Wine yeast selection in the Iberian Peninsula: Saccharomyces and non-Saccharomyces as drivers of innovation in Spanish and Portuguese wine industries.

Yeast selection for the wine industry in Spain started in 1950 for the understanding of the microbial ecology, and for the selection of optimal strains to improve the performance of alcoholic fermentation and the overall wine quality. This process has been strongly developed over the last 30 years, firstly on Saccharomyces cerevisiae, and, lately, with intense activity on non-Saccharomyces. Several thousand yeast strains have been isolated, identified and tested to select those with better performance and/or specific technological properties. The present review proposes a global survey of this massive ex-situ preservation of eukaryotic microorganisms, a reservoir of biotechnological solutions for the wine sector, overviewing relevant screenings that led to the selection of strains from 12 genera and 22 species of oenological significance. In the first part, the attention goes to the selection programmes related to relevant wine-producing areas (i.e. Douro, Extremadura, Galicia, La Mancha and Uclés, Ribera del Duero, Rioja, Sherry area, and Valencia). In the second part, the focus shifted on specific non-Saccharomyces genera/species selected from different Spanish and Portuguese regions, exploited to enhance particular attributes of the wines. A fil rouge of the dissertation is the design of tailored biotechnological solutions for wines typical of given geographic areas.

The impact of active dry yeasts in commercial wineries from the Denomination of Origen "Vinos de Madrid", Spain.

This paper has studied the success of implantation for 16 commercial active dry yeasts (ADYs) during industrial fermentation (30) and the impact of these yeasts during spontaneous fermentations (19) in 10 wineries from the Denomination of Origin "Vinos de Madrid" over two consecutive years. Yeasts strains were identified by molecular techniques, pulsed field electrophoresis and microsatellite analysis. According to these techniques, all the ADYs were different with the exceptions of two strains, L2056 and Rh, which showed the same karyotype and loci size. The results showed that inoculating fermentations with ADYs did not ensure their dominance throughout the fermentation; the implantation level of ADYs was above 80% in only 9 of the 30 commercial fermentations studied; while in 16 fermentations, the dominance of the inoculated ADYs was below 50%. The type of vinification with the best implantation results overall were those associated with red wine fermentations. ADYs affected spontaneous fermentations, although their impact was observed to decrease in the second year of the study. Therefore, specific adaptation studies are necessary before using commercial yeasts during the fermentation process. At the same time, a study was carried out on the frequency of commercial strains in IMIDRA's yeast collection, made up of strains isolated from spontaneous fermentations of the different areas and cellars since the beginning of the Denomination of Origin "Vinos de Madrid" in 1990. Six different ADYs were found with a frequency of less than 5%.

Temporal Variation in the Abundance and Timing of Daily Activity of Chagas Disease Vector Triatoma gerstaeckeri (Stål, 1859) in a Natural Habitat in the Lower Rio Grande Valley, South Texas.

Chagas disease caused by Trypanosoma cruzi is a burden to millions of people in South and Central America. A sylvatic life cycle of the parasite exists in the Southern United States, but recent studies indicate an active peri-domestic life cycle of T. cruzi in Texas. The United States-Mexico border region in Texas displays areas of high poverty and sub-standard housing conditions which are important risk factors for a potential spill-over transmission to a domestic life cycle including humans. The objectives of the study were to examine short- and long-term temporal variation in vector activity and to evaluate the effect of different combinations of attractants on the capture of potential triatomine vectors. We collected local triatomine vectors (all of them identified as Triatoma gerstaeckeri) from a natural habitat in South Texas during the course of a year. The exact time of collection was recorded to examine the timing of flight activity of the triatomine vector. We also conducted a comparative study of the efficiency of 2 commonly used attractants (light and CO2) and the combination of those on the capture rate of Tr. gerstaeckeri. Our study indicates a short season of dispersal of Tr. gerstaeckeri (April/May) and it suggests a unimodal distribution of activity peaking between 2 and 3 hr after sunset. Ultra-violet light served as the main attractant of Tr. gerstaeckeri while CO2 from dry ice did not significantly contribute to the collection of vectors. The pronounced timing of activity in Tr. gerstaeckeri reported in this study contributes to our understanding of the epidemiology of T. cruzi in wildlife and its potential as a Chagas disease vector to humans in the Rio Grande Valley, South Texas.

