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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↗

Effects of alkylphosphates and nitrous oxide on microbial degradation of polycyclic aromatic hydrocarbons.

We conducted a series of liquid-culture experiments to begin to evaluate the abilities of gaseous sources of nitrogen and phosphorus to support biodegradation of polycyclic aromatic hydrocarbons (PAHs). Nutrients examined included nitrous oxide, as well as triethylphosphate (TEP) and tributylphosphate (TBP). Cultures were established using the indigenous microbial populations from one manufactured gas plant (MGP) site and one crude oil-contaminated drilling field site. Mineralization of phenanthrene was measured under alternative nutrient regimes and was compared to that seen with ammoniacal nitrogen and PO(4). Parallel cultures were used to assess removal of a suite of three- to five-ring PAHs. In summary, the abilities of the different communities to degrade PAH when supplemented with N(2)O, TEP, and TBP were highly variable. For example, in the MGP soil, organic P sources, especially TBP, supported a considerably higher degree of removal of low-molecular-weight PAHs than did PO(4); however, loss of high-molecular-weight compounds was impaired under these conditions. The disappearance of most PAHs was significantly less in the oil field soil when organophosphates were used. These results indicate that the utility of gaseous nutrients for PAH bioremediation in situ may be limited and will very likely have to be assessed on a case-by-case basis.

Bacteria↗

[Effects of specific microbial biocides on N transformation in soil with glucose amendment].

In an incubation test of soil with glucose amendment, two kinds of nitrogenous fertilizer and three kinds of specific microbial biocides were applied, and the contents of soil NH(4+)-N, NO3(-)-N, glucosamine and muramic acid were measured to differentiate the relative contribution and timing characteristics of soil microbes in nitrogen immobilization. The results showed that penicillin and streptomycin decreased the transformation rate of NH(4+)-N markedly, with more significant effects than actidione. The amount ratio of glucosamine to muramic acid after applying penicillin and streptomycin rapidly increased first, and tended to equilibrium then. With the application of actidione, the transformation rate of NO3(-)-N decreased continuously, and the synthesis of glucosamine was inhibited, while penicillin and streptomycin had no significant effects on them. At the early stage of incubation, bacteria could rapidly immobilize both NH(4+)-N and NO3(-)-N, with NH(4+)-N preferred, while at the later stage of incubation, fungi were the dominant contributor to nitrogen transformation, and had much stronger ability of utilizing NO3(-)-N than bacteria.

Anti-Bacterial Agents↗

Gene replacement in Staphylococcus carnosus and Staphylococcus xylosus.

A system for high-efficiency gene replacement in Staphylococcus carnosus and Staphylococcus xylosus has been developed, that is based on temperature-sensitive Escherichia coli-Staphylococcus shuttle vectors for fragment delivery and erythromycin resistance cassettes to facilitate selection of genomic copies of disrupted genes. The approach was tested by constructing a phosphotransferase-deficient mutant of S. carnosus and an S. xylosus mutant strain unable to utilize sucrose. Allelic replacements were observed at rather high frequencies, ranging from approximately 10% for the ptsI gene in S. carnosus up to 50% for the scrB gene in S. xylosus. These differences most likely reflect the length of homology rather than strain-specific variations in recombination efficiencies. Apart from the staphylococcal species tested in this study, the system appears to be applicable in other staphylococci.

Alleles↗

Meteorite organics in planetary environments: hydrothermal release, surface activity, and microbial utilization.

Up to 50% of the organics in the Murchison meteorite, possibly including some of the polymer, is released in high temperature and pressure aqueous environments, to 350 degrees C and 250 bar, that simulate submarine volcanic, hydrothermal or impact-induced conditions. Meteorite organics of prebiotic significance, such as nonanoic acid, glycine, and pyrene survive the hydrothermal conditions. The released material is surface active with surface pressures up to 19.8 x 10(-3) N m-1, and exhibits an extended surface tension isotherm which suggests a mixture of amphiphilic components. One component, nonanoic acid, is shown to form vesicles. The materials extracted under mild conditions, at 120 degrees C, are nutrients for the humic acid bacterium Pseudomonas maltophilia and efficient nutrients for the oligotroph Flavobacterium oryzihabitans, demonstrating the capability of microorganisms to metabolize extraterrestrial organics.

