interval, 100 µL aliquots were transferred to microtubes containing Müeller-Hinton broth and incubated at 30ºC for 24 h. The biofilm formed was quantified by the crystal violet method. The adherence rate was calculated using the difference in the absorbance of the treatment, compared to the control. There was a reduction in adhesion capacity from 40.8% in the first 20 minutes to approximately 11% after 60 minutes. The reduction in the adhesion rate accompanied the decrease in planktonic cells, determined from the measurement of the absorbance of the supernatant at 620 nm. The results indicated action by bacteriostasis.

Molecules with antimicrobial properties, such as those found in non-antibiotic pharmaceuticals, may pressure bacteria to adapt to the environment. In this work, five of the most sold non-antibiotic pharmaceuticals in Brazil and in Egypt in 2020 (losartan 50 mg, acetylsalicylic acid 100 mg, diclofenac 50 mg, paracetamol 750 mg and metformin 500 mg and dipyrone 100 mg/mL), were tested against the wild strain Pseudomonas aeruginosa TGC04, previously isolated by our group and which has been characterized over the years. We evaluated inhibition of biofilm adhesion over 48 h using the crystal violet test. Exposure of P. aeruginosa TGC04 to the compounds promoted a similar moderate reduction in adherence for all the pharmaceuticals tested, amounting to about 60%. This indicated that the cells were also moderately adherent. However, although all non-antibiotic pharmaceuticals did not produce a biocidal effect on the wild strain, intense antibiofilm activity was observed, except for paracetamol.

Os biofilmes são agregados microbianos complexos que representam risco à saúde porque inúmeros patógenos são capazes de colonizar superfícies bióticas e abióticas, conferindo a estes microrganismos, maior resistência a antibióticos e outros agentes antimicrobianos. Além disso, os biofilmes são responsáveis por perdas na indústria, bem como pelo processo de biodeterioração de diferentes materiais. Visando formas de inibir ou erradicar biofilmes, diferentes métodos são propostos, incluindo a associação entre eles. Esta revisão foi elaborada por discentes de Pós-Graduação em Biologia Celular e Molecular da UFPB, como trabalho final da Disciplina “Biofilmes microbianos” e traz exemplos e perspectivas de tratamentos físicos, químicos e biológicos no controle destas estruturas.

Pyocyanin is an active redox phenazine of intense blue color and specific to Pseudomonas aeruginosa. The synthesis of the molecule confers different benefits to the bacterium. Pyocyanin can control its growth and persistence in environments with high nutritional pressures, forming biofilms. As well, synthesis of pyocyanin enables P. aeruginosa tolerate and uptake highly toxic compounds such as polycyclic aromatic hydrocarbons (PAHs), considered the most dangerous compounds among all molecules found in crude oil and petroderivatives. Additionally, pyocyanin increases the bioavailability of PAHs and its metabolites are used to synthetize crucial molecules for the biodegradation of other PAHs. On the other hand, oil hydrocarbons can serve as oxygen vectors during the synthesis of pyocyanin, contributing to the sustainability of the biodegradation process. This review is a compilation of recent advances reported in the literature about the relationship between pyocyanin expression and the hydrocarbonoclastic activity of P. aeruginosa. This characteristic, for the pyocyanin-deficient strains, is important for the degradation of PAHs, a topic that has been unevenly studied.

Pseudomonas aeruginosa is a ubiquitous fluorescent, rod-shaped pseudomonad, with a high metabolic capacity, and potential for application in processes to remove recalcitrant compounds such as petroleum hydrocarbons (PHC) from the environment. The bacterium persists in sites with highly selective pressures such as those contaminated by PHC. One of the bacterium's strategies is to colonize biofilms which enhance its protection from toxic compounds and favor oil uptake. It is the most prevalent microbe at sites impacted by PHC owing to the use of aliphatic hydrocarbons to form biofilms and other metabolites crucial for the uptake and degradation of crude oil. P. aeruginosa could be useful in biofilm-mediated bioremediation; however, it has been poorly explored in the last ten years. This systematic study addresses recent research on the application of P. aeruginosa/pseudomonads biofilms in bioremediation The studies come from Asia and Africa and emphasized the formation of biofilm by P. aeruginosa and other pseudomonads as crucial elements in the detoxification process of the environment.

Efluentes líquidos gerados por laboratórios de pesquisa não recebem a devida atenção em países em desenvolvimento, representando riscos em potencial em termos de contaminação de corpos hídricos superficiais e subterrâneos. Esta minirrevisão foi elaborada por discentes de Pós-Graduação em Biotecnologia, como trabalho final da Disciplina que trata de impactos ambientais e estratégias de transformação dos resíduos por atividade biológica. O documento disserta especialmente sobre o impacto destes resíduos em águas marinhas, bem como a perturbação dos serviços ecossistêmicos.

Pyocyanin is a pigment produced by 95% of Pseudomonas aeruginosa strains and exhibits antimicrobial properties that can be used for different purposes. In this work, PMMA-based nanoparticles that were encapsulated into 200 µg/mL of pyocyanin (Np-Pyo) were produced by the nanoprecipitation method. They were evaluated with respect to antifouling activity against Aspergillus sp. and Penicillium sp. With an encapsulation efficiency of 56%, the NpPyo remained stable for 90 days. Their characteristics were satisfactory for the following parameters: average size (616.90±38.30 nm; blank: 282.58±22.89 nm), polydispersion index (0.51±0.01; blank: 0.45±0.78), zeta potential (-5.13±0.41 mV; blank: -6.44±1.12 mV) and pH (6.18±0.03; blank: 6.42±0.01). The in vitro biofilm formation assay was performed on dolomite coupons measuring 1 cm2, on which the formulation was applied. There were tested conditions with and without immersion for 72h at 30 ºC. In the tests with the immersed coupons, there was fungal colonization; this was, however, lower than that observed in the control. A. niger decreased by 3 log units. No growth was observed on the coupons that were not immersed. The results were promising and demonstrated viability as a means of antifouling protection, particularly on dry surfaces.

Este trabalho avaliou a degradação de gasolina em um solo argiloso utilizando um consórcio bacteriano anaeróbio. Foram preparados 11 microcosmos contendo 300 g do solo esterilizado, contaminado com gasolina 5% (v/m) aos quais a degradação do combustível foi avaliada por 30 dias, variando-se a concentração de fertilizante (0; 25 e 50 mg/Kg de solo) e etanol (0,0; 12,5 e 25,0 mL/Kg de solo). Os microcosmos receberam 30 g do inóculo composto do solo contendo o consórcio formado por bactérias heterotróficas, redutoras de sulfato e hidrocarbonoclásticas. Foram testadas duas condições de umidade, 20 e 40%. Descontadas as perdas abióticas, o consórcio removeu entre 20-60% da gasolina e a associação do bioaumento com o bioestímulo se mostrou a melhor estratégia. A adição de fertilizante foi a única variável com efeito significativo no processo. Independente do teor de umidade, a remoção do contaminante demonstrou um padrão similar, entretanto concentração do consórcio foi maior quando o teor de água foi 40%. Embora o teste de ecotoxicidade indicar a presença de metabólitos tóxicos, os resultados sugerem que a remoção de gasolina em condições anaeróbicas pode ser efetiva na ausência de um cossolvente, desde que sejam atendidas as condições nutricionais mínimas requeridas pelos microrganismos.

