Quality Indicators and Heat Damage of Dried and Cooked Gluten Free Spaghetti.

The quality and safety indicators of commercial dried gluten free (GF) pasta were analyzed to investigate, for the first time, the real nutritional intake through the chemical composition and the heat damage during processing by quantification of furosine. Eight samples of GF spaghetti were compared with wheat spaghetti. Dried and cooked GF pasta had lower protein and ash content than wheat spaghetti. GF samples composed solely by corn flour had higher optimal cooking time. Samples with emulsifier showed lower losses during cooking. Considering their composition, no trend could be established to explain textural behavior. Samples constituted merely by corn showed the highest resilience and elasticity. Spaghetti constituted only from corn and rice showed the highest firmness. The furosine content in dried samples ranged between 19 and 134 mg FUR/100 g proteins and in cooked samples ranged between 48 to 360 mg FUR/100 g proteins. Furosine content of GF pasta was in general lower than in wheat pasta, and those differences were even enlarged when comparing them after cooking. The results of PCA indicated it was possible to discriminate GF pasta regarding their technological and nutritional behavior.

Glycemic Response to Corn Starch Modified with Cyclodextrin Glycosyltransferase and its Relationship to Physical Properties.

Corn starch was modified with cyclodextrin glycosyltransferase (CGTase) below the gelatinization temperature. The porous granules with or without CGTase hydrolysis products may be used as an alternative to modified corn starches in foods applications. The amount and type of hydrolysis products were determined, containing mainly ß-cyclodextrin (CD), which will influence pasting behavior and glycemic response in mice. Irregular surface and small holes were observed by microscopic analysis and differences in pasting properties were observed in the presence of hydrolysis products. Postprandial blood glucose in mice fed gelatinized enzymatically modified starch peaked earlier than their ungelatinized counterparts. However, in ungelatinized enzymatically modified starches, the presence of ß- CD may inhibit the orientation of amylases slowing hydrolysis, which may help to maintain lower blood glucose levels. Significant correlations were found between glycemic curves and viscosity pattern of starches.

What drives wild boar density and population growth in Mediterranean environments?

Accurate prediction of fluctuations of wildlife local number of individuals is crucial for effective population management to minimise human-wildlife conflicts. Climate, habitat, food availability, and density dependence are among the main factors influencing mammalian population dynamics. In southern Europe, precipitation and temperature, particularly during summer have been suggested as key factors affecting wild boar (Sus scrofa L.). However, there is uncertainty regarding the role of these factors and the mechanisms driving population fluctuations. This study utilized long-term data of wild boar populations from 14 study sites collected for 23 years in Catalonia, Spain, to analyse the factors that drive population density and growth rate. Generalized Additive Mixed Models (GAMM) explained respectively, 94 % and 65 % of the density and growth rate variability. Spring precipitation in both current and previous year, female weight, and forest cover (particularly above 60 %) were directly associated with higher wild boar densities and population growth rates. The interaction between crop cover and total annual precipitation also played a significant role in determining population density. Higher densities were linked to lower population growth in the following year, likely due to a density-dependent process. These results suggest that the expected decrease in rainfall linked with global warming may limit the availability of natural resources and potentially slow wild boar population growth. Nevertheless, wild boar can exploit alternative anthropogenic food sources, potentially leading to an increase of human-wildlife conflicts. Therefore, incorporating management policies aimed at restricting wild boar access to human food sources is key for controlling their reproductive output. Additionally, landscape management strategies targeted at diminishing refuge and resource availability in regions experiencing high wild boar impact are essential for contributing to sustainable coexistence between wild boars and human populations.

Prediction of denaturing tendency of organic solvents in mixtures with water by measurement of naphthalene solubility.

Addition of miscible organic solvents to water increases the solubility of naphthalene. The logarithm of the solubility is linearly dependent on the co-solvent concentration, in an intermediate range. The relative solubilising effects of different solvents correlate well with their known tendency to denature proteins (using literature data for trypsin, cytochrome c, chymotrypsinogen, chymotrypsin, laccase and myoglobin). This is expected if denaturation occurs when the hydrophobic effect has been reduced by a characteristic extent for a given protein. Naphthalene solubility predicts denaturation as well as does the denaturation capacity model.

Changes in peripheral blood leukocyte populations in pigs with natural postweaning multisystemic wasting syndrome (PMWS).

