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The xylanolytic and amylolytic yeasts were qualitatively determined by Cong red xylan agar and soluble starch agar plates, respectively. The most xylanase and α-amylase inducible strain (AUN-02) was selected and identified using PCR amplification of 26S rRNA gene and sequence analysis. The comparison of the alignment results and phylogenetic analysis of the sequences of the isolated yeast to published rRNA gene sequences in GenBank, confirmed the identification of the isolate as Pichia membranifaciens. Xylanase and α-amylase production by isolated P. membranifaciens were investigated at different pH values (4-8), temperature degrees (20-45°C), incubation time (1-7 days) and various substrates.A higher production of xylanase (38.8 U/mL) and a-amylase (28.7 U/mL) was obtained after 4 days of fermentation of P. membranifaciens. Higher activity of xylanase (36.83 U/mL) and a-amylase (27.7 U/mL) was obtained in the fermentation of P. membranifaciens in a culture medium adjusted to pH 7.0. The optimum temperature showed maximum xylanase and a-amylase activity (42.6 and 32.5 units/mL, respectively) was estimated at 35 °C. The xylanase and a-amylase activities of P. membranifaciens were estimated and compared for the different substrates tested. The strain revealed 100% relative activity of xylanase and a-amylase on beechwood and potato starch, respectively. The affinity of enzymes towards substrate was estimated using Km values. The Km values of xylanase and α-amylase increased in the order of pH’s 7.0, 6.0 and 4.5 (0.85, 1.6 and 3.4 mg xylan/mL and 0.22, 0.43 and 2.8 mg starch/mL, respectively). the yeast P. membranifaciensis is suitable for produce neutral xylanase and α-amylase enzymes. So, it could be used as a promising strain for production of these enzymes in industrial field.

The aim of this study was to assess the potential of bacteria Escherichia coli ATCC 35218, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923 and yeast Pichia membranifaciens ZIM 2417 to adhere to wooden surfaces such as poplar (Populus sp.), Norway spruce (Picea abies), European beech (Fagus sylvatica), beech coated with the commercial Belinka oil food contact and disinfectant P3-oxonia active 150, and investigate their survival on the beech wood surface under different relative humidities (RH; 65%, 75%, 85%, 98%) and temperatures (10 °C, 20 °C, 27 °C/37 °C). To extend the research goals, the scanning electron microscopy (SEM) analysis was also performed. The adhesion was determined by the number of colony-forming units per mm2 of sample (CFU/mm2). Results showed that all tested bacteria and yeast were able to adhere to the wooden surfaces, although differences were observed according to strains and type of wood. It was evident that number of adhered cells of S. aureus was lower on spruce (3.62 × 103 CFU/mm2) compared to poplar and beech (1.09 × 105 and 2.11 × 104 CFU/mm2, respectively). Furthermore, oil and disinfectant promoted the adhesion of P. aeruginosa (155.93 and 130.50%, respectively) on the beech surfaces, while they had a strong inhibitory effect on the other tested microorganisms E. coli (87.44 and 88.44%, respectively), S. aureus (91.24 and 96.80%, respectively) and P. membranifaciens (92.45 and 87.24%, respectively). These findings are consistent with SEM micrographs. The current data also indicated that relative humidity and temperature significantly affected the adhesion of tested bacteria and yeast. The highest degree of adhesion was observed at a relative humidity of 98% and temperature of 20 and 37 °C for bacteria, or 20 and 27 °C for yeast. Thus, the knowledge of how these microorganisms adhere to wooden surfaces and which factors affect this phenomenon proves to be of great importance in order to avoid their colonization.

In the present study, a total of eight sourdough samples were collected from three different bakeries at two different times in Turkey. Also, laboratory-scale sourdough production was conducted by daily back-slopping for 7 days. Microbiological and chemical properties of the sourdoughs were investigated. Yeast species in the sourdoughs were identified by subjecting all presumptive yeast cultures to internal transcribed spacer region amplification of the 5.8S rRNA gene, restriction fragment length polymorphism analysis using Hae III, Hha I, and Hinf I endonucleases, and sequence analysis of the D1/D2 domain of the 26S rDNA gene. A total of seven profiles were determined according to the restriction fragments. Totally, 148 yeast isolates were identified at the species level (≥400 bp, 99% identity) as Saccharomyces cerevisiae (106), Kazachstania bulderi (11), Pichia fermentans (nine), Pichia membranifaciens (eight), Kazachstania servazzii (seven), Kazachstania unispora (four), and Hanseniaspora valbyensis (three). Although collected sourdoughs were produced without using baker's yeast, S. cerevisiae was the most frequently isolated yeast species. This can be related to the contamination of the bakery environment with commercial baker's yeast during the production of other bakery products. The pH and acidity levels of the collected sourdough samples ranged from 3.71 to 3.96 and 6.78 to 23.93 mL 0.1 N NaOH/10 g dough, respectively. Mean values of the content of maltose + sucrose, glucose, fructose, lactic acid, and acetic acid were 2.43, 1.57, 2.67, 7.30, and 1.40 g/kg, respectively. Due to the artisan and region-dependent handling of the sourdough, different biochemical patterns were observed among the collected samples.

