Debaryomyces Hansenii Research Articles

Bacterial-fungal interactions promote parallel evolution of global transcriptional regulators in a widespread Staphylococcus species.

Experimental studies of microbial evolution have largely focused on monocultures of model organisms, but most microbes live in communities where interactions with other species may impact rates and modes of evolution. Using the cheese rind model microbial community, we determined how species interactions shape the evolution of the widespread food- and animal-associated bacterium Staphylococcus xylosus. We evolved S. xylosus for 450 generations alone or in co-culture with one of three microbes: the yeast Debaryomyces hansenii, the bacterium Brevibacterium aurantiacum, and the mold Penicillium solitum. We used the frequency of colony morphology mutants (pigment and colony texture phenotypes) and whole-genome sequencing of isolates to quantify phenotypic and genomic evolution. The yeast D. hansenii strongly promoted diversification of S. xylosus. By the end of the experiment, all populations co-cultured with the yeast were dominated by pigment and colony morphology mutant phenotypes. Populations of S. xylosus grown alone, with B. aurantiacum, or with P. solitum did not evolve novel phenotypic diversity. Whole-genome sequencing of individual mutant isolates across all four treatments identified numerous unique mutations in the operons for the SigB, Agr, and WalRK global regulators, but only in the D. hansenii treatment. Phenotyping and RNA-seq experiments highlighted altered pigment and biofilm production, spreading, stress tolerance, and metabolism ofS. xylosus mutants.Fitness experiments revealed antagonistic pleiotropy, where beneficial mutations that evolved in the presence of the yeast had strong negative fitness effects in other biotic environments. This work demonstrates that bacterial-fungal interactions can have long-term evolutionary consequences within multispecies microbiomes by facilitating the evolution of strain diversity.

Co-Fermentation of Agri-Food Residues Using a Co-Culture of Yeasts as a New Bioprocess to Produce 2-Phenylethanol.

Whey is a dairy residue generated during the production of cheese and yogurt. Whey contains mainly lactose and proteins, contributing to its high chemical oxygen demand (COD). Current environmental regulations request proper whey disposal to avoid environmental pollution. Whey components can be transformed by yeast into ethanol and biomolecules with aroma and flavor properties, for example, 2-phenyethanol (2PE), highly appreciated in the industry due to its organoleptic and biocidal properties. The present study aimed to valorize agri-food residues in 2PE by developing suitable bioprocess. Cheese whey was used as substrate source, whereas crab headshells, residual soy cake, and brewer's spent yeast (BSY) were used as renewable nitrogen sources for the yeasts Kluyveromyces marxianus and Debaryomyces hansenii. The BSYs promoted the growth of both yeasts and the production of 2PE in flask fermentation. The bioprocess scale-up to 2 L bioreactor allowed for obtaining a 2PE productivity of 0.04 g2PE/L·h, twofold better productivity results compared to the literature. The bioprocess can save a treatment unit because the whey COD decreased under the detection limit of the analytical method, which is lower than environmental requirements. In this way, the bioprocess prevents environmental contamination and contributes to the circular economy of the dairy industry.

Application of beekeeping products in the technology of raw smoked sausages

The safety of fermented sausage products is due to the elimination of a number of risks of biological origin, mainly of a microbiological nature. One of the most promising directions for the development of barrier technologies is the use of starter cultures and the biologically active substances produced by them in the production of fermented meat products. The research aims to study the influence of starter cultures and honey on a complex of physico-chemical, structuralmechanical, organoleptic indicators and nutritional value of raw smoked sausages. The article presents changes in the physical and chemical parameters of raw smoked sausages when using starter cultures in combination with different types of honey. When performing the work, 4 types of honey were used: from sunflower, phacelia, honeydew honey and from medicinal herbs. At the first stage of research, the proportion of honey application and the type of starter crops were selected. The next stage of the study was to determine the impact of the starter cultures included in the product BIOSTART PLUS 50 (lactic acid bacteria Lactobacillus Sake and yeasts Debaryomyces Hansenii) and honey on moisture loss during fermentation of raw smoked sausages, physicochemical parameters of finished products. Determination of moisture content in samples was carried out by drying in a drying cabinet at a temperature of 103±2°C. Conducted studies prove that the use of the product BIOSTART PLUS 50 (lactic acid bacteria Lactobacillus Sakei and yeasts Debaryomyces Hansenii) and honey allows shortening the ripening and drying technological process. A comparative analysis of manufactured raw-smoked sausages in terms of nutritional value makes it possible to state that manufactured raw-smoked sausages with the addition of honey meet the requirements of the standard. The results of the set of studies carried out in this work showed that the use of a composition of lactic acid bacteria and yeast with the use of honey as a source of carbohydrates will allow intensifying the technological process and improve the quality of finished products