Every animal needs safe water for drinking. The quality of this water is essential for welfare. This study discusses on the quality of water furnished to about 50 domestic animals based on the categorization of species practiced by residents at São Rafael slum, located in the city of João Pessoa, Brazil. The drinking water comes from three sources: municipal supply system, bottled mineral water and directly from the Jaguaribe River. The water perceived as safest was the same as offered to pets. Domestic animal water samples were tested and rated as not recommended in more than 65% of the samples. The density of thermotolerant coliforms ranged between 0 and 11,000 cells/100 mL. One third of the samples considered unsuitable for animal watering were also outside the recommended pH. In addition, a differential behavior was observed in the choice of water source according to the caretaker's perception of safety. Dogs and cats, considered family members, received the same water as consumed by the family. Even these, 38.5% of the samples, were rated as not recommended and represented a risk to the health of all those who consumed it. As well, a need for good practices of hygiene in water management and cleaning of water dishes, tanks and troughs was seen.

Antimicrobial activity of pyocyanin against competing organisms of Pseudomonas aeruginosa is related to the oxidative stress that the compound promotes in susceptible cells. The objective of this work was to produce, extract and verify the activity of pyocyanin in planktonic and sessile forms from clinical strains, Staphylococcus aureus UFPEDA 02 and Escherichia coli UFPEDA 224. About 600 µg/mL of pyocyanin were obtained. The planktonic cells were highly sensitive. The MIC determined for S. aureus UFPEDA 02 and E. coli UFPEDA 224 were 18.75 and 37.5 µg/mL, respectively. The pyocyanin demonstrated biocidal effect against S. aureus UFPEDA 02. On the other hand, pyocyanin was not active in either sessile strain. The presence of the pigment allowed a greater adherence of the strains, forming more robust biofilms compared to the control. S. aureus UFPEDA 02 and E. coli UFPEDA 224 presented moderate and high hydrophobicity, respectively. Glass and dolomite surfaces were tested in the in vitro biofilm test. Both strains formed the biofilm better on the dolomite surface, obtaining a cell concentration (MPN/cm2) in the order of 3 log units after 48h of incubation.

Carvacrol has been recognized as an efficient growth inhibitor of food pathogens. However, carvacrol oil is poorly water-soluble and can be oxidized, decomposed or evaporated when exposed to the air, light, or heat. To overcome these limitations, a carvacrol nanoemulsion was developed and its antimicrobial activity against food pathogens evaluated in this study. The nanoemulsion containing 3% carvacrol oil, 9% surfactants (HLB 11) and 88% water, presented good stability over a period of 90 days. In general, the carvacrol nanoemulsion (MIC: 256 µg ml-1 for E. coli and Salmonella spp., 128 µg ml-1 for Staphylococcus aureus and Pseudomonas aeruginosa) exhibited improved antimicrobial activity compared to the free oil. The carvacrol nanoemulsion additionally displayed bactericidal activity against Escherichia coli, P. aeruginosa and Salmonella spp. Therefore, the results of this study indicated that carvacrol oil nanoemulsions can potentially be incorporated into food formulations, wherein their efficacy for the prevention and control of microbial growth could be evaluated.

The trans-isomer of resveratrol is a naturally occurring bioactive compound, and it is important because of its biological properties, which are beneficial to health. In recent years, interest in the antimicrobial activity of this molecule has grown; little is known, however, about its antibiofilm activity. This systematic review focused on the advances in research on the topic, in the decade up to 2021. With most studies coming from Asia, research on trans-resveratrol, alone or associated, have shown potential for exploration by the food industry. Low concentrations of the compound exhibit activity against biofilm formation by pathogenic bacteria whose inhibition mechanisms are multifactorial and involve quorum sensing. Few bacterial species, however, have been investigated up to now.

Esta nota técnica descreve um rápido, simples e confiável método detecção do potencial de assimilação e transformação de herbicidas por Pseudomonas aeruginosa utilizando o indicador redox 2,6-diclorofenol indofenol (DCPIP). Isolados hidrocarbonoclásticos selvagens de P. aeruginosa foram cultivados em meio mineral adicionado com dois herbicidas amplamente empregados no cultivo de cana-de-acúcar, ametrina e hexazinona. Dentro de 120h de incubação à 30ºC, a degradação ocorreu entre 48 e 96h. Observou-se também que quanto maior concentração de piocianina produzida pelo isolado, mais cedo o indicador detectou a degradação do herbicida. Os resultados indicaram que P. aeruginosa isolados de ambientes contaminados por moléculas complexas, podem ser uma opção como agente de biorremediação de outros compostos tóxicos do ambiente, incluindo as triazinas. A presença de piocianina demonstra um maior potencial.

Patulin (PAT) is a natural product isolated from several species of fungi. Here, we evaluated the effect of PAT (62.5–4,000 ng/ml) in lipopolysaccharide (LPS)-activated murine peritoneal macrophages. Cell viability assay showed that PAT at concentrations up to 250 ng/ml did not affect macrophage viability. PAT (250 ng/ml) significantly reduced LPS-induced nitric oxide production (by 98.4%), inducible nitric oxide synthase (iNOS) expression (by 83.5%), and iNOS messenger ribonucleic acid expression (by 100.0%). Moreover, PAT significantly reduced LPS-induced interleukin-1β (by 80.6%), cluster of differentiation (CD) 69 (by 63.1%), and Toll-like receptor (TLR) 4 (by 91.9%) protein expression. Finally, PAT significantly reduced LPS-triggered phosphorylation of all mitogen-activated protein kinases (MAPK) assessed: extracellular signal-regulated kinase (ERK; by 89.5%), c-Jun N-terminal kinase (JNK; by 77.5%), and p38 (by 72.3%). Taken together, these data suggest that PAT downregulates acute inflammatory response, inhibiting nitric oxide production by suppressing CD69-TLR4/ERK-JNK-p38 MAPKs/Nos2/iNOS signaling pathway.

Light energy is known to be used to combat microbial growth. Photodynamic antimicrobial chemotherapy (PACT) has the potential to use different naturally-occurring compounds, such as photosensitizers. Curcumin is an example of a molecule of interest in different areas under different optics. This systematic review surveys the aims and scope of research on curcumin-mediated PACT published between January 2011 and December 2020. The search was carried out in MEDLINE, PubMed, EMBASE and Periódicos CAPES databases employing the keywords “Photodynamic antimicrobial chemotherapy”, “photosensitizer”, “curcumin” and the descriptor “Light-Emitting Diode”. It was observed that in the last decade little material meeting these criteria was published. Brazilian institutions concentrated most of their studies on cytotoxic activity. The most recent work, however, focused on antibiofilm activity. Gram-positive bacteria are more sensitive to curcumin-mediated PACT over a short wavelength range. Different concentrations and exposure time of the photosensitizer were evaluated, but the amount of information is still insufficient to establish the best treatment condition as the number of tested pathogens is still poor.