Table olive brines, inoculated with six different starters of lactic acid bacteria (LAB) or spontaneously fermented, have been used as isolating source of killer yeasts throughout the fermentation process (120 d). Killer yeast isolates were identified and evaluated for technological and probiotic traits. Although the count of yeast population did not markedly vary among the different vessels and over time, the killer yeast phenotype was mainly present in yeast strains isolated from spontaneous fermentation; the number of killer isolates decreased over fermentation time. Killer phenotype was found in species identified as Pichia kluyveri, Zygoascus hellenicus, Wickerhamomyces anomalus, Pichia membranifaciens, Candida boidinii, Candida diddensiae and Saccharomyces cerevisiae. Among all tested isolates, W. anomalus strains evidenced the widest spectrum of enzymatic activities and the highest β-glucosidase and phtytase activity. These strains evidenced also the best growth at low pH and increasing bile salt concentration, when grown at 37 °C, as well as the most constant viability index (%) during in vitro digestion.

In this study, yeasts isolated from filter membranes used for the quality control of bottled wines were identified and tested for their resistance to some cleaning agents and potassium metabisulphite, adhesion to polystyrene and stainless-steel surfaces, and formation of a thin round biofilm, referred to as a MAT. A total of 40 strains were identified by rRNA internal transcribed spacer (ITS) restriction analysis and sequence analysis of D1/D2 domain of 26S rRNA gene. Strains belong to Pichia manshurica (12), Pichia kudriavzevii (9), Pichia membranifaciens (1), Candida sojae (6), Candida parapsilosis (3), Candida sonorensis (1), Lodderomyces elongisporus (2), Sporopachydermia lactativora (3), and Clavispora lusitaniae (3) species. Regarding the adhesion properties, differences were observed among species. Yeasts preferred planktonic state when tested on polystyrene plates. On stainless-steel supports, adhered cells reached values of about 6 log CFU/mL. MAT structures were formed only by yeasts belonging to the Pichia genus. Yeast species showed different resistance to sanitizers, with peracetic acid being the most effective and active at low concentrations, with minimum inhibitory concentration (MIC) values ranging from 0.08% (v/v) to 1% (v/v). C. parapsilosis was the most sensible species. Data could be exploited to develop sustainable strategies to reduce wine contamination and establish tailored sanitizing procedures.

Konya Kuflu Tulum cheese is a well-known variety of Turkish mold-ripened cheeses produced by cutting the mature Tulum cheese into pieces to allow the filamentous fungi to grow on its surface in the cool and humid atmosphere in cellars or caves. The aim of the present study was to determine the fungal flora of Kuflu cheese using 54 filamentous fungi and 8 yeasts that were isolated from 26 cheese samples. Internal transcribed spacer (ITS) sequencing indicated that 53 of the mold isolates were Penicillium roqueforti and 1 was Cladosporium cladosporioides. The yeasts were identified as Pichia membranifaciens, Candida zeylanoides, Debaryomyces hansenii, and Geotrichum candidum. Morphological examination of the P. roqueforti isolates on various media revealed similar phenotypes among all but two isolates; however, (GTG)5 fingerprinting analysis indicated that the isolated P. roqueforti strains were highly similar in all but one case, which displayed a different pattern. To our knowledge, this study represents the first to conduct molecular methods for identification of fungi associated with Konya Kuflu Tulum cheese. In addition, the morphological and genetic diversities of the P. isolates are presented. Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [119O314] This study was funded by the Scientific and Technological Research Council of Turkey (TUBITAK) , grant number 119O314 to Banu Metin.

