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Papers by Adelaide Almeida

Research paper thumbnail of Wetting behaviour of femtosecond laser textured Ti–6Al–4V surfaces

Applied Surface Science, 2013

Research paper thumbnail of The Role of Porphyrinoid Photosensitizers for Skin Wound Healing

International Journal of Molecular Sciences

Microorganisms, usually bacteria and fungi, grow and spread in skin wounds, causing infections. T... more Microorganisms, usually bacteria and fungi, grow and spread in skin wounds, causing infections. These infections trigger the immune system and cause inflammation and tissue damage within the skin or wound, slowing down the healing process. The use of photodynamic therapy (PDT) to eradicate microorganisms has been regarded as a promising alternative to anti-infective therapies, such as those based on antibiotics, and more recently, is being considered for skin wound-healing, namely for infected wounds. Among the several molecules exploited as photosensitizers (PS), porphyrinoids exhibit suitable features for achieving those goals efficiently. The capability that these macrocycles display to generate reactive oxygen species (ROS) gives a significant contribution to the regenerative process. ROS are responsible for avoiding the development of infections by inactivating microorganisms such as bacteria but also by promoting cell proliferation through the activation of stem cells which re...

Research paper thumbnail of Photodynamic Action against Wastewater Microorganisms and Chemical Pollutants: An Effective Approach with Low Environmental Impact

Research paper thumbnail of Evaluation of resistance development and viability recovery by a non-enveloped virus after repeated cycles of aPDT

Antiviral Research, 2011

Nowadays, the emergence of drug resistant microorganisms is a public health concern. The antimicr... more Nowadays, the emergence of drug resistant microorganisms is a public health concern. The antimicrobial photodynamic therapy (aPDT) has an efficient action against a wide range of microorganisms and can be viewed as an alternative approach for treating microbial infections. The aim of this study was to determine if a model target virus (T4-like bacteriophage), in the presence of the tricationic porphyrin 5,10,15-tris(1-methylpyridinium-4-yl)-20-(pentafluorophenyl)porphyrin tri-iodide (Tri-Py(+)-Me-PF), can develop resistance to aPDT and recover its viability after photodynamic treatments. To assess the development of aPDT resistance after repeated treatments, a suspension of T4-like bacteriophage was irradiated with white light (40 Wm(-2)) for 120 min in the presence of 5.0 μM of Tri-Py(+)-Me-PF (99.99% of inactivation) and new phage suspensions were produced from the surviving phages, after each cycle of light exposure. The procedure was repeated ten times. To evaluate the recovery of viral viability after photoinactivation, a suspension of T4-like bacteriophage was irradiated with white light for 120 min in the presence of 5.0 μM of Tri-Py(+)-Me-PF on five consecutive days. In each day, an aliquot of the irradiated suspension was plated and the number of lysis plaques was counted after 24, 48, 72, 96 and 120 h of dark incubation at 37 °C. The profile of bacteriophage photoinactivation did not change after ten consecutive cycles and no recovery of viability was detected after five accumulated cycles of photodynamic treatment. The results suggest that aPDT represents a valuable and promising alternative therapy to treat viral infections, overcoming the problem of microbial resistance.

Research paper thumbnail of Bacterial biomass production in an estuarine system: high variability of leucine conversion factors and changes in bacterial community structure during incubation

Aquatic Microbial Ecology, 2011

Page 1. AQUATIC MICROBIAL ECOLOGY Aquat Microb Ecol Vol. 62: 299–310, 2011 doi: 10.3354/ame01472 ... more Page 1. AQUATIC MICROBIAL ECOLOGY Aquat Microb Ecol Vol. 62: 299–310, 2011 doi: 10.3354/ame01472 Published online February 8 INTRODUCTION Heterotrophic bacterioplankton play a key role in the biological transfer of carbon via the microbial loop (Ghiglione et al. ...