International audience; This work aimed to evaluate the effect of acid mist on microbial communities developed on mortar with same composition from the wall of the National Museum in Rio de Janeiro. Five autochthonous microbial groups were inoculated on coated with acrylic paint and uncoated mortar surfaces and submitted to acid mist or water mist for 100 days. A heterogeneous biofilm was well developed in uncoated coupons under acid mist. Molecular analysis revealed that the bacterial communities were structured according to the incubation condition. The impact of coating on bacterial community structure was more important than the acid mist effect. The eukaryotic community structure was not affected by the acid mist, whereas the impact of coating by paint has been observed. The alpha diversity indexes also varied according to the treatment. H’ and D were higher for uncoated conditions, particularly when submitted to acid mist. Actinobacteria, Chloroflexi, Cyanobacteria, Firmicutes and Proteobacteria were dominant. Additionally, eukaryotic community was not affected by the acid mist and paint and Tremellomycetes dominated.

PURPOSE: Certain ocular resident or pathogenic microbes may remain viable in the presence of multi-purpose disinfectant solutions (MPDSs), subsequently developing biofilms inside contact lens storage cases (CLSCs) which pose a risk of infection to wearers. This study evaluated the formation of ocular microbiota biofilms exposed to three top selling MPDS. METHODS: Crystal violet assay was carried out for the verification of biofilm formation. The in vitro assays evaluated Pseudomonas aeruginosa UFPEDA 416 and Staphylococcus aureus UFPEDA 02 exposure of 48 h to MPDS, as well as the use of 40 KHz ultrasound at the beginning and with 24 h immersion in the MPDS. Subsequently, in vivo assays evaluated the formation of microbial biofilms on the CLSC walls containing silicone-hydrogel contact lenses immersed in MPDS from 15 healthy volunteer patients, who had been wearing the lenses for 7 days. RESULTS: Biofilms were inhibited by 26%–98% in the in vitro assays, with a statistically significant difference only for P. aeruginosa UFPEDA 416 exposed to diluted MPDS. Most inhibitions occurred moderately and weakly. In addition, adherent cells were detected in more than 90% of the tests. Biofilm was not inhibited in more than one third of the results, nor was it disturbed, especially with the ultrasound treatments. The average of obtained optical densities at 590 nm was between 0.6 and 0.8 in the in vivo assays. The results were similar between the CLSC right and left wells. There was a correlation between microbial biofilm formation and the type of MPDS tested, with statistical difference between the three treatments. CONCLUSION: MPDS promoted a partial inhibition of microbial biofilm formation but only one MPDS proved to be more effective in vitro and in vivo. This study, however, could not distinguish the effect of possible errors in the good hygiene practices of the users.

The more pyocyanin produced by the hydrocarbon degrading Pseudomonas aeruginosa strains, the more hydrocarbons would be assimilated and transformed. To evaluate this assumption,we asssed the potential use of nine wild types of pyocyanin-producing P. aeruginosain bioremediation. They exhibit concentrations of pyocyanin ranging from 0.08 to 28.68 μg/mL.Hydrocarbonoclastic activity in the presence of pyocyanin was determined by two protocols. First, ahigh correlation was found between the synthesis of pyocyanin and the emulsification index of lubricating oil. Second, two strains were tested for their ability to degrade anthracene and pyrene in soil by the concentration of pigment produced.Microcosms were filled with 250g of sterile sandy soil, supplemented with glycerin and then contaminated with a 20 mg/kg of mixture of the two compounds. The volume of the inoculum suspension (≈108 CFU/mL) was equivalent to 5% of the soil mass contained in the microcosms. Static incubation lasted 60 days at 25°C. A roughly 60-fold difference between the pigment concentrations produced by the two strains resulted in an increase of 65 and 45% in the pyrene and anthracene biodegradation, also indicating that the molecule served as a co-substrate of pyrene degradation.

The addition of co-substrates as adjuvants to the bioremediation process enables almost complete removal of the polycyclic aromatic hydrocarbons (PAH) in the soil. The aim of this work was to associate biostimulation and bioaugmentation, represented respectively by the addition of residues from the processing of three oilseeds, and by the addition of consortia formed by Pseudomonas aeruginosa and Burkholderia cepacia strains, in the removal of phenanthrene and pyrene from a soil contaminated by a lubricating oil mixture containing approximately 50 mg/kg PAH. Three consortia were prepared from antimicrobial activity tests and each was stimulated with cotton, peanut or sesame cakes (20 and 40 mg/kg). The biodegradation tests were carried out on polyethylene reactors filled with 200 g of sandy soil. After 60 days incubation at room temperature, between 65 and 80% of the phenanthrene and pyrene was removed with preferential degradation of the three consortia by pyrene. In all cases, the most successful condition was obtained when the cake contents were added, independent of amount of contaminant content (40 or 80 mL/kg), emphasizing the importance of the use of co-substrate and reuse of agro-industrial wastes in bioremediation. Phytotoxicity assays demonstrated that Zea mays was the best indicator of soil fertility after treatment.

The biodegradation of paclobutrazol (PBZ) was evaluated during 49 days using glycerol and agro-industrial wastes (biodiesel-derived glycerol, peanut cake and sesame cake) as additional carbon sources and unsaturated soil with a history of PBZ application. Biodiesel-derived peanut cake was the best agro-industrial waste, and this was used in the following four strategies. Four biodegradation strategies were performed using 100, 80, 50 and 20 % of soil without history with the addition of 0, 20, 50 and 80 % of soil with history, respectively. The addition of soil with history (50 or 80 %) and the selected waste (peanut cake) favored biodegradation of soil without history, with rates about 95 %. The peanut cake waste proved to be an adequate source of carbon and nitrogen, and the soil with history, proved to be an efficient reservoir of microorganisms capable of biodegrading PBZ.

Pseudomonas aeruginosa é conhecida por sua versatilidade metabólica e extrema capacidade de adaptação a diferentes ambientes, inclusive aquáticos. Para desinfecção de águas, o cloro e agentes que contêm cloro continuam sendo os mais usados no mundo. O objetivo deste trabalho foi avaliar a resistência ao cloro de linhagens de P. aeruginosa, isoladas de amostras de águas de diversos ambientes. Foram testados diferentes tempos de contato (1, 5, 10, 20, 30 e 40 minutos) e soluções aquosas de cloro, com concentrações definidas com base na legislação vigente no país para água potável: 0,5; 1,0 e 2,0 ppm. O teste de resistência ao cloro foi desenvolvido por meio da exposição direta das bactérias às soluções. Os resultados revelaram que P. aeruginosa, isoladas de diferentes fontes de água, têm a habilidade de sobreviver a diferentes concentrações de cloro. Na concentração de 1 ppm, a maioria das linhagens não foi inibida. As linhagens mais resistentes ao cloro também apresentaram relação de multirresistência à maioria dos antibióticos testados.The nutritional versatility and the adaptability of Pseudomonas aeruginosa to different environments, including water, are well known. Chlorine and other chlorine agents are used as water disinfecting all around the world. The aim of this work was to evaluate the possible chlorine resistance amongst P. aeruginosa strains isolated from different aquatic sources by using different contact time (1, 5, 10, 20, 30 and 40 minutes) in solutions with known chlorine concentrations according current legislation in the country to potable water: 0.5; 1.0 and 2.0 ppm. The chlorine resistance test was done by direct exposure of P. aeruginosa under a solution with known chlorine concentration. Results showed that P. aeruginosa strains isolated from different aquatic sources are able to survive in different chlorine concentrations. At 1 ppm, most of them were not inhibited. It was also observed that the most resistant strains were antibiotic multiresistant.

