Evaluation of the in vitro biological activities of extracts from carob tree and mediterranean oaks

Dissertação mest., Engenharia Biológica, Universidade do Algarve, 2009

Chemical and microbial changes during the natural fermentation of strawberry tree (Arbutus unedo L.) fruits

Aguardente de medronho is the name given in Portugal to a spirit made from the fermented fruit of Arbutus unedo (strawberry tree), a plant grown in the Mediterranean region. In order to gain a better understanding of the fermentation process, as it is performed in the farms, a natural fermentation with wild microbiota was carried out during 36 days, and some physicochemical and microbiological parameters were studied. The microbial parameters analyzed were total viable, lactic and acetic acids bacteria, and yeast counts. The physicochemical parameters monitored were sugars, minerals, ethanol, organic acids and pH. Yeasts were the main responsible for the fermentation of the fruits, as the lactic and acetic acids bacteria are absent. As the fermentation progressed, the sugars increased during the first 2 days and gradually decreased along the fermentation period. Maintaining the good quality of the product could contribute to the preservation and valorization of traditional resources that are of great importance to prevent their disappearance.

Effect of nitrogen and potassium fertilisation on vegetative growth and flowering of mature carob trees (Ceratonia siliqua): variations in leaf area index and water use indices

This work aimed to assess how potassium (K) and nitrogen (N) fertilisation may affect the use of precipitation in terms of vegetative and flowering response of 15-year-old carob trees during a 3-year experiment. A field trial was conducted in 1997, 1998 and 1999 in Algarve (Southern Portugal) in a calcareous soil. Four fertilisation treatments were tested: no fertiliser (control); 0.8 kg N/tree (N treatment); 1 kg K 2 O/tree (K treatment) and 0.8 kg N/tree plus 1 kg K 2 O/tree (NK treatment). No irrigation was applied during the experimental period. Branch length increments were measured every month throughout the growing season and inflorescence number was registered once per year. There was a strong seasonal effect on vegetative growth, since low levels of precipitation (115 mm) during October 1998–March 1999 suppressed the increment in branch length. N supplied to the trees (N and NK treatments) tended to increase water use indices in terms of vegetative growth. No response to K alone was observed in trees fertilised only with K. The number of inflorescences increased throughout the experimental period, particularly for N and NK treatments, and a reduction of the precipitation amount during April, May and June, may also enhance flowering. This knowledge could be important when making decisions concerning fertilisation under dry conditions. The results reported here indicate that tree growth (expressed as the branch growth) and flower production under dry-farming conditions, may be achieved by applying 0.8 kg of N (as ammonium nitrate) per tree during the growing season. However, N uptake and use depends on soil water availability.

Evolution of matrix and bone gamma-carboxyglutamic acid proteins in vertebrates

The evolution of calcified tissues is a defining feature in vertebrate evolution. Investigating the evolution of proteins involved in tissue calcification should help elucidate how calcified tissues have evolved. The purpose of this study was to collect and compare sequences of matrix and bone γ-carboxyglutamic acid proteins (MGP and BGP, respectively) to identify common features and determine the evolutionary relationship between MGP and BGP. Thirteen cDNAs and genes were cloned using standard methods or reconstructed through the use of comparative genomics and data mining. These sequences were compared with available annotated sequences (a total of 48 complete or nearly complete sequences, 28 BGPs and 20 MGPs) have been identified across 32 different species (representing most classes of vertebrates), and evolutionarily conserved features in both MGP and BGP were analyzed using bioinformatic tools and the Tree-Puzzle software. We propose that: 1) MGP and BGP genes originated from two genome duplications that occurred around 500 and 400 million years ago before jawless and jawed fish evolved, respectively; 2) MGP appeared first concomitantly with the emergence of cartilaginous structures, and BGP appeared thereafter along with bony structures; and 3) BGP derives from MGP. We also propose a highly specific pattern definition for the Gla domain of BGP and MGP. Previous Section Next Section BGP1 (bone Gla protein or osteocalcin) and MGP (matrix Gla protein) belong to the growing family of vitamin K-dependent (VKD) proteins, the members of which are involved in a broad range of biological functions such as skeletogenesis and bone maintenance (BGP and MGP), hemostasis (prothrombin, clotting factors VII, IX, and X, and proteins C, S, and Z), growth control (gas6), and potentially signal transduction (proline-rich Gla proteins 1 and 2). VKD proteins are characterized by the presence of several Gla residues resulting from the post-translational vitamin K-dependent γ-carboxylation of specific glutamates, through which they can bind to calcium-containing mineral such as hydroxyapatite. To date, VKD proteins have only been clearly identified in vertebrates (1) although the presence of a γ-glutamyl carboxylase has been reported in the fruit fly Drosophila melanogaster (2) and in marine snails belonging to the genus Conus (3). Gla residues have also been found in neuropeptides from Conus venoms (4), suggesting a wider prevalence of γ-carboxylation.

