Orientador: José Roberto Guimarães / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Civil, Arquitetura e Urbanismo / Made available in DSpace on 2018-08-27T20:18:28Z (GMT). No. of bitstreams: 1 Oliveira_AmandaMarchiDuartede_M.pdf: 2576445 bytes, checksum: b72f8b00f5d7ccd5b5b4282cff3876ce (MD5) Previous issue date: 2015 / Resumo: O antimicrobiano lomefloxacina (LOM) foi detectado em matrizes aquosas em vários países. Sua introdução contínua no ambiente pode apresentar risco potencial para organismos aquáticos e terrestres, devido à presença de atividade antimicrobiana. A fim de degradar a LOM e outros contaminantes emergentes, utiliza-se os processos oxidativos avançados (UV/H2O2, O3 em pH 11) devido à geração de radicais hidroxila altamente reativos e não seletivos. O objetivo deste estudo foi avaliar a degradação da LOM por fotólise, peroxidação, peroxidação assistida por radiação ultravioleta e ozonização em diferentes valores de pH, além de avaliar a atividade antimicrobiana residual, a toxicidade aguda e também identificar e sugerir os produtos formados durante os processos de degradação. Soluções de 500 ?g L-1 de LOM foram submetidas aos processos fotolítico (? = 254 nm), peroxidação (CH2O2 = 0,335; 1,675 e 6,77 mmol L-1); UV/H2O2 (CH2O2 de 0,067 a 6,77 mmol L-1) e ozonização (Dose O3 = 5,5 a 54,34 mg L-1) em diferentes valores de pH (3, 7 e 11). Após ensaio de degradação em escala laboratorial, realizou-se a extração em fase sólida e injeção em HPLC para avaliar o percentual de degradação. Foram avaliadas também a remoção da atividade antimicrobiana, com o organismo-teste Escherichia coli K12 (ATCC 23716), e a toxicidade com o organismo-teste Vibrio fischeri (Microtox). Com os resultados do presente trabalho foi possível sugerir que a peroxidação foi ineficaz para a degradação da lomefloxacina: menos que 15% da concentração do fármaco foi reduzida, mesmo após rtempo de ensaio (60 minutos); esse processo também foi ineficaz na redução da atividade antimicrobiana. Ambos os processos de fotólise e UV/H2O2 foram capazes de degradar mais de 95% de LOM em 60 minutos (Dose de radiação = 5267 mJ cm-2). Em relação à degradação da LOM, não se observou diferença quando utilizado UV (95,7%) ou UV/H2O2 (concentração de H2O2 variando de 0,067 a 6,77 mmol L-1) (96,9%). No entanto, o processo UV/H2O2 com 6,77 mmol L-1 de H2O2 foi o mais eficaz na redução da atividade antimicrobiana (96,3%). Em todos os processos avaliados, não foi verificado a remoção completa da atividade antimicrobiana. A ozonização em pH 3 apresentou eficiência quanto à degradação, redução da atividade antimicrobiana e, principalmente da toxicidade aguda, sendo a inibição da V. fischeri de 12,8%. Em todos os processos observou-se incremento na toxicidade, sugerindo a formação de produtos intermediários de degradação mais tóxicos que o composto original. Os processos avaliados demonstraram alto potencial para serem incorporados ou utilizados em estações de tratamento de água que contenham a LOM e/ou compostos químicos emergentes / Abstract: The antimicrobial drug lomefloxacin (LOM) has been detected in the aqueous matrices in several countries. Its continuous introduction into the environment is a potential risk to aquatic and terrestrial organisms due to the presence of antimicrobial activity. Advanced oxidation processes (UV/H2O2, O3 pH 11) due to hydroxyl radicals that were highly reactive and non-selective were used to degrade emerging contaminants and were evaluated to degraded LOM aqueous solutions. The aim of this study was to evaluate the degradation of LOM by photolysis, peroxidation, peroxidation assisted by ultraviolet radiation and ozonation at different pH values; moreover, the residual antimicrobial activity, the acute toxicity, and the degradation byproducts formed during degradation processes were evaluated. Aqueous solutions of 500 ?g L-1 LOM were underwent to UV radiation (? = 254 nm), peroxidation (CH2O2 = 0.335; 1.675 e 6.77 mmol L-1), UV/H2O2 (CH2O2 0.067 to 6.77 mmol L-1) and the ozonation (Dose O3 = 5.5 to 54.34 mg L-1) at different pH values (3, 7 and 11). LOM degradation was evaluated by high performance liquid chromatography (HPLC) with UV detection. Samples were prepared prior to measurements by solid phase extraction (SPE). Antimicrobial activity was determined by evaluating growth inhibition of Escherichia coli K12 (ATCC 23716) and the acute toxicity of Vibrio fischeri test organism (Microtox). Peroxidation did not effectively degrade LOM, reducing less than 15% of the initial drug concentration (500 ?g L-1) after 60 min of testing; this process was also ineffective to reduce antimicrobial activity. Both photolysis and UV/H2O2 (6.7 10-2 mmol L-1 H2O2) were able to degrade more than 95% of LOM in 60 min of testing (radiation dose = 5267 mJ cm-2). However, the UV/ H2O2 (CH2O2 = 6.77 mmol L-1) was more effective to reduce the antimicrobial activity (96.3%). Antimicrobial activity of the solutions was not totally eliminated in any of the processes evaluated. The ozonation at pH 3 was efficient to degradation, reduction of antimicrobial activity and, especially of acute toxicity, with an inhibition of 12.8% of V. fischeri. In all cases there was an increase in toxicity, suggesting the formation of toxic intermediates more degradation than the parent compound. The evaluated processes showed high potential to be incorporated or used in water treatment plants that contain the LOM and/or emerging contaminants / Mestrado / Saneamento e Ambiente / Mestra em Engenharia Civil

