Shear-Thinning and Designable Responsive Supramolecular DNA Hydrogels Based on Chemically Branched DNA.

A novel supramolecular DNA hydrogel system was designed based on a directly synthesized chemically branched DNA. For the hydrogel formation, a self-dimer DNA with two sticky ends was designed as the linker to induce the gelation of B-Y. By programing the linker sequence, thermal and metal-ion responsiveness could be introduced into this hydrogel system. This supramolecular DNA hydrogel shows shear-thinning, designable responsiveness, and good biocompatibility, which will simplify the hydrogel composition and preparation process of the supramolecular DNA hydrogel and accelerate its biomedical applications.

A refined medium to enhance the antimicrobial activity of postbiotic produced by Lactiplantibacillus plantarum RS5.

Postbiotic RS5, produced by Lactiplantibacillus plantarum RS5, has been identified as a promising alternative feed supplement for various livestock. This study aimed to lower the production cost by enhancing the antimicrobial activity of the postbiotic RS5 by improving the culture density of L. plantarum RS5 and reducing the cost of growth medium. A combination of conventional and statistical-based approaches (Fractional Factorial Design and Central Composite Design of Response Surface Methodology) was employed to develop a refined medium for the enhancement of the antimicrobial activity of postbiotic RS5. A refined medium containing 20 g/L of glucose, 27.84 g/L of yeast extract, 5.75 g/L of sodium acetate, 1.12 g/L of Tween 80 and 0.05 g/L of manganese sulphate enhanced the antimicrobial activity of postbiotic RS5 by 108%. The cost of the production medium was reduced by 85% as compared to the commercially available de Man, Rogosa and Sharpe medium that is typically used for Lactobacillus cultivation. Hence, the refined medium has made the postbiotic RS5 more feasible and cost-effective to be adopted as a feed supplement for various livestock industries.

Hydrogen sulphide production by Saccharomyces cerevisiae UCD 522 in a synthetic grape juice medium deficient of thiamin (vitamin B1 ) and/or pyridoxine (vitamin B6 ).

The impacts of thiamin and pyridoxine along with YAN on alcoholic fermentation and hydrogen sulphide production by Saccharomyces cerevisiae were studied. Using a synthetic grape juice medium, three fermentation trials were conducted; (i) 2 × 3 factorial design with thiamin (0, 0·2, or 0·5 mg l-1 ) and YAN (60 or 250 mg l-1 ) as variables, (ii) 2 × 3 factorial design with pyridoxine (0, 0·25, or 0·5 mg l-1 ) and YAN (60 or 250 mg l-1 ) as variables, and (iii) 3 × 3 factorial design with thiamin (0, 0·2 or 0·5 mg l-1 ) and pyridoxine (0, 0·25 or 0·5 mg l-1 ) as variables in media containing 60 mg l-1 YAN. Although the progress of fermentations was affected by thiamin or pyridoxine, YAN had a larger impact than either vitamin. H2 S production was significantly lower with increasing amounts of thiamin in those fermentations under low YAN (60 mg l-1 ) while even lower amounts (<30 µg l-1 ) were produced under high YAN (250 mg l-1 ) with or without the vitamin. The highest amounts of H2 S were synthesized in those fermentations without any pyridoxine (>110 µg l-1 ), with the lowest production in media with pyridoxine and high YAN (<20 µg l-1 ). SIGNIFICANCE AND IMPACT OF THE STUDY: Concentrations of thiamin, pyridoxine and yeast assimilable nitrogen (YAN) influenced the synthesis of hydrogen sulphide (H2 S) by Saccharomyces cerevisiae in a synthetic grape juice medium. With a few exceptions, an increase in the concentration of either vitamin or YAN resulted in less H2 S released. This is the first report to demonstrate that both thiamin and pyridoxine along with YAN affected H2 S production, emphasizing the need to assess yeast nutrients to lower risks of off-odours during fermentation.

Carotenoid production in sugarcane juice and synthetic media supplemented with nutrients by Rhodotorula rubra l02

ABSTRACT In order for the use of biological carotenoids to become feasible, it is necessary to have adequate low cost sources and improved methods of cultivation. The aim of this study was to evaluate the effect of supplementation with nitrogen, phosphorus, zinc, and magnesium, on the biomass and carotenoid volumetric production by yeast Rhodotorula rubra L02 using a complex medium (sugarcane juice) and synthetic media (sucrose and maltose) as substrates. The experimental design used for each substrate was randomized in blocks with 16 treatments and 3 repetitions. The treatments were compound for 15 different combinations of nutrients (N; Mg; Zn; P, N + Mg; N + Zn; N + P; Mg + Zn; Mg + P; Zn + P; N + P + Zn; N + P + Mg; N + Zn + Mg; P + Zn + Mg; N + Zn + Mg + P) alone and combined, and a control. The results were submitted to analysis of variance and Tukey test at 5% significance level. Among the treatments evaluated, the highest production of dry biomass, with both maltose and sucrose, was observed for Mg (1.60 g/L and 1.94 g/L, respectively). Additionally, another treatment that stood out in terms of biomass production was the control treatment with maltose (1.54 g/L). After the incubation time, killer activity was not observed since there was no formation of inhibition halo around the L02 yeast.