Neste estudo, foram investigadas 13 linhagens de Pseudomonas aeruginosa oriundas de diferentes fontes de ambientes aquaticos em Recife-PE: pocos de cemiterios, agua mineral, agua tratada do sistema de abastecimento e efluentes de aterro sanitario. Foram empregadas as classes de antibioticos mais utilizadas no tratamento contra P. aeruginosa : agentes beta-lactâmicos, aminoglicosideos, carbepenemas e quinolonas. Todas as linhagens selvagens mostraram-se sensiveis aos antibioticos testados.

Patulin (PAT) is a fungi-derived natural product with immunosuppressive properties, but the signaling pathways involved in the effect of PAT remain undefined. This work aimed to evaluate the effect of PAT in lipopolysaccharide (LPS)-activated murine peritoneal macrophages. For this purpose, murine peritoneal macrophages were stimulated with LPS and treated with PAT at different concentrations (62.5-4,000 ng/ml). Cell viability assay showed that PAT at concentrations up to 250 ng/ml did not affect macrophage viability. Then, nitric oxide response was evaluated using the nontoxic PAT concentrations (62.5; 125; and 250 ng/ml). As a result, PAT, at concentration of 250 ng/ml, significantly reduced nitric oxide production (by 98.4%), inducible nitric oxide synthase (iNOS) expression (by 83.5%), and iNOS messenger ribonucleic acid expression (by 100.0%) in the cells stimulated with LPS. Next, we investigated whether PAT interferes with interleukin (IL)-1β, IL-6, IL-10, and tumor necrosis factor (TNF)-α release. PAT significantly reduced LPS-induced IL-1β release (by 80.6%), without interfering with the production of IL-6, IL-10, and TNF-α. Moreover, PAT significantly reduced cluster of differentiation (CD) 69 (by 63.1%) and Toll-like receptor (TLR) 4 (by 91.9%) protein expression in the presence of LPS, without altering the gene expression of these proteins. Finally, to investigate PAT mechanism of action, mitogen-activated protein (MAP) kinase phosphorylation analysis was performed. PAT significantly reduced LPS-triggered phosphorylation of all MAP kinases assessed: extracellular signal-regulated kinase (ERK; by 89.5%), c-Jun N-terminal kinase (JNK; by 77.5%), and p38 (by 72.3%). Taken together, these data suggest that PAT downregulates acute inflammatory response, inhibiting nitric oxide production by suppressing CD69-TLR4/ERK-JNK-p38 MAP kinases/Nos2/iNOS signaling pathway in murine peritoneal macrophages. Thus, PAT could become a promising pharmacological tool for the control of inflammation, capable of completely inhibiting the nitric oxide pathway. Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES A patulina (PAT) é um produto natural derivado de fungos com propriedades imunossupressoras. No entanto, as vias de sinalização envolvidas no efeito da PAT permanecem indefinidas. Este trabalho teve como objetivo avaliar o efeito da PAT em macrófagos peritoneais murinos ativados com lipopolissacarídeo (LPS). Para isso, macrófagos peritoneais murinos foram estimulados com LPS e tratados com a PAT em diferentes concentrações (62,5-4.000 ng/ml). O ensaio de viabilidade celular mostrou que a PAT em concentrações de até 250 ng/ml não afetou a viabilidade dos macrófagos. Então, foi avaliada a produção de óxido nítrico usando as concentrações não tóxicas de PAT (62,5; 125 e 250 ng/ml). Como resultado, a PAT, na concentração de 250 ng/ml, reduziu significativamente a produção do óxido nítrico (98,4%), a expressão da óxido nítrico sintase induzida (iNOS; 83,5%) e a expressão do ácido ribonucleico mensageiro da iNOS (100,0%) nas células estimuladas com o LPS. Em seguida, foi investigado se a PAT interfere na produção de interleucina (IL) 1β, IL-6, IL-10 e fator de necrose tumoral (TNF)-α. A PAT reduziu significativamente a produção da IL-1β (80,6%) induzida pelo LPS, sem interferir na produção da IL-6, IL-10 e TNF-α. Além disso, a PAT reduziu significativamente a expressão das proteínas grupamento de diferenciação (CD) 69 (63,1%) e receptor do tipo Toll (TLR) 4 (91,9%) na presença do LPS, sem alterar a expressão gênica dessas proteínas. Por fim, para investigar o mecanismo de ação da PAT, foi realizada a análise da fosforilação das proteínas ativadas por mitógenos (MAP) quinases. A PAT reduziu significativamente a fosforilação induzida pelo LPS de todas as MAP quinases avaliadas: quinase regulada por sinal extracelular (ERK; 89,5%), quinase N-terminal c-Jun (JNK; 77,5%) e p38 (72,3%). Em conjunto, esses dados inéditos sugerem que a PAT regula negativamente a resposta inflamatória aguda, inibindo a produção de óxido nítrico ao suprimir a via de sinalização CD69-TLR4/MAP quinases ERK-JNK-p38/Nos2/iNOS em macrófagos peritoneais murinos. Assim, a PAT pode se tornar uma ferramenta farmacológica promissora para o controle da inflamação, capaz de inibir completamente a via do óxido nítrico.

