Yeast communities of the cactusPilosocereus arrabidae and associated insects in the Sandy Coastal Plains of Southeastern Brazil

The yeast communities from necrotic tissues, decaying flowers and fruits, and from larval feeding sites of the mothSigelgaita sp. in the cactusPilosocereus arrabidae were surveyed in three restinga ecosystems in Southeastern Brazil. Insects associated with these substrates were sampled to verify the vectoring of yeasts. The cactusPilosocereus arrabidae was shown to have four different yeast communities associated with it. Necrotic stems had a diverse yeast community with the prevalent speciesPichia barkeri, Candida sonorensis, Pichia cactophila, Geotrichum sp.,Myxozyma mucilagina andSporopachydermia sp. A, representing about 80% of the total isolates.Pichia sp. A and aCandida domercqii-like species represented more than 90% of the yeast isolates from decaying flowers. Fruits had a heterogeneous yeast community with typical fruit yeasts of the genusKloeckera, basidiomicetous anamorphs of the genusCryptococcus, the black yeastAureobasidium pullulans, Pichia sp. A, aCandida domercqii-like species, and some cactophilic yeasts, especiallyClavispora opuntiae. The feeding site ofSigelgaita sp. larvae hadClavispora opuntiae as the prevalent species. Insect vectors are suggested as one the most important factors influencing the composition of these yeast communities.     

Selection of non-conventional yeasts and their use in immobilized form for the bioremediation of olive oil mill wastewaters

The yeast population dynamics in olive wastewaters (OMW), sampled in five mills from Salento (Apulia, Southern Italy), were investigated. Three hundred yeasts were isolated in five industrial mills and identified by molecular analysis. Strains belonging to Geotrichum, Saccharomyces, Pichia, Rhodotorula and Candida were detected. Five G. candidum strains were able to grow in OMW as the sole carbon source and to reduce phenolics, chemical oxygen demand (COD) and antimicrobial compounds. One G. candidum isolate was selected for whole-cell immobilization in calcium alginate gel. The COD and phenolic reduction obtained with immobilized cells showed a 2.2- and 2-fold increase compared to the removal obtained with free cells, respectively. The immobilization system enhanced yeast oxidative activity by avoiding the presence of microbial protease in treated OMW. To our knowledge, this is the first report on G. candidum whole-cell immobilization for OMW bioremediation.     

Yeasts from phylloplane and their capability to produce indole-3-acetic acid

Yeasts were isolated from the phylloplane of various plant species collected from seven provinces in Thailand. A total of 114 yeast strains and 10 strains of a yeast-like fungus were obtained by enrichment isolation from 91 out of 97 leaf samples (93.8?%). On the basis of the D1/D2 domain of the large subunit rRNA gene sequence similarity, 98 strains were identified to be of 36 yeast species in 18 genera belonging to Ascomycota viz. Candida, Clavispora, Cyberlindnera, Debaryomyces, Hanseniaspora, Hyphopichia, Kazachstania, Kluyveromyces, Kodamaea, Lachancea, Metschnikowia, Meyrozyma, Pichia, Starmerella, Torulaspora and Wickerhamomyces, and to Basidiomycota viz. Sporidiobolus and Trichosporon. Three strains were found to represent two novels Candida species which were previously described as C. sirachaensis and C. sakaeoensis. Ten strains of yeast-like fungus were identified as Aureobasidium pullulans of the phylum Ascomycota. Ascomycetous yeast species accounted altogether for 98.0?% of the 98 strains. The prevalent species was Candida tropicalis with a low frequency of isolation (14.3?%). Diversity of yeasts other than ballistoconidium-forming yeast in phylloplane in a tropical country in Asia has been reported for the first time. All strains obtained were accessed for the capability to produce IAA and result revealed that 39 strains in 20 species, one strain each of an undescribed and a novel species, and two unidentified strains showed the capability of producing IAA when cultivated in yeast extract peptone dextrose broth supplemented with 0.1?% l-tryptophan. All five strains of Candida maltosa produced relatively high concentrations of IAA.     