International audience; This study aimed at investigating the influence of the process environment and raw materials as sources of microorganisms during Nyons black table olive fermentations. Fermented olives and/or brine from spoiled fermentation tanks were analyzed and compared to good quality samples from fermentations collected during 3 consecutive harvest years. Fresh olives, salt and different process environment samples were also analyzed. Microbial diversity of all samples was analyzed using 16S and ITS2 amplicon sequencing and SourceTracker tool was used to investigate links between environment, raw materials and fermentation samples. First, comparison of microbial diversity in control and most spoiled fermentations revealed striking differences in bacterial composition with an overall higher diversity in spoiled fermentations especially for lactic acid bacteria with Lentilactobacillus buchneri, Lentilactobacillus parafarraginis dominating in brine and Pediococcus parvulus, Pediococcus ethanolidurans dominating in olive fruits. Fungal communities were similar in composition although higher abundances of Pichia membranifaciens and Penicillium carneum/roqueforti were observed in spoiled samples. Secondly, process environment samples were characterized by high bacterial and fungal diversity, especially compared to fresh olive fruits. Overall, dominant fungal species in control fermentations were also found in most environmental samples revealing a "house mycobiota". SourceTracker analysis further highlighted the contribution of brine and water from the optical sorter as a source of fungi. Most interestingly, spoilage fungi and most bacteria were retrieved in brine and environmental samples while others such as P. ethanolidurans were only found in environmental samples indicating that the studied spoilage originated from a fermentation deviation rather than a punctual contamination. Taken altogether, these results highlighted the positive and negative influence of the process environment and emphasized the relevance of studying it to better understand microbial vectors occurring during food fermentations, especially natural ones.

The aim of the present study was to assess the microecosystem of 13 homemade spontaneously fermented wheat sourdoughs from different regions of Greece, through the combined use of culture-dependent (classical approach, clustering by Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) and identification by PCR species-specific for Lactiplantibacillus plantarum, and sequencing of the 16S-rRNA and 26S-rRNA gene, for Lactic Acid Bacteria (LAB) and yeasts, respectively) and independent approaches [DNA- and RNA-based PCR-Denaturing Gradient Gel Electrophoresis (DGGE)]. The pH and Total Titratable Acidity (TTA) values ranged from 3.64&ndash, 5.05 and from 0.50&ndash, 1.59% lactic acid, respectively. Yeast and lactic acid bacteria populations ranged within 4.60&ndash, 6.32 and 6.28&ndash, 9.20 log CFU/g, respectively. The yeast: LAB ratio varied from 1:23&ndash, 1:10,000. A total of 207 bacterial and 195 yeast isolates were obtained and a culture-dependent assessment of their taxonomic affiliation revealed dominance of Lb. plantarum in three sourdoughs, Levilactobacillus brevis in four sourdoughs and co-dominance of these species in two sourdoughs. In addition, Companilactobacillusparalimentarius dominated in two sourdoughs and Fructilactobacillussanfranciscensis and Latilactobacillus sakei in one sourdough each. Lactococcus lactis, Lb. curvatus, Leuconostoc citreum, Ln. mesenteroides and Lb. zymae were also recovered from some samples. Regarding the yeast microbiota, it was dominated by Saccharomyces cerevisiae in 11 sourdoughs and Pichia membranifaciens and P. fermentans in one sourdough each. Wickerhamomyces anomalus and Kazachstania humilis were also recovered from one sample. RNA-based PCR-DGGE provided with nearly identical results with DNA-based one, in only one sample the latter provided an additional band. In general, the limitations of this approach, namely co-migration of amplicons from different species to the same electrophoretic position and multiband profile of specific isolates, greatly reduced resolution capacity, which resulted in only partial verification of the microbial ecology detected by culture-dependent approach in the majority of sourdough samples. Our knowledge regarding the microecosystem of spontaneously fermented Greek wheat-based sourdoughs was expanded, through the study of sourdoughs originating from regions of Greece that were not previously assessed.

