Learning from 80 years of studies: a comprehensive catalogue of non-Saccharomyces yeasts associated with viticulture and winemaking.

Non-Saccharomyces yeast species are nowadays recognized for their impact on wine´s chemical composition and sensorial properties. In addition, new interest has been given to the commercial exploitation of non-Saccharomyces starter cultures in the wine sector. However, over many years, these yeast species were considered sources of contamination in wine production and conservation, mainly due to the high levels of volatile acidity obtained. The present manuscript systematizes 80 years of literature describing non-Saccharomyces yeast species isolated from grapes and/or grape musts. A link between each reference, the accepted taxonomic name of each species and their geographical occurrence is presented, compiling information for 293 species, in a total of 231 citations. One major focus of this work relates to the isolation of non-Saccharomyces yeasts from grapevines usually ignored in most sampling studies, also as isolation from damaged grapes. These particular niches are sources of specific yeast species, which are not identified in most other explored environments. These yeasts have high potential to be explored for important and diversified biotechnological applications.

Clavispora santaluciae f.a., sp. nov., a novel ascomycetous yeast species isolated from grapes.

During a study of yeast diversity in Azorean vineyards, four strains were isolated which were found to represent a novel yeast species based on the sequences of the internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) and of the D1/D2 domain of the large subunit (LSU) rRNA gene, together with their physiological characteristics. An additional strain isolated from Drosophila suzukii in Italy had identical D1/D2 sequences and very similar ITS regions (five nucleotide substitutions) to the Azorean strains. Phylogenetic analysis using sequences of the ITS region and D1/D2 domain showed that the five strains are closely related to Clavispora lusitaniae, although with 56 nucleotide differences in the D2 domain. Intraspecies variation revealed between two and five nucleotide differences, considering the five strains of Clavispora santaluciae. Some phenotypic discrepancies support the separation of the new species from their closely related ones, such as the inability to grow at temperatures above 35 °C, to produce acetic acid and the capacity to assimilate starch. Neither conjugations nor ascospore formation were observed in any of the strains. The name Clavispora santaluciae f.a., sp. nov., is proposed to accommodate the above noted five strains (holotype, CBS 16465T; MycoBank no., MB 835794).

Starmerella vitis f.a., sp. nov., a yeast species isolated from flowers and grapes.

A novel yeast species of Starmerella vitis f.a. sp. nov. is proposed to accommodate five strains isolated from flowers, grapes and an insect in the Azores, Canada, Hungary, Palau and Taiwan. As the strains were genetically distinct, we used parsimony network analysis based on ITS-D1/D2 sequences to delineate the species in a statistically objective manner. According to sequence comparisons and phylogenetic analysis, the novel species is most closely related to Starmerella lactis-condensi. The two species cannot be distinguished by conventional physiological tests. The type strain of Starmerella vitis f.a., sp. nov. is CBS 16418T; Mycobank number MB 835251.

Differentiation of Saccharomyces cerevisiae populations from vineyards of the Azores Archipelago: Geography vs Ecology.

Aiming to elucidate the roles that ecology and geography play in shaping the differentiation of fermentative grape-associated Saccharomyces cerevisiae populations, several locations on six islands of the Azores Archipelago were surveyed. A total of 249 strains were isolated from spontaneous fermentations of grape samples from several varieties of two distinct grapevine species (Vitis vinifera L. and Vitis labrusca L.), in vineyards that are under regular cultivation or in abandoned vineyards. Strains were genetically analyzed using a set of nine microsatellite loci, and also phenotypically characterized using relevant physiological/biotechnological tests. Results showed that genetic divergence among populations of the same island was lower than from populations from different islands. Phenotypic comparison of the populations from each of the islands revealed significant differences between them. Strains isolated from the islands with more intensive viticultural activity - Pico, Terceira and Graciosa - showed higher levels of SO2 tolerance, possibly resulting from selection by human activity. The percentage of strains producing low levels of H2S was higher in S. Jorge (60%). Our findings were supported both by genetic and phenotypic data and provide clear evidence for the prevailing role of the geography over ecology in the differentiation of S. cerevisiae populations in the Azores Archipelago.

Association between Grape Yeast Communities and the Vineyard Ecosystems.

The grape yeast biota from several wine-producing areas, with distinct soil types and grapevine training systems, was assessed on five islands of Azores Archipelago, and differences in yeast communities composition associated with the geographic origin of the grapes were explored. Fifty-seven grape samples belonging to the Vitis vinifera grapevine cultivars Verdelho dos Açores (Verdelho), Arinto da Terceira (Arinto) and Terrantez do Pico (Terrantez) were collected in two consecutive years and 40 spontaneous fermentations were achieved. A total of 1710 yeast isolates were obtained from freshly crushed grapes and 1200 from final stage of fermentations. Twenty-eight species were identified, Hanseniaspura uvarum, Pichia terricola and Metschnikowia pulcherrima being the three most representative species isolated. Candida carpophila was encountered for the first time as an inhabitant of grape or wine-associated environments. In both sampling years, a higher proportion of H. uvarum in fresh grapes from Verdelho cultivar was observed, in comparison with Arinto cultivar. Qualitatively significant differences were found among yeast communities from several locations on five islands of the Archipelago, particularly in locations with distinctive agro-ecological compositions. Our results are in agreement with the statement that grape-associated microbial biogeography is non-randomly associated with interactions of climate, soil, cultivar, and vine training systems in vineyard ecosystems. Our observations strongly support a possible linkage between grape yeast and wine typicality, reinforcing the statement that different viticultural terroirs harbor distinctive yeast biota, in particular in vineyards with very distinctive environmental conditions.