Carbon↗

Growth of Lactobacillus paracasei ssp. paracasei on tofu whey.

The liquid by-product of the soybean product tofu, tofu whey (TW), was used as a growth medium for the production of Lactobacillus paracasei ssp. paracasei LG3 cultures. The TW used in this study contained stachyose, raffinose, sucrose, fructose and glucose, but the strain used could only utilize the three latter. The lactobacilli population obtained in MRS broth was three times higher than that in TW alone, and supplementation of TW was thus examined. Of 19 mixtures of yeast extracts (YE), peptones and potato extracts examined, the best nitrogen sources were YE and tryptone. The addition of YE, salts (phosphates, citrates, Mg and Mn), glucose as well as Tween to TW tripled the populations to 2.9 x 10(9) cfu/ml, which was as high as that obtained in MRS broth. Growth of L. paracasei LG3 in cow rehydrated skim milk was inferior to that in TW.

Carbohydrate Metabolism↗

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↗

Effects of oxygen on biodegradation of benzoate and 3-chlorobenzoate in a denitrifying chemostat.

A mixed microbial culture degraded a mixture of benzoate (863 mg/L), 3-chlorobenzoate (3-CB) (69.7 mg/L), and pyruvate (244 mg/L) under denitrifying conditions in a chemostat. Biodegradation under denitrifying conditions was stable, complete (effluent concentrations below detection limits), and proceeded without the production of toxic intermediates like chlorocatechols. The addition of oxygen at mass input rates of 6.2%, 15.5%, and 43.9% of the mass input rate of chemical oxygen demand (COD) (337 mg COD/h) did not induce the synthesis of aerobic biodegradation pathways and thus did not disrupt biodegradation. Rather, the oxygen was used as a terminal electron acceptor, displacing a stoichiometric amount of nitrate, leading to microaerobic conditions (dissolved oxygen concentration <0.050 mg/L) in which oxygen utilization and denitrification occurred simultaneously. The reduction of nitrate occurred fully to N(2) gas with no accumulation of nitrite, nitrous oxide, or nitric oxide, although the ability of the culture to transfer electrons to the nitrogen oxides decreased as the oxygen input was increased. The anoxic benzoate uptake capability was unaffected by the increase in oxygen addition, but the anoxic 3-CB uptake capability increased, as did the level of benzoyl-CoA reductase in the cells.

Anti-Infective Agents↗

Microbial diversity in sediments of saline Qinghai Lake, China: linking geochemical controls to microbial ecology.

Saline lakes at high altitudes represent an important and extreme microbial ecosystem, yet little is known about microbial diversity in such environments. The objective of this study was to examine the change of microbial diversity from the bottom of the lake to sediments of 40 cm in depth in a core from Qinghai Lake. The lake is saline (12.5 g/L salinity) and alkaline (pH 9.4) and is located on the Qinghai-Tibetan Plateau at an altitude of 3196 m above sea level. Pore water chemistry of the core revealed low concentrations of sulfate and iron (<1 mM), but high concentrations of acetate (40-70 mM) and dissolved organic carbon (1596-5443 mg/L). Total organic carbon and total nitrogen contents in the sediments were approximately 2 and <0.5%, respectively. Acridine orange direct count data indicated that cell numbers decreased from 4 x 10(9) cells/g at the water-sediment interface to 6 x 10(7) cells/g wet sediment at the 40-cm depth. This change in biomass was positively correlated with acetate concentration in pore water. Phospholipid fatty acid (PLFA) community structure analyses determined decrease in the proportion of the Proteobacteria and increase in the Firmicutes with increased depth. Characterization of small subunit (SSU) rRNA genes amplified from the sediments indicated a shift in the bacterial community with depth. Whereas the alpha-, beta-, and gamma-Proteobacteria and the Cytophaga/Flavobacterium/Bacteroides (CFB) were dominant at the water-sediment interface, low G + C gram-positive bacteria (a subgroup of Firmicutes) became the predominant group in the anoxic sediments. Both PLFA and the sequence data showed similar trend. The Proteobacteria, CFB, and gram-positive bacteria are present in other saline lakes, but the presence of Actinobacteria and Acidobacteria/Holophaga in significant proportions in the Qinghai Lake sediments appears to be unique. The archaeal diversity was much lower, and clone sequences could be grouped in the Euryarchaeota and Crenarchaeota domains. The archaeal clones were not related to any known cultures but to sequences previously found in methane-rich sediments. Acetate-utilizing methanogens were isolated from sediment incubations, and alpha- and gamma-proteobacterial isolates were obtained from a water sample from the lake-bottom (23 m). Our data collectively showed that the observed diversity and shift in the community structure with depth was correlated with geochemical parameters (the redox state and availability of electron acceptor and donor). Heterotrophic methanogenesis is possibly adominant metabolic process in the Qinghai Lake sediments. These results reinforce the importance of geochemical controls on microbial ecology in saline and alkaline lake environments.