Changes in microbial population during fermentation of feedlot waste with corn

A new process for recycling feedlot waste involves the fermentation of liquid from this waste combined with corn. Changes in the flora of this silage-like fermentation were followed. The fermentation was dominated by lactobacilli and yeasts, which initially constitute 1% or less of the natural flora. The species of yeasts and lactics involved were characterized. The fermentation has two phases. A single heterolactic species multiplied rapidly for the first 24 h until it represented 95% of the lactobacilli and more than 90% of the total microflora. It displaced the betabacterium predominant among lactics of the original waste; the acid produced killed coliforms and other organisms in feedlot waste; and the acetic acid produced probably caused the death of the dominant native yeast Trichosporon cutaneum (de Beurm., Gougerot et Vaucher) Ota. The peak lactobacillus count remained constant (about 2 × 10⁹ organisms/g [wet weight]) throughout the rest of the fermentation. Homolactics dominated the later phase and yeasts increased to 9.5 × 10⁷ organisms/g (wet weight). At 6 days, a stable mixture of three lactobacilli was present, one streptobacterium, one thermobacterium, and one betabacterium. Similarly, yeasts stabilized as a mixture of two Candida sp. and one Pichia sp. The dominant species of lactics were characterized. Information on the sequence of microorganisms provides a basis for enhanced protein synthesis in the fermentation.     

Species diversity of yeast in oral colonization of insulin-treated diabetes mellitus patients

The aim of this study was to investigate oral yeast colonization, antifungal susceptibility and strain diversity in insulin-dependent diabetes mellitus patients (175), as well as to evaluate the influence of dental prostheses. Oral rinse samples were cultured on selective media, in order to isolate, count and identify the yeasts recovered. More than half of the diabetic subjects (53%) carried significant amounts of Candida cells in the buccal cavity and these organisms were recovered at higher densities in diabetics wearing dentures. A total of 93 yeast strains were isolated from these patients, including: Candida spp. (n = 89); Pichia (n = 02); Trichosporon (n = 1), and Geotrichum (n = 1). C. albicans represented 56% of these strains, non-albicans Candida 39.8%, and other genera of yeast 4.3%. C. albicans was prevalent, followed by C. parapsilosis, C. tropicalis, C. glabrata, C. krusei, C. rugosa and C. guilliermondii. Agar disk-diffusion tests of the susceptibility of non-albicans Candida and other genera of yeast to fluconazole showed resistance in 21.9%, mainly in C. rugosa (100%), C. glabrata (57%) and C. krusei (50%). Local oral factors, such as the presence of dentures, in association with diabetes, seemed to have the effect of increasing the amount and variety of Candida species in the oral cavities, mainly those with lower drug susceptibilities.     

Wastewater treatment systems harbor specific and diverse yeast communities

Yeasts, as a group of single-celled fungi, are widely distributed in nature and play important roles in biotechnological applications. However, how the types of wastewater and the treatment processes influence yeast populations is not clear. In this study, both cultivation and culture-independent methods were used to investigate the distribution and diversity of yeasts in three typical full-scale plants processing biopharmaceutical, papermaking and municipal wastewater. Cultivable yeasts were very abundant ranging from 10² to 10⁵ cfu g⁻¹ sludge, and highly diverse with 48 taxons belonging to 21 different genera, thus exceeding the yeast richness reported for marine and sugar-rich soil habitats. Genera Rhodotorula, Candida, Trichosporon, Pichia and some unidentified Ascomycetes were the most frequent populations cultivated. However, the compositions of yeast community structures in the plants were dissimilar and were shaped primarily by the type of wastewater treated but also by processing conditions. Culturing yeasts is a powerful way to complement culture-independent approaches to study their diversity, since significant more yeast species, especially quite a few possible novel species were recorded and isolated by cultivation method.     

