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Quorum sensing in Saccharomyces cerevisiae brewing strains: effects of 2-phenylethanol on proteomic, lipidomic, and metabolomic profile.

Quorum sensing (QS) is a known mechanism by which microbial populations adjust gene expression and coordinate community-wide social behaviors based on the proximate population density. This regulatory system has garnered significant interest in both scientific research and the food industry. However, a central question remains whether industrial strains of Saccharomyces cerevisiae, the yeast species predominantly utilized in brewing, employ quorum signalling mechanisms similar to those observed in laboratory strains and other fungi. Despite the potential relevance of microbial social behavior regulators to brewing practices, studies examining QS in Saccharomyces spp. are limited. In this investigation, three industrial brewing strains of S. cerevisiae were cultivated on SLAD (nitrogen-restrictive) and SHAD (nitrogen-sufficient) agar media supplemented with 200 μM of the aromatic alcohol 2-phenylethanol (2-PE) over 72 h at 24°C. Subsequent analyses of the harvested biomass included proteomic, lipidomic, and metabolomic assessments. Results indicated that two of the industrial strains showed minimal differences in their profiles upon exposure to 2-PE, while the third strain exhibited significant differences. These findings imply that the impact of the QS molecule 2-PE on the proteome, lipidome, and metabolome of industrial S. cerevisiae may be strain-specific rather than universally applicable to the species.

Quorum Sensing

Microbial utilization of Pinus radiata bark.

A screening program using suspensions of ground bark in mineral salts media, or extracts prepared from ground bark by treating with hot water, sulfuric acid, ammonium hydroxide, or sodium hydroxide, yielded more than 200 pure cultures of fungi, yeasts, and bacteria. Only 38 of these have good growth on liquid bark media. All were filamentous fungi, although many bacteria and yeasts were among the cultures that failed to give appreciable growth. Species of Penicillium, Scopulariopsis, Aspergillus, Trichoderma, Cladosporium, and Fusarium were among the most actively growing cultures. Cell biomass yields, as measured by cell nitrogen determination, were too low for economic production of single cell protein.

Bacteria

Relationship between ruminal ammonia and nonprotein nitrogen utilization by ruminants. III. Influence of intraruminal urea infusion on ruminal ammonia concentration.

In three trials, we studied the effect of incremental amounts of intraruminally infused urea on mean ruminal ammonia concentration of steer fed at 2-h intervals. Basal rations contained these percentages of crude protein and total digestible nutrients (dry matter basis); Trial I, 11.1 and 81; Trial II, 6.0 and 54; Trial III, 6.5 and 58. Mean ruminal ammonia concentration reached 5 mg ammonia nitrogen/100 ml rumen fluid at crude protein equivalents of 12.0, 9.3, and 9.4% in I, II, and III. Once ruminal ammonia began to accumulate, there was a linear relationship between intake of urea and mean concentration of amino acids of plasma, serving as an indirect measure of amino acid absorption from the intestine, was not increased by increased intake of urea in III. Results of this experiment support the concept from in vitro data that microbial protein synthesis is unaffected by ruminal ammonia concentration in excess of 5 mg ammonia nitrogen/100 ml rumen fluid.

Amino Acids

Influence of nitrate and molybdenum on sulfur utilization by rumen microorganisms.

Twenty-four hours in vitro incubations were used to study the effect of nitrate and molybdenum on sulfur utilization by rumen microorganisms. Sulfur was added as sodium sulfate or sulfide at .1, .2, .3, or .4% of the substrate dry matter. Cellulose digestion was an indicator of microbial growth. The addition of .1 to .4% sulfate or sulfide sulfur increased cellulose digestion over the conrol, the two sulfur sources being equal in promoting cellulose digestion. No differences in cellulose digestion were found between .1 and .4% added sulfur. However, the addition of .4 of .8% nitrate-nitrogen depressed cellulose digestion and increased the requirement for both sulfate and sulfide. Depression was greater with .8% nitrate-nitrogen. In the presence of nitrate, sulfide was superior to sulfate in promoting cellulose digestion. When 4 or 8 ppm molybdenum were added to the incubations, increasing concentrations of both sulfate and sulfide were required to obtain maximum cellulose digestion. Molybdenum additions increased both the sulfate and sulfide requirement for maximum cellulose digestion.