Tradicionalmente, no tratamento de feridas graves o antimicrobiano tópico deve ser aplicado uma a duas vezes ao dia na área lesada para reduzir as infecções. No entanto, os pacientes geralmente sofrem desconforto com a aplicação de drogas tópicas e com a troca de curativos. Nesse contexto, o desenvolvimento de sistemas curativos biodegradáveis contendo fármaco antimicrobiano são fundamentais para a promoção de ações terapêuticas, reduzindo as infecções causadas por microrganismos e podendo substituir os curativos convencionas que não possuem polímeros biodegradáveis em sua composição. Para ir de encontro com essa necessidade, filmes de PBAT e PBAT/gentamicina foram preparados por meio de duas abordagens. Na primeira abordagem filmes de PBAT obtidos por casting foram funcionalizados por radiação UV durante 90 minutos na presença de O2 atmosférico para posterior inserção de gentamicina na superfície através de soluções tampões contendo EDC e gentamicina. A superfície das amostras foi analisada pelo WCA, FTIR-ATR e XPS. A análise antimicrobiana dos filmes também foi realizada. Os resultados para os filmes de PBAT por casting mostram que o tempo de irradiação produziu superfícies altamente hidrofílicas com grupos oxigenados enxertados na superfície do polímero (C=O e C-OH). A análise antimicrobiana indicou que a gentamicina não ligou covalentemente com o PBAT, porém observou-se a presença de nitrogênio na superfície das amostras pela análise de XPS Na segunda abordagem foram preparadas fibras de PBAT e PBAT/gentamicina a partir da eletrofiação. As soluções utilizadas no processo de eletrofiação foram preparadas com diferentes concentrações de gentamicina e passaram por análises de condutividade e viscosidade. As fibras de PBAT e PBAT/gentamicina foram caracterizadas quanto a suas propriedades físico-químicas, morfológicas, térmicas, antimicrobianas e biológicas. Nos resultados obtidos para as fibras de PBAT e PBAT/gentamicina observou-se que não houve mudança nos espectros após a mistura dos materiais, uma vez que as bandas de absorção foram preservadas e não houve o aparecimento de novos picos. A presença da gentamicina modificou a morfologia das amostras, diminuindo o diâmetro das fibras. O grau de intumescimento foi alterado com o acréscimo de gentamicina e houve modificação da morfologia das fibras após 48 horas de imersão em PBS, porém as fibras mantiveram-se presentes. A gentamicina aumentou o grau de molhabilidade das fibras PBAT. As fibras de PBAT/gentamicina possuem atividade antimicrobiana frente às cepas de E. coli e não apresentaram citotoxicidadede acordo com o padrão ISO 10993-5. Estes resultados reforçam que o PBAT pode ser usado como um sistema curativo com incorporação de fármaco, mostrando ser uma alternativa interessante de curativo/adesivo para uso tópico. / Traditionally in the treatment of serious wounds, topical antimicrobial should be applied 1–2 times daily to the injured area to reduce infections. However, patients often suffer from discomfort with the application of topical drugs and with the exchange of dressings. In this context, the development of biodegradable curative systems containing antimicrobial drug are fundamental for the promotion of therapeutic actions, reducing the infections caused by microorganisms and being able to substitute conventional dressings that there aren’t biodegradable polymers in their composition. To meet this need, PBAT and PBAT/gentamicin films were prepared using two approaches. In the first approach PBAT films obtained by casting were functionalized by UV radiation for 90 minutes in the presence of atmospheric O2 for subsequent insertion of gentamicin on the surface through buffer solutions containing EDC and gentamicin. The surface of the samples was analyzed by the WCA, FTIR-ATR and XPS. Antimicrobial analysis of the films was also performed. The results for the PBAT films by casting show that the irradiation time produced highly hydrophilic surfaces with oxygenated groups grafted onto the polymer surface (C=O and C-OH). Antimicrobial analysis indicated that gentamycin did not bind covalently with PBAT, but nitrogen was observed on the surface of the samples by XPS analysis In the second approach, PBAT and PBAT/gentamicin fibers were prepared from the electrospun. The solutions used in the electrospun process were prepared with different concentrations of gentamicin and underwent conductivity and viscosity analyzes. The PBAT and PBAT/gentamicin fibers were characterized as their physical-chemical, morphological, thermal, antimicrobial and biological properties. In the results obtained for the PBAT and PBAT/gentamicin fibers, it was observed that there was no change in the spectra after the materials were mixed, since the absorption bands were preserved and there were no new peaks. The presence of gentamicin modified the morphology of the samples, reducing the diameter of the fibers. The degree of swelling was altered with the addition of gentamicin and there was a change in the morphology of the fibers after 48 hours of immersion in PBS, but the fibers remained present. Gentamicin increased the degree of wettability of PBAT fibers. PBAT/gentamicin fibers have antimicrobial activity against E. coli strains and did not present cytotoxicity according to ISO 10993-5 standard. These results reinforce that PBAT can be used as a curative system with drug incorporation, proving to be an interesting dressing/adhesive alternative for topical use.