Debaryomyces hansenii supplementation in low fish meal diets promotes growth, modulates microbiota and enhances intestinal condition in juvenile marine fish

BackgroundThe development of a sustainable business model with social acceptance, makes necessary to develop new strategies to guarantee the growth, health, and well-being of farmed animals. Debaryomyces hansenii is a yeast species that can be used as a probiotic in aquaculture due to its capacity to i) promote cell proliferation and differentiation, ii) have immunostimulatory effects, iii) modulate gut microbiota, and/or iv) enhance the digestive function. To provide inside into the effects of D. hansenii on juveniles of gilthead seabream (Sparus aurata) condition, we integrated the evaluation of the main key performance indicators coupled with the integrative analysis of the intestine condition, through histological and microbiota state, and its transcriptomic profiling.ResultsAfter 70 days of a nutritional trial in which a diet with low levels of fishmeal (7%) was supplemented with 1.1% of D. hansenii (17.2 × 105 CFU), an increase of ca. 12% in somatic growth was observed together with an improvement in feed conversion in fish fed a yeast-supplemented diet. In terms of intestinal condition, this probiotic modulated gut microbiota without affecting the intestine cell organization, whereas an increase in the staining intensity of mucins rich in carboxylated and weakly sulphated glycoconjugates coupled with changes in the affinity for certain lectins were noted in goblet cells. Changes in microbiota were characterized by the reduction in abundance of several groups of Proteobacteria, especially those characterized as opportunistic groups. The microarrays-based transcriptomic analysis found 232 differential expressed genes in the anterior-mid intestine of S. aurata, that were mostly related to metabolic, antioxidant, immune, and symbiotic processes.ConclusionsDietary administration of D. hansenii enhanced somatic growth and improved feed efficiency parameters, results that were coupled to an improvement of intestinal condition as histochemical and transcriptomic tools indicated. This probiotic yeast stimulated host-microbiota interactions without altering the intestinal cell organization nor generating dysbiosis, which demonstrated its safety as a feed additive. At the transcriptomic level, D. hansenii promoted metabolic pathways, mainly protein-related, sphingolipid, and thymidylate pathways, in addition to enhance antioxidant-related intestinal mechanisms, and to regulate sentinel immune processes, potentiating the defensive capacity meanwhile maintaining the homeostatic status of the intestine.

Perspectives on the Probiotic Potential of Indigenous Moulds and Yeasts in Dry-Fermented Sausages.

The role of indigenous fungi in the appropriate development of sensory properties and the safety of dry-fermented sausages has been widely established. Nonetheless, their applications as probiotic agents have not been elucidated in such products yet, despite their promising functional features. Thus, it should be interesting to evaluate the probiotic potential of native Debaryomyces hansenii isolates from dry-fermented sausages and their application in the meat industry, because it is the most frequently isolated yeast species from these foodstuffs and its probiotic effects for animals as well as its possible probiotic activity for human beings have been demonstrated. Within the functional ability of foodborne yeasts, anti-inflammatory, antioxidant, antimicrobial, antigenotoxic, and immunomodulatory properties have been reported. Similarly, the use of dry-fermented sausages as vehicles for probiotic moulds remains a challenge because the survival and development of moulds in the gastrointestinal tract are still unknown. Nevertheless, some moulds have been isolated from faeces possibly from their spores as a form of resistance. Additionally, their beneficial effects on animals and humans, such as the decrease in lipid content and the anti-inflammatory activity, have been reported, although they seem to be more related to their postbiotic capacity due to the generated bioactive compounds with profunctional attributes than to their role as probiotics. Therefore, further studies providing knowledge useful for generating dry-fermented sausages with improved functionality are fully necessary.