Animals

Determinants in microbial colonization of the murine gastrointestinal tract: pH, temperature, and energy-yielding metabolism of Torulopsis pintolopesii.

Torulopsis pintolopesii is an indigenous yeast that colonizes the secreting epithelia in the stomachs of mice and rats. A wild-type strain of this microbe was isolated and identified. To attempt to learn characteristics of the yeast that are advantageous to it in colonizing its natural habitat in vivo, we examined some aspects of its nutrition and energy-yielding metabolism and some environmental conditions that influence its growth in vitro. The yeast appeared to be limited in the compounds it can utilize as carbon and nitrogen sources. It grew best at 37 degrees C and did not grow at 23 or 43 degrees C. It grew optimally at neutral pH but could grow aerobically at pH values as low as 2.0 and anaerobically at pH values as low as 3.4. As assessed by measurements of growth rates and yield coefficients, it grew better aerobically than anaerobically. When grown aerobically, it had a cyanide-sensitive system for taking up O(2) and tested positively for cytochrome c oxidase activity. A petite mutant strain isolated from the wild-type strain had a growth rate and yield coefficient when incubated aerobically that were essentially the same as those of the wild-type parent grown anaerobically. Likewise similar to the wild-type parent grown anaerobically, the petite strain, though incubated aerobically, did not take up O(2). Yeast-free mice associated with either the wild-type or the petite mutant strain were colonized at essentially the same rates and to similar final population levels by both strains. The yeast's capacity to respire may be of little advantage to it in its natural environment. By contrast, its abilities to grow best at 37 degrees C and to grow at low pH values are undoubtedly advantageous characteristics in this respect. The limitations in its carbon and nitrogen nutrition are difficult to evaluate as ecological factors in its colonization of the natural habitat.

Animals

[Contribution to the problem of microbially induced urea transformation in soil. I. On the ability of urea utilization by soil micro-organisms (author's transl)].

The present publication is the first of a series on the enzymatic urea tranformation in soil. With about 2,000 pure cultures of micro-organisms it was possible to prove the very good urea utilization by the soil micro-organisms (bacteria, actinomycetes, and fungi). Above all the fungi showed an excellent utilization of urea, while bacteria and actinomycetes were somewhat poorer. Contrary to this is the urease activity of these organisms, and that is the reason why fungi in soil may be regarded as short-time accumulators for urea nitrogen and must not be suppressed by inhibitors.

Actinomycetales

Bacterial enzymes in uremia management.

The theoretical background for a conservative therapeutic treatment of uremia is described, with illustrative results from preliminary clinical trials in 10 patients and 10 normal reference subjects. The proposed treatment focuses upon the patient's gastrointestinal tract--the normal site for metabolism of both exogenous (dietary) and endogenous (recycled) protein--enabling it to behave like the rumen of the cow. The objective is to induce the uremic's organism to utilize its own "waste" substances. The patient swallows enterosoluble capsules containing specifically adapted enzymes (immobilized or free) from apathogenic soil microorganisms. These are pre-adapted to convert urea, creatinine, uric acid, guanidino derivatives, and other nonprotein nitrogen compounds (NPN). The enzymes utilize many other substances, in particular ammonia, potassium, phosphorus, and several other factors potentially dangerous for the uremic. The enzymes apparently cleave vasoconstrictatory peptides in the intestines. In the course of the therapy, renoparenchymal hypertension decreased significantly, and increased again when the regimen was interrupted. The results from the present studies are in full accord with the information published in the relevant fields. The time appears ripe for large-scale trials of the therapeutic regimen outlined, especially as many commercial microbial enzymes already have a long history of safe use in food processing.

Animals

Studies on the mode of action of 5-fluorocytosine in Aspergillus species.