Bioprospecting of microbes allows the use of their metabolites, such as biodegradable molecules in substitution of not eco-friendly compounds, for example of pigments as good natural substitutes for synthetic dyes. This work aimed to produce, to extract and to characterize pyocyanin from two wild isolates of Pseudomonas aeruginosa, TGC02 and TGC04, and to apply the pigment as dyeing agent in two fibers. King A broth added with 2% (w/v) of malt bagasse were used to produce about 30 ml of pyocyanin (585 μg/ml) with a high degree of purity. Kinetic parameters of maximum cell production, maximum pigment production and yield coefficient were determined. In addition, cell concentration and growth rate were determined. The dyeing of 50 and 100 μg/mL of pyocyanin in cotton and polyester/elastane fibers was done using the exhaustion method, using sodium bicarbonate as mordant. Both isolates were similar in terms of microbial parameters. The cell population was increased up to 9.5 times from the inoculum, producing a maximum concentration of pyocyanin about 1.0 μg/mL.h-1, and increased significantly after 36 h and seemed to modulate the microbial growth. The yield coefficient was up to 240 μg/mL.UA-1. Pyocyanin was more functional to the cotton fiber and produced two shades of blue, without undergoing significant changes after washing and drying. Additionally, the highest percentages of pigment absorption and the lowest percentages of exhaustion were recorded. The opposite was found in the polyester/elastane fiber. The adsorption of the pigment was low, influencing the color resulted, whose perception in white light was a very pale lilac-bluish color. Additionally, the shades poorly varied as a function of the concentration of pyocyanin applied. In conclusion, pyocyanin may be used as alternative biocolorant on cotton fibers. Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES A bioprospecção de microrganismos permite a utilização de produtos do metabolismo para o benefício da sociedade, empregando por exemplo, moléculas biodegradáveis em substituição a substâncias não ecologicamente corretas, tais como pigmentos bacterianos como substitutos naturais para corantes sintéticos empregados no tingimento de fibras. Este trabalho teve por objetivo produzir, extrair e caracterizar piocianina de dois isolados selvagens de Pseudomonas aeruginosa, TGC02 e TGC04, e empregar o pigmento em duas fibras têxteis. Caldo King A suplementado com bagaço de malte 2% (m/v) foi utilizado para a produção do pigmento e produziu-se cerca de 30 mL de solução de piocianina (585 μg/mL), com alto grau de pureza. Foram determinados parâmetros cinéticos de produção máxima de células, produção máxima de pigmento e coeficiente de rendimento. Além disso determinou-se a concentração celular e taxa de crescimento. O tingimento com 50 e 100 μg/mL de piocianina nas fibras de algodão e poliéster/elastano ocorreu pelo método da exaustão, utilizando bicarbonato de sódio como mordente. O desempenho de TGC02 e TGC04 foi similar. Houve incremento da população em até 9,5 vezes a partir do inóculo, produzindo uma concentração máxima de piocianina cerca de 1,0 μg/mL.h-1, aumentando significativamente a partir de 36h de bioprocesso, permanecendo crescente e parecendo modular a população celular. O coeficiente de rendimento foi de até 240 μg/mL.UA-1. A piocianina foi mais funcional ao algodão, produzindo duas tonalidades de azul, sem sofrer alterações significativas após lavagem e secagem. Adicionalmente, foram registrados os maiores percentuais de absorção do pigmento e os menores percentuais de exaustão. O oposto foi verificado na fibra de poliéster/elastano. A adsorção do pigmento foi baixa, influenciando na cor obtida, cuja percepção à luz branca foi de um lilás-azulado muito pálido. Os tons variaram em função da concentração aplicada. Concluiu-se que a piocianina pode ser uma alternativa de agente de coloração para a fibra de algodão.

Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2022-02-22T12:36:44Z No. of bitstreams: 1 PDF - Isabela Motta Felício.pdf: 1225450 bytes, checksum: d36e7e458695173b20597b71e7dcccfb (MD5) Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2022-02-22T14:00:44Z (GMT) No. of bitstreams: 1 PDF - Isabela Motta Felício.pdf: 1225450 bytes, checksum: d36e7e458695173b20597b71e7dcccfb (MD5) Made available in DSpace on 2022-02-22T14:01:19Z (GMT). No. of bitstreams: 1 PDF - Isabela Motta Felício.pdf: 1225450 bytes, checksum: d36e7e458695173b20597b71e7dcccfb (MD5) Previous issue date: 2020-03-04 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES In recent decades, microbial resistance has become a serious risk to public health, a phenomenon resulting from the indiscriminate use of antibiotic antibiotics by health institutions, the population, in agriculture and agriculture, or by gene recombination or mutation between species. With bacterial resistance, food pathogens are a major concern for the food industry, consumers and safety authorities. The use of bioactive products derived from secondary metabolism of vegetables is a viable alternative, as is the case of Carvacrol oil (CV), a phenolic monoterpene with proven antimicrobial activity. However, characteristics, such as its hydrophobia, oxidation facility, instability in the presence of light, heat and humidity, can limit its application as a food additive. With this, the use of emulsified systems, such as nanoemulsions (NEs) appear as an alternative, in order to increase the stability of these compounds and increase their solubility. Furthermore, the choice of nanoemulsified systems as antimicrobial agents is important, as they interact with microbial cell membranes, causing them to stop growing. Thus, the objective of this work was the development, characterization and evaluation of the antimicrobial activity against the bacteria Staphylococcus aureus UFPEDA02, Escherichia coli UFPEDA224, Pseudomonas aeruginosa UFPEDA416, Salmonella enteritidis ATCC13076 and Salmonella typhimurium ATCC14028 of a NE containing CV. To obtain a stable NE, a study of HLB identification of the CV by the microemultocrit method was first carried out, so at the end an HLB value of 11 was found for the CV used in this study. Then, a study was carried out to obtain a NE with greater stability, for that, it varied the proportions of CV (3 and 5%), surfactants (a mixture of Tween 80® and Span 80®) (2, 5 and 9%) and water (86, 88, 90, 92 and 95%). From the prepared NEs, just the one with 3% oil, 9% surfactant and 88% water (NECV) remained stable for up to 90 days. The high energy emulsification methodology was used to obtain the systems, where the NEs went through four cycles of homogenization in the sonicator and ultrasound bath. The amount of oil in the NECV system was performed using analytical methodology developed in the UV-VIS spectrophotometer, which obtained an encapsulation efficiency of 99.45% ± 0.019 (equivalent to 2.98 mg CV per mL of NE). Then, it was characterized as for the macroscopic aspect, presenting liquid aspect with white color. During the 90 days, no creaming formation or phase separation was observed, which NECV remained with an expected average droplet size (D90 = 164.5 ± 0.69 nm), PDI (D90 = 0.118 ± 0.024), zeta potential (D90 = -14.6 ± 0.529 mV) and pH (D90 = 5.52 ± 0.011). After this, the antibacterial activity of the isolated compound and the nanoemulsified system was evaluated by the microdilution method, in which Rezasurin was used as a visual indicator of cellular metabolism. In terms of antibacterial activity, NECV (MIC: 256 µg / mL for E. coli and Samonella spp., 128 µg / mL for S. aureus and P. aeruginosa) obtained a better result when compared to the free compound against strains S. aureus, P. aeruginosa and S. typhimurium. Furthermore, there was a bactericidal effect against E. coli, P. aeruginosa and Salmonella spp. The results obtained indicated that NECV can be incorporated as a food additive to prevent and control microbial growth. Nas últimas décadas, a resistência microbiana frente aos antibacterianos tornou-se um sério risco a saúde coletiva, fenômeno esse decorrente do uso indiscriminado de antibióticos antibióticos por instituições de saúde, população, na agropecuária e na agricultura, ou de recombinação gênica ou mutação entre as espécies Com a resistência bacteriana patógenos alimentares são uma grande preocupação para a indústria de alimentos, consumidores e autoridades de segurança. O uso de bioativos derivados de metabolismo secundário dos vegetais correspondem uma alternativa viável, como é o caso do óleo de Carvacrol (CV), um monoterpeno fenólico com atividade antimicrobiana comprovada. No entanto, características, como sua hidrofobia, facilidade de oxidação, instabilidade na presença de luz, calor e umidade, podem limitar sua aplicação como aditivo alimentar. Com isso, o uso de sistemas emulsionados, como as nanoemulsão (NEs) surgem como uma alternativa, afim de aumentar a estabilidade desses compostos e aumentar sua solubilidade. Além do mais, a escolha de sistemas nanoemulsionados como agentes antimicrobianos é importante, pois esses interagem com as membranas das células microbianas, fazendo com que interrompa o crescimento das mesmas. Dessa forma, o objetivo desse trabalho foi o desenvolvimento, caracterização e avaliação da atividade antimicrobiana frente as bactérias Staphylococcus aureus UFPEDA02, Escherichia coli UFPEDA224, Pseudomonas aeruginosa UFPEDA416, Salmonella enteritidis ATCC13076 e Salmonella typhimurium ATCC14028 de uma NE contendo CV. Para obtenção de uma NE estável, primeiramente foi realizado um estudo de identificação de HLB do CV pelo método de microemultócrito, assim ao final encontrou-se um valor de HLB de 11 para o CV empregado neste estudo. Em seguida, foi realizado um estudo para se obter uma NE com uma maior estabilidade, para isso variou-se as proporções de CV (3 e 5%), surfactantes (uma mistura de Tween 80® e Span 80®) (2, 5 e 9%) e água (86, 88, 90, 92 e 95%). Das NEs preparadas, apenas a com 3% de óleo, 9% de surfactante e 88% de água (NECV) permaneceu estável por até 90 dias. A metodologia de alta energia de emulsificação foi utilizada para obtenção dos sistemas, onde as NEs passaram por quatro ciclos de homogeneização no aparelho de sonicação e banho de ultrassom. A quantidade de óleo no sistema NECV foi realizada utilizando metodologia analítica desenvolvida no espectrofotômetro de UV-VIS, a qual obteve uma eficiência de encapsulação de 99,45% ± 0, 019 (equivalente a 2,98 mg de CV por mL de NE). Em seguida, foi caracterizado quanto ao aspecto macroscópico, apresentando aspecto líquido com cor branca. Durante os 90 dias, não foi observada formação de cremação ou separação de fases, na qual a NECV se manteve com um tamanho médio de gotícula (D90 = 164.5 ±0.69 nm), PDI (D90 = 0.118 ± 0.024), potencial zeta (D90 = -14.6 ± 0.529 mV) e pH (D90 = 5.52 ± 0.011) esperado. Posteriormente, foi avaliada a atividade antibacteriana do composto isolado e do sistema nanoemulsionado pelo método da microdiluição, na qual foi usado a Rezasurina como indicador visual de metabolismo celular. Da atividade antibacteriana, a NECV (CIM: 256 µg / mL para E. coli e Samonella spp., 128 µg / mL para S. aureus e P. aeruginosa) obteve melhor resultado quando comparado ao composto livre frente as linhagens S. aureus, P. aeruginosa e S. typhimurium. Além disso, houve efeito bactericida contra E. coli, P. aeruginosa e Salmonella spp. Os resultados obtidos indicaram que a NECV pode ser incorporada como aditivo alimentar para prevenir e controlar o crescimento microbiano.