Yeast Biodiversity in Vineyard Environments Is Increased by Human Intervention.

One hundred and five grape samples were collected during two consecutive years from 33 locations on seven oceanic islands of the Azores Archipelago. Grape samples were obtained from vineyards that were either abandoned or under regular cultivation involving common viticultural interventions, to evaluate the impact of regular human intervention on grape yeast biota diversity in vineyards. A total of 3150 yeast isolates were obtained and 23 yeast species were identified. The predominant species were Hanseniaspora uvarum, Pichia terricola, Starmerella bacillaris and Issatchenkia hanoiensis. The species Barnettozyma californica, Candida azymoides and Pichia cecembensis were reported in grapes or wine-associated environments for the first time. A higher biodiversity was found in active vineyards where regular human intervention takes place (Shannon index: 1.89 and 1.53 in the first and second years, respectively) when compared to the abandoned ones (Shannon index: 0.76 and 0.31). This finding goes against the assumptions that human intervention can destroy biodiversity and lead to homogeneity in the environment. Biodiversity indices were considerably lower in the year with the heaviest rainfall. This study is the first to report on the grape yeast communities from several abandoned vineyards that have undergone no human intervention.

Tratamento da mordida aberta anterior com miniplacas / Treatment of anterior open bite with miniplates

Este artigo reporta o caso clínico de um paciente do gênero feminino, com 53 anos, portadora de má oclusão Classe II subdivisão esquerda, com mordida aberta anterior esquelética. O tratamento foi realizado com a intrusão dos dentes posteriores superiores e inferiores, por meio da ancoragem esquelética com quatro miniplacas. Os resultados demonstraram a intrusão dos molares superiores e inferiores, distalização dos molares superiores do lado esquerdo, rotação anti-horária da mandíbula, com diminuição da altura facial inferior, correção da mordida aberta anterior e da Classe II subdivisão esquerda.

This article reports the clinical case of a 53 years old female patient, with Angle Class II subdivision left, with skeletal anterior open bite. The treatment was performed with the intrusion of maxillary and mandibular posterior teeth, by skeletal anchorage with four miniplates. The results showed the intrusion of upper and lower molars distalization of upper molars on the left, counterclockwise rotation of the mandible, with decreased lower facial height, correction of anterior open bite, and Class II subdivision left.

Bacteria isolated from a sugarcane agroecosystem: their potential production of polyhydroxyalcanoates and resistance to antibiotics

In this investigation, a sugarcane agroecosystem at a coastal tableland, in the northeast of Brazil, was screened to obtain bacteria strains able to synthesize poly-ß-hydroxyalkanoates (PHA), using sucrose as the main carbon source. The potential to synthesize PHA was tested qualitatively by Sudan Black staining of colonies growing in different carbon sources: sucrose, glucose, fructose, propionate and cellulose. In a typical sugarcane crop management system, the plantation is burned before harvesting and vinasse, a by-product of alchohol production, is used in a fertirrigation system causing, probably, selective pressures on the microbiota of natural environments. Eighty-two bacteria strains, belonging to 16 different genera and 35 different species, were isolated. The data showed that 11 strains (ca 13 per cent), nine of which belonging to the genus Pseudomonas, presented a strong Sudan Black staining in several carbon sources tested and, simultaneously, showed multiple resistance to antibiotics. Resistance to antibiotics is an advantageous feature for the biotechnological production of PHAs. The total number of isolates with multiple resistance to antibotics was 73, and 38 (per cent) of them belong to the genus, Pseudomonas. Among the isolates ca, 86 (per cent) and 43 (per cent) grew in the presence of 10-100U/ml of penicillin and/or 100-300 mg/ml of virginiamycin, respectively. These antiotics are utilized in the alcohol distillery we investigated. The results suggest that some agroecosystem environments could be considered as habitats where bacteria are submitted to nutritional unbalanced conditions, resulting in strains with potential ability to produce PHAs, and also, to an increase in the microbial diversity.

Nuestras últimas experiencias en la microcirugía del segmento anterior / Our latest experiences in microsurgery of the anterior segment

Se presentan los resultados de las últimas experiencias de los autores con relación al empleo de la microcirugía del segmento anterior del ojo. Para ello utilizaron un microscopio quirúrgico de la casa Opton, combinado en ocasiones con la telelupa binocular de la casa Carl Zeiss. Se analizan las técnicas empleadas, las complicaciones trans y posoperatorias, el material de sutura, el tipo de iridectomía (cuando se realizó), y el estado ocular al mes de operado. Se tuvieron en cuenta en el estudio las afecciones generales que presentaba cada paciente