Impact of the timing and the nature of nitrogen additions on the production kinetics of fermentative aromas by Saccharomyces cerevisiae during winemaking fermentation in synthetic media.

During alcoholic fermentation, many parameters, including the nitrogen composition of the must, can affect aroma production. The aim of this study was to examine the impact of several types of nitrogen sources added at different times during fermentation. Nitrogen was added as ammonium or a mixture of amino acids at the beginning of fermentation or at the start of the stationary phase. These conditions were tested with two Saccharomyces cerevisiae strains that have different nitrogen requirements. The additions systematically reduced the fermentation duration. The aroma production was impacted by both the timing of the addition and the composition of the nitrogen source. Propanol appeared to be a metabolic marker of the presence of assimilable nitrogen in the must. The production of ethyl esters was slightly higher after the addition of any type of nitrogen; the production of higher alcohols other than propanol was unchanged, and acetate esters were overproduced due to the overexpression of the genes ATF1 and ATF2. Finally the parameter affecting the most the synthesis of beneficial aromas was the addition timing: The supply of organic nitrogen at the beginning of the stationary phase was more favorable for the synthesis of beneficial aromas.

Carotenoid production in sugarcane juice and synthetic media supplemented with nutrients by Rhodotorula rubra l02.

In order for the use of biological carotenoids to become feasible, it is necessary to have adequate low cost sources and improved methods of cultivation. The aim of this study was to evaluate the effect of supplementation with nitrogen, phosphorus, zinc, and magnesium, on the biomass and carotenoid volumetric production by yeast Rhodotorula rubra L02 using a complex medium (sugarcane juice) and synthetic media (sucrose and maltose) as substrates. The experimental design used for each substrate was randomized in blocks with 16 treatments and 3 repetitions. The treatments were compound for 15 different combinations of nutrients (N; Mg; Zn; P, N+Mg; N+Zn; N+P; Mg+Zn; Mg+P; Zn+P; N+P+Zn; N+P+Mg; N+Zn+Mg; P+Zn+Mg; N+Zn+Mg+P) alone and combined, and a control. The results were submitted to analysis of variance and Tukey test at 5% significance level. Among the treatments evaluated, the highest production of dry biomass, with both maltose and sucrose, was observed for Mg (1.60g/L and 1.94g/L, respectively). Additionally, another treatment that stood out in terms of biomass production was the control treatment with maltose (1.54g/L). After the incubation time, killer activity was not observed since there was no formation of inhibition halo around the L02 yeast.

Three mitochondrial transporters of Saccharomyces cerevisiae are essential for ammonium fixation and lysine biosynthesis in synthetic minimal medium.

The nuclear genes of Saccharomyces cerevisiae YHM2, ODC1 and ODC2 encode three transporters that are localized in the inner mitochondrial membrane. In this study, the roles of YHM2, ODC1 and ODC2 in the assimilation of nitrogen and in the biosynthesis of lysine have been investigated. Both the odc1Δodc2Δ double knockout and the yhm2Δ mutant grew similarly as the YPH499 wild-type strain on synthetic minimal medium (SM) containing 2% glucose and ammonia as the main nitrogen source. In contrast, the yhm2Δodc1Δodc2Δ triple knockout exhibited a marked growth defect under the same conditions. This defect was fully restored by the individual expression of YHM2, ODC1 or ODC2 in the triple deletion strain. Furthermore, the lack of growth of yhm2Δodc1Δodc2Δ on 2% glucose SM was rescued by the addition of glutamate, but not glutamine, to the medium. Using lysine-prototroph YPH499-derived strains, the yhm2Δodc1Δodc2Δ knockout (but not the odc1Δodc2Δ and yhm2Δ mutants) also displayed a growth defect in lysine biosynthesis on 2% glucose SM, which was rescued by the addition of lysine and, to a lesser extent, by the addition of 2-aminoadipate. Additional analysis of the triple mutant showed that it is not respiratory-deficