Browsing by Author "Domingues, Fernanda"
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- Chemical Profiling and Evaluation of Antioxidant and Anti-Microbial Properties of Selected Commercial Essential Oils: A Comparative StudyPublication . Luís, Ângelo; Duarte, Ana Paula; Pereira, L.; Domingues, FernandaBackground: The last decades have seen an increased awareness by the scientific community of the extent of resistance to conventional antibiotics, particularly with respect to the emerging multidrug-resistant pathogenic microbes. Additionally, natural antioxidants have received significant attention among food professionals and consumers because of their assumed safety and potential therapeutic value. The aim of this work was to assess the antioxidant activities of eight selected commercial essential oils (EOs), together with the evaluation of their antibacterial and anti-quorum sensing properties. Methods: The chemical profiling of the EOs was performed using gas chromatography-mass spectrometry (GC-MS) analysis. The antioxidant properties of the EOs were evaluated using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and by β-carotene bleaching test. Disc diffusion assays were employed to evaluate the anti-bacterial and anti-quorum sensing activities of the EOs. Results: It was observed that EOs from three Eucalyptus species are rich in eucalyptol. Generally, linalool is abundant in EOs from four Lavandula species. The oil of Cymbopogon citratus is the one with the best capacity to scavenge the DPPH free radicals and presented great antibacterial activity. Conclusions: The geographical origins of the plant species are determinant factors in the EO composition and in the corresponding biological activities.
- Insect-based chitin and chitosan from whole body sources and rearing by-products: extraction, physicochemical, structural and bioactivity characterisationPublication . Navarro, Pedro; Ribeiro, José Carlos Reis ; Luís, Ângelo Filipe Santos ; Domingues, Fernanda; Anjos, Ofélia; Cunha, LuisFractionation of edible insects and their rearing by-products can lead to expanded industrial applications and extraction of value-added products. The main goal of this study was to extract chitin and synthesise chitosan from three different insect sources – Tenebrio molitor larvae, adult Acheta domesticus and A. domesticus rearing by-products (legs and wings). Furthermore, the physicochemical, structural, and bioactive properties of insectderived chitin and chitosan were characterised and compared to commercial crustacean-based samples. Chitin was isolated from defatted sources through chemical hydrolysis followed by decolourisation and was deacetylated with a strong alkaline solution to synthesise chitosan. Notably, A. domesticus rearing by-products yielded the highest chitin and chitosan content. Chitin and chitosan derived from insects exhibited physicochemical and structural characteristics consistent with the α-polymorphic form, similar to the commercial samples. Differences were found in surface morphology, with insect-derived samples presenting large and irregular flakes and porosity, while the crustacean-derived samples presented irregularly arranged fibres and a more regular and smoother surface. As for antioxidant activity, although all chitosan samples demonstrated poor DPPH radical scavenging activity, this study showed for the first time that insect-derived chitosan presents lipid peroxidation inhibition ability. All chitosan samples presented antimicrobial activity against different pathogenic bacteria, with K. pneumoniae being the most susceptible strain. Nevertheless, there is potential for enhancement of the biological properties through modifications on the molecular weight and deacetylation degree. This research introduces the potential of cricketrearing by-products as sustainable sources of chitosan with functional bioactive properties.
- Natural Transformation as a Mechanism of Horizontal Gene Transfer in Aliarcobacter butzleriPublication . Bonifácio, Marina; Mateus, Cristiana; Alves, Ana R.; Maldonado, Emanuel; Duarte, Ana Paula; Domingues, Fernanda; Oleastro, Mónica; Ferreira, SusanaAliarcobacter butzleri is an emergent enteropathogen, showing high genetic diversity, which likely contributes to its adaptive capacity to different environments. Whether natural transformation can be a mechanism that generates genetic diversity in A. butzleri is still unknown. In the present study, we aimed to establish if A. butzleri is naturally competent for transformation and to investigate the factors influencing this process. Two different transformation procedures were tested using exogenous and isogenic DNA containing antibiotic resistance markers, and different external conditions influencing the process were evaluated. The highest number of transformable A. butzleri strains were obtained with the agar transformation method when compared to the biphasic system (65% versus 47%). A. butzleri was able to uptake isogenic chromosomal DNA at different growth phases, and the competence state was maintained from the exponential to the stationary phases. Overall, the optimal conditions for transformation with the biphasic system were the use of 1 µg of isogenic DNA and incubation at 30 ◦C under a microaerobic atmosphere, resulting in a transformation frequency ~8 × 10−6 transformants/CFU. We also observed that A. butzleri favored the transformation with the genetic material of its own strain/species, with the DNA incorporation process occurring promptly after the addition of genomic material. In addition, we observed that A. butzleri strains could exchange genetic material in co-culture assays. The presence of homologs of well-known genes involved in the competence in the A. butzleri genome corroborates the natural competence of this species. In conclusion, our results show that A. butzleri is a naturally transformable species, suggesting that horizontal gene transfer mediated by natural transformation is one of the processes contributing to its genetic diversity. In addition, natural transformation can be used as a tool for genetic studies of this species.
- Quorum sensing inhibition evaluation method: An experiment-based microbiology laboratory coursePublication . Luís, Ângelo Filipe Santos ; Domingues, FernandaBacteria have developed a cell-to-cell communication system called quorum sensing (QS), allowing them to regulate group behavior and synchronize the expression of virulence factors, responsible for increasing their infection capacity and resistance to antimicrobials. Although the control of microbial infections through the inhibition of microbial growth has traditionally been the basis of antimicrobial chemotherapy, the emergence of antimicrobial resistance has led to the search for new microbial control strategies, namely through the inhibition of QS. Among the agents studied to inhibit this bacterial communication are essential oils (EO), which are considered very effective QS inhibitors. When searching for new QS inhibitor agents, it is essential to have a cheap and easy-to-perform method that allows the evaluation of this activity. Chromobacterium violaceum is a Gram-negative bacterium that has been widely used as a model organism in QS research laboratories because it produces the violet-colored pigment violacein, which is regulated by QS and is an easily observable and quantifiable characteristic marker. The objective of this work is to describe a method to evaluate the inhibition of the QS using Cymbopogon martinii EO as a potential inhibitory agent for violacein production by C. violaceum, which can be applied in the Microbiology laboratory course as a part of the programs of several science degrees. The proposed method is inexpensive and does not require specific equipment, enabling its easy implementation by the laboratory team and professors.