The mode of action of 5-fluorocytosine (5-FC) was studied in three isolates of pathogenic Aspergillus with varying degrees of susceptibility to the drug. Distribution studies showed that susceptibility or resistance to 5-FC was not dependent on uptake of the drug. While only a small percentage of the total 5-FC taken up was found in the RNA fraction of the cells, most remained in the acid-soluble intracellular pool. 5-FC, 5-fluorouracil (5-FU) and 5-fluorodeoxyuridine monophosphate (5-Fd-UMP) were among metabolites identified in the pool. In addition, fluoroorotic acid appeared to be a major constituent of the metabolites derived from 5-FC. The aspergilli also were capable of utilizing cytosine as a nitrogen source and this is suggested as a possible mechanism of resistance. A dual mode of action for 5-FC in the aspergilli is proposed. This consists of, first, incorporation of 5-FU into RNA and, second, inhibition of DNA synthesis by production of 5-FdUMP.

Aspergillus fumigatus

Deciphering the effects of sulfonamide antibiotics on denitrification from a metagenomic perspective: Inhibition of nitrite reduction and succession patterns of functional microorganisms.

Limited research has thoroughly elucidated the impact mechanisms of antibiotics on the denitrification process at the genomic and gene levels, which has hindered the optimization and development of nitrogen removal technology for antibiotic-containing swine wastewater. Lab-scale sequencing batch reactors were constructed in this study to treat synthetic wastewater containing different sulfonamides and nitrate. Investigations were carried out on denitrification performance, microbial community diversity, denitrifier succession patterns, and functional gene distribution. The stress of sulfonamides inhibited the nitrite reduction process, transforming complete denitrification into partial denitrification and causing significant nitrite accumulation. The average nitrogen removal efficiency in the treatment groups decreased from 81.0% ± 2.2-40.1% ± 6.1%. Alicycliphilus and Thauera were identified as the key taxa, accounting for 32.2% and 16.9% of all potential denitrifying bacteria, respectively. Although metagenome-assembled genomes (MAGs) from Thauera were enriched with genes encoding nitrate reductases (nap, nar) and nitrite reductases (nir), this genus preferentially utilized nitrate as an electron acceptor, resulting in the preferential nitrate reduction and subsequent nitrite accumulation. In contrast, Alicycliphilus MAGs developed tolerance to the sulfonamides stress during later stages, with concomitant enrichment of associated functional genes. They replaced Thauera to reemerge as the dominant group, thereby restoring complete denitrification. This study provides new insights into the regulatory mechanisms governing complete versus partial denitrification in nitrogen removal from antibiotic-containing wastewater.

Denitrifier succession

Simultaneous removal of nitrogen, Cu2+, and bisphenol A in a hydrogel-biochar-AQDS immobilized bioreactor with added bicarbonate: Performance and metagenomic insights.

As the complexity of industrial wastewater pollution continues to increase, the simultaneous removal of nitrogen, metal contaminants, and persistent organic pollutants under low carbon conditions has become a key challenge for biological treatment systems. To address the operational instability and dependence on carbon sources observed in immobilized systems when exposed to copper (Cu2+) and bisphenol A (BPA), the Pseudoalteromonas japonicus strain LY0623 was integrated into a hydrogel-biochar-AQDS composite carrier to construct a multifunctional immobilized biofilm system. Notably, under conditions containing only NaHCO3, the R4 system achieved an NH4+-N removal rate of 89%. Under conditions where Cu2+ and BPA coexist, the R4 system achieved removal of NH4+-N (89%), NO3--N (100%), Cu2+ (85%), and BPA (88%). Sediment characterization confirmed that Cu2+ was immobilized through adsorption, complexation, and microbiologically induced carbonate precipitation (MICP). Metagenomic analysis further indicated that the Pseudomonadota phylum remained the dominant phylum, while functional pathways associated with inorganic carbon assimilation, HNAD nitrogen metabolism, endogenous carbon transformation, biomineralization, electron transfer, and aromatic compound degradation were preserved. By combining ammonia oxidation driven energy production, inorganic carbon utilization, redox mediated processes, and biomineralization, this study provides a highly promising low carbon strategy for treating industrial wastewater containing mixed pollutants.