The objective of the present study was to analyze, by flow cytometry, changes in PBMC subsets in pigs having postweaning multisystemic wasting syndrome (PMWS), a new condition associated to porcine circovirus type 2 (PCV2) infection. Thirteen acutely PMWS affected pigs were selected from a farm seronegative to porcine reproductive and respiratory syndrome virus (PRRSV) and to Aujeszky's disease virus (ADV); 11 clinically healthy pigs were selected from a high health farm with no history of PMWS and free of the major swine pathogens, and used as a control group. All pigs were necropsied, and tissue samples were fixed in formalin; blood with EDTA anticoagulant was used to perform the flow cytometric analysis. PBMC were incubated with mAb against porcine CD3, CD4, CD8, CD25, CD45, IgM, SWC3, and SLA-Class II. Flow cytometric analysis showed substantial changes in leukocyte subsets in the peripheral blood of PMWS-affected pigs, which were characterized by an increase of monocytes, a reduction of T (mainly CD4(+)) and B-lymphocytes, and the presence of low-density immature granulocytes. Altogether, these changes would suggest an inability of acutely PMWS-affected pigs to mount an effective immune response.

Use of fungal phytase to improve breadmaking performance of whole wheat bread.

The possible use of phytase as a breadmaking improver has been tested in whole wheat breads by adding different amounts of fungal phytase. The effect of phytase addition on the fermentation stage and the final bread quality was analyzed. The phytase addition shortened the fermentation period, without affecting the bread dough pH. Regarding the whole wheat bread, a considerable increase of the specific bread volume, an improvement of the crumb texture, and the width/height ratio of the bread slice were obtained. An in vitro assay revealed that the improving effect of phytase on breadmaking might be associated with the activation of alpha-amylase, due to the release of calcium ions from calcium-phytate complexes promoted by phytase activity. As a conclusion, phytase offers excellent possibilities as a breadmaking improver, with two main advantages: first, the nutritional improvement produced by decreasing phytate content, and second, all the benefits produced by alpha-amylase addition can be obtained by adding phytase, which promotes the activation of endogenous alpha-amylase.

Experimental approach to optimize the use of alpha-Amylases in breadmaking.

alpha-Amylases from different origins (wheat, malted barley, fungi, and bacteria) are used extensively to improve breadmaking. However, the enzyme activities, in addition to the differences associated with their origins, are strongly affected by the process conditions and the presence of other compounds in the medium. The activity of different alpha-amylases was tested under different conditions (pH and temperature), and in the presence of some bread ingredients (salt and sugar), some breadmaking additives (ascorbic acid and sodium propionate), and some metabolites (organic acids and saccharides) generated during the fermentation step, to envisage the behavior of these alpha-amylases during the breadmaking process. The alpha-amylase activities were affected to a different extent by the addition of these compounds depending on the enzyme origin. In general, the alpha-amylases from cereals (wheat and malted barley) were less sensitive to the presence of some ingredients, additives, and metabolites. These results show the great variation of the alpha-amylase activity with the process conditions and the importance of its knowledge in the selection of the appropriate alpha-amylase for a specific breadmaking process.

Additional stabilization of penicillin G acylase-agarose derivatives by controlled chemical modification with formaldehyde.

We have tested the effect of chemical modifications with formaldehyde on the activity/stability of immobilized derivatives of the enzyme penicillin G acylase (PGA). These derivatives were previously stabilized through enzyme-support multipoint covalent attachment. We carried out very different chemical treatments of our derivatives by testing the effect of different variables which control the intensity and the nature of these amine-formaldehyde reactions. The variables tested were: formaldehyde concentration, pH, time, and temperature. We also developed a colorimetric titration of the free amine groups on immobilized PGA in order to evaluate the extension of the reaction between formaldehyde and the amine groups of the enzyme. As a consequence of these studies, we have been able to get additional stabilizations of our previously stabilized-immobilized derivatives: e.g. a factor of 24-fold was achieved in terms of stabilization against irreversible thermal inactivation. The integrated effect of additional chemical modification plus previous multipoint covalent attachment has allowed us to prepare PGA derivatives which are 50,000 more thermostable than native PGA as well as most of the commercial PGA derivatives.

Enzyme reaction engineering: synthesis of antibiotics catalysed by stabilized penicillin G acylase in the presence of organic cosolvents.