Comparative evaluation of the flavour-promoting role of autochthonous yeast strains on dry sausages

The taste and odour profiles of dry sausages separately inoculated with four autochthonous yeasts (Debaryomyces hansenii SH4, Candida zeylanoides DQ7, Torulaspora delbrueckii HG7 and Pichia kudriavzevii MDJ1) were investigated. The inoculation of yeast increased the scores of fermented, floral and fruity odour and decreased the scores of sour taste. Notably, D. hansenii SH4 and C. zeylanoides DQ7 inoculated sausages had higher umami free amino acid contents than control sausage, as well as showed the highest umami and salty taste scores of 3.64 and 5.42, respectively. Meanwhile, the inoculation of D. hansenii SH4, Pichia kudriavzevii MDJ1 and C. zeylanoides DQ7 could increase the contents of ester, alcohol and aldehyde by 1.31, 1.32 and 1.38 times from dry sausages, respectively. Partial least squares regression analysis revealed that nine taste compounds and 34 odour compounds were mainly responsible for sensory differences between dry sausages. This study provided new insights into the relationship between chemical compounds and sensory characteristics of dry sausage, demonstrating that four autochthonous yeasts contribute significantly to the flavour formation of dry sausage.

Short-Term Changes in Aroma-Related Volatiles in Meat Model: Effect of Fat and D. hansenii Inoculation.

This study assessed the effect of replacing pork lard with coconut oil and Debaryomyces hansenii inoculation on the biotransformation of amino acids into volatile compounds in a meat model system. Yeast counts, solid-phase microextraction, and gas chromatography/mass spectrometry were used to assess yeast growth and volatile production, respectively. Yeast growth was confirmed until 28 d, although the volatile profile changed until 39 d. Forty-three volatiles were quantified, and their odor activity values (OAVs) were calculated. The presence of fat and yeasts contributed to differences in volatiles. In pork lard models, a delayed formation of lipid-derived aldehyde compounds was observed, whereas in coconut oil models, the generation of acid compounds and their respective esters was enhanced. Yeast activity affected amino acid degradation, which produced an increase in branched-chain aldehydes and alcohols. The aroma profile in the coconut models was influenced by hexanal, acid compounds, and their respective esters, whereas in pork lard models, aroma was affected by methional (musty, potato) and 3-methylbutanal (green, cocoa). The yeast inoculation contributed to the generation of 3-methylbutanoic acid (cheesy) and phenylethyl alcohol (floral). The type of fat and yeast inoculation produced a differential effect on the aroma.

The effect of using different yeast species on the composition of carbohydrates and volatile aroma compounds in kefir drinks

Kefir is a fermented dairy product known for its high content of lactic acid bacteria and various types of yeast. The yeast microorganisms present in kefir cultures play a crucial role in shaping the distinctive characteristics of kefir. This study aimed to investigate the impact of different yeast species commonly used in industrial kefir production on the carbohydrate and volatile aroma components of kefir during the storage period. To this end, kefir cultures with various yeast species, including Debaryomyces hansenii (DH), Kluyveromyces marxianus (KM), Candida colliculosa (CC), Yarrowia lipolytica (YL), Saccharomyces bouardii (SB), Saccharomyces cerevisiae (SC), Kluyveromyces lactis (KL), and Geotricum candidum (GC), were prepared and utilized in kefir production. The study findings revealed significant effects of different yeast species on the composition of carbohydrate and volatile aroma components in kefir (p < 0.05), with Klyveromyces spp. having particularly notable effects. It was observed that kefir drinks containing ethanol exhibited negligible lactose content, while their ethanol content was high. Kefir drinks containing minimal Klyveromyces marxianus contained glucose and galactose, whereas those containing Yarrowia lipolitica exhibited the lowest ethanol content.

Biotransformation of gallate esters by a pH-stable tannase of mangrove-derived yeast Debaryomyces hansenii.

Introduction: Tannase is a crucial enzyme that finds wide applications in the pharmaceutical industry, feed processing, and beverage manufacturing. Although extensive studies have been conducted on tannases from fungi and bacteria, reports on tannases exhibiting favorable pH stability are relatively limited. Methods: In this study, a tannin-degrading strain Debaryomyces hansenii was screened to induce tannase production, and the corresponding tannase coding gene TANF was successfully cloned and expressed in Yarrowia lipolytica. SDS-PAGE analysis revealed that the purified TanF tannase had a molecular weight of approximately 70kDa. Results and Discussion: The enzyme demonstrated optimal activity at 40°C and retained over 80% of its activity in the range of 35°C-60°C. Of particular interest, TanF exhibited remarkable enzyme activity at pH 5.0 and retained more than 70% of its relative activity across a wide pH range of 3.0-8.0. Furthermore, TanF exhibited broad substrate specificity for gallate esters. The final gallic acid production by TanF from tannic acid achieved 18.32g/L. Therefore, the excellent properties TanF has been demonstrated to be an efficient tool for the preparation of gallic acid.

Yeast community succession in cow dung composting process

Yeast complexes in the composting process of cow dung prepared to fertilize the soil for growing vegetables and fruits were studied. The average abundance of yeasts changed during the four temperature stages of the composting process. The highest abundance of yeasts, 1.38 × 104 cfu/g, was observed in the second stage of heating from 20 to 40 °C; the lowest was studied in the stage with the highest temperature (65 °C), 1.68 × 103 cfu/g. A total of 19 yeast species were observed and identified: 11 ascomycetes and 8 basidiomycetes, belonging to five subphyla of Fungi: Saccharomycotina (10), Pezizomycotina (1), Agaricomycotina (5), Pucciniomycotina (2), and Ustilaginomycotina (1). The greatest diversity of yeasts was found in the initial (20 °C) and second (heating up to 40 °C) temperature stages of composting (Aureobasidium pullulans (yeast-like fungus), Candida parapsilosis, Candida saitoana, Candida santamariae, Candida tropicalis, Curvibasidium cygneicollum, Cutaneotrichosporon moniliforme, Debaryomyces fabryi, Debaryomyces hansenii, Filobasidium magnum, Kazachstania sp., Moesziomyces bullatus, Naganishia globosa, Papiliotrema flavescens, Rhodotorula mucilaginosa, Scheffersomyces insectosa, Torulaspora delbrueckii, Vanrija musci), and the lowest in the stage of maximum heating (65 °C) (C. parapsilosis, C. tropicalis, Cyberlindnera jadinii).The opportunistic yeasts C. parapsilosis and C. tropicalis were obtained not only in the initial, second and third temperature stages of the composting process, but also in mature compost in the final stage prepared for soil application. This study shows that the cow dung, used in the farm studied did not meet the microbiological safety criteria. The reduction of opportunistic yeast species was not achieved with the composting method used. The likelihood of these species entering agricultural products via compost and soil and developing as endophytes in the internal tissues of fruits is very high. Since some strains of opportunistic Candida species from cow dung exhibited virulent characteristics (they produced hydrolytic enzymes and were resistant to antifungal compounds), additional phenotypic and genetic studies of the compost strains and their comparison with clinical isolates should be pursued.

Autochthonous Cultures to Improve Safety and Standardize Quality of Traditional Dry Fermented Meats

Traditional dry fermented meat products are obtained artisanally in many countries, where they represent a gastronomic heritage well distinguished from industrial counterparts. This food category is most often obtained from red meat, a food commodity that is under attack because of evidence of increased risk of cancer and degenerative diseases with high consumption. However, traditional fermented meat products are intended for moderate consumption and gastronomic experience, and, as such, their production must be continued, which would also help safeguard the culture and economy of the geographical areas of origin. In this review, the main risks attributed to these products are considered, and how these risks are reduced by the application of autochthonous microbial cultures is highlighted by reviewing studies reporting the effects of autochthonous lactic acid bacteria (LAB), coagulase negative staphylococci (CNS), Debaryomyces hansenii and Penicillium nalgiovense on microbiological and chemical safety and on sensory attributes. The role of dry fermented sausages as a source of microorganisms that can be beneficial to the host is also considered. From the results of the studies reviewed here it appears that the development of autochthonous cultures for these foods can ensure safety and stabilize sensory characteristics and has the capacity to be extended to a larger variety of traditional products.

Sodium and lithium exert differential effects on the central carbon metabolism of Debaryomyces hansenii through the glyoxylate shunt regulation.