Research paper thumbnail of Evaluation of resistance development and viability recovery by toxigenic and non-toxigenic Staphylococcus aureus strains after repeated cycles of high hydrostatic pressure

Food Microbiology, Sep 28, 2014

In this work, the development of resistance and the recovery of growth after several consecutive ... more In this work, the development of resistance and the recovery of growth after several consecutive cycles of high hydrostatic pressure (HPP) were for the first time evaluated in different strains of Staphylococcus aureus. Three strains of this important and highly resilient to HPP foodborne pathogen were used: a non-enterotoxigenic ATCC 6538 strain, treated with 600 MPa for 30 min at 20 °C, and the toxigenic strains 2153 MA (with enterotoxin A) and 2065 MA (with the enterotoxins A, G and I), treated with 600 MPa for 15 min at 20 °C. After the first treatment, surviving colonies were used to produce new bacterial cultures. This procedure was repeated nine times more for each bacterium or until total inactivation occurred. The inactivation profile of non-enterotoxic strain and the two enterotoxic strains did not change after consecutive cycles, but the toxic strain with three enterotoxins was completely inactivated after the fourth cycle. The three strains did not recover their viability after 14 days. The results indicate that HPP effectively inactivates non-toxigenic and toxigenic strains of S. aureus after a single treatment. The surviving bacteria did not develop resistance after 10 cycles of pressurization and did not recover their viability after 14 days of incubation.

Research paper thumbnail of Baptista I 2014 Final Food Microbiology

Research paper thumbnail of Involvement of oxidative stress in UV-induced impairment of bacterial activity and culturability

Research paper thumbnail of Effects of UV-B Radiation on the Structural and Physiological Diversity of Bacterioneuston and Bacterioplankton

Applied and Environmental Microbiology, 2012

Research paper thumbnail of Effect of tributyltin (TBT) in the metabolic activity of TBT-resistant and sensitive estuarine bacteria

Environmental Toxicology, 2012

The effect of tributyltin (TBT) on growth and metabolic activity of three estuarine bacteria with... more The effect of tributyltin (TBT) on growth and metabolic activity of three estuarine bacteria with different TBT resistance profiles was investigated in an organic-rich culture medium (TSB) and in phosphate buffered saline (PBS) buffer. Exposure to TBT was assessed by determining its effect on growth (OD(600 nm) measurement), bacterial productivity (leucine incorporation), viability (CFU counts), aggregation and cell size (from Live/Dead analysis), ATP and NADH concentrations. TBT exposure resulted in decrease of bacterial density, cell size, and metabolic activity. In addition, cell aggregates were observed in the TBT-treated cultures. TBT strongly affected bacterial cell metabolism and seemed to exert an effect on its equilibrium, interfering with cell activity. Also, TBT toxicity was lower when cells were grown in TSB than in PBS, suggesting that a nutrient-rich growth medium can protect cells from TBT toxicity. This study contributes to our understanding of the TBT-resistant cell behavior reflected in its physiology and metabolic activity. This information is of utmost importance for further studies of TBT bioremediation.

Research paper thumbnail of A microcosm approach to evaluate the degradation of tributyltin (TBT) by Aeromonas molluscorum Av27 in estuarine sediments

Environmental Research, 2014

Research paper thumbnail of The UV responses of bacterioneuston and bacterioplankton isolates depend on the physiological condition and involve a metabolic shift

FEMS Microbiology Ecology, 2012

Research paper thumbnail of Baptista et al 2014 proof Food Microbiology

Research paper thumbnail of Contribution of chemical water properties to the differential responses of bacterioneuston and bacterioplankton to ultraviolet-B radiation

FEMS Microbiology Ecology, 2014

The surface microlayer (SML) is characterized by different physicochemical properties from underl... more The surface microlayer (SML) is characterized by different physicochemical properties from underlying waters (UW). However, whether these differences in abiotic factors underlie the distinct sensitivity of bacterioneuston (i.e. SML bacteria) and bacterioplankton to environmental stressors remains to be addressed. We investigated the contribution of abiotic factors to the UV-B sensitivity of bacterioneuston and bacterioplankton. Nutrients (especially nitrogen and phosphate) emerged as important determinants of bacterial UV-B sensitivity. The role of particles, nutrients, and dissolved organic components on bacterial UV-B sensitivity was further evaluated using dilution cultures. Filtered samples were twofold more UV sensitive than unfiltered samples, suggesting a UV-protective effect of particles. High nutrient concentrations attenuated bacterial UV-B sensitivity (up to 40%), compared with unamended conditions, by influencing bacterial physiology and/or community composition. Suspending cells in natural water, particularly from the SML, also attenuated UV-B sensitivity (up to 23%), compared with suspension in an artificial mineral solution. Bioassays using Pseudomonas sp. strain NT5I1.2B revealed that chemical water properties influence UV-induced oxidative damage. UV-B sensitivity was associated with high cell-specific activities. The chemical environment of the SML and UW influences UV-B effects on the corresponding bacterial communities. Maintaining low cell activities might be advantageous in stressful environments, like the SML.