SYNTHESIS AND BILOGICAL SIGNIFICANCE OF 1,2,4-OXADIAXOLIDIN-5-ONE Chimdi kalu, Austin Miller and Dr. Abbas G. Shilabin Department of Chemistry, East Tennessee State University, Johnson City, TN 37614. ABSRTACT Due to the challenge posed by microbial resistance to broad spectrum of antibiotics, there has been a great need to synthesize of a novel compound which has a different mechanism of action on microbial activity. 1,2,4-oxadiaxolidin-5-One constitute an important class of compound with tremendous potential as pharmaceutical and otherwise biologically relevance substance due to the fact that its five member ring is a configurationally stable building block. This unit is found in other compound like alkaloids, with vast medical application. This study describes the synthesis of 1,2,4-oxadiaxolidin-5-one in two-step procedure using nitroethane and benzaldehyde as starting materials to produce nitrone, which in turn undergoes 1,3- dipolar cycloaddition with phenyl isocyanate to give 1,2,4-oxadiaxolidin-5-one. The product was characterized using proton NMR and GC-MS. There is an ongoing investigation on the summary of some important inhibitory activity against class A β-lactamase by 1,2,4-oxadiaxolidin-5-one heterocyclic core structure to provide effective antimicrobial β-lactamase inhibitors, hence, solving the problem of microbial resistance to currently used antimicrobial drugs.

The aim of this diploma thesis was to determine the optimal extraction conditions for obtaining an extract from the plant Plectranthus amboinicus. Plectranthus amboinicus is an aromatic herb of the Lamiaceae family containing a large number of bioactive compounds. Because of this, it has a number of biological effects – antimicrobial, anti-inflammatory, antioxidant, analgesic, etc. To maintain the maximum of sensory and nutritionally valuable components of P. amboinicus, a simple maceration of fresh chopped herb was chosen. Maceration was performed under the following conditions: 40 minutes at 40 ° C, solvent 40% ethanol, weight 20 g of herb per 100 ml of solvent. The extract prepared under these optimal conditions was comprehensively characterized in terms of the content of total polyphenols, their concentration was determined to 0,08 ± 0,02 mgGEA·ml-1, which corresponds to 0,40 ± 0,07 mgGEA·g-1 of the plant. The antioxidant activity of the extract was 241,24 ± 29,24 µgTEAC·ml-1 and the radical scavenging activity of ABTS•+ was determined to be 49,29 ± 5,97 %. The antimicrobial activity was verified by the agar well diffusion method. Two types of bacteria were selected for the determination – gram-positive Bacillus cereus, gram-negative Escherichia coli and yeast Candida glabrata. The results showed that the extracts have the highest antimicrobial activity against B. cereus. There were 64 volatile aromatic compounds identified in the extract, with monoterpenes and sesquiterpenes dominating. The obtained results confirm that P. amboinicus, or its prepared extract, has a good potential for use as a new and non-traditional flavor in various foods and delicacies, which can also increase their nutritional value.