French PDO Nyons black table olives are produced according to a traditional slow spontaneous fermentation in brine. The manufacture and unique sensorial properties of these olives thus only rely on the autochthonous complex microbiota. This study aimed at unraveling the microbial communities and dynamics of Nyons olives during a 1.5-year-long spontaneous fermentation to determine the main microbial drivers and link microbial species to key metabolites. Fermentations were monitored at a local producer plant at regular time intervals for two harvests and two olive types (organically and conventionally grown) using culture-dependent and metabarcoding (ITS2 for fungi, V3-V4 region for bacteria) approaches. Olives and brines were also sampled for volatiles, organic acids and phenolic compounds. No major differences in microbiota composition were observed according to olive type or harvest period. Throughout the fermentation, yeasts were clearly the most dominant. ITS2 sequencing data revealed complex fungal diversity dominated by Citeromyces nyonsensis, Wickerhamomyces anomalus, Zygotorulaspora mrakii, Candida boidinii and Pichia membranifaciens species. Bacterial communities were dominated by the Celerinatantimonas genus, while lactic acid bacteria remained scarce. Clear shifts in microbial communities and biochemical profiles were observed during fermentation and, by correlating metabolites and microbiota changes, four different phases were distinguished. During the first 7 days, phase I, a fast decrease of filamentous fungal and bacterial populations was observed. Between days 21 and 120, phase II, W. anomalus and C. nyonsensis for fungi and Celerinatantimonas diazotrophica for bacteria dominated the fermentation and were linked to the pH decrease and citric acid production. Phase III, between 120 and 183 days, was characterized by an increase in acids and esters and correlated to increased abundances of Z. mrakii, P. membranifaciens and C. boidinii. During the last months of fermentation, phase IV, microbial communities were dominated by P. membranifaciens and C. boidinii. Both species were strongly correlated to an increase in fruity esters and alcohol abundances. Overall, this study provides an in-depth understanding about microbial species succession and how the microbiota shapes the final distinct olive characteristics. It also constitutes a first step to identify key drivers of this fermentation.

Hundreds of sourdoughs have been investigated in the last decades. However, many studies used a culture-dependent and/or culture-independent microbiological approach [mainly based on denaturing gradient gel electrophoresis (DGGE) of PCR amplicons], seldomly combined with a metabolite target analysis, to characterize the microbial species communities of the sourdoughs examined. Moreover, attention was mainly paid on lactic acid bacteria (LAB) and yeast species. In the present study, distinct household-scale (including an artisan lambic brewery) and artisan bakery-scale backslopped sourdoughs (17 in total), obtained from different regions (Belgium, France, United Kingdom, and USA), were examined through a multiphasic approach, encompassing a culture-dependent analysis [targeting LAB, acetic acid bacteria (AAB), and yeasts], different culture-independent techniques [rRNA-PCR-DGGE, metagenetics, and metagenomics (four bakery sourdoughs)], and metabolite target analysis. It turned out that the microbial species diversity of the sourdoughs was influenced by the house microbiota of the producer. Further, when the producer made use of different flours, the sourdoughs harbored similar microbial communities, independent of the flour used. AAB were only present in the Belgian sourdoughs, which might again be related to the processing environment. Fructilactobacillus sanfranciscensis (formerly known as Lactobacillus sanfranciscensis) was the prevalent LAB species of the eight sourdoughs produced by two of the three bakeries of different countries analyzed. These sourdoughs were characterized by the presence of either Saccharomyces cerevisiae or Kazachstania humilis. Moreover, the presence of Fl. sanfranciscensis was positively correlated with the production of mannitol and negatively correlated with the presence of other LAB or AAB species. Sourdoughs produced in an artisan lambic brewery were characterized by the presence of the yeast species Dekkera anomala and Pichia membranifaciens. One household sourdough was characterized by the presence of uncommon species, such as Pediococcus parvulus and Pichia fermentans. Metagenomic sequencing allowed the detection of many more LAB and AAB species than the other methods applied, which opened new frontiers for the understanding of the microbial communities involved during sourdough production processes.

Integrated metabolomic and metagenomic techniques were used to study the metabolite dynamics and phytochemistry of a soy whey-based beverage bio-transformed by water kefir consortium. The UPLC- MS/MS and HPLC-ESI-MS metabolite quantification and the OPLS-DA result showed that the kefir consortium induced a significant change in the metabolite composition and altered the phytochemistry of the fermented beverage. Bioactive peptide analogues, flavonoids, and glycerophospholipids including N-acetyl-L-phenylalanine, acetyl-DL-leucine; tephcalostan, wogonin, pelargonin, genistein, daidzein, and glycerophosphoserines (PS), glycerophosphoethanolamines (PE) respectively were synthesized while flavonoid glycosides and soyasaponins were degraded in the novel beverage. Furthermore, the beverage showed high ACE inhibitory and DPPH radical scavenging activity of 92.31% and 87.51% respectively. Lactobacillus, Saccharomyces cerevisiae, and Pichia membranifaciens were the predominant microbial groups in the new beverage as revealed by the metagenomic sequence analysis. The study thus provides discrete data evidence that kefir consortium is a viable starter for transforming soy whey into a bioactive beverage.