Entomofauna associated with the olive tree in southern Portugal

Since 2004 several studies have been carried out in order to identify the main insect species that usually inhabiting the olive ecosystem. The field trials have taken place in two olive groves, one situated in Olhão and the other one in Loulé, both in Algarve and also under Integrated Pest Management (IPM). The sampling techniques used differ according to their purpose (sticky traps, pheromone traps, pitfall traps and samples of aerial parts of the trees such as inflorescences, leaves, fruits and branches). Results showed that the main insect pests of olive tree in southern Portugal were the olive fruit fly Bactrocera oleae Gmelin (Diptera: Tephritidae) and the olive moth Prays oleae Bernard (Lepidoptera: Hyponeumetidae). Other insect pests were also found in our olive groves namely the olive psyllid Euphyllura olivina Costa (Homoptera: Psyllidae), the olive dark beetle Phloeotribus scarabaeoides Bernard (Coleoptera: Curculionidae), the mediterranean black scale Saissetia oleae (Olivier) (Homoptera: Coccidae) and the olive thrip Liothripes oleae Costa (Thysanoptera: Phlaeothripidae). Concerning the auxiliary insects that were found in our olives groves they belong to the following orders and families: Diptera (Syrphidae), Coleoptera (Carabidae, Coccinelidae and Staphylinidae), Hemiptera (Anthocoridae and Miridae), Neuroptera (Chrysopidae) and Hymenoptera (Braconidae, Encyrtidae, Eulophidae, Formicidae and Trichogrammatidae).

Yeast diversity in the Mediterranean strawberry tree (Arbutus unedo L.) fruits' fermentations

In the Mediterranean region the fruits of the strawberry tree (Arbutus unedo L.) may be fermented and distilled to produce a traditional beverage very much appreciated in Southern Europe. The aim of the present work was to study the diversity of the yeast population and the killer activity of the isolates identified as Saccharomyces cerevisiae, obtained during solid state industrial fermentations of the arbutus berries. The identification of the isolates was performed by the 5.8S rRNA-ITS region restriction analysis and by sequencing the D1/D2 region of the large subunit of the rRNA gene. At the start of the fermentations, various non-Saccharomyces species were detected including Aureobasidium pullulans, Dothichiza pithyophila, Dioszegia zsoltii, Hanseniaspora uvarum and yeasts belonging to the genera Metschnikowia, Cryptococcus and Rhodotorula. However, as the biological processes progressed the number of different species decreased with S. cerevisiae and Pichia membranaefaciens becoming dominant at advanced stages of the must fermentation that is characterized by high concentrations of ethanol. Forty three isolates identified as S. cerevisiae were tested for killer activity against two sensitive reference strains and Zygosaccharomyces bailii. Their killer sensitivity in relation to five killer referenced toxins (K2, K5, K8, K9 and K10) was also studied. Out of the isolates analyzed, 95.3% were sensitive and 4.7% were tolerant against the killer toxins tested. Only three isolates revealed killer activity against one sensitive strain and two of them against the spoiler yeast Z. bailii. The microbiota obtained revealed an interesting potential to be used as starter cultures to overcome unpredictable uncontrolled fermentations of the arbutus fruits as well as in other applications of biotechnological interest. (C) 2012 Elsevier Ltd. All rights reserved.