Acridine Orange↗

[Absorption and utilization of amino acids infused into the cecum of growing swine. 3. Studies with 15N- and 14C-labeled isoleucine].

In 3 experiments growing female pigs, (live weight 35-56 kg) received continuous infusions of 15N-labelled or 15N- and 14C-doubly labelled isoleucine by means of caecal cannulae. The whereabouts of the infused isoleucine were investigated. Between 8.9 and 12.3% of the infused 15N-excess (15N') was excreted in faeces mainly in the forms of bacteria protein, between 0.7 and 0.9% only of the infused amount unchanged as 15N-isoleucine. The highest quota is microbially decomposed in the colon and absorbed in the form of NH3 or amines. Between 45 and 70% of the infused 15N are excreted in urine, mainly as urea (85-95% of the 15N-amount in urine). Virtually no 15N from the infused isoleucine is incorporated in the organ and tissue proteins (exception: colon wall). These findings are corroborated by the 14C-activity measuring as no intact 14C labelled isoleucine could be detected in the tissue protein. These results show that intact isoleucine is not absorbed in the colon. The nitrogen originating from the absorbed NH3 is nearly completely excreted in urine and thus not available to the biosynthesis of body protein.

Animals↗

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↗

Recent advances in rumen microbial ecology and metabolism: potential impact on nutrient output.

Feedstuffs consumed by ruminants are all initially exposed to fermentative activity in the rumen prior to gastric and intestinal digestion. The extent and type of transformation of feedstuffs thus determines the productive performance of the host. Research on rumen microbial ecology and metabolism is essentially a study of the interactions between the host, microorganisms present, substrates available, and end products of digestion. Furthermore, the interactions of the normal microbial flora with the host can be manipulated to improve the efficiency of nutrient utilization in ruminant animals. Three important areas of ruminal fermentation will be reviewed, N metabolism, fiber degradation, and biotransformation of toxic compounds. The extent of protein degradation and the rate of uptake of resultant peptides and ammonia are extremely important factors in determining the efficiency of N utilization by rumen bacteria and, therefore, the relative amounts of microbial or bypass protein available to the host. Strategies aimed at identifying and characterizing rate-limiting enzymes of cellulolytic bacteria are essential in elucidating mechanisms involved in ruminal fiber degradation. Results obtained with ruminococci will be described. The detoxification of phytotoxins by passage through the gastrointestinal tract of ruminants is a process deserving special attention and several examples will be presented. Opportunities for manipulation of rumen fermentation are good. However, successful manipulation and full exploitation depend on a through understanding of the mechanisms involved.

Animal Feed↗

[Absorption and utilization of amino acids infused into the cecum of growing pigs. 1. Measurement of N-balance for utilization of lysine and isoleucine; isoleucine requirement for growing pigs].

