Maria Leonor Faleiro

Antimicrobial activity of essential oils isolated from Portuguese endemic species of Thymus

Aims: Thymus species are wild species mostly found in the arid lands of Portugal. Possible antimicrobial properties of Thymus essential oils have been investigated. The chemical composition of the essential oils and the antimicrobial activity of Thymus mastichina (L) L. subsp. mastichina, T. camphoratus and T. lotocephalus from different regions of Portugal were analysed.

Stress response of Listeria monocytogenes isolated from cheese and other foods

The responses to pH and sodium chloride of four strains of Listeria monocytogenes isolated from Portuguese cheese, with a sodium chloride concentration of about 2% (w/v) and a pH value from 5.1 to 6.2, were studied. Two isolates from meat and two clinical isolates related to food-borne listeriosis, in which the implicated food product had about 2-3.5% (w/v) sodium chloride, also were studied. The effect of temperature on pH and sodium chloride sensitivity was also determined. The results show that natural isolates vary in response to these stresses and the data were often at variance with previously published data. Strains varied in sensitivity to low pH and to high sodium chloride concentration but the cheese isolates tended to be more resistant. A lower temperature was associated with a decrease in resistance to low pH and to sodium chloride. All strains showed an acid tolerance response induction when grown at pH 5.5 and although the time required for maximum induction of the response varied between strains, 2 h of acid adaptation, at least, was necessary which is longer than previously reported. Some strains showed an osmotolerance response after incubation in 3.5% (w/v) sodium chloride. Osmoadaptation, in addition to inducing an osmotolerance response, also induced cross-protection against acid shock conditions (pH 3.5). The acid tolerance response also induced a cross-protection against osmotic shock conditions (20% (w/v) sodium chloride). In some cases there was a relationship between the degree of resistance and adaptation, but usually the behaviour of a particular strain was independent of the conditions from which it was isolated.

Portuguese Thymbra and Thymus Species Volatiles: Chemical Composition and Biological Activities

Thymbra capitata and Thymus species are commonly known in Portugal as thyme and they are currently used as culinary herbs, as well as for ornamental, aromatizing and traditional medicinal purposes. The present work reports on the state of the art on the information available on the taxonomy, ethnobotany, cell and molecular biology of the Portuguese representatives of these genera and on the chemotaxonomy and antibacterial, antifungal and antioxidant activities of their essential oils and other volatile-containing extracts.

Marked intra-strain variation in response of Listeria monocytogenes dairy isolates to acid or salt stress and the effect of acid or salt adaptation on adherence to abiotic surfaces

During food processing, and particularly in cheese manufacturing processes, Listeria monocytogenes may be exposed routinely to environments of low pH or high salt concentration. It has been suggested that these environmental conditions may contribute to bacterial adherence to abiotic surfaces and increased resistance to disinfection. In this study strains isolated from the environment of artisanal cheese-making dairies were used to investigate the behaviour of L. monocytogenes in response to acid and salt stress and clear differences between strains was observed. In planktonic culture, strains varied in resistance to low pH or high NaCl concentration and in the occurrence of an adaptive response to moderate acid or NaCl. There was dislocation in responses to salt and acid. Strains resistant, or adaptive, to acid were not resistant or adaptive to NaCl. The reverse also was observed. Exposure to moderate acid did not promote adherence to polystyrene but survival, at low pH or high NaCl concentration, of cells adherent to stainless steel was increased, even for strains that had no adaptive response planktonically, but the detail of these observations varied between strains. In contrast to acid adaptation, with some strains salt adaptation enhanced adherence of L. monocytogenes to polystyrene but this was not true for all strains. For some strains salt- or acid adaptation may enhance the survival of sessile cells exposed to hypochlorite disinfection.

Anaerobic bio-removal of uranium (VI) and chromium (VI): Comparison of microbial community structure

Several microbial communities, obtained from uranium contaminated and non-contaminated samples, were investigated for their ability to remove uranium (VI) and the cultures capable for this removal were further assessed on their efficiency for chromium (VI) removal. The highest efficiency for removal of both metals was observed on a consortium from a non-contaminated soil collected in Monchique thermal place, which was capable to remove 91% of 22 mg L(-1) U(VI) and 99% of 13 mg L(-1) Cr(VI). This study revealed that uranium (VI) removing communities have also ability to remove chromium (VI), but when uranium (VI) was replaced by chromium (VI) several differences in the structure of all bacterial communities were observed. TGGE and phylogenetic analysis of 16S rRNA gene showed that the uranium (VI) removing bacterial consortia are mainly composed by members of Rhodocyclaceae family and Clostridium genus. On the other hand, bacteria from Enterobacteriaceae family were detected in the community with ability for chromium (VI) removal. The existence of members of Enterobacteriaceae and Rhodocyclaceae families never reported as chromium or uranium removing bacteria, respectively, is also a relevant finding, encouraging the exploitation of microorganisms with new abilities that can be useful for bioremediation.

Mechanism of uranium (VI) removal by two anaerobic bacterial communities.