Bisphenol A Compounds

Metagenome-based diversity and functional analysis of culturable microbes in sugarcane.

UNLABELLED: Sugarcane is a key crop for sugar and energy production, and understanding the diversity of its associated microbes is crucial for optimizing its growth and health. However, there is a lack of thorough investigation and use of microbial resources in sugarcane. This study conducted a comprehensive analysis of culturable microbes and their functional features in different tissues and rhizosphere soil of four diverse sugarcane species using metagenomics techniques. The results revealed significant microbial diversity in sugarcane's tissues and rhizosphere soil, including several important biomarker bacterial taxa identified, which are reported to engage in several processes that support plant growth, such as nitrogen fixation, phosphate solubilization, and the production of plant hormones. The Linear discriminant analysis Effect Size (LEfSe) studies identified unique microbial communities in different parts of the same sugarcane species, particularly Burkholderia, which exhibited significant variations across the sugarcane species. Microbial analysis of carbohydrate-active enzymes (CAZymes) indicated that genes related to sucrose metabolism were mostly present in specific bacterial taxa, including Burkholderia, Pseudomonas, Paraburkholderia, and Chryseobacterium. This study improves understanding of the diversities and functions of endophytes and rhizosphere soil microbes in sugarcane. Moreover, the approaches and findings of this study provide valuable insights for microbiome research and the use of comparable technologies in other agricultural fields. IMPORTANCE: This work utilized metagenomics techniques for conducting a comprehensive examination of culturable microbes and their functional characteristics in various tissues and rhizosphere soil of four distinct sugarcane species. This study enhances comprehension of the diversity and functions of endophytes and rhizosphere soil microbes in sugarcane. Furthermore, the methodologies and discoveries of this work offer new perspectives for microbiome investigation and the use of similar technologies in other agricultural fields.

Saccharum

Multi-omics revealed the effects of rumen to blood path on early lactation performance in transition dairy cows.

BACKGROUND: The transition period is vitally important to the life cycle of dairy cows. However, the function of the microbiota during both pre- and post-partum and their relationship with ruminal, plasma, and milk metabolites still require systematic investigation. To address this, the 7 highest- and 7 lowest-performing animals among a cohort of 100 dairy cows were selected based on their postpartum energy-corrected milk yield. Rumen fluid and plasma samples were collected during both pre- and post-partum periods, whereas milk samples were obtained postpartum. Shotgun metagenomics of rumen contents in addition to metabolomics of rumen, plasma, and milk samples were performed to evaluate the associations between ruminal microbes and early lactation performance in transition dairy cows. RESULTS: Compared with prepartum cows, postpartum high-yield cows had greater concentrations of ruminal volatile fatty acids and plasma total bile acid. Moreover, plasma urea nitrogen and most amino acids, peptides, and their derivatives in plasma and milk were increased in postpartum high-yield cows, relative to postpartum low-yield cows. Metagenomic analysis revealed that the relative abundances of several species within the Prevotella, Succinimonas, Succinatimonas, and Methanosphaera increased, while other bacteria belong to Alistipes and Bacteroides, and archaeal Methanobrevibacter species decreased in postpartum cows, particularly in postpartum high-yield cows. Co-occurrence network and correlation analysis suggested that Prevotella and Succinatimonas were negatively correlated to Alistipes, Bacteroides, and Methanobrevibacter, potentially contributing to the nutritionally efficient phenotype of postpartum high-yield cows. A metabolic pathway analysis of our metagenomic data revealed that postpartum high-yield cows possessed more microbial genes involved in starch utilization and amino acid synthesis, while a wide range of microbial genes involved in cellulose utilization, acetogenesis, and amino acid degradation were found in prepartum cows with low-yield in postpartum. A structural equation model analysis showed that the increased relative abundances of Prevotella tf.2-5 and Succinatimonas CAG_777 were related to greater concentrations of plasma chenodeoxycholic acid glycine conjugate, milk 5-Methoxytryptophan, and energy-corrected milk yield. Finally, pan-genomic analysis confirmed that Alistipes, Bacteroides, and Methanobrevibacter possess genetic conservation of both hydrogenases and dehydrogenases, which may contribute to energy loss in the rumen via hydrogen dissipation. CONCLUSION: In summary, our findings provide a fundamental understanding of how microbiome-dependent mechanisms contribute to early lactation performance in dairy cows during the transition period. The increased abundance of Prevotella, Succinimonas, and Succinatimonas in postpartum cows suggest that they are important microbes during the transition period and may help in coping with metabolic challenges, while improving nutrient utilization efficiency during this period. Our study underscores the importance of the ruminal microbiome during the transition period and highlights the need for rumen-based nutritional intervention strategies to improve production efficiency in ruminants. Video Abstract.