The aim of this study was to investigate the lactic acid bacteria (LAB) and yeast community from home-made sauerkraut collected from Southwest China through culture-dependent and culture-independent technology. Forty-eight samples of home-made sauerkraut were collected from households at three different locations in Southwest China. The pH, total acidity and salt contents among these fermented vegetables were 3.69 ± 0.42, 0.86 ± 0.43 g/100 ml, and 3.86 ± 2.55 g/100 ml, respectively. The number of lactic acid bacteria (LAB) and yeasts were 7.25 ± 1.05 log10 colony-forming units (CFU)/ml and 3.74 ± 1.01 log CFU/ml, respectively. A total of 182 LAB and 81 yeast isolates were identified. The dominant isolates were Lactobacillus plantarum, L. brevis, Pediococcus ethanolidurans, Pichia membranifaciens, P. fermentans and Kazachstania bulderi. Denaturing gradient gel electrophoresis (DGGE) showed that L. plantarum, uncultured Lactobacillus sp, P. ethanolidurans, and K. exigua were the predominant microflora. Our studies demonstrated that the DGGE technique combined with a culture-dependent method is very effective for studying the LAB and yeast community in Chinese traditional fermentation vegetables. The results will give us an understanding of LAB and yeast community of Chinese sauerkraut and improve the knowledge of LAB and yeast community of Chinese sauerkraut.

During the present study 80 samples of kareish (semi-soft cheese with 20% fats) and soft cheese (40 % fats) were collected Assiut City during June to December 2015. The fungal content of these samples was evaluated using Dichloran Rose Bengal Chloramphenicol (DRBC) and Yeast Extract Malt extract (YM) agar media. The fungal count per sample ranged from 7 colonies/g in soft cheese to 44800 colonies/g in kareish cheese. the total number of fungal species in kareish cheese was slightly lower on DRBC than on YM (24 and 29 species respectively). The number of fungal species per sample fluctuated between 1-8 species with the highest being recovered from kareish cheese. Aspergillus was the most prevalent genus contaminating 45 -85 % of the samples with the highest being recovered from Kareish Cheese. Penicillium came next contaminating 20 - 47.5% of samples with white soft cheese being the most affected product. Candida, Clavispora, Klyveromyces and Pichia contaminated 2.5-65 % of samples and the highest incidence was that of Clavispora in kareish cheese. Aspergillus niger, A. fumigatus, A. flavus, Penicillium chrysogenum, P. aurantiogriseum and P. brevicompactum were the commonest species in cheese samples. Some yeast fungi were identified by sequencing of rRNA gene. Among the 19 species of yeasts Candida tropicalis, Clavispora lusitaniae, Pichia kudriavzevii, Pichia membranifaciens and kluyveromyces lactis were the most common. Testing the natural occurrence of aflatoxins revealed that AFM1 contaminated 55% and 50 % of kareish and soft cheese respectively whereas AFM2 was found only in 10 % of kareish cheese samples.

The pellicle-forming yeast could cause the quality deterioration of wine. In this study, a pellicle-forming strain Hmp-1 was isolated from the spoilage blackberry wine, and identified as Pichia membranifaciens based on the morphology and partial nucleotide sequence of 26S rDNA. The effects of fermentation conditions (ethanol, sulfur dioxide, sugar, and temperature) on the growth of P. membranifaciens strain Hmp-1 and Saccharomyces cerevisiae strain FM-S-115 (a strain used for the blackberry wine fermentation) were investigated, respectively. The results indicated that Hmp-1 had lower resistance to these factors compared to FM-S-115, and the growth of Hmp-1 was completely inhibited by 10% (v/v) or 50 mg L–1 SO2 during the fermentation of blackberry wine. These results suggested that Hmp-1 could effectively be controlled by increasing ethanol or SO2 concentration during the fermentation and storage of blackberry wine. Furthermore, the analysis based on gas chromatography-mass spectrometry (GC-MS) showed that Hmp-1 remarkably decreased kinds of volatile compounds in blackberry wine, especially aldehydes and esters. In addition, some poisonous compounds were detected in the blackberry wine fermented by FM-S-115 and Hmp-1. These results suggested that Hmp-1 was a major cause leading to the quality deterioration of blackberry wine.