Debaryomyces hansenii is a halotolerant/halophilic yeast usually found in salty environments. The yeast accumulated sodium at high concentrations, which improved growth in salty media. In contrast, lithium was toxic even at low concentrations and its presence prevented cell proliferation. To analyse the responses to both cations, metabolite levels, enzymatic activities and gene expression were determined, showing that NaCl and LiCl trigger different cellular responses. At high concentrations of NaCl (0.5 or 1.5 M) cells accumulated higher amounts of the intermediate metabolites glyoxylate and malate and, at the same time, the levels of intracellular oxoglutarate decreased. Additionally, 0.5 M NaCl increased the activity of the enzymes isocitrate lyase and malate synthase involved in the synthesis of glyoxylate and malate respectively and decreased the activity of isocitrate dehydrogenase. Moreover, transcription of the genes coding for isocitrate lyase and malate synthase was activated by NaCl. Also, cells accumulated phosphate upon NaCl exposure. None of these effects was provoked when LiCl (0.1 or 0.3 M) was used instead of NaCl. Lithium induced accumulation of higher amounts of oxoglutarate and decreased the concentrations of glyoxylate and malate to non-detectable levels. Cells incubated with lithium also showed higher activity of the isocitrate dehydrogenase and neither increased isocitrate lyase and malate synthase activities nor the transcription of the corresponding genes. In summary, we show that sodium, but not lithium, up regulates the shunt of the glyoxylic acid in D. hansenii and we propose that this is an important metabolic adaptation to thrive in salty environments.

Bioefficacy of bacteria and yeast bioagents on disease suppression and quality retention of stored Kinnow mandarin fruits

Kinnow mandarin is a high value citrus crop in the fruit industry. However, the fruits are vulnerable to a variety of phytopathogens and suffer huge quantitative and qualitative losses. Pathogenic filamentous fungi like Penicillium digitatum, Penicillium italicum, and Geotrichum candidum are mainly responsible for postharvest losses. Alternative to synthetic fungicides, the bacteria and yeast-based biocontrol approaches are the most efficient means for controlling postharvest diseases. The antagonistic potential of five biocontrol agents such as Debaryomyces hansenii, Lactobacillus plantarum, Metschnikowia pulcherima, Pichia guilliermondi and Rhodotorula minuta var. minuta was assessed under both in vitro and in vivo conditions against the mycelial growth of postharvest pathogens using dual culture method. The results revealed that R. minuta var. minuta was the most promising antagonist with 71.98%, 76.18% and 67.46% mean mycelial growth inhibition of P. digitatum, P. italicum and G. candidum respectively, over untreated potato dextrose agar (PDA) plates after 192 h of incubation under in vitro. Further, under in vivo, the fruits treated with R. minuta var. minuta did not impair the quality and sensory attributes during ambient storage. Hence, R. minuta var. minuta proved an eco-friendly and safer substitute to synthetic fungicides for postharvest disease management of Kinnow mandarin fruits.

Changes in quality and microbiome composition of strawberry fruits following postharvest application of Debaryomyces hansenii, a yeast biocontrol agent

Microbial antagonists provide a new perspective on biological control by altering microbial diversity to control postharvest diseases of fruit. In this study, the biological control effect of Debaryomyces hansenii on postharvest diseases of strawberry and the influence on strawberry quality were determined. At the same time, the effect of D. hansenii on the composition of surface microbiome of strawberry fruit was analyzed by high-throughput sequencing technology. The results showed that D. hansenii treatment can inhibit the natural decay rate of strawberries, and the treatment group maintained a higher ascorbic acid content than the control group. However, there was no difference between the control and treatment groups in firmness, weight loss rate, soluble solids, titratable acidity, and other quality indicators. On the surface of strawberries treated with D. hansenii, the diversity of fungal community was changed, the composition and structure of the community were also altered (except the abundance of Debaryomyces spp.). Strawberries treated with D. hansenii could alter bacterial and fungal populations, inhibit plant pathogens, and lower the frequency of postharvest diseases in strawberries. This study helps to understand the microbial community structure on the surface of strawberries and guides the synthesis of strawberry postharvest disease complex biocontrol strains to achieve stable and sustainable postharvest disease control strategies of the fruits.

Isolation of Debaryomyces hansenii and selection of an optimal strain to improve the quality of low-grade beef rump (middle gluteal) during dry aging.

The objective of this study was to evaluate the effect of Debaryomyces hansenii isolated from dry-aged beef on the tenderness and flavor attributes of low-grade beef during dry aging. Five D. hansenii strains were isolated from dry-aged beef samples. The rump of low-grade beef was inoculated with individual D. hansenii isolates and subjected to dry aging for 4 weeks at 5°C and 75% relative humidity. Microbial contamination levels, meat quality attributes, and flavor attributes in the dry-aged beef were measured. Of the five isolates, the shear force of dry-aged beef inoculated with SMFM201812-3 and SMFM201905-5 was lower than that of control samples. Meanwhile, all five isolates increased the total free amino acid, glutamic acid, serine, glycine, alanine, and leucine contents in dry-aged beef. In particular, the total fatty acid, palmitic acid, and oleic acid contents in samples inoculated with D. hansenii SMFM201905-5 were higher than those in control samples. These results indicate that D. hansenii SMFM201905-5 might be used to improve the quality of beef during dry aging.

Effect of the Types of Starter on Microbiological and Physicochemical Properties of Dry-Cured Ham.

This study analyzed the microbiological (Lactobacillus spp., Staphylococcus spp., mold, yeast, aerobic bacteria) and physicochemical properties [pH, salinity, water activity, volatile basic nitrogen (VBN), and thiobarbituric acid reactive substances]. The starters were used by mixing Debaryomyces hansenii separated from Korean Doenjang (D) and fermented sausage (S). The starter was inoculated with dry-cured ham and aged for 6 weeks at 20°C and 25°C, respectively. The aerobic bacteria, Lactobacillus spp., and Staphylococcus spp. of D, S, and DS treatment showed significantly higher values at 25°C than at 20°C. Among them, S25 treatment showed a high tendency. At week 6, the mold of the S25 treatment was significantly higher than the S20 treatment, and the yeast was higher in 25°C than 20°C (p<0.05). The pH of all treatment groups increased with the aging period. Compared with that at 25°C, the pH was significantly higher at 20°C (p<0.05). The water activity showed a significant decrease as the aging period increased, and the treatment of D25, S20, and DS20 showed a significantly higher value at week 6 (p<0.05). Compared with that at 20°C, the VBN content was higher at 25°C. At week 6, the VBN contents of the C20, S25, and DS25 groups were higher than those of the other treatment groups. Therefore, inoculation of D. hansenii separated from fermented sausage produced in Korean starter at 25°C is expected to improve the safety of harmful microorganisms and physiochemical properties in dry-cured ham.

A method for reducing the concentrations of Fusarium graminearum trichothecenes in durum wheat grain with the use of Debaryomyces hansenii

Fusarium head blight (FHB), caused mainly by Fusarium graminearum, is one of the most dangerous diseases of durum wheat. This hemibiotrophic pathogen transitions from the biotrophic phase, during which it penetrates host tissues and secretes trichothecenes, to the necrotrophic phase which leads to the destruction of host tissues. Yeasts applied to spikes often reduce mycotoxin concentrations, but the underlying mechanisms have not been fully elucidated. Therefore, the aim of this study was to analyze the concentrations trichothecenes in durum wheat grain and changes in the F. graminearum transcriptome under the influence the Debaryomyces hansenii antagonistic yeast strain. Debaryomyces hansenii cells adhered to and formed cell aggregates/biofilm on the surface of spikes and pathogenic hyphae. Biological control suppressed the spread of F. graminearum by 90 % and decreased the content of deoxynivalenol (DON) in spikes by 31.2 %. Yeasts significantly reduced the expression of pathogen's genes encoding the rpaI subunit of RNA polymerase I and the activator of Hsp90 ATPase, but they had no effect on mRNA transcript levels of genes encoding the enzymes involved in the biosynthesis of trichothecenes. The yeast treatment reduced the number of F. graminearum operational taxonomic units (OTUs) nearly five-fold and increased the number of D. hansenii OTUs more than six-fold in the spike mycobiome. The mechanisms that suppress infections should be explored to develop effective biological methods for reducing the concentrations mycotoxins in wheat grain.

Changes in the gut mycobiome in pediatric patients in relation to the clinical activity of Crohn's disease

Numerous studies have shown that in Crohn's disease (CD), the gut microbiota is of great importance in the induction and maintenance of inflammation in the gastrointestinal tract. Until recently, studies have focused almost exclusively on bacteria in the gut. Lately, more attention has been paid to the role of intestinal fungi. To study the gut mycobiome analysis of pediatric patients with CD (in different stages of disease activity) compared to healthy children. Fecal samples were collected from patients: With active, newly diagnosed CD (n = 50); active but previously diagnosed and treated CD (n = 16); non-active CD and who were in clinical remission (n = 39) and from healthy volunteers (n = 40). Fungal DNA was isolated from the samples. Next, next generation sequencing (MiSeq, Illumina) was performed. The composition of mycobiota was correlated with clinical and blood parameters. Candida spp. were overrepresented in CD patients, while in the control group, the most abundant genus was Saccharomyces. In CD patients, the percentage of Malassezia was almost twice that of the control (P < 0.05). In active CD patients, we documented a higher abundance of Debaryomyces hansenii (D. hansenii) compared to the non-active CD and control (P < 0.05) groups. Moreover, statistically significant changes in the abundance of Mycosphaerella, Rhodotorula, and Microidium were observed. The analyses at the species level and linear discriminant analysis showed that in each group it was possible to distinguish a specific species characteristic of a given patient population. Moreover, we have documented statistically significant correlations between: D. hansenii and patient age (negative); C. zeylanoides and patient age (positive); C. dubliniensis and calprotectin (positive); C. sake and calprotectin (positive); and C. tropicalis and pediatric CD activity index (PCDAI) (positive). Mycobiome changes in CD patients, and the positive correlation of some species with calprotectin or PCDAI, give strong evidence that fungi may be of key importance in the development of CD.