Animals

Rumen bypass and protection of proteins and amino acids.

Potent rumen microbial proteases and deaminases rapidly degrade protein and amino acids which are soluble in the rumen liquid phase. Because protein sources vary in their solubility, the degree of degradation in the rumen is variable. Methods of decreasing protein and amino acid degradation in the rumen include heat treatment, chemical treatment, encapsulation, use of amino acid analogs, selective manipulation of balances of rumen metabolic pathways, and esophageal groove closure. It is important that procedures do not interfere with ruminal metabolism or post-ruminal digestion. Bypassing the rumen changes sites in the digestive tract of nutrient digestion and absorption and provides a mechanism for supplementing outflow of nutrients from the rumen. A feasible approach to production of animal protein from ruminants would be utilization of nonprotein nitrogen for rumen protein production, maximization of rumen bypass of dietary protein, and supplementation with rumen nondegradable amino acids.

Amino Acids

Nitrogen utilization within equine large intestine.

The distribution of nitrogen in ligated segments of the stomach and intestine of two groups of ponies has been examined at sacrifice 2, 4, 8, and 12 h following the final 12-h scheduled feeding of diets designed to provide either 1) ample protein, or 2) limited protein plus supplemental urea as the major nitrogen source. Concentrations and total quantities of total N, NH3, urea plus NH3, and alpha-amino N were determined, and protein N was calculated by difference. Liquid marker (PEG) distribution rate constants and N concentrations were used to calculate rates of entry and exit by digesta flow for the large intestinal compartments. These values, together with measured accumulations of each constituent, allowed assessment of transitory net appearance and net disappearance within each compartment due to processes other than flow along the tract. The data suggested cyclic fluctuations of microbial populations within the large intestine, particularly the dorsal and ventral colon. These segments also were implicated as major sites of microbial protein synthesis and degradation. the former process appeared more evident in animals consuming the urea-supplemented diet.

Amino Acids

Efficiency of use of nitrogen from dried microbial cells after a period of N deprivation in growing pigs.

1. Semi-synthetic diets, with dried microbial cells (Pruteen) as the nitrogen source, were used to measure N retention in 50 kg pigs which had been given only sufficient N (5.3 g/d) to maintain N equilibrium for the previous 12 d. Control pigs were given 33.2 gN/d. 2. Metabolic faecal N losses were 1.62 g/d (1.2 g/kg dry matter eaten) and endogenous urinary losses were 3.90 g/d. 3. Realimentation of N-deprived pigs was achieved with diets providing 20.0, 33.2 and 67.4 g N/d and daily rates for N retention were 11.2, 17.8 and 25.9 respectively; the corresponding value for control pigs was 15.0 g. 'Catch-up' protein growth was demonstrated in pigs given both 33.2 and 67.4 g N/d. In the former instance, this was associated with an increase in the efficiency of utilization of dietary N. 4. The biological value of the protein in Pruteen was 0.85, and it appeared that under conditions of increased demand for N the pig could utilize some of the nucleic acid-N fraction of this protein source.

Amino Acids

Transcriptomic insights into exogenous fatty acid-enhanced halotolerance in Zygosaccharomyces rouxii.