The phenotypic characterization of yeast strains to stresses inherent to wine fermentation in warm climates.

AIMS: Climate change is exerting an increasingly profound effect on grape composition, microbiology, chemistry and the sensory aspects of wine. Identification of autochthonous yeasts tolerant to stress could help to alleviate this effect. METHODS AND RESULTS: Tolerance to osmotic pressure, ethanol and pH of 94 Saccharomyces cerevisiae strains and 29 strains non-Saccharomyces from the warm climate region DO 'Vinos de Madrid' (Spain) using phenotypic microarray and their fermentative behaviour were studied. The screening highlighted 12 strains of S. cerevisiae isolated from organic cellars with improved tolerance to osmotic stress, high ethanol concentrations and suitable fermentative properties. Screening of non-Saccharomyces spp. such as Lanchacea thermotolerans, Torulaspora delbrueckii, Schizosaccharomyces pombe and Mestchnikowia pulcherrima also highlighted tolerance to these stress conditions. CONCLUSIONS: This study confirmed the adaptation of native strains to the climatic conditions in each area of production and correlated these adaptations with good fermentation properties. Screening has revealed that identifying yeast strains adapted to fermentation stresses is an important approach for making quality wines in very warm areas. SIGNIFICANCE AND IMPACT OF THE STUDY: The results have special relevance because it is a pioneering study that has approached the problem of climate change for wines from a microbiological aspect and has analysed the situation in situ in wineries from a warm climate zone. Resistant strains were found with good biological properties; studying these strains for their stress response mechanisms during fermentation will be of interest to the wine making industry.

Climate-induced range shifts and possible hybridisation consequences in insects.

Many ectotherms have altered their geographic ranges in response to rising global temperatures. Current range shifts will likely increase the sympatry and hybridisation between recently diverged species. Here we predict future sympatric distributions and risk of hybridisation in seven Mediterranean ischnurid damselfly species (I. elegans, I. fountaineae, I. genei, I. graellsii, I. pumilio, I. saharensis and I. senegalensis). We used a maximum entropy modelling technique to predict future potential distribution under four different Global Circulation Models and a realistic emissions scenario of climate change. We carried out a comprehensive data compilation of reproductive isolation (habitat, temporal, sexual, mechanical and gametic) between the seven studied species. Combining the potential distribution and data of reproductive isolation at different instances (habitat, temporal, sexual, mechanical and gametic), we infer the risk of hybridisation in these insects. Our findings showed that all but I. graellsii will decrease in distributional extent and all species except I. senegalensis are predicted to have northern range shifts. Models of potential distribution predicted an increase of the likely overlapping ranges for 12 species combinations, out of a total of 42 combinations, 10 of which currently overlap. Moreover, the lack of complete reproductive isolation and the patterns of hybridisation detected between closely related ischnurids, could lead to local extinctions of native species if the hybrids or the introgressed colonising species become more successful.

Genetic diversity in commercial wineries: effects of the farming system and vinification management on wine yeasts.

AIMS: Analysis of the diversity and distribution of wine yeasts isolated from organically and conventionally grown grapes, and during the subsequent fermentation with or without starter cultures in six different commercial wineries. METHODS AND RESULTS: PCR-RFLP screening of isolates revealed the involvement of ten different species. Saccharomyces cerevisiae, scarcely isolated from grapes, was the dominant species during the latter phases of fermentation, identifying 108 different genotypes by means of SSR analysis. Species and strains' diversity and presence were strongly influenced by the farming system used to grow the grapes and the system of vinification. CONCLUSIONS: Organic farming management was more beneficial in terms of diversity and abundance than the conventional one. Induced fermentation generated a great replacement of native yeasts. Although winery-resident yeasts resulted to be predominant in the process, some noncommercial strains originally in the vineyard were found in final stages of the fermentation, confirming that autochthonous strains of S. cerevisiae are capable to conduct the fermentation process up to its end. SIGNIFICANCE AND IMPACT OF THE STUDY: The study of natural yeast communities from commercial vineyards and wineries is an important step towards the preservation of native genetic resources. Our results have special relevance because it is the first time that the real situation of the yeast ecology of alcoholic fermentation in commercial wineries belonging to the relevant wine-producing Appellation of Origin 'Vinos de Madrid' is shown.