Certain ocular resident or pathogenic microbes may remain viable in the presence of multipurpose disinfectant solutions (MPDS), subsequently developing biofilms inside contact lens storage cases (CLSC) which pose a risk of infection to wearers. This study evaluated the formation of ocular microbiota biofilms exposed to three top selling MPDS. To this, crystal violet assay was carried out for the verification of biofilm formation. The in vitro assays evaluated Pseudomonas aeruginosa UFPEDA 416 and Staphylococcus aureus UFPEDA 02 exposure of 48h to MPDS, as well as the use of 40 KHz ultrasound at the beginning and with 24h immersion in the MPDS. Subsequently, in vivo assays evaluated the formation of microbial biofilms on the CLSC walls containing silicone-hydrogel contact lenses immersed in MPDS from 15 healthy volunteer patients, who had been wearing the lenses for 7 days. Tests for clinical evaluation of function and tear production were also performed in all volunteers. Biofilms were inhibited by 26-98% in the in vitro assays, with a statistically significant difference only for P. aeruginosa UFPEDA 416 exposed to diluted MPDS. Most inhibitions occurred moderately and weakly. Additionally, adherent cells were detected in more than 90% of the tests. Biofilm was not inhibited in more than one third of the results, nor was it disturbed, especially with the ultrasound treatments. The average of obtained optical densities at 590 nm were between 0.6-0.8 in the in vivo assays. Results were similar between the CLSC right and left wells. There was a correlation between microbial biofilm formation and the type of MPDS tested, with statistical difference between the three treatments. The evaluation of function and tear production showed no changes and there was no relationship with the formation of biofilms. MPDS promoted a partial inhibition of microbial biofilm formation but only one MPDS proved to be more effective in vitro and in vivo. This study, however, could not distinguish the effect of possible errors in the good hygiene practices of the users. Nenhuma Alguns microrganismos da superfície ocular, residentes ou patógenos, podem permanecer viáveis na presença das soluções multiuso usadas para desinfecção de lentes de contato, desenvolvendo, posteriormente, biofilmes nos estojos de armazenamento das lentes e representando risco de infecções para os usuários. Este estudo avaliou a ação biocida de três marcas comerciais de soluções multiuso (SM) utilizadas no Nordeste do Brasil e a formação de biofilmes em estojos de armazenamento de lentes de contato gelatinosas. Para a verificação da formação do biofilme utilizou-se o teste do cristal violeta. Os ensaios in vitro, para análise do efeito biocida das soluções multiuso, avaliaram a exposição por 48h de Pseudomonas aeruginosa UFPEDA 416 e Staphylococcus aureus UFPEDA 02 às três SM, como também a ação de ultrassom a 40 KHz no início e após 24h de imersão nas SM. Posteriormente, ensaios in vivo avaliaram a formação de biofilmes microbianos nas paredes dos estojos de armazenamento, contendo lentes de contato gelatinosas (LCG) de silicone-hidrogel imersas em soluções multiuso, de 15 pacientes voluntários sadios que as usaram LCG durante sete dias. Também foram realizados exames para avaliação clínica da função e produção lacrimal em todos os voluntários. Em mais de um terço dos ensaios o biofilme não foi inibido, tão pouco perturbado, especialmente nos tratamentos com ultrassom. Nos testes in vivo, as médias obtidas das densidades ópticas a 590nm foram entre 0.6-0.8. Os resultados foram similares entre os poços direito e esquerdo dos estojos de armazenamento das LCG. Houve correlação entre a formação de biofilme microbiano e o tipo de SM testada, com diferença estatística entre os três tratamentos. Nos ensaios in vitro os biofilmes foram inibidos entre 26-98%, com diferença estatisticamente significativa apenas com P. aeruginosa UFPEDA 416 quando exposta às SM diluídas. Adicionalmente, células aderentes foram detectadas em mais de 90% dos testes. A avaliação da função e produção lagrimal não demonstrou alterações significativas. Embora as SM tenham promovido uma inibição parcial da formação dos biofilmes microbianos nos ensaios in vitro e in vivo.