In N-balance experiments with growing pigs (40-60 kg live weight) investigations were made whether lysine or isoleucine that is infused into the caecum can be absorbed there and to what extent these amino acids in that case can be utilised by the animal for protein synthesis. The pigs either received basic rations with insufficient lysine or isoleucine resp. (negative control group) or the amounts of lysine and isoleucine lacking to meet the requirement were supplemented with the feed (positive control group) or continuously infused into the caecum with the help of caecal infusion cannulae (test group). In the experiments with lysine the animals in the negative control group and in the test group showed considerably lower N-balances than the positive control group. There were no differences as regards the apparent digestibility of lysine between the positive control group and the test group. The urine of the test group contained distinctly more NH3. This shows clearly that lysine that is infused into the caecum cannot be utilised by the pigs, it is, on the contrary, microbially decomposed, the nitrogen is chiefly absorbed as NH3 and excreted in urine. As regards isoleucine, the deficit brought about with the basic ration was insufficient in order to achieve significant differences between the N-balance values of the groups so that unambiguous statements on the absorption and utilisation of isoleucine infused into the caecum cannot be made. Concerning apparent digestibility of isoleucine and the NH3 content of the urine, the results of the isoleucine experiments were similar to those in the lysine experiments. According to our N-balance experiments the isoleucine requirement of pigs indicated in relevant literature as 5.6 g/kg dry matter of the feed is by far too high. It should be limited to 3.5 or a maximum of 4.0 g isoleucine per kg dry matter of the feed.

Ammonia↗

Effects of increasing dietary protein on nutrient utilization in heifers.

Four prepubertal Holstein heifers, average age 146.0 +/- 3.6 d and 152.8 +/- 8.6 kg of body weight (BW), fitted with rumen cannulae were used to evaluate increasing levels of dietary protein with a constant metabolizable energy intake on rumen ammonia N concentrations, N balance, total tract apparent digestibility, and urinary excretion of purine derivatives in a 4 x 4 Latin square design with 20 d periods. Heifers were fed dietary crude protein levels (CP) of 11.9, 16.7, 18.1, and 20.1% CP with similar amounts of metabolizable energy (ME) (2.6 Mcal per kg of dry matter) at 2.0% BW as dry matter intake. Resulting protein to ME ratios (CP:ME) were 45.0, 63.3, 69.4, and 77.3 g of dietary CP per Mcal of ME. Rumen ammonia N and plasma urea nitrogen increased as CP level increased. Rumen pH, volatile fatty acids, and the acetate-to-propionate ratio were not affected with increasing CP levels. Greater urinary N excretion resulted in an increase in total N excretion with increasing CP levels. Apparent dry matter digestibility was similar for all diets, while apparent total tract N digestibility was lowest for the 11.9% CP level. Microbial N calculated from urinary excretion of purine derivatives increased with increasing CP levels. Holstein heifers between 153 and 196 kg consuming a diet at 2.0% BW as dry matter intake containing a CP level of 16.7% achieved a better synergistic relationship of dietary protein to energy (CP:ME of 63.3 g of CP per Mcal of ME) than the diets lower or higher in CP that were studied.

Acetates↗

Effect of increasing degradable intake protein on intake and digestion of low-quality, tallgrass-prairie forage by beef cows.

Five ruminally and duodenally fistulated Angus x Hereford cows were used in a 5 x 5 Latin square to monitor intake, ruminal fermentation responses, and site and extent of digestion associated with providing increasing amounts of supplemental degradable intake protein (DIP). Cows had ad libitum access to low-quality, tallgrass-prairie forage (1.9% CP, 77% NDF) that was fed twice daily. The supplemental DIP (sodium caseinate; 90% CP) was infused intraruminally at 0630 and 1830 immediately before feeding forage. Levels of DIP were 0, 180, 360, 540, and 720 g/d. Each period consisted of 14 d of adaptation and 6 d of sampling. Forage OM intake increased quadratically (P < .01) with increasing supplemental DIP reaching a peak at the 540 g/d level. True ruminal OM and NDF digestion increased with the addition of 180 g/d supplemental DIP, but exhibited only moderate and somewhat variable responses when greater amounts of supplemental DIP were infused (cubic, P < or = .03). Microbial N flow and efficiency increased linearly (P < .01) with increasing supplemental DIP. However, a quadratic effect (P < .01) was observed for total duodenal N flow, which was maximized at 540 g/d supplemental DIP. A linear (P = .02) treatment effect was observed for ruminal fluid dilution rate. Total ruminal VFA and ammonia concentrations increased (P < .01) in response to DIP supplementation. In conclusion, increasing supplemental DIP generally improved forage utilization; intake of digestible OM was maximized when it contained approximately 11% DIP.