Characterisation of the diversity and physiology of cellobiose-fermenting yeasts isolated from rotting wood in Brazilian ecosystems

We investigated the yeast species associated with rotting wood samples obtained from Brazilian ecosystems, with a special focus on cellobiose-fermenting species. About 647 yeast strains were isolated from rotting wood samples collected from the areas of Atlantic rainforest, Cerrado, and Amazonian forest. Eighty-six known species and 47 novel species of yeasts were isolated. Candida boidinii, Cyberlindnera subsufficiens, Meyerozyma guilliermondii, Schwanniomyces polymorphus, Candida natalensis, and Debaryomyces hansenii were the most frequently isolated species. Among the cellobiose-fermenting yeasts, 14 known and three novel yeast species were identified. Scheffersomyces queiroziae, Sc. amazonensis, Yamadazyma sp.1, Hanseniaspora opuntiae, C. jaroonii, and Candida tammaniensis were the main ethanol-producing yeasts. These species also produced an intracellular β-glucosidase responsible for cellobiose hydrolysis. In fermentation assays using a culture medium containing 50 g L^?1 cellobiose, ethanol production was observed in all cases; Sc. queiroziae and Sc. amazonensis showed the highest yield, efficiency, and productivity. Candida jaroonii and Yamadazyma sp.1 strains also showed high efficiency in cellobiose fermentation, while C. tammaniensis and H. opuntiae strains produced low amounts of ethanol. This study shows the potential of rotting wood samples from Brazilian ecosystems as a source of yeasts, including new species as well as those with promising biotechnological properties.     

Patterns of Human Oral Yeast Species Distribution on Hainan Island in China

Infections by yeast strains of the genus Candida are among the most prevalent fungal infections of humans. These yeasts are common residents of the oral mucosa and other body surfaces. Since most yeast infections are due to endogenous strains and that species of Candida differ in virulence properties and in intrinsic susceptibilities to antifungal drugs, understanding the human commensal yeast flora can help designing effective treatment and prevention strategies against yeast infections. Here, we report the patterns of yeast species distributions in the oral cavities of 1,799 people from Hainan Island in southern China. Based on sequence information at the fungal barcode locus ITS regions, 368 of the 415 obtained oral yeast strains were identified as belonging to 26 yeast species, while the remaining 47 strains all showed significant sequence divergence to the currently described species. The four most common yeast species were C. albicans (42 %), C. tropicalis (20 %), C. glabrata (5.5 %), and C. parapsilosis (4.1 %) and 10 of the 26 yeast species were represented by only one strain each. Our analyses identified that the gender of hosts and ethnical background showed no contribution to oral yeast species distributions. However, the health status, place of birth, current residency, and the age of hosts all showed significant contributions to the distributions of the four dominant yeast species. We compared our results with those reported previously and discussed the potential mechanisms for the observed differences in oral yeast species distributions.     

Species Distribution and Virulence Factors of Candida spp. Isolated from the Oral Cavity of Kidney Transplant Recipients in Brazil

Although yeasts belonging to the genus Candida are frequently seen as commensals in the oral cavity, they possess virulence attributes that contribute for pathogenicity. The aims of the present study were to study the prevalence of Candida spp. isolated from the oral cavity of renal transplant recipients and to analyze strains virulence factors. We isolated a total of 70 Candida strains from 111 transplant recipients, and Candida albicans was the most prevalent species (82.86 %). Oral candidiasis was diagnosed in 14.4 % kidney transplant patients, while 11 isolates (15.7 %) corresponded to non-Candida albicans Candida (NCAC) species. C. albicans adhered to a higher extension than NCAC strains. Some isolates of Candida tropicalis were markedly adherent to human buccal epithelial cells and highly biofilm-forming strains. Regarding proteinase activity, Candida orthopsilosis was more proteolytic than Candida metapsilosis. Candida glabrata and Candida dubliniensis showed very low ability to form biofilm on polystyrene microtiter plates. We have demonstrated here diverse peculiarities of different Candida species regarding the ability to express virulence factors. This study will contribute for the understanding of the natural history and pathogenesis of yeasts belonging to the genus Candida in the oral cavity of patients who were submitted to kidney transplant and are under immunosuppressive therapies.     

