Microbiological and physicochemical aspects of “salpicão”, a traditional dry sausage produced in the northeast of Portugal

Counts of total viable mesophilic bacteria (TVC), lactic acid bacteria (LAB), Microccocaceae, Enterobacteriaceae, Salmonella spp. and Listeria monocytogenes, in traditional Portuguese dry sausages from two industrial producers, were compared in batter and final product. During the production process, the TVC increased significantly, most likely due to the multiplication of fermentative flora. Enterobacteriaceae decreased from batter to final product while the S. aureus increased. Great variability was verified in detection of L. monocytogenes both between batches and industrial producers

Development of an historical landscape photography database to support landscape change analysis in the Northeast of Portugal

Repeat photography is an efficient, effective and useful method to identify trends of changes in the landscapes. It was used to illustrate long-term changes occurring in the landscapes. In the Northeast of Portugal, landscapes changes is currently driven mostly by agriculture abandonment and agriculture and energy policy. However, there is a need to monitoring changes in the region using a multitemporal and multiscale approach. This project aimed to establish an online repository of oblique digital photography from the region to be used to register the condition of the landscape as recorded in historical and contemporary photography over time as well as to support qualitative and quantitative assessment of change in the landscape using repeat photography techniques and methods. It involved the development of a relational database and a series of web-based services using PHP: Hypertext Preprocessor language, and the development of an interface, with Joomla, of pictures uploading and downloading by users. The repository will make possible to upload, store, search by location, theme, or date, display, and download pictures for Northeastern Portugal. The website service is devoted to help researchers to obtain quickly the photographs needed to apply RP through a developed search engine. It can be accessed at: http://esa.ipb.pt/digitalandscape/.

Study of antioxidant, antiproliferative and apoptosis-inducing properties of wild mushrooms from the Northeast of Portugal

Mushrooms are known as a powerful source of bioactive compounds including antioxidants, inhibitors of human tumour cell lines growth, inducers of apoptosis and enhancers of immunity. Indeed, many pre-clinical studies have been conducted in human tumour cell lines and in some cases a number of compounds isolated from mushrooms have followed to clinical trials. The Northeast of Portugal is one of the European regions with higher wild mushrooms diversity. However, to our knowledge, no studies had been conducted so far to verify their bioactivities. The main aim of this work was the evaluation of the bioactive properties (antioxidant properties and growth inhibitory potential on human tumour cell lines) of wild edible mushrooms collected in the Northeast of Portugal. Once properly identified, methanolic, ethanolic and boiling water extracts were prepared from thirty eight wild mushroom species collected in that region. Chemical characterization was obtained by high performance liquid chromatography (HPLC) coupled to a photodiode array detector (DAD) or to a refraction index detector (RI). Antioxidant activity assays were carried out in those extracts, including evaluation of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging capacity, reducing power and inhibition of β-carotene bleaching. Extract-induced cell growth inhibition was assessed with the sulforhodamine B assay in four human tumour cell lines (NCI-H460 - lung cancer, MCF-7 -breast cancer, HCT-15 -colon cancer and AGS - gastric cancer). The effects on cell cycle profile and apoptosis were evaluated by flow cytometry and the effect on the expression levels of proteins related to cell cycle and apoptosis was further investigated by Western blotting. Three wild edible mushroom species revealed growth inhibitory activity in the studied human tumour cell lines: Clitocybe alexandri ethanolic extract, Lepista inversa methanolic extract and Suillus collinitus methanolic extract. C. alexandri ethanolic extract induced an S-phase cell cycle arrest and increased the percentage of apoptotic cells, in the NCI-H460 cell line. The analysed mushroom species also provided interesting antioxidant potential, mainly the boiling water extract of L. inversa which showed the highest DPPH radical scavenging activity, reducing power and β-carotene bleaching inhibition. S. collinitus methanolic extract induced a slight increase in the number of cells in G1, with a concomitant decrease in the percentage of cells in the S phase of the cell cycle and an increase in the percentage of apoptotic cells, in the MCF-7 cell line. The combined use of the S. collinitus methanolic extract and etoposide caused a greater decrease in the percentage of cell growth, when compared to either of them used individually, indicating the potential benefit of this combination. The tested extracts were chemically characterized and protocatechuic, p-hydroxybenzoic, p-coumaric and cinnamic acids were the main compounds identified on the phenolic (methanolic and ethanolic) extracts, while mannitol, trehalose and arabinose were the main sugars found in the polysaccharidic (boiling water) extracts after hydrolysis. The individual compounds identified in the extracts were submitted to a screening of tumour cells growth inhibitory activity, but only the phenolic acids and a related compound, cinnamic acid, presented activity. This compound was found to be the most potent one regarding cell growth inhibition in the NCI-H460 cell line. The effect of the individual and combined treatment with the identified compounds was also evaluated. Cinnamic and protochatequic acids caused a statistically significantly reduction in the number of viable cells. In addition, p-hydroxybenzoic acid did not show any significantly reduction in the viable cell number. Nevertheless, it was verified that the concomitant use of the three compounds provided the strongest decrease in the viable cell number, suggesting a possible concomitant effect of those compounds. Overall, the present work has contributed to further understand the bioactive potential of wild edible mushrooms from the Northeast of Portugal. This study allowed to identify some species with antioxidant or tumour cell growth inhibitory potential.