Research paper thumbnail of Can Volatile Organic Metabolites Be Used to Simultaneously Assess Microbial and Mite Contamination Level in Cereal Grains and Coffee Beans?

PLoS ONE, 2013

A novel approach based on headspace solid-phase microextraction (HS-SPME) combined with comprehen... more A novel approach based on headspace solid-phase microextraction (HS-SPME) combined with comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-ToFMS) was developed for the simultaneous screening of microbial and mite contamination level in cereals and coffee beans. The proposed approach emerges as a powerful tool for the rapid assessment of the microbial contamination level (ca. 70 min versus ca. 72 to 120 h for bacteria and fungi, respectively, using conventional plate counts), and mite contamination (ca. 70 min versus ca. 24 h). A full-factorial design was performed for optimization of the SPME experimental parameters. The methodology was applied to three types of rice (rough, brown, and white rice), oat, wheat, and green and roasted coffee beans. Simultaneously, microbiological analysis of the samples (total aerobic microorganisms, moulds, and yeasts) was performed by conventional plate counts. A set of 54 volatile markers was selected among all the compounds detected by GC×GC-ToFMS. Principal Component Analysis (PCA) was applied in order to establish a relationship between potential volatile markers and the level of microbial contamination. Methylbenzene, 3-octanone, 2-nonanone, 2-methyl-3-pentanol, 1-octen-3-ol, and 2-hexanone were associated to samples with higher microbial contamination level, especially in rough rice. Moreover, oat exhibited a high GC peak area of 2-hydroxy-6-methylbenzaldehyde, a sexual and alarm pheromone for adult mites, which in the other matrices appeared as a trace component. The number of mites detected in oat grains was correlated to the GC peak area of the pheromone. The HS-SPME/GC×GC-ToFMS methodology can be regarded as the basis for the development of a rapid and versatile method that can be applied in industry to the simultaneous assessment the level of microbiological contamination and for detection of mites in cereals grains and coffee beans.

Research paper thumbnail of Phage Therapy as an Approach to Prevent Vibrio anguillarum Infections in Fish Larvae Production

Research paper thumbnail of Chapter 3 da tese

Research paper thumbnail of Poster Microbiotec 2013 fagos Aeromonas

Research paper thumbnail of Chapter 4 da tese

Research paper thumbnail of Chapter 6 da tese

Research paper thumbnail of Wetting behaviour of femtosecond laser textured Ti–6Al–4V surfaces

Applied Surface Science, 2013

Research paper thumbnail of The Role of Porphyrinoid Photosensitizers for Skin Wound Healing

International Journal of Molecular Sciences

Microorganisms, usually bacteria and fungi, grow and spread in skin wounds, causing infections. T... more Microorganisms, usually bacteria and fungi, grow and spread in skin wounds, causing infections. These infections trigger the immune system and cause inflammation and tissue damage within the skin or wound, slowing down the healing process. The use of photodynamic therapy (PDT) to eradicate microorganisms has been regarded as a promising alternative to anti-infective therapies, such as those based on antibiotics, and more recently, is being considered for skin wound-healing, namely for infected wounds. Among the several molecules exploited as photosensitizers (PS), porphyrinoids exhibit suitable features for achieving those goals efficiently. The capability that these macrocycles display to generate reactive oxygen species (ROS) gives a significant contribution to the regenerative process. ROS are responsible for avoiding the development of infections by inactivating microorganisms such as bacteria but also by promoting cell proliferation through the activation of stem cells which re...

Research paper thumbnail of Photodynamic Action against Wastewater Microorganisms and Chemical Pollutants: An Effective Approach with Low Environmental Impact

Research paper thumbnail of Evaluation of resistance development and viability recovery by a non-enveloped virus after repeated cycles of aPDT