Yeasts associated with pollinating bees and flower nectar

A study of the yeast flora of 328 honey stomachs from 7 different pollinating bee species, and 342 flower nectar samples of 9 different flower species yielded 766 yeast isolates composed of 16 genera and 47 species. Most of the yeast species from both the sources belonged to the genusCandida, while the most frequently isolated yeasts wereDekkera intermedia from honey stomach andCandida blankii from flower nectar. Among the honey bees,Xylocopa sp., and among flowers,Citrus medica, yielded the highest number of yeast species. Nineteen species of yeasts belonging to 9 genera were common to both the sources.     

Yeast Diversity and Persistence in Botrytis-Affected Wine Fermentations

Culture-dependent and -independent methods were used to examine the yeast diversity present in botrytis-affected (“botrytized”) wine fermentations carried out at high (~30°C) and ambient (~20°C) temperatures. Fermentations at both temperatures possessed similar populations of Saccharomyces, Hanseniaspora, Pichia, Metschnikowia, Kluyveromyces, and Candida species. However, higher populations of non-Saccharomyces yeasts persisted in ambient-temperature fermentations, with Candida and, to a lesser extent, Kluyveromyces species remaining long after the fermentation was dominated by Saccharomyces. In general, denaturing gradient gel electrophoresis profiles of yeast ribosomal DNA or rRNA amplified from the fermentation samples correlated well with the plating data. The direct molecular methods also revealed a Hanseniaspora osmophila population not identified in the plating analysis. rRNA analysis also indicated a large population (>10⁶ cells per ml) of a nonculturable Candida strain in the high-temperature fermentation. Monoculture analysis of the Candida isolate indicated an extreme fructophilic phenotype and correlated with an increased glucose/fructose ratio in fermentations containing higher populations of Candida. Analysis of wine fermentation microbial ecology by using both culture-dependent and -independent methods reveals the complexity of yeast interactions enriched during spontaneous fermentations.     

Application of response surface methodology for the optimization of arabinose biotransformation to arabitol by Candida parapsilosis

L-arabitol, a polyol with applications in the food and pharmaceutical industries, is secreted by different yeasts, e.g., Candida spp., Pichia spp., and Debaryomyces spp. The process of its biotechnological production is highly dependent on the physical and chemical conditions of culture. The aim of this study was to use statistical response surface methodology (RSM) to optimize the biotransformation of L-arabinose to arabitol by Candida parapsilosis, a yeast species able to assimilate pentoses. Batch cultures of the yeast were prepared following a Plackett-Burman design for seven variables. Following this, rotation speed, temperature, and L-arabinose concentration were chosen for a central composite design (CCD) experiment, which was carried out to optimize the production L-arabitol. The results showed that the optimal levels for the three factors were: rotation speed 150 rpm, temperature 28°C, and L-arabinose concentration 32.5 g/l. The predicted concentration of arabitol after two days of incubation of C. parapsilosis under the above conditions was 14.3 g/l. The value of R²=0.8323 suggested that this model was well-fitted to the experimental data, and this was confirmed during a verification experiment.     

Diversity and physiological characterization of d-xylose-fermenting yeasts isolated from the brazilian amazonian forest

Background

This study is the first to investigate the Brazilian Amazonian Forest to identify new D-xylose-fermenting yeasts that might potentially be used in the production of ethanol from sugarcane bagasse hemicellulosic hydrolysates.

Methodology/Principal Findings

A total of 224 yeast strains were isolated from rotting wood samples collected in two Amazonian forest reserve sites. These samples were cultured in yeast nitrogen base (YNB)-D-xylose or YNB-xylan media. Candida tropicalis, Asterotremella humicola, Candida boidinii and Debaryomyces hansenii were the most frequently isolated yeasts. Among D-xylose-fermenting yeasts, six strains of Spathaspora passalidarum, two of Scheffersomyces stipitis, and representatives of five new species were identified. The new species included Candida amazonensis of the Scheffersomyces clade and Spathaspora sp. 1, Spathaspora sp. 2, Spathaspora sp. 3, and Candida sp. 1 of the Spathaspora clade. In fermentation assays using D-xylose (50 g/L) culture medium, S. passalidarum strains showed the highest ethanol yields (0.31 g/g to 0.37 g/g) and productivities (0.62 g/L·h to 0.75 g/L·h). Candida amazonensis exhibited a virtually complete D-xylose consumption and the highest xylitol yields (0.55 g/g to 0.59 g/g), with concentrations up to 25.2 g/L. The new Spathaspora species produced ethanol and/or xylitol in different concentrations as the main fermentation products. In sugarcane bagasse hemicellulosic fermentation assays, S. stipitis UFMG-XMD-15.2 generated the highest ethanol yield (0.34 g/g) and productivity (0.2 g/L·h), while the new species Spathaspora sp. 1 UFMG-XMD-16.2 and Spathaspora sp. 2 UFMG-XMD-23.2 were very good xylitol producers.