BACKGROUND: High salinity restricts microbial growth during brine-based food fermentation. Although exogenous unsaturated fatty acids improve the salt tolerance of Zygosaccharomyces rouxii, the associated transcriptional mechanisms remain unclear. This study investigated the transcriptomic response of Z. rouxii CGMCC 3791 to palmitoleic acid (C16:1) under high salt conditions. RESULTS: Cells were cultured in yeast extract peptone dextrose (YPD) containing 120&#x2009;g&#x2009;L-1 NaCl, with or without 20&#x2009;&#x3bc;g&#x2009;mL-1 C16:1. They were analyzed by RNA sequencing. Principal component analysis clearly separated the two treatments. Using q&#x2009;<&#x2009;0.05 and |log2 fold change|&#x2009;>&#x2009;1, 23 differentially expressed genes were identified - three upregulated and 20 downregulated. INO1, MLS1, POX1, MEP2, and SOD5 were among the major responsive genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that peroxisome-associated functions, lipid metabolism, oxidative stress responses, nitrogen utilization, and mitogen-activated protein kinase (MAPK) signaling were the principal C16:1-responsive processes. CONCLUSION: Exogenous C16:1 elicited a focused transcriptional adjustment rather than broad transcriptome-wide reprogramming in salt-stressed Z. rouxii. The results indicated that peroxisome-linked lipid processes and redox regulation were candidate mechanisms underlying fatty-acid-associated halotolerance and provided targets for improving the robustness of high-salt food fermentation. &#xa9; 2026 Society of Chemical Industry.

Zygosaccharomyces rouxii

A nutritional and taxonomic survey of Arthrobacter soil isolates.

One hundred and sixty Arthrobacter soil isolates, four groups of 40 from each of four soil types, and 17 Arthrobacter named strains were characterized on the basis of 203 different nutritional tests performed on each culture. One hundred and twenty-one compounds were examined as sole sources of carbon, 44 compounds as sole sources of nitrogen, 22 carbohydrates for the production of acid, and 16 tests were done for tolerance to various dyes. The four groups of soil arthrobacters differed markedly in acid production from carbohydrates, and in utilization of aliphatic amino acids and aromatic hydrocarbons as sole sources of carbon. The named strains did not exhibit as much nutritional diversity as any of the four groups of soil isolates. The results of the nutritional tests were subjected to a computerized taxonomic analysis. Eighty-four of the isolates were contained in nine separate clusters with 87 to 94% similarity; 39 isolates were contained in five mixed but recognizable clusters with 84 to 86% similarity; 60 isolates were randomly mixed with no recognizable clusters and similarity of 72 to 83%. All clusters were composed of nearly equal numbers of isolates from each of the four soils; the named strains were not found associated with any of the clusters of soil isolates. The results indicated that the diversity of the genus Arthrobacter is not represented by the named species already described, and the possibility of using a series of standardized tests to categorize soil arthrobacters into readily identifiable nutritional groups is discussed.

Acids

Fluorescent pseudomonads capable of growth at 41 degrees C but distinct from Pseudomonas aeruginosa.

One hundred and twenty-seven apyocyanogenic fluorescent Pseudomonas strains capable of growth at 41 degrees C, but differing from Pseudomonas aeruginosa, were typed serologically and tested for pyocin production, antibiotic susceptibility, selected biochemical reactions, and utilization of selected substrates. Results were compared with those from 40 apyocyanogenic and 14 pyocyanin-producing strains of P. aeruginosa. Unidentified fluorescent Pseudomonas (UFP) strains generally were not agglutinated by P. aeruginosa antisera and showed little or no pyocin activity. In contrast to P. aeruginosa strains, UFP strains usually failed to oxidize D-gluconate or reduce nitrate to nitrogen gas. They could not use D-gluconate or D-mannitol as sole carbon source and were susceptible to kanamycin. The cellular fatty acid compositions of major UFP groups resembled those of the alcaligenes-stutzeri groups.

Anti-Bacterial Agents