Tetradenia spp. is herb naturally growing in Africa and is part of family called Lamiaceae. The herb is used in traditional medicine to cure malaria, angina or as treatment for tropical skin diseases. Maceration was chosen from number of extraction methods, to obtain bioactive compounds from this plant. Optimization was performed with aim to retrieve as much of bioactive compounds as possible. Optimal conditions chosen were: temperature 60 °C, extraction time 40 minutes, solvent 40% ethanol and solid-solvent ratio 1:5. In total 63 aromatic active compounds were found in optimal extract. Terpenes were the most abundant (88,60 %), from them oxygenated monoterpenes (55,65 %) and monoterpenes (29,52 %). Compounds with highest percentage quantity were camphor (51,76 %), cis--ocimene (8,17 %), 3-octyl acetate (7,71 %), trans--ocimene (6,76 %), -terpinene (4,93 %), o-cymene (3,81 %), terpinolene (1,33 %), limonene (1,33 %), camphene (1,19 %) and borneol (1,20 %). Further, total phenolic content was 0,31 ± 0,02 mgGAEml-1, which equals 1,52 ± 0,11 mgGAEg-1 of plant. Antioxidant activity was 417,56 ± 32,84 gTEACml-1 and percentage of radical scavenging activity ABTS•+ was 88,99 ± 7,00 %. Finally antimicrobial activity was determined against bacteria Bacillus cereus, Escherichia coli and yeast Candida glabrata. Against E. coli and C. glabrata there was no inhibition exhibited. Against B. cereus there was seen mild inhibition activity. The results confirm that Tetradenia spp. contain a number of interesting aroma active compounds, which shows antioxidant and antimicrobial activity. The herb has a good potential as natural source of nutritious and sensory active compounds to be used into various foods.

This master thesis deals with preparation of extracts from fruits of aronia, optimization of conditions for extraction and subsequent characteristics of the obtained optimal extract. In the theoretical part information about the botanical characteristics of Aronia melanocarpa and about its utilization are mentioned. Furthermore, the chemical composition of aronia fruits is described and the main biologically active substances contained in the plant are described. At the end of the theoretical part extraction and separation techniques, determination methods of antioxidant, antimicrobial activity and determination of aromatic active compounds are described. The experimental part describes the optimization of maceration so that the macerate contains as much polyphenolic substances as possible, as well as aromatic active substances. 40% ethanol, at a temperature of 60 ° C, with a weight solvent ratio of 1: 5, for 40 minutes proved to be optimal for the maceration of aronia fruits. The total polyphenol content of the optimal extract was determined to be 5237.2 ± 40.7 mgGAEkg-1 and the antioxidant activity by TEAC to be 512.5 ± 5.04 gml-1. 24 aromatic active compounds were identified in the extract, with the largest percentage being monoterpenes and aldehydes. Furthermore, it was found that the extract did not show antimicrobial activity against the microorganisms used (Bacillus cereus, Escherichia coli and Candida glabrata). The obtained results confirm that aronia extracts are an important source of many biologically active compounds and can be used in food as a natural source of nutritionally and sensory valuable substances.

The main goal of this thesis was to analyse the amount of beta glucans in selected mushroom species. The overall characterization of the composition and content of bioactive compounds in mushrooms was studied too. The theoretical part was focused on description of Fungi, especially the Basidiomycota phyla. The overview of fungal morphology and chemical composition was elaborated. Further, theoretical basis of analytical methods used was introduced. In the experimental part, extracts from selected species of mushrooms (shiitake, oyster mushroom, white champignon, brown champignon and jew’s ear) were prepared. Overall group parameters such as polyphenols, flavonoids, antioxidant activity were determined by spectrophotometry. The amount of beta-glucans in the fungi was measured after enzyme hydrolysis using commercial kit. The content and composition of fatty acids was measured by gas spectrometry. Using emission spectrometry (ICP/OES) the presence of heavy metals was analysed in fungal extracts. Cytotoxicity was tested by MTT test on human cell lines. In the last part the antimicrobial activity against Gram-positive bacterium Micrococcus luteus, Gram-negative Serratia marcescens and yeast Candida labrata was tested. In the thesis, a complex characterization of composition and biological effect of some mushrooms was given.