An alternative to the reference method for testing the radiochemical purity of 99Tcm-tetrofosmin.

The recommended method for controlling the radiochemical purity (RCP) of 99Tcm-labelled tetrofosmin is thin-layer chromatography (TLC) with silica gel as the stationary phase and acetone:dichloromethane (35:65, v/v) as the mobile phase. However, this test is time-consuming and uses a toxic substance. We propose an alternative method to control RCP, based on a solid-phase extraction method (SPE) using a Sep Pak C18 cartridge. We used both methods to analyse 35 samples and to determine the percentages of colloid, radiopharmaceutical and free 99Tcm-pertechnetate. Normal labelling processes were modified to obtain a wide range of values. The range of RCP values obtained was 63.4-96.5% (median = 93.8%) by the standard TLC method and 70.1-96.4% (median = 94.1%) by the SPE method. A significant difference was observed only for the percentage of free 99Tcm-pertechnetate (P < 0.05). The agreement between the two methods, using 90% RCP as the limit, gave a kappa index = +1 (P < 0.001), indicating total agreement between them. The proposed method is useful to control RCP because it yields results that are in good agreement with those of the standard method, and because it is safer and less time-consuming.

Serum zinc, copper, insulin and lipids in Alzheimer's disease epsilon 4 apolipoprotein E allele carriers.

BACKGROUND: Copper (Cu) and zinc (Zn) have been implicated in the development of Alzheimer's disease (AD) and, in this regard, Cu and Zn serum concentrations have been analysed but with inconclusive results. Serum insulin, glucose and cholesterol concentrations have been related to the apolipoprotein E genotype in non-AD populations. DESIGN: In this study, we have analysed the relationship between serum Cu, Zn, insulin, glucose and lipid parameters (cholesterol, triglycerides, apoA and apoB apolipoproteins) in AD and AD epsilon 4 apolipoprotein E carriers by multivariate analysis using logistic regression, including the variables that showed a significance of P < 0.05 in the bivariate analysis. RESULTS: The results obtained show that epsilon 4 apoE allele is an independent AD risk factor (OR = 6. 67, 95% CI = 2.59-17.16). In AD epsilon 4 apoE allele carriers, we found significantly higher Zn, Cu and insulin serum concentrations. Non-demented control subjects with at least one epsilon 4 apoE allele had the lowest serum insulin concentrations. There was no significant association between epsilon 4 apolipoprotein E allele and lipid parameters in the sample studied. CONCLUSIONS: In AD we have found a significant association between higher serum Zn, Cu and insulin concentrations and the presence of an epsilon 4 apoE allele, but only greater serum Zn concentration appears to be an independent risk factor associated with the development of AD.

Calculation of airborne radioactivity in a Technegas lung ventilation unit.

Airborne contamination by 99Tcm has been monitored in the Nuclear Medicine Department in our hospital to assess the risk of internal contamination to occupational workers exposed to Technegas studies. An air sampler fitted with a membrane filter was used. The optimum time for air absorption for obtaining the maximum activity in the filter was calculated. Maximum activity in the membrane filter ensures minimum uncertainty, which is especially important when low-level activities are being measured. The optimum time depends on air absorption velocity, room volume and filter efficiency for isotope collection. It tends to 1/lambda (lambda = disintegration constant for 99Tcm) for large volume and low velocity. Room activity with the air pump switched on was related to filter activity, and its variation with time was studied. Free activity in air for each study was approximately 7 x 10(-4) the activity used, and the effective half-life of the isotope in the room was 13.9 min (decay and diffusion). For a typical study (630 MBq), the effective dose to staff was 0.01 microSv when in the room for 10 min.

Comparison and variation of different methodologies for the detection of autoantibodies to nuclear antigens (ANA).