Antiviral Research, 2011

Nowadays, the emergence of drug resistant microorganisms is a public health concern. The antimicr... more Nowadays, the emergence of drug resistant microorganisms is a public health concern. The antimicrobial photodynamic therapy (aPDT) has an efficient action against a wide range of microorganisms and can be viewed as an alternative approach for treating microbial infections. The aim of this study was to determine if a model target virus (T4-like bacteriophage), in the presence of the tricationic porphyrin 5,10,15-tris(1-methylpyridinium-4-yl)-20-(pentafluorophenyl)porphyrin tri-iodide (Tri-Py(+)-Me-PF), can develop resistance to aPDT and recover its viability after photodynamic treatments. To assess the development of aPDT resistance after repeated treatments, a suspension of T4-like bacteriophage was irradiated with white light (40 Wm(-2)) for 120 min in the presence of 5.0 μM of Tri-Py(+)-Me-PF (99.99% of inactivation) and new phage suspensions were produced from the surviving phages, after each cycle of light exposure. The procedure was repeated ten times. To evaluate the recovery of viral viability after photoinactivation, a suspension of T4-like bacteriophage was irradiated with white light for 120 min in the presence of 5.0 μM of Tri-Py(+)-Me-PF on five consecutive days. In each day, an aliquot of the irradiated suspension was plated and the number of lysis plaques was counted after 24, 48, 72, 96 and 120 h of dark incubation at 37 °C. The profile of bacteriophage photoinactivation did not change after ten consecutive cycles and no recovery of viability was detected after five accumulated cycles of photodynamic treatment. The results suggest that aPDT represents a valuable and promising alternative therapy to treat viral infections, overcoming the problem of microbial resistance.

Research paper thumbnail of Bacterial biomass production in an estuarine system: high variability of leucine conversion factors and changes in bacterial community structure during incubation

Aquatic Microbial Ecology, 2011

Page 1. AQUATIC MICROBIAL ECOLOGY Aquat Microb Ecol Vol. 62: 299–310, 2011 doi: 10.3354/ame01472 ... more Page 1. AQUATIC MICROBIAL ECOLOGY Aquat Microb Ecol Vol. 62: 299–310, 2011 doi: 10.3354/ame01472 Published online February 8 INTRODUCTION Heterotrophic bacterioplankton play a key role in the biological transfer of carbon via the microbial loop (Ghiglione et al. ...

Research paper thumbnail of Evaluation of resistance development and viability recovery by toxigenic and non-toxigenic Staphylococcus aureus strains after repeated cycles of high hydrostatic pressure

Food Microbiology, Sep 28, 2014

In this work, the development of resistance and the recovery of growth after several consecutive ... more In this work, the development of resistance and the recovery of growth after several consecutive cycles of high hydrostatic pressure (HPP) were for the first time evaluated in different strains of Staphylococcus aureus. Three strains of this important and highly resilient to HPP foodborne pathogen were used: a non-enterotoxigenic ATCC 6538 strain, treated with 600 MPa for 30 min at 20 °C, and the toxigenic strains 2153 MA (with enterotoxin A) and 2065 MA (with the enterotoxins A, G and I), treated with 600 MPa for 15 min at 20 °C. After the first treatment, surviving colonies were used to produce new bacterial cultures. This procedure was repeated nine times more for each bacterium or until total inactivation occurred. The inactivation profile of non-enterotoxic strain and the two enterotoxic strains did not change after consecutive cycles, but the toxic strain with three enterotoxins was completely inactivated after the fourth cycle. The three strains did not recover their viability after 14 days. The results indicate that HPP effectively inactivates non-toxigenic and toxigenic strains of S. aureus after a single treatment. The surviving bacteria did not develop resistance after 10 cycles of pressurization and did not recover their viability after 14 days of incubation.

Research paper thumbnail of Baptista I 2014 Final Food Microbiology

Research paper thumbnail of Involvement of oxidative stress in UV-induced impairment of bacterial activity and culturability

Research paper thumbnail of Effects of UV-B Radiation on the Structural and Physiological Diversity of Bacterioneuston and Bacterioplankton

Applied and Environmental Microbiology, 2012

Research paper thumbnail of Effect of tributyltin (TBT) in the metabolic activity of TBT-resistant and sensitive estuarine bacteria

Environmental Toxicology, 2012

The effect of tributyltin (TBT) on growth and metabolic activity of three estuarine bacteria with... more The effect of tributyltin (TBT) on growth and metabolic activity of three estuarine bacteria with different TBT resistance profiles was investigated in an organic-rich culture medium (TSB) and in phosphate buffered saline (PBS) buffer. Exposure to TBT was assessed by determining its effect on growth (OD(600 nm) measurement), bacterial productivity (leucine incorporation), viability (CFU counts), aggregation and cell size (from Live/Dead analysis), ATP and NADH concentrations. TBT exposure resulted in decrease of bacterial density, cell size, and metabolic activity. In addition, cell aggregates were observed in the TBT-treated cultures. TBT strongly affected bacterial cell metabolism and seemed to exert an effect on its equilibrium, interfering with cell activity. Also, TBT toxicity was lower when cells were grown in TSB than in PBS, suggesting that a nutrient-rich growth medium can protect cells from TBT toxicity. This study contributes to our understanding of the TBT-resistant cell behavior reflected in its physiology and metabolic activity. This information is of utmost importance for further studies of TBT bioremediation.