Ecological interactions enable the transfer of molecular and genetic information between microorganisms, being crucial for their establishment and development in a variety of environments. The literature reports that Pseudomonas aeruginosa, a cosmopolitan bacillar bacterium, is able to interact with the environment and other microorganisms, including yeasts, through its metabolites, highlighting pyocyanin, a pigment linked to its virulence. The aim of this work was to verify cellular alterations promoted by pyocyanin exposure in four yeasts: Candida albicans ATCC 76485, C. parapsilosis ATCC 22019, C. tropicalis ATCC 13803 and C. krusei ATCC 6258. For this purpose, in vitro tests were carried out on the action of pyocyanin in the: Minimal Inhibition Concentration (MIC) and Minimum Fungicide Concentration (CFM); cell viability; damage to the cell wall and membrane; surface adhesion and disturbance of mature biofilm. The pyocyanin MIC was 600 μg/mL for all tested strains. On the other hand, CFM was 1.200 μg/mL for C. albicans ATCC 76485, C. parapsilosis ATCC 22019 and > 1.200 μg/mL for C. tropicalis ATCC 13803 and C. krusei ATCC 6258. Pyocyanin promoted damage to the cell wall and did not interfere with the membrane. The cell viability after 48h of pigment exposure was 7.69% (C. albicans ATCC 76485), 13.62% (C. tropicalis ATCC 13803), 10.83% (C. parapsilosis ATCC 22019) and 2.19 % (C. krusei ATCC 6258). In addition, pyocyanin interfered in the adhesion of C. tropicalis ATCC 13803, whereas in the other strains, cell adhesion and stablished biofilm were not affected. The results suggest that the interaction between Candida spp. and P. aeruginosa may be antagonistic, however, yeasts may exhibit mechanisms to ensure their permanence when the pigment is present Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES As interações ecológicas permitem as transferências de informações moleculares e genéticas entre microrganismos e são cruciais para o estabelecimento e seu desenvolvimento em uma variedade de ambientes. A literatura reporta que Pseudomonas aeruginosa, uma bactéria bacilar cosmopolita, é capaz de interagir com o ambiente e outros microrganismos, incluindo leveduras, por meio de seus metabólitos, destacando a piocianina, pigmento atrelado à virulência da espécie. O objetivo deste trabalho foi verificar alterações celulares promovidas pela exposição de piocianina em quatro leveduras: Candida albicans ATCC 76485, C. parapsilosis ATCC 22019, C. tropicalis ATCC 13803 e C. krusei ATCC 6258. Para isso, ensaios in vitro analisaram a ação da piocianina pela determinação das Concentrações, Inibitória Mínima (CIM) e Fungicida Mínima (CFM) do pigmento; percentual de viabilidade celular; danos à parede e membrana; e sobre a adesão à superfície e perturbação do biofilme maduro. A CIM da piocianina foi estabelecida em 600 μg/mL para todas linhagens testadas. Por outro lado, as CFM foram 1.200 μg/mL para C. albicans ATCC 76485, C. parapsilosis ATCC 22019 e >1.200 μg/mL para C. tropicalis ATCC 13803 e C. krusei ATCC 6258. A piocianina promoveu dano à parede celular e não interferiu na membrana. A viabilidade celular após 48h de exposição ao pigmento foi de 7,69% (C. albicans ATCC 76485), 13,62% (C. tropicalis ATCC 13803), 10,83% (C. parapsilosis ATCC 22019) e 2,19% (C. krusei ATCC 6258). Em complemento, a piocianina interferiu na adesão de C. tropicalis ATCC 13803, enquanto nas demais linhagens, a adesão das células e o desenvolvimento do biofilme, não foram afetados. Os resultados sugerem que a interação entre P. aeruginosa produtoras de piocianina e Candida spp. pode ser antagônica, contudo, as leveduras podem exibir mecanismos para garantir sua permanência quando o pigmento está presente

The secretion of metabolites with antimicrobial activity is one of the strategies employed by bacteria in response to negative stimuli promoted during interspecies competition. In its stationary phase Pseudomonas aeruginosa and Escherichia coli can synthesize diffusible exometabolites whose action is to mutually inhibit the exposed cells, ensuring the balance of both populations in a given place. The former may have an advantage due to the production of pyocyanine. However, the excretion of indol and acetate by E. coli can balance this advantage. This study aimed to detect the influence of indol and acetate on the synthesis of pyocyanin and adhesion of P. aeruginosa, as well as the action of pyocyanin on viable E. coli cells, as well as its adhesion. For this purpose, in vitro tests were conducted to determine the Minimum Inhibitory Concentration (MIC), extraction and quantification of pyocyanin and adhesion by the violet crystal method. The synthesis of piocyanin by Pseudomonas aeruginosa was reduced by up to 50% in the presence of indol. On the other hand, no change in pigment production was observed in the presence of acetate, alone or associated with concentrations lower than 0.5 mM, reflecting an increase of more than 20% in piocyanine production. Indol was also effective in reducing the adhesion of Pseudomonas aeruginosa cells compared to control. For the E. coli strains, the MIC of pyocyanin was 37.5 µg/mL, also reducing cell adhesion. It was observed that despite the inhibition having occurred, no exometabolite promoted biocidal effect in the evaluated strains. The results contribute to the understanding of the ecological mechanisms that occur during the competition between these two bacterial species in aqueous medium. Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES A secreção de metabólitos com atividade antimicrobiana é uma das estratégias empregadas por bactérias como resposta aos estímulos negativos promovidos durante a competição interespécies. Em sua fase estacionária Pseudomonas aeruginosa e Escherichia coli podem sintetizar exometabólitos difusíveis cuja ação é inibir mutualmente as células expostas, garantindo o equilíbrio de ambas as populações num determinado sítio. A primeira pode levar vantagem em razão da produção de piocianina. Entretanto, a excreção de indol e acetato pela E. coli pode equilibrar esta vantagem. Este trabalho teve o objetivo de detectar a influência do indol e acetato sobre a síntese de piocianina e adesão da P. aeruginosa, bem como a ação da piocianina sobre as células viáveis de E. coli, bem como de sua adesão. Para isso, foram conduzidos ensaios in vitro de determinação da Concentração Inibitória Mínima (CIM), de extração e quantificação de piocianina e da adesão, pelo método do cristal violeta. A síntese de piocianina por Pseudomonas aeruginosa foi reduzida em até 50% na presença do indol. Por outro lado, não foi observada alteração na produção do pigmento na presença de acetato, isoladamente ou associado com concentrações menores que 0,5 mM, refletindo um aumento de mais de 20% da produção de piocianina. O indol também se mostrou eficaz na redução da adesão de células de Pseudomonas aeruginosa, comparadas ao controle. Para as linhagens de E. coli, a CIM de piocianina foi 37,5 µg/mL, reduzindo também a adesão celular. Observouse que apesar da inibição ter ocorrido, nenhum exometabólito promoveu efeito biocida nas linhagens avaliadas. Os resultados contribuem para a auxiliar a compreensão dos mecanismos ecológicos que ocorrem durante a competição entre estas duas espécies bacterianas em meio aquoso.