Background and Aims Fungicides used against the mould Botrytis cinerea can also non‐specifically affect indigenous yeasts important for the development of complex wine aroma. Our aim was to determine the minimal inhibitory concentration of the fungicides iprodione, pyrimethanil and fludioxonil plus cyprodinil for 109 yeasts of the grape berry microbiota. Methods and Results We measured the respiration activity of yeasts, changes in growth kinetics and adenosine triphosphate production. Fungicide tolerance may be a conserved feature within the yeast's genera, however, some heterogeneity between isolates remains. The viability and growth of 15 beneficial isolates were inhibited by fungicides at a residue concentration below the maximum permitted residue limits. These isolates belonged to the yeast species: Metschnikowia pulcherrima, Pichia kluyveri, Hanseniaspora uvarum and Bulleromyces albus. Furthermore, isolates of typical spoilage yeasts, such as Dekkera bruxellensis, Meyerozyma guilliermondii and Pichia membranifaciens, were found to be tolerant of a concentration of fungicides greater than that recommended for application by the suppliers. Conclusions Fungicide residues hindered the viability of desirable isolates and promoted the growth of several spoilage isolates and thus may have a negative impact on wine aroma. Significance of the Study For spontaneous fermentations the application of fungicides in the vineyard could add to the seasonal fluctuations of aroma and to the development of off‐flavours.

Vishniacozyma victoriae and Pichia membranifaciens were selected in a previous work for their biocontrol effectiveness against Penicillium expansum and Botrytis cinerea, the causal agents of blue and grey mold of pear fruits. The objective of this study was to determine the effects of different nutrient or additives on the growth of the two antagonist yeasts and of the two fungal pathogens in order to develop a rational selection based on nutritional profiles to be used in biocontrol enhancement of decay in pear fruits. Twenty-six different nutrient sources including 18 amino acids, five sugars, three inorganic nitrogen and one iron source were tested in vitro for their effect on the growth of the two yeasts and the two pathogens. Nutrients that promoted the growth of the yeasts and inhibited the growth of pathogens were applied with the antagonist to wounded fruits to evaluate their effect on enhancing biocontrol in cold storage. In general, the effect of each additive was specific to the yeasts and pathogens used in the in situ assays. The combination of the yeast and some additives resulted in a significantly higher activity with respect to the single treatments applied separately, producing synergistic effects. The effect of the exogenous application of CaCl 2 and chitosan together with the antagonist yeasts was also evaluated against the two pathogens. The most effective mixtures were the CaCl 2 with the two antagonist yeasts. Our research demonstrates that manipulating the chemical environment by adding several amino acid and/or Cl 2 Ca results in improved antagonistic activity of Vishniacozyma victoriae and Pichia membranifaciens against two postharvest diseases of pear fruits. Fil: Gramisci, Betina Rocío. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigación y Desarrollo en Ingeniería de Procesos, Biotecnología y Energías Alternativas. Universidad Nacional del Comahue. Instituto de Investigación y Desarrollo en Ingeniería de Procesos, Biotecnología y Energías Alternativas; Argentina Fil: Lutz, María Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte; Argentina. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias; Argentina Fil: Lopes, Christian Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigación y Desarrollo en Ingeniería de Procesos, Biotecnología y Energías Alternativas. Universidad Nacional del Comahue. Instituto de Investigación y Desarrollo en Ingeniería de Procesos, Biotecnología y Energías Alternativas; Argentina. Universidad Nacional del Comahue. Facultad de Ciencias Agrarias; Argentina Fil: Sangorrin, Marcela Paula. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte. Instituto de Investigación y Desarrollo en Ingeniería de Procesos, Biotecnología y Energías Alternativas. Universidad Nacional del Comahue. Instituto de Investigación y Desarrollo en Ingeniería de Procesos, Biotecnología y Energías Alternativas; Argentina

37 Páginas.-- 2 Tablas.-- 3 Figuras.-- 5 Tablas suplementarias, Knowledge of the microbial biodiversity through the different stages of black ripe olive processing is minimal. This study applied a metagenomic approach to evaluate the bacterial and fungal communities present in samples of two olive cultivars collected from an industrial producer. High-throughput sequencing and analysis of the V2-V3 region of the bacterial 16S rRNA genes showed that genus Acetobacter and the lactic acid bacteria (LAB) Lactobacillus and Oenococcus were the most abundant during most of the process. Other LAB genera (Enterococcus, Lactococcus, Weisella, Leuconostoc and Streptococcus), as well as Enterobacteriaceae and Vibrio were also detected, increasing their abundance in the last step before packaging. Some genera, such as Pseudoalteromonas, Alteromonas, Marinomonas and Oenococcus, had never been previously detected in table olives. The fungal community was characterized by the presence of Pichia membranifaciens, Kregervanrija fluxuum and members of the family Dispodascaceae. Although black ripe olives are sterilized before trading, the consequences that these findings may have on the quality of this product are discussed., This research was supported by the Spanish Government (Projects AGL2013-42739-R and AGL2016-76820-R), and partially financed by the European Union (European Regional Development Funds, ERDF). The authors gratefully thank Mrs. Alejandra Expósito for her skilful experimental work in the laboratory.