Animals↗

Kinetic modeling of a mixed culture of Pseudomonas denitrificans and Bacillus subtilis under aerobic and anoxic operating conditions.

The kinetics of biological denitrification have been studied and several models, with varying degree of complexity, to be used for design purposes have been presented in the recent years. However, most of these kinetic studies were performed with mixed (and not well defined) microbial systems, such as activated sludge. In the present work, kinetic experiments were carried out in order to study the dynamic characteristics of a defined mixed culture of the denitrifiers Pseudomonas denitrificans and Bacillus subtilis under anoxic and aerobic conditions in a defined synthetic medium involving a mixture of organic substrates, in the presence of nitrates and/or nitrites. Denitrification was assumed to occur by the consecutive reduction of nitrates to nitrites and then to nitrogen gas without accumulation of intermediate gaseous products. The behavior of these defined mixed cultures was predicted using a kinetic model based on the kinetic models that have already been developed for each bacterium separately and the predictions were compared with the results from mixed culture experiments. The overall mathematical model that was developed and validated in the present work is capable of describing the behavior of the mixed culture in the above conditions, i.e. the nitrates and nitrites reduction kinetics, the cell growth, and the organic carbon utilization rates.

Bacillus subtilis↗

Alcaligenes eutrophus hydrogenase genes (Hox).

Mutants of Alcaligenes eutrophus H16 lacking catalytically active soluble hydrogenase (Hos-) grew very slowly lithoautotrophically with hydrogen. Mutants devoid of particulate hydrogenase activity (Hop-) were not affected in growth with hydrogen. The use of Hos- and Hop- mutants as donors of hydrogen-oxidizing ability in crosses with plasmid-free recipients impaired in both hydrogenases (Hox-) resulted in transconjugants which had inherited the plasmid and the phenotype of the donor. This indicates that the structural genes which code for the hydrogenases reside on plasmid pHG1. The Hox function of one class of Hox- mutants could not be restored by conjugation. These mutants exhibited a pleiotropic phenotype since they were unable to grow with hydrogen and also failed to grow heterotrophically with nitrate (Hox- Nit-). Nitrate was scarcely utilized as electron acceptor or as nitrogen source. Hox- Nit- mutants did not act as recipients but could act as donors of the Hox character. Transconjugants derived from those crosses were Hox+ Nit+, indicating that the mutation which leads to the Hox- Nit- phenotype maps on the chromosome. Apparently, the product of a chromosomal gene is involved in the expression of plasmid-encoded Hox genes. We observed that the elimination of plasmid pHG1 coincided with the occurrence of multiple resistances to various antibiotics. Since Hox+ transconjugate retained the antibiotic-resistant phenotype, we conclude that this property is not directly plasmid associated.

Alcaligenes↗

In situ identification of azo dye inhibition effects on nitrifying biofilms using microelectrodes.

In this study, the inhibitory effects of acid orange 7 (AO7), a common azo dye, on nitrification in biofilms were investigated in situ using microelectrodes. Biofilms were obtained from laboratory rotating drum biofilm reactor after the nitrification process reached a pseudo-steady state. Dissolved oxygen, pH, NH4+, NO3-, and redox potential microelectrodes, with tip diameters ranging from 3-15 microm, were used to monitor the spatial distribution and change of microbial activities within nitrifying biofilms. It was found that at lower concentration (1 mg/L), AO7 had only a slight impact on the NH4+-N concentration profiles. The ammonium consumption rate decreased as higher AO7 concentrations (15 mg/L and 25 mg/L) were exposed to the biofilms. A similar trend was observed for the NO3(-)-N microprofiles. The nitrate production rate decreased as the AO7 concentration in the bulk solution increased. The dissolved oxygen and pH microprofiles also showed oxygen and alkalinity utilization, but at lower rates throughout the biofilms when the nitrification process was inhibited. No significant redox potential differences were observed in the biofilms after AO7 was applied.

Azo Compounds↗