Interest in the assessment of autoantibody specificity stems from the need for an autoantibody marker capable of predicting clinical events in autoimmune disorders. However, the multiplicity of epitopes present on autoantigenic particles, the quantitative and qualitative heterogeneity of autoantibodies, as well as the nature of the tests, mean that each of the assays used in their determination have different characteristics. The aim of this study was to compare the specificities of different ANAs using four commercial assays. The routine method used for the detection of ANA is indirect immunofluorescence on Hep-2 cells. The assays used were: counterimmunoelectrophoresis (CIE), enzyme-linked immunosorbent assay (ELISA), and two immunoblotting assays. Kappa statistic was applied to evaluate the consistency between tests. Kappa index is a measure of agreement between categorical data. Kappa has a maximum of 1.00 when the agreement is perfect, a value of zero indicates no agreement better than chance, and negative values show worse than chance agreement. For SS-B antibodies, there was a good concordance between all four methods used (Kappa 0.66-0.74). For anti RNP antibodies, the results for CIE/ELISA (Kappa 0.60) were consistent as were the two immunoblot methods (Kappa 0.69). For anti Scl-70 (topoisomerase I) antibody, results from the ELISA and CIE methods were totally consistent (Kappa 1.00). In spite of the high prevalence of anti SS-A/Ro antibodies, the agreement between the methods was poor, without statistical significance. Finally, for Sm antibodies, more consistent results were obtained between CIE/ELISA (Kappa 0.51) and between one of the immunoblotting methods and ELISA (Kappa 0.54). In conclusion, CIE concurs mostly with ELISA for anti-RNP, Scl-70, Sm and SS-B antibodies, but with some disagreement for SS-A antibodies.

[Genotype and phenotype of apolipoprotein E in patients with Alzheimer's disease in Castille and Leon]. / Genotipo y fenotipo de la apolipoproteína E en enfermos de Alzheimer en Castilla y León.

We determined the variants of the apolipoprotein E (apo-E) in a sample taken from a castellano-leonesa population with Alzheimer's disease (AD). The prevalence of the allele E-4 is 0.33 in AD and 0.12 in the control group. This results allow us to estimate that the risk of suffering from the disease is higher when the patients have an allele E-4. A significant correlation was found between the presence of the allele E4 and the AD (p < 0.005).

[Thrombophlebitis of the cavernous sinus secondary to polysinusitis. Report of a new case]. / Tromboflebitis del seno cavernoso secundaria a polisinusitis. Aportación de un nuevo caso.

Cavernous sinus thrombophlebitis syndrome is uncommon nowadays. Despite the wide range of antimicrobial agents it carries a high mortality. The disease frequently arises from infection of face, mouth, nose and paranasal sinuses, reaching the cavernous sinus by venous spread. Involvement of adjacent anatomic structures accounts for most of the clinical findings. Meningeal involvement, bilateral orbital edema and ocular cranial nerves palsy are very helpful in the differential diagnosis. Streptococcus spp., Staphylococcus spp., Haemophilus influenzae and anaerobic organisms are considered to be the most frequent etiological agents. In the present paper we report a case of cavernous sinus thrombophlebitis following left side polysinusitis. The patient presented with the main clinical features of the syndrome along with Streptococcus equisimilis and Fusobacterium necrophorum bacteremia. Urgent surgery of infected sinuses and appropriate antibiotic therapy led to a favorable outcome. The patient made and excellent progress and was discharged completely recovered two weeks after admission.

Development, Distribution, and Host Studies of the Fungus Macrobiotophthoira vermicola (Entomophthorales).

The life cycle and host range of Macrobiotophthora vermicola were studied. Secondary spores produced from forcibly ejected primary spores adhered to the cuticle of Cruznema tripartitum, germinated, and penetrated the cuticle within 30 minutes. New primary spores were produced within 24 hours of initial spore adhesion. In a host range study, species of Rhabditidae, Diplogasteridae, and Aphelenchoidea were hosts, but not species of Bunonematidae, Tripylidae, Cephalobida, or Tylenchina. Numbers of second-stage Meloidogyne incognita juveniles were not decreased when added to soil seeded with infected C. tripartitum. In six Tennessee soybean fields, Macrobiotophthora vermicola was the most commonly encountered nematode-destroying fungus, followed by a sterile, nonseptate fungus and Arthrobotrys conoides. Nematophagous fungi were isolated more frequently from silt loam soils than from clay soils. Addition of C. tripartitum to soil extract plates as a bait nematode did not increase isolations of nematophagous fungi.