Conclusions/Significance

This study demonstrates the promise of using new D-xylose-fermenting yeast strains from the Brazilian Amazonian Forest for ethanol or xylitol production from sugarcane bagasse hemicellulosic hydrolysates.     

Yeasts and Yeast-Like Fungi Associated with Tree Bark: Diversity and Identification of Yeasts Producing Extracellular Endoxylanases

A total of 239 yeast strains was isolated from 52 tree bark samples of the Medaram and Srisailam forest areas of Andhra Pradesh, India. Based on analysis of D1/D2 domain sequence of 26S rRNA gene, 114 strains were identified as ascomycetous; 107 strains were identified as basidiomycetous yeasts; and 18 strains were identified as yeast-like fungi. Among the ascomycetous yeasts, 51% were identified as members of the genus Pichia, and the remaining 49% included species belonging to the genera Clavispora, Debaryomyces, Kluyveromyces, Hanseniaspora, Issatchenkia, Lodderomyces, Kodamaea, Metschnikowia, and Torulaspora. The predominant genera in the basidiomycetous yeasts were Cryptococcus (48.6%), Rhodotorula (29%), and Rhodosporidium (12.1%). The yeast-like fungi were represented by Aureobasidium pullulans (6.7%) and Lecythophora hoffmanii (0.8%). Of the 239 yeast strains tested for Xylanase, only five strains of Aureobasidium sp. produced xylanase on xylan-agar medium. Matrix-assisted laser desorption ionization-time of flight analysis and N-terminal amino-acid sequence of the xylanase of isolate YS67 showed high similarity with endo-1-4-β-xylanase (EC 3.2.1.8) of Aureobasidium pullulans var. melanigenum. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Yeasts in Antarctic soils

The yeast flora of 126 soil samples from the Ross Dependency of Antarctica was examined and compared with that of eight samples from East Greenland. Fifty-two Antarctic samples contained yeasts, in numbers ranging from 5 to over 100,000/g; species isolated belonged to the generaDebaryomyces, Cryptococcus, Candida, Trichosporon andRhodotorula. Nearly all isolates ofCandida were obligate psychrophils; they belonged toCandida scottii, C. nivalis, C. gelida andC. frigida. Duplicate samples, taken at the same site within a few yards of one another, contained the same size and kind of yeast populations. Although not all samples which contained plants (algae, lichens or mosses) contained yeasts, almost all samples which contained yeasts contained plants. There was no correlation between yeast population and the pH of the sample, nor between yeast populations and the presence of vertebrate animals. The samples from East Greenland, which were from an area sufficiently warm and moist to support growth of higher plants, all contained yeasts, in numbers from 200 to 56,000/g. Species isolated were similar to those from Antarctic material.     

Fungal diseases of the respiratory tract

The proportion ofCandida and non-Candida species in the clinical material from patients. with respiratory-tract diseases was determined.C. albicans was isolated in 102 cases. An additional 89 strains of yeasts, isolated in association with respiratory diseases, belonged to 10 non-albicans Candida spp. andCryptococcus spp. The prevailing species, which occurred in 47 cases, wasC. parapsilosis. C. tropicalis, C. glabrata, andC. guilliermondii were isolated in 12, 10, and 9 cases, respectively. Four strains ofC. krusei and three strains ofC. lusitaniae and one strain each ofC. freyschussii, C. robusta, C. zeylanoides, andCryptococcus neoformans were also isolated.     