By using very active and very stable penicillin G acylase (PGA)--agarose derivatives we have studied the industrial design of equilibrium-controlled synthesis of lactamic antibiotics. In the presence of high concentrations of organic cosolvents we have carried out the direct enzymatic condensation of phenylacetic acid and 6-aminopenicillanic acid to yield the model antibiotic penicillin G. We have mainly studied the integrated effect of different variables that define the reaction medium on a number of parameters of industrial interest:time course of antibiotic synthesis, highest synthetic yields, stability of the catalyst, and solubility and stability of substrates and products. The main variables tested were the nature and concentration of the organic cosolvent, pH, and temperature. The effects of the variables tested on different parameters were quite different and sometimes opposite. Hence, the optimal experimental conditions for antibiotic synthesis catalysed by PGA were established, as a compromise solution, in order to obtain good values for every parameter of industrial interest. These conditions seem to be important parameters for scale-up (e.g. we have been able to reach more than 95% of synthetic yields with productivities around 0.5 tons of model antibiotic per year per liter of catalyst).

Preparation of activated supports containing low pK amino groups. A new tool for protein immobilization via the carboxyl coupling method.

A method for the preparation of new aminated agarose gels containing monoaminoethyl-N-aminoethyl structures, MANA-agarose gels, has been developed. These gels contain primary amino groups with a very low pK value (6.8). In addition to that, we have been able to prepare very highly activated gels (e.g., 10% agarose gels containing up to 200 mu Eq of primary amines per milliliter). These two properties make these activated supports suitable for performing novel and interesting methods for protein immobilizations via very mild carbodiimide activation of carboxy groups. For example, very effective coupling reactions can be performed at pH 5.0-6.0 in the presence of low concentrations of activating agent, e.g., 1 mM. By using a model industrial enzyme, beta-galactosidase from Aspergillus oryzae, we have been able to demonstrate the excellent prospects of these novel activated supports.

Experimental inoculation of conventional pigs with tissue homogenates from pigs with post-weaning multisystemic wasting syndrome.

This report describes the experimental inoculation of conventional pigs with a tissue homogenate obtained from two pigs affected with postweaning multisystemic wasting syndrome (PMWS). Eight 2-month-old pigs were inoculated by the intranasal route, and two pigs were left as uninfected controls. Clinical signs, rectal temperatures and body weights were recorded. Pigs were necropsied at days 14 or 21 post-inoculation, and tissue samples were taken for histopathology and porcine circovirus (PCV) in-situ hybridization. Although only mild clinical signs of disease were observed, lesions of PMWS were seen, and PCV was shown to have been successfully transmitted to six of the eight pigs. Seroconversion of all inoculated pigs to PCV-2, but not to PCV-1, was also detected, suggesting that the PCV nucleic acid detected by in-situ hybridization in inoculated pigs corresponded to PCV-2. In conclusion, this report shows that PCV-2 is transmissible to pigs, and the inoculation of tissue homogenates containing the virus results in the development of PMWS-like lesions.

Pathological, immunohistochemical, and in-situ hybridization studies of natural cases of postweaning multisystemic wasting syndrome (PMWS) in pigs.

Fifteen pigs from five farms on which there had been a previous clinical and histopathological diagnosis of postweaning multisystemic wasting syndrome (PMWS) were investigated. At necropsy, enlargement of lymph nodes was the most obvious lesion; other lesions were non-collapsed lungs, ulceration of the gastric pars oesophagica, and cranioventral pulmonary consolidation. Microscopical lesions attributable to PMWS were found in lymphoid organs (including lymph nodes, tonsil, Peyer's patches and spleen), liver, kidney and lungs. Varying degrees of lymphocellular depletion, affecting both lymphoid follicles and parafollicular zones, and progressive multifocal to diffuse infiltration of lymphoid tissue by large histiocytic cells were the characteristic lesions. Syncytial cells were seen frequently, especially in lymphoid organs. A prominent finding was the presence of sharply demarcated, spherical, basophilic, cytoplasmic inclusions in histiocytic cells. The lymphoid lesions were suggestive of immunosuppression. Non-lymphoid lesions included interstitial pneumonia, periportal mononuclear inflammatory infiltration of the liver in varying degrees, and interstitial nephritis. Porcine circovirus (PCV) antigen and nucleic acid were regularly found in lymphoid organs, lung, liver and, to a lesser degree, kidney. Target cells for PCV replication included monocyte/macrophage lineage and antigen-presenting cells. To a lesser extent, epithelial cells such as renal tubular, bronchial and bronchiolar cells, endothelial cells, hepatocytes and lymphocytes were also labelled. One pig did not show PCV nucleic acid; sequence differences among different viral isolates are discussed as the probable cause of this lack of labelling by the in-situ hybridization PCV-specific probe.

Identification of porcine circovirus in tissues of pigs with porcine dermatitis and nephropathy syndrome.