The aim of this diploma thesis was to prove the antimicrobial actvivity of the selected syntetic preservatives used in cosmetics, such as methylparaben, butylparaben, phenoxyethanol, bronopol, imidazolidinyl urea and methylisothaizolinone, in different concentrations (up to the highest permitted concentration and in increased concentrations), and of some essential oils, which are possible to use in cosmetics (essential oil from Eucalyptus globulus, Pimpinella anisum, Lavandula officinalis, Rosmarinus officinalis, Foeniculum vulgare, Mentha piperita, Juniperus communis and Pinus sylvestris). The effect was investigated in every preservative system alone, in order to evaluate the spectrum and the rate of each one. As indicator microorganism was chosen Bacillus subtilis and Penicillium chrysogenum. The used methods was disk difusion method and modify difusion method. From synthetic preservatives had the best effects bronopol, imidazolidinyl urea and methylisothiazolinone. From chosen natural antimicrobails had the effect against both indicator microorganisms the Mentha piperita, Foeniculum vulgare and Juniperus communis extracts.

No description available.

>Magister Scientiae - MSc / The high rate of rediscovery of known compounds has led to a decline in the discovery of novel natural products. The high biodiversity of organisms growing in extreme conditions such as oceans has led to the increased interest by researchers for their use as a source of novel natural products. Marine bacteria are known for their extensive biosynthetic capacity to produce diverse natural products, which are suitable for various biotechnology applications such as in agriculture, for treatment of fungal plant pathogens, and as antibiotics, for treatment of bacterial infections. This study aimed at discovering novel secondary metabolites from marine bacteria previously associated with novel marine invertebrate species endemic to the South African coast. The methodologies used in this study included a bioassay guided fractionation coupled to genome sequencing and mining. For the bioassay guided fractionation approach, the study first focused on screening marine bacteria for antimicrobial activity when cultured on 4 different media, against fungal strains previously shown to be virulent olive trunk pathogens. In parallel, the bacterial isolates with the most inhibitory activity against the fungal pathogens were also screened for antimicrobial activity against 4 indicator strains including Gram-negative Escherichia coli 1699 (E. coli), Pseudomonas putida, and Gram-positive Staphylococcus epidermidis ATCC14990, and Bacillus cereus ATCC10702. One of the marine bacterial isolates, PE6-126, showed diverse antimicrobial activity including antibacterial and antifungal activity against the tested strains. The genome sequencing data revealed that this isolate was B. cereus based on the average nucleotide identity (ANI) (>99%) to reference strains. antiSMASH analysis of the genome revealed nine predicted secondary metabolite clusters including bacteriocins (2), non-ribosomal peptide synthetase (NRPS) (2), siderophore (1), sactipeptide (1), betalactone (1), linear azol(in)e-containing peptides (LAP) - bacteriocin (1) and a terpene (1). Some of these pathways had low to no sequence similarity to known pathways, indicating the potential of these pathways to produce novel compounds. One of the pathways showed very high sequence similarity to the thuricin CD pathway in Bacillus thuringiensis. Considering that thuricin CD has been reported to have antimicrobial activity against B. cereus (ATCC1072), it was hypothesised that it could also be produced by PE6-126. However, the antimicrobial extract from PE6-126 was tested for sensitivity to proteinase K and heat treatment, which thuricin CD is known to be sensitive to. The results revealed that the antimicrobial activity was not lost after treatment, implying that a different metabolite could be responsible for the anti-B. cereusactivity. In addition, PE6-126 initially displayed antimicrobial activity against a multi-drug resistant E. coli 1699, suggesting some of the antimicrobial compound/(s) produced by this strain could potentially be novel. The bioassay-guided fractionation approach coupled to Liquid Chromatography Mass Spectrometry (LC-MS) did not lead to identification of the antimicrobial compound/(s), therefore it remains a question whether the secondary metabolite pathways predicted by antiSMASH lead to the production of the active compound/(s).The results from this study showed that even well studied species have the potential to synthesize as yet undescribed compounds, based on the novelty of some of the pathways. This study highlights the importance of employing a genome-guided approach in drug discovery, as there may be many novel compounds to discover from biosynthetic pathways that have not yet been characterised. Further research is needed to identify the antimicrobial compound/(s) produced by PE6-126.