A total of 105 yeast isolates was obtained from 15 sourdough samples collected from different regions in China and subjected to random amplified polymorphic DNA (RAPD) analysis. Six species were identified including Pichia membranifaciens, which has not previously been reported in Chinese sourdoughs. Different species of yeast were used in single-culture fermentation to make Chinese steamed bread (CSB). The volatiles of the CSB were captured by solid-phase microextraction method, separated and identified by gas chromatography-mass spectrometry. In total, 41 volatile compounds were found in all the steamed breads. All CSBs showed a similar volatile profile; however, significant differences in the quantity of some volatile compounds were seen among the CSB fermented by different yeast species. A partial least squares discriminant analysis showed that the CSBs could be separated by their characteristic volatile profiles. The study suggested that the aromatic properties of CSB are determined by the yeast used.

Coffee rust is a fungal disease that has affected every coffee-producing region in the world. Given that the effectivity of the protectant and systemic fungicides applied routinely to control the spread of the causative agent of the disease (Hemileia vastatrix) has gradually diminished, besides are harmful to mammals and ecosystems, the objective of this work was to search for a mixture of harmless natural compounds with the potential to be applied in the field. So, a yeast strain producing a battery of long-chain carboxylic acids (CA) with fungicide properties was isolated from soil of coffee crop and identified as Pichia membranifaciens by ITS sequencing. Culture conditions of the yeast were optimized and the CA in the solution were characterized by Gas Chromatography-Mass Spectrometry (GC-MS) as ethyl formate (55.5 g L-1), octadecenoic acid (3.5 g L-1), propionic acid (7.2 g L-1), 3-(octadecanoyl)-propionic acid (7.2 g L-1) and methyl acetate (8.4 g L-1). Randomized field studies were conducted in three different locations in Chiapas, México. Five treatments were tested including three concentrations of the CA solution (389, 584 and 778 ppm) and copper oxychloride (5 000 ppm) as conventional control. The initial coffee rust incidence averages varied between sites: Maravillas (3-9%), Santo Domingo (10-16%) and Búcaro (16-22%). The treatments of CA solution proved to be effective at slowing down the progress of the rust disease even for the sites where initial incidence was high. Likewise, the CA solution reduced the viability of H. vastatrix spores, as assessed by fluorescence microscopy. RESUMO A ferrugem do cafeeiro é uma doença que tem afetado as regiões produtoras de café no mundo. Considerando que os funguicidas protetores e sistemáticos que são aplicados rotinariamente para controlar a difusão do agente causador da doença (Hemileia vastatrix) são nocivos para os humanos, animais e ecossistemas, o objetivo deste trabalho é determinar um componente natural com potencial de uso. Uma cepa de levedura produzindo uma bateria de ácidos carboxílicos com ação funguicida foi isolada do solo e identificada como Pichia membranifaciens por sequenciamento ITS. Foram optimizadas as condições de cultura e os ácidos carboxílicos presentes na semi purificada sobrenadante (solução CA) foram caracterizados por cromatografia de gás espectometia de massa (GC-MS) como formato de etilo (55.5 g L-1), ácido octadecenóico (3.5 g L-1), ácido propiônico (7.2 g L-1), 3-(octadecanoil)-ácido propiônico 7.2 g L-1) e acetato de metilio (8.4 g L-1). Estudos do campo foram conduzidos em três locais na região de Chiapas, Mexico. Cinco tratamentos foram testados incluindo três concentrações da solução CA (389, 584 e 778 ppm) e o funguicida comercial contendo oxicloreto de cobre (500 ppm). As médias iniciais de incidência da ferrugem do cafeeiro variou em função do local: Maravilla (3-9%), Santo domingo (10-16%) e Búcaro (16-22%). A solução CA provou ser efetiva retardando o progresso da doença, mesmo nos locais onde a incidência inicial foi elevada. A solução CA reduziu a viabilidade dos esporos de H. Vastatrix, observado por meio de fluorescência.