Thirty-three pigs affected by porcine dermatitis and nephropathy syndrome, 30 from Spain and three from the USA, were investigated in order to detect porcine circovirus (PCV) in their tissues. A standard in situ hybridisation technique using a specific DNA 317-bp probe based on a well-conserved sequence of PCV (which recognises both PCV-1 and PCV-2) was applied to formalin-fixed, paraffin-embedded tissues. Twenty-eight of the 30 Spanish pigs and all three American pigs had PCV in at least one tissue. Viral nucleic acid was detected mainly in lymphoid organs, and especially the lymph nodes. The viral genome was also found, in order of decreasing quantity, in Peyer's patches, tonsil, lung, spleen, kidney, liver, and skin. Viral nucleic acid was located mainly within the cytoplasm of monocyte/macrophage lineage cells, including follicular dendritic cells, macrophages, histiocytes and Kupffer cells. No viral nucleic acid was found in damaged glomeruli or arteriolar walls. In frozen samples available from three Spanish pigs, the virus was identified as type 2 by using the polymerase chain reaction and restriction fragment length polymorphism. Most of the pigs from which serum was available were seropositive against porcine respiratory and reproductive syndrome virus (PRRSV), and PRRSV antigen was detected in the lung of two of the Spanish pigs. These results suggested that PCV is present in tissues of almost all pigs affected by PDNS, and PCV has to be considered as a possible agent involved in the pathogenesis of the syndrome.

Clinical and pathological observations on pigs with postweaning multisystemic wasting syndrome.

The aim of this work was to characterise the lesions and agents present in clinically normal and clinically affected pigs on a farm during an outbreak of postweaning multisystemic wasting syndrome (PMWS), and to evaluate the diagnostic techniques for detecting porcine circovirus type 2 (PCV-2) and other microorganisms. Four pigs in the early stage and 11 pigs in the late stage of the disease, and eight clinically normal pigs were necropsied. Samples of lymphoid tissue and serum were also obtained from 12 slaughter pigs from the same farm. The tissues were examined histopathologically, and in situ hybridisation, serology and PCR were used to detect porcine circovirus type 1 (PCV-1) and/or PCV-2 in tissues and/or sera. The presence of porcine reproductive and respiratory syndrome virus (PRRSV), Aujeszky's disease virus (ADV) and porcine parvovirus (PPV) were also investigated. Characteristic microscopical lesions of PMWS were observed in the lymphoid tissues of the pigs in all three necropsied groups; the lesions were most common and severe in the pigs in the early stage of the disease, less so in the pigs in the late stage of the disease, and least in the clinically normal pigs. PCV-2 infection was detected in all the necropsied pigs by in situ hybridisation and PCR. Only three pigs had the PCV-1 genome in serum or lymph node tissue. In contrast, the slaughter pigs had no microscopical lesions and no PCV-2 nucleic acid in their serum or tissues, and only one of them had the pCV-1 genome in its serum. Immunohistochemical, serological and PCR studies revealed that PRRSV and ADV were also present on the farm during the outbreak.

Functionality of porous starch obtained by amylase or amyloglucosidase treatments.

Porous starch is attracting very much attention for its absorption and shielding ability in many food applications. The effect of two different enzymes, fungal α-amylase (AM) or amyloglucosidase (AMG), on corn starch at sub-gelatinization temperature was studied as an alternative to obtain porous starch. Biochemical features, thermal and structural analyses of treated starches were studied. Microscopic analysis of the granules confirmed the enzymatic modification of the starches obtaining porous structures with more agglomerates in the case of AMG treated starches. Several changes in thermal properties and hydrolysis kinetics were observed in enzymatically modified starches. Hydration properties were significantly affected by enzymatic modification being greater influenced by AMG activity, and the opposite trend was observed in the pasting properties. Overall, results showed that enzymatic modification at sub-gelatinization temperatures really offer an attractive alternative for obtaining porous starch granules to be used in a variety of foods applications.

Ultrasonic study of wheat flour properties.

In this work, the wheat flour properties are investigated using ultrasound techniques. Moreover, the flour samples were also characterized by means of well established techniques such as protein content, Alveograph and Mixolab®. A set of 35 dough samples, made of wheat flours with diverse physical and quality properties, were studied. The obtained results shown that ultrasound measurements can detect changes in the dough consistency induced by proteins and also by gelatinization of the starch. Furthermore, ultrasound measurements can be related to parameters indicative of the proteolytic degradation or softening of the dough due to protease activity. Thus, ultrasound can be considered a low cost and rapid tool, complementary to conventional test, for wheat flour characterization.