Environments contaminated by polycyclic aromatic hydrocarbons are challenges that can be minimized with the use of microorganisms with degradation potential. The objective of this work was to evaluate the degradation capacity of anthracene and pyrene by a Pseudomonas aeruginosa isolate and to correlate with laccase and manganese peroxidase activity. Three isolates of Pseudomonas aeruginosa (TGC-02, 03 and 07) and one positive control with Aspergillus niger were evaluated. The experiments were divided into two steps: screening with P. aeruginosa isolates was initially screened for emulsification and drop collapse tests. The higher hydrocarboclastic activity was observed for the TGC-02 isolate, which degraded more than 30% of the contaminants tested. Subsequently, the biodegradation assays of the anthracene and pyrene contaminants by the selected TGC-02 isolate and compared to an A. niger isolate were conducted. The assays lasted 25 days at room temperature in microcosms containing 30 mL of distilled water, contaminated with 50 mg / L of pyrene or anthracene, under static incubation. For each point of analysis, pH, biomass, cell viability, total proteins and enzymatic activity were measured. The content of the contaminants was determined by gas chromatography coupled to flame spectrometry after 25 days of the bioprocess. No specific activity was observed for laccase or manganese peroxidase in the tested isolates, and it was not possible to correlate the degradation capacity with the specific activity of these enzymes. However, when analyzing the total protein results, it was observed that during the 25 days there was an increase in the concentrations of total proteins in presence of the contaminants, especially for P. aeruginosa TGC-02. The TGC-02 and A. niger isolates cultured in the presence of the contaminants presented viability during the 20 days of the bioprocess. Discounting the abiotic losses, the positive control A. niger degraded 28.11% and 30.70% of the pyrene and anthracene, respectively. P. aeruginosa TGC-02 presented better results, reducing pyrene and anthracene in 37.83% and 33.01%, respectively. A. niger is a fungus known as a degradation agent of aromatic compounds, however, in this work P. aeruginosa proved to be more efficient in the degradation of pyrene and anthracene in the liquid medium in 25 days. The results demonstrate the potential of bioremediation of P. aeruginosa for the removal of pyrene and anthracene, apparently independent of the production of manganese peroxidase and laccase. Nenhuma Ambientes contaminados por hidrocarbonetos policíclicos aromáticos são desafios que podem ser minimizados com o uso de microrganismos com potencial de degradação. O objetivo deste trabalho foi avaliar a capacidade de degradação do antraceno e pireno por um isolado de Pseudomonas aeruginosa e correlacionar com a atividade da lacase e manganês peroxidase. Foram avaliados três isolados de Pseudomonas aeruginosa (TGC-02, 03 e 07) e um controle positivo com Aspergillus niger. Os experimentos foram divididos em duas etapas: inicialmente realizou-se uma triagem com os isolados de P. aeruginosa, submetidas a testes de emulsificação e de colapso de gota. A maior atividade hidrocarbonoclástica foi observada para o isolado TGC-02, que degradou mais de 30% dos contaminantes testados. Posteriomente, foram conduzidos os ensaios de biodegradação dos contaminantes antraceno e pireno pelo isolado selecionado TGC-02 e comparado com uma isolados de A. niger. Os ensaios duraram 25 dias, à temperatura ambiente, em microcosmos contendo 30 mL de água destilada, contaminada com 50 mg/L do pireno ou antraceno, sob incubação estática. Para cada ponto de análise, foi mensurado pH, biomassa, viabilidade celular, proteínas totais e atividade enzimática. O teor dos contaminantes foi determinado por cromatografia gasosa acoplada à espectrometria de chama após 25 dias do bioprocesso. Não foi observado atividade específica para lacase ou manganês peroxidase nos isolados testados, não sendo possível correlacionar a capacidade de degradação com a atividade específica destas enzimas. Entretanto, ao analisar os resultados de proteínas totais, observou-se que ao longo dos 25 dias houve aumento nas concentrações de proteínas totais em presença dos contaminantes, especialmente para P. aeruginosa TGC-02. O isolado TGC-02 e A. niger cultivadas em presença dos contaminantes apresentaram viabilidade ao longo de 20 dias do bioprocesso. Descontadas as perdas abióticas, O controle positivo A. niger degradou 28,11% e 30,70% do pireno e antraceno, respectivamente. Já P. aeruginosa TGC-02 apresentou melhores resultados, reduzindo pireno e antraceno em 37,83% e 33,01%, respectivamente. O A. niger é um fungo conhecido como agente de degradação de compostos aromáticos, entretanto, neste trabalho a P. aeruginosa se mostrou mais eficiente na degradação de pireno e antraceno do meio líquido em 25 dias. Os resultados demonstramo potencial de biorremediação da P. aeruginosa para remoção de pireno e antraceno, aparentemente independente da produção de manganês peroxidase e lacase.

FACEPE O paclobutrazol (PBZ) é o regulador vegetal utilizado no manejo da produção da mangueira na maioria dos pomares sob as condições semiáridas do Nordeste. Apesar da sua eficiência na produtividade, estudos demonstram que o PBZ permanece ativo no solo por muito tempo e isso pode afetar o desenvolvimento das próximas colheitas, reduzindo o vigor vegetativo. A biodegradação do paclobutrazolem solo não saturado e com histórico de aplicação,foi avaliada durante 49 dias usando glicerol e resíduos industriais (glicerol do biodiesel, torta de amendoim e torta de gergelim) como fontes de adicionais de carbono. Os resíduos agroindustriais (torta de amendoime e torta de gergelim) além de carbono, também são fontes de nitrogênio. Foram realizados estudos cinéticos do consumo de PBZ, além da contagem dos micro-organismos totais, do perfil do pH e da fitotoxicidade no início e final dos experimentos. Adicionalmente, estudos investigativos acerca de produtos da biodegradação foram realizados, com análises de espectroscopia de infravermelho. A torta de amendoim foi o melhor resíduo agroindustrial, sendo usado nas quatro estratégias realizadas na próxima etapa. Nessas quatro estratégias de biodegradação foram realizados experimentos usando 100, 80, 50 e 20 % m/m do solo sem histórico com a adicão de 0, 20, 50 e 80 % m/m de solo com histórico, respectivamente. A adição de solo com histórico (50 ou 80% m/m) e a torta de amendoim favoreceram a biodegradação dosolo sem histórico, cuja biodegradação foi maior que 95%. O resíduoselecionado apresentou-se como uma adequada fonte de carbono e de nitrogênio, e, o solo com histórico um eficiente reservatório de micro-organismos capazes de degradar o PBZ. Paclobutrazol (PBZ) is a growth regulator used in the management of mango production in most orchards in semi-arid conditions of the Northeast. Despite its efficiency in productivity, studies show that the PBZ remains active in the soil for a long time and this may affect the development of the next crop, reducing plant vigor. The biodegradation of paclobutrazol (PBZ) was evaluated during 49 days using glycerol and agro-industrial wastes (biodieselderived glycerol, peanut cake and sesame cake) as additional carbon sources and unsaturated soil with a history of PBZ application. Kinetic studies were performed of PBZ consumption beyond count of total microorganisms, pH profile and phytotoxicity at the beginning and end of the experiments. Additionally, investigative studies on the caracterization of biodegradation samples were performed with infrared spectroscopy analysis. Biodieselderived peanut cake was the best agro-industrial waste, andthis was used in the following four strategies. Four biodegradation strategies were performed using 100 %, 80 %, 50 % and 20 % of soil without history with the addition of 0 %, 20 %, 50 % and 80 % of soil with history, respectively. The addition of soil with history (50 or 80%) and the selected waste (peanut cake) favored biodegradation of soil without history, with rates about 95 %. The peanut cake waste proved to be an adequate source of carbon and nitrogen, and the soil with history, proved to be an efficient reservoir of microorganisms capable of biodegrading PBZ.