Development of plastic-degrading microbial consortia by induced selection in microcosms.

The increase in the production of highly recalcitrant plastic materials, and their accumulation in ecosystems, generates the need to investigate new sustainable strategies to reduce this type of pollution. Based on recent works, the use of microbial consortia could contribute to improving plastic biodegradation performance. This work deals with the selection and characterization of plastic-degrading microbial consortia using a sequential and induced enrichment technique from artificially contaminated microcosms. The microcosm consisted of a soil sample in which LLDPE (linear low-density polyethylene) was buried. Consortia were obtained from the initial sample by sequential enrichment in a culture medium with LLDPE-type plastic material (in film or powder format) as the sole carbon source. Enrichment cultures were incubated for 105 days with monthly transfer to fresh medium. The abundance and diversity of total bacteria and fungi were monitored. Like LLDPE, lignin is a very complex polymer, so its biodegradation is closely linked to that of some recalcitrant plastics. For this reason, counting of ligninolytic microorganisms from the different enrichments was also performed. Additionally, the consortium members were isolated, molecularly identified and enzymatically characterized. The results revealed a loss of microbial diversity at each culture transfer at the end of the induced selection process. The consortium selected from selective enrichment in cultures with LLDPE in powder form was more effective compared to the consortium selected in cultures with LLDPE in film form, resulting in a reduction of microplastic weight between 2.5 and 5.5%. Some members of the consortia showed a wide range of enzymatic activities related to the degradation of recalcitrant plastic polymers, with Pseudomonas aeruginosa REBP5 or Pseudomonas alloputida REBP7 strains standing out. The strains identified as Castellaniella denitrificans REBF6 and Debaryomyces hansenii RELF8 were also considered relevant members of the consortia although they showed more discrete enzymatic profiles. Other consortium members could collaborate in the prior degradation of additives accompanying the LLDPE polymer, facilitating the subsequent access of other real degraders of the plastic structure. Although preliminary, the microbial consortia selected in this work contribute to the current knowledge of the degradation of recalcitrant plastics of anthropogenic origin accumulated in natural environments.

Anti-Inflammatory, Barrier Maintenance, and Gut Microbiome Modulation Effects of Saccharomyces cerevisiae QHNLD8L1 on DSS-Induced Ulcerative Colitis in Mice.

The use of probiotics has been considered as a new therapy option for ulcerative colitis (UC), and yeast has recently received widespread recommendation for human health. In this study, the probiotic characteristics of four yeast strains, Saccharomyces boulardii CNCMI-745, Kluyveromyces marxianus QHBYC4L2, Saccharomyces cerevisiae QHNLD8L1, and Debaryomyces hansenii QSCLS6L3, were evaluated in vitro; their ability to ameliorate dextran sulfate sodium (DSS)-induced colitis was investigated. Among these, S. cerevisiae QHNLD8L1 protected against colitis, which was reflected by increased body weight, colon length, histological injury relief, decreased gut inflammation markers, and intestinal barrier restoration. The abundance of the pathogenic bacteria Escherichia-Shigella and Enterococcaceae in mice with colitis decreased after S. cerevisiae QHNLD8L1 treatment. Moreover, S. cerevisiae QHNLD8L1 enriched beneficial bacteria Lactobacillus, Faecalibaculum, and Butyricimonas, enhanced carbon metabolism and fatty acid biosynthesis function, and increased short chain fatty acid (SCFAs) production. Taken together, our results indicate the great potential of S. cerevisiae QHNLD8L1 supplementation for the prevention and alleviation of UC.