The mechanism of uranium (VI) removal by two anaerobic bacterial consortia, recovered from an uncontaminated site (consortium A) and other from an uranium mine (consortium U), was investigated. The highest efficiency of U (VI) removal by both consortia (97%) occurred at room temperature and at pH 7.2. Furthermore, it was found that U (VI) removal by consortium A occurred by enzymatic reduction and bioaccumulation, while the enzymatic process was the only mechanism involved in metal removal by consortium U. FTIR analysis suggested that after U (VI) reduction, U (IV) could be bound to carboxyl, phosphate and amide groups of bacterial cells. Phylogenetic analysis of 16S rRNA showed that community A was mainly composed by bacteria closely related to Sporotalea genus and Rhodocyclaceae family, while community U was mainly composed by bacteria related to Clostridium genus and Rhodocyclaceae family.

Salvia officinalis L. essential oils: effect of hydrodistillation time on the chemical composition, antioxidant and antimicrobial activities

Salvia officinalis L. oils were isolated from the plants commercial dried aerial parts, by hydrodistillation, with different distillation times. The essential oils were analysed by gas chromatography and gas chromatography–mass spectrometry. The antioxidant ability was measured using a free radical scavenging activity assay using 2,2-diphenyl-1-picrylhydrazyl (DPPH), a thiobarbituric acid reactive substances (TBARS) assay, a deoxyribose assay for the scavenging of hydroxyl radical, an assay for site-specific actions and a 5-lipoxygenase assay. Antibacterial activity was determined by the agar diffusion method. 1,8-Cineole, α-pinene and camphor were the dominant components of all the essential oils. The different hydrodistillation times did not affect the oil yield nor the relative amount of the oil components. The time of hydrodistillation influenced the antioxidant activity. With the DPPH method, the oils isolated for 2 and 3 h were stronger free radical scavengers, while with the TBARS method, the highest antioxidant values were obtained in the oils isolated for 30 min, 2 and 3 h. Hydroxyl radical scavenging and lipoxygenase activity assays showed the best results with oils isolated for 1 and 3 h. With the deoxyribose method, sage oils at concentrations <1000 mg L−1 showed better activity than mannitol. The essential oil of S. officinalis showed very weak antimicrobial activity.

Effect of uranium (VI) on two sulphate-reducing bacteria cultures from a uranium mine site

This work was conducted to assess the impact of uranium (VI) on sulphate-reducing bacteria (SRB) communities obtained from environmental samples collected on the Portuguese uranium mining area of Urgeiriça. Culture U was obtained from a sediment, while culture W was obtained from sludge from the wetland of that mine. Temperature gradient gel electrophoresis (TGGE) was used to monitor community changes under uranium stress conditions. TGGE profiles of dsrB gene fragment demonstrated that the initial cultures were composed of SRB species affiliated with Desulfovibrio desulfuricans, Desulfovibrio vulgaris and Desulfomicrobium spp. (sample U), and by species related to D. desulfuricans (sample W). A drastic change in SRB communities was observed as a result of uranium (VI) exposure. Surprisingly, SRB were not detected in the uranium removal communities. Such findings emphasize the need of monitoring the dominant populations during bio-removal studies. TGGE and phylogenetic analysis of the 16S rRNA gene fragment revealed that the uranium removal consortia are composed by strains affiliated to Clostridium genus, Caulobacteraceae and Rhodocyclaceae families. Therefore, these communities can be attractive candidates for environmental biotechnological applications associated to uranium removal.

Listeria monocytogenes dairy isolates show a different proteome response to sequential exposure to gastric and intestinal fluids.

The gastrointestinal system poses different stresses to the foodborne pathogen, Listeria monocytogenes, including the low pH of the stomach and the presence of bile and the high osmolality of the intestinal fluid. The present study evaluated how previous exposure of three L. monocytogenes dairy isolates (C882 and T8, serovar 4b isolates and A9 serovar 1/2a or 3b isolate) to a cheese-simulated medium (p H5.5 and 3.5% NaCl [w/v], adapted cultures) affected subsequent survival in a simulated gastrointestinal system. Listerial cultures exposed to the cheese-simulated medium at pH7.0, with no added NaCl, were considered non-adapted. To investigate the main events involved in listerial survival during the gastric and intestinal subsequent challenge, a proteomic approach was used. All L. monocytogenes strains were able to survive the deleterious effects of the gastrointestinal fluids and no significant differences were observed between adapted and non-adapted cells. However the L. monocytogenes strains showed a different protein pattern in response to the gastrointestinal stress. Data indicated that synthesis of stress related proteins is more pronounced in non-adapted cells. Although, a significant number of enzymes involved in glycolysis and energy production were also consistently over-produced by the three strains. These findings provided new insights into the means used by L. monocytogenes to overcome the gastrointestinal system and allow the pathogen to move to the next phase of the infectious process.

Arbutus unedo L.: Chemical and Biological Properties

Arbutus unedo L. (strawberry tree) has a circum-Mediterranean distribution, being found in western, central and southern Europe, north-eastern Africa (excluding Egypt and Libya) and the Canary Islands and western Asia. Fruits of the strawberry tree are generally used for preparing alcoholic drinks (wines, liqueurs and brandies), jams, jellies and marmalades, and less frequently eaten as fresh fruit, despite their pleasing appearance. An overview of the chemical composition of different parts of the plant, strawberry tree honey and strawberry tree brandy will be presented. The biological properties of the different parts of A. unedo and strawberry tree honey will be also overviewed.