Patterns of Mycobacterium tuberculosis-complex excretion and characterization of super-shedders in naturally-infected wild boar and red deer

Wild boar (Sus scrofa) and red deer (Cervus elaphus) are the main maintenance hosts for bovine tuberculosis (bTB) in continental Europe. Understanding Mycobacterium tuberculosis complex (MTC) excretion routes is crucial to define strategies to control bTB in free-ranging populations, nevertheless available information is scarce. Aiming at filling this gap, four different MTC excretion routes (oronasal, bronchial-alveolar, fecal and urinary) were investigated by molecular methods in naturally infected hunter-harvested wild boar and red deer. In addition MTC concentrations were estimated by the Most Probable Number method. MTC DNA was amplified in all types of excretion routes. MTC DNA was amplified in at least one excretion route from 83.0% (CI95 70.8-90.8) of wild ungulates with bTB-like lesions. Oronasal or bronchial-alveolar shedding were detected with higher frequency than fecal shedding (p < 0.001). The majority of shedders yielded MTC concentrations <10(3) CFU/g or mL. However, from those ungulates from which oronasal, bronchial-alveolar and fecal samples were available, 28.2% of wild boar (CI95 16.6-43.8) and 35.7% of red deer (CI95 16.3-61.2) yielded MTC concentrations >10(3) CFU/g or mL (referred here as super-shedders). Red deer have a significantly higher risk of being super-shedders compared to wild boar (OR = 11.8, CI95 2.3-60.2). The existence of super-shedders among the naturally infected population of wild boar and red deer is thus reported here for the first time and MTC DNA concentrations greater than the minimum infective doses were estimated in excretion samples from both species.

Bacterial cellulose as a novel substrate for retinal pigment epithelium cells transplantation in age-related macular degeneration

Tese de Doutoramento em Engenharia Química e Biológica.

The role of ascorbate peroxidase, guaiacol peroxidase, and polysaccharides in cassava (Manihot esculenta Crantz) roots under postharvest physiological deterioration

Abstract This study aimed to investigate the role of ascorbate peroxidase (APX), guaiacol peroxidase (GPX), polysaccharides, and protein contents associated with the early events of postharvest physiological deterioration (PPD) in cassava roots. Increases in APX and GPX activity, as well as total protein contents occurred from 3 to 5 days of storage and were correlated with the delay of PPD. Cassava samples stained with periodic acid-Schiff (PAS) highlighted the presence of starch and cellulose. Degradation of starch granules during PPD was also detected. Slight metachromatic reaction with toluidine blue is indicative of increasing of acidic polysaccharides and may play an important role in PPD delay. Principal component analysis (PCA) classified samples according to their levels of enzymatic activity based on the decision tree model which showed GPX and total protein amounts to be correlated with PPD. The Oriental (ORI) cultivar was more susceptible to PPD.

Removal of ochratoxin A from contaminated white and red wines using oenological fining agents

Mycotoxins are toxic secondary metabolites produced by certain moulds, being ochratoxin A (OTA) one of the most relevant. Its chemical structure is a dihydro-isocoumarin connected at the 7-carboxy group to a molecule of L--phenylalanine via an amide bond. OTA contamination of wines might be a risk to consumer health, thus requiring treatments to achieve acceptable standards for human consumption [1]. According to the Regulation No. 1881/2006 of the European Commission, the maximum limit for OTA in wine is 2 µg/kg [2]. Therefore, the aim of this work was to know the effect of different fining agents on OTA removal, as well as their impact on white and red wine physicochemical characteristics. To evaluate their efficiency, 11 commercial fining agents (mineral, synthetic, animal and vegetable proteins) were used to get new approaches on OTA removal from white and red wines. Trials were performed in wines artificially supplemented (at a final concentration of 10 µg/L) with OTA. The most effective fining agent in removing OTA (80%) from white wine was a commercial formulation that contains gelatine, bentonite and activated carbon. Removals between 10-30% were obtained with potassium caseinate, yeast cell walls and pea protein. With bentonites, carboxymethylcellulose, polyvinylpolypyrrolidone and chitosan no considerable OTA removal was verified. In red wine, removals between 6-19% were obtained with egg albumin, yeast cell walls, pea protein, isinglass, gelatine, polyvinylpolypyrrolidone and chitosan. The most effective fining agents in removing OTA from red wine were an activated carbon (66%) followed again by the commercial formulation (55%), being activated carbon a well-known adsorbent of mycotoxins. These results may provide useful information for winemakers, namely for the selection of the most appropriate oenological product for OTA removal, reducing wine toxicity and simultaneously enhancing food safety and wine quality.

Beeswax organogels: Influence of gelator concentration and oil type in the gelation process

This work focused on how different types of oil phase, MCT (medium chain triglycerides) and LCT (long chain triglycerides), exert influence on the gelation process of beeswax and thus properties of the organogel produced thereof. Organogels were produced at different temperatures and qualitative phase diagrams were constructed to identify and classify the type of structure formed at various compositions. The microstructure of gelator crystals was studied by polarized light microscopy. Melting and crystallization were characterized by differential scanning calorimetry and rheology (flow and small amplitude oscillatory measurements) to understand organogels' behaviour under different mechanical and thermal conditions. FTIR analysis was employed for a further understanding of oil-gelator chemical interactions. Results showed that the increase of beeswax concentration led to higher values of storage and loss moduli (G, G) and complex modulus (G*) of organogels, which is associated to the strong network formed between the crystalline gelator structure and the oil phase. Crystallization occurred in two steps (well evidenced for higher concentrations of gelator) during temperature decreasing. Thermal analysis showed the occurrence of hysteresis between melting and crystallization. Small angle X-ray scattering (SAXS) analysis allowed a better understanding in terms of how crystal conformations were disposed for each type of organogel. The structuring process supported by medium or long-chain triglycerides oils was an important exploit to apprehend the impact of different carbon chain-size on the gelation process and on gels' properties.