Research paper thumbnail of A microcosm approach to evaluate the degradation of tributyltin (TBT) by Aeromonas molluscorum Av27 in estuarine sediments

Environmental Research, 2014

Research paper thumbnail of The UV responses of bacterioneuston and bacterioplankton isolates depend on the physiological condition and involve a metabolic shift

FEMS Microbiology Ecology, 2012

Research paper thumbnail of Baptista et al 2014 proof Food Microbiology

Research paper thumbnail of Contribution of chemical water properties to the differential responses of bacterioneuston and bacterioplankton to ultraviolet-B radiation

FEMS Microbiology Ecology, 2014

The surface microlayer (SML) is characterized by different physicochemical properties from underl... more The surface microlayer (SML) is characterized by different physicochemical properties from underlying waters (UW). However, whether these differences in abiotic factors underlie the distinct sensitivity of bacterioneuston (i.e. SML bacteria) and bacterioplankton to environmental stressors remains to be addressed. We investigated the contribution of abiotic factors to the UV-B sensitivity of bacterioneuston and bacterioplankton. Nutrients (especially nitrogen and phosphate) emerged as important determinants of bacterial UV-B sensitivity. The role of particles, nutrients, and dissolved organic components on bacterial UV-B sensitivity was further evaluated using dilution cultures. Filtered samples were twofold more UV sensitive than unfiltered samples, suggesting a UV-protective effect of particles. High nutrient concentrations attenuated bacterial UV-B sensitivity (up to 40%), compared with unamended conditions, by influencing bacterial physiology and/or community composition. Suspending cells in natural water, particularly from the SML, also attenuated UV-B sensitivity (up to 23%), compared with suspension in an artificial mineral solution. Bioassays using Pseudomonas sp. strain NT5I1.2B revealed that chemical water properties influence UV-induced oxidative damage. UV-B sensitivity was associated with high cell-specific activities. The chemical environment of the SML and UW influences UV-B effects on the corresponding bacterial communities. Maintaining low cell activities might be advantageous in stressful environments, like the SML.

Research paper thumbnail of Can Volatile Organic Metabolites Be Used to Simultaneously Assess Microbial and Mite Contamination Level in Cereal Grains and Coffee Beans?

PLoS ONE, 2013

A novel approach based on headspace solid-phase microextraction (HS-SPME) combined with comprehen... more A novel approach based on headspace solid-phase microextraction (HS-SPME) combined with comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry (GC×GC-ToFMS) was developed for the simultaneous screening of microbial and mite contamination level in cereals and coffee beans. The proposed approach emerges as a powerful tool for the rapid assessment of the microbial contamination level (ca. 70 min versus ca. 72 to 120 h for bacteria and fungi, respectively, using conventional plate counts), and mite contamination (ca. 70 min versus ca. 24 h). A full-factorial design was performed for optimization of the SPME experimental parameters. The methodology was applied to three types of rice (rough, brown, and white rice), oat, wheat, and green and roasted coffee beans. Simultaneously, microbiological analysis of the samples (total aerobic microorganisms, moulds, and yeasts) was performed by conventional plate counts. A set of 54 volatile markers was selected among all the compounds detected by GC×GC-ToFMS. Principal Component Analysis (PCA) was applied in order to establish a relationship between potential volatile markers and the level of microbial contamination. Methylbenzene, 3-octanone, 2-nonanone, 2-methyl-3-pentanol, 1-octen-3-ol, and 2-hexanone were associated to samples with higher microbial contamination level, especially in rough rice. Moreover, oat exhibited a high GC peak area of 2-hydroxy-6-methylbenzaldehyde, a sexual and alarm pheromone for adult mites, which in the other matrices appeared as a trace component. The number of mites detected in oat grains was correlated to the GC peak area of the pheromone. The HS-SPME/GC×GC-ToFMS methodology can be regarded as the basis for the development of a rapid and versatile method that can be applied in industry to the simultaneous assessment the level of microbiological contamination and for detection of mites in cereals grains and coffee beans.

Research paper thumbnail of Phage Therapy as an Approach to Prevent Vibrio anguillarum Infections in Fish Larvae Production

Research paper thumbnail of Chapter 3 da tese

Research paper thumbnail of Poster Microbiotec 2013 fagos Aeromonas

Research paper thumbnail of Chapter 4 da tese

Research paper thumbnail of Chapter 6 da tese