Yeast cell factories for fine chemical and API production

This review gives an overview of different yeast strains and enzyme classes involved in yeast whole-cell biotransformations. A focus was put on the synthesis of compounds for fine chemical and API (= active pharmaceutical ingredient) production employing single or only few-step enzymatic reactions. Accounting for recent success stories in metabolic engineering, the construction and use of synthetic pathways was also highlighted. Examples from academia and industry and advances in the field of designed yeast strain construction demonstrate the broad significance of yeast whole-cell applications. In addition to Saccharomyces cerevisiae, alternative yeast whole-cell biocatalysts are discussed such as Candida sp., Cryptococcus sp., Geotrichum sp., Issatchenkia sp., Kloeckera sp., Kluyveromyces sp., Pichia sp. (including Hansenula polymorpha = P. angusta), Rhodotorula sp., Rhodosporidium sp., alternative Saccharomyces sp., Schizosaccharomyces pombe, Torulopsis sp., Trichosporon sp., Trigonopsis variabilis, Yarrowia lipolytica and Zygosaccharomyces rouxii.     

Yamadazyma ubonensis f.a., sp. nov., a novel xylitol-producing yeast species isolated in Thailand

Three hundred and thirty-seven xylose-utilizing yeast strains were isolated from various natural samples. Among these, 68 strains produced xylitol in the range of 0.1–0.69 g xylitol/g xylose. Thirty-nine xylitol-producing strains were identified to be Candida tropicalis. Ten strains were found belonging to 14 known species in the genus Candida, Cyberlindnera, Meyerozyma, Pichia, Wickerhamomyces, Yamadazyma and Cryptococcus. Two strains were identified to be two Candida species and two strains (DMKU-XE142^T and DMKU-XE332) were found to be a novel species. Strain DMKU-XE142^T was isolated from tree bark and DMKU-XE332 was obtained from decaying plant leaf collected in Thailand. On the basis of morphological, biochemical, physiological and chemotaxonomic characteristics and sequence analysis of the D1/D2 region of the large subunit rRNA gene (LSU) and the internal transcribed spacer (ITS) region, the two strains were determined to represent a novel Yamadazyma species although formation of ascospores was not observed. The sequences of the D1/D2 region of the LSU rRNA gene and the ITS region of the two strains were identical but differed from Yamadazyma phyllophila, the closest species in terms of pairwise sequence similarity of the D1/D2 region, by 1.7 % nucleotide substitutions and 3.5 % nucleotide substitutions in the ITS region. The name Yamadazyma ubonensis f.a., sp. nov. is proposed (type strain is DMKU-XE142^T = BCC 61020^T = CBS 12859^T).     

Production of ethanol and xylanolytic enzymes by yeasts inhabiting rotting wood isolated in sugarcane bagasse hydrolysate

Yeasts associated with rotting wood from four Atlantic Rain forest sites in Brazil were investigated using a culture medium based on sugarcane bagasse hydrolysate. A total of 330 yeast strains were isolated. Pichia manshurica, Candida pseudolambica, and Wickerhamomyces sp. 3 were the most frequently isolated species. Fourteen novel species were obtained in this study. All isolates were tested for their ability to ferment d-xylose and to produce xylanases. In the fermentation assays using d-xylose (30 g L⁻¹), the main ethanol producers were Scheffersomyces stipitis (14.08 g L⁻¹), Scheffersomyces sp. (7.94 g L⁻¹) and Spathaspora boniae (7.16 g L⁻¹). Sc. stipitis showed the highest ethanol yield (0.42 g g⁻¹) and the highest productivity (0.39 g L⁻¹h⁻¹). The fermentation results using hemicellulosic hydrolysate showed that Sc. stipitis was the best ethanol producer, achieving a yield of 0.32 g g⁻¹, while Sp. boniae and Scheffersomyces sp. were excellent xylitol producers. The best xylanase-producing yeasts at 50 °C belonged to the species Su. xylanicola (0.487 U mg⁻¹) and Saitozyma podzolica (0.384 U mg⁻¹). The results showed that rotting wood collected from the Atlantic Rainforest is a valuable source of yeasts able to grow in sugarcane bagasse hydrolysate, including species with promising biotechnological properties.