Graphical abstract, Highlights • Effects of ultrasound on wine microorganisms: yeasts and Lactiplantibacillus plantarum. • The antimicrobial effect of ultrasound depends on the total energy. • Ultrasound could affect vitality (by reversibly delaying growth) or viability (by irreversibly reducing growth). • Different behaviors have been observed in different wine microorganisms, showing low and high susceptibility., Ultrasound is one of the most promising non-thermal an emerging technique in food technology. The objective of the present work was to evaluate the effect of different ultrasonic treatments on the most important wine microbiota (Saccharomyces and non-Saccharomyces yeasts and lactic acid bacteria). Two stages were carried out: the assessment step, where six different ultrasonic treatments (with varying power, time, and pulses) were used on Saccharomyces cerevisiae, Brettanomyces spp., and Lactiplantibacillus plantarum; and the validation step, where two chosen ultrasonic treatments were used on Zigosaccharomyces bailli, Brettanomyces spp., Saccharomyces cerevisiae, Saccharomyces bayanus, Pichia membranifaciens, Schizosaccharomyces pombe, and Hanseniaspora osmophila. The most sensitive microorganism was Brettanomyces spp., and the most resistant was Lactiplantibacillus plantarum. Ultrasonic treatments had varying effects on vitality (delay of growth or maximum OD reduction) and on viability (reduction of microbial growth).

Rapid yeast identification is of particular importance in monitoring wine fermentation and assessing strain application in winemaking. We used MALDI-TOF MS analysis supported by 26 S rRNA gene sequence analysis and Saccharomyces-specific PCR testing to differentiate reference and field strains recovered from organic wine production facilities in Waipara, New Zealand, in which Pinot Noir wine was produced by spontaneous fermentations in the vineyard and in the winery. Strains were isolated from each of four key stages of each ferment to evaluate changes in taxonomic diversity. MALDI-TOF MS analysis was confirmed as an excellent yeast identification method, with even closely related Saccharomyces species readily distinguished. A total of 13 indigenous species belonging to eight genera were identified from Pinot Noir ferments, with taxonomic diversity generally reducing as fermentation progressed. However, differences between the taxa recovered were observed between the vineyard and winery ferments, despite the grapes used being from the same batch. Furthermore, some consistent proteomic differences between strains of S. cerevisiae, Hanseniasporum uvarum, Candida californica, Pichia membranifaciens and Starmerella bacillaris correlated with the different fermentation systems used. The high speed, low cost, taxonomic resolution and ability to characterise subtle changes in phenotype that may result from variations in environmental conditions makes MALDI-TOF analysis an attractive tool for further and wider applications in the wine industry. Such applications may include monitoring wine fermentation to actively support the consistency of high-quality wine products, and potentially for the development of such products too.

Interspecific hybridization is a common mechanism enabling genetic diversification and adaptation; however, the detection of hybrid species has been quite difficult. The identification of microbial hybrids is made even more complicated, as most environmental microbes are resistant to culturing and must be studied in their native mixed communities. We have previously adapted the chromosome conformation capture method Hi-C to the assembly of genomes from mixed populations. Here, we show the method's application in assembling genomes directly from an uncultured, mixed population from a spontaneously inoculated beer sample. Our assembly method has enabled us to de-convolute four bacterial and four yeast genomes from this sample, including a putative yeast hybrid. Downstream isolation and analysis of this hybrid confirmed its genome to consist of Pichia membranifaciens and that of another related, but undescribed, yeast. Our work shows that Hi-C-based metagenomic methods can overcome the limitation of traditional sequencing methods in studying complex mixtures of genomes. Copyright © 2017 John Wiley & Sons, Ltd.

Aims To determine the effect of three different freezing temperatures on post-freeze-drying survival rates of wine yeasts and lactic acid bacteria (LAB). To know if a similar freeze-drying protocol can be used for both micro-organisms. Methods and Results Cells from liquid culture media were recovered and concentrated in appropriate lyoprotectants. Aliquots of each strain were frozen at −20, −80 and −196°C before vacuum drying. Viable cell counts were done before freezing and after freeze-drying. Survival rates were calculated. Freezing temperatures differently affected yeast and bacteria survival. The highest survival rates were obtained at −20 and −80°C for yeasts, but at −196°C for LAB. Major differences in survival rates were recorded among freeze-dried yeasts, but were less drastic for LAB. Yeasts Pichia membranifaciens, Starmerella bacillaris and Metschnikowia pulcherrima, and LAB Lactobacillus paracasei, Pediococcus parvulus and Lactobacillus mali, were the most tolerant species to freeze-drying, regardless of freezing temperature. Conclusions Yeast and LAB survival rates differed for each tested freezing temperature. For yeasts, −20°C ensured the highest post-freeze-drying viability and −196°C for LAB. Significance and Impact of the Study Freezing temperature to freeze-dry cells is a crucial factor for ensuring good wine yeast and LAB survival. These results are important for appropriately preserving micro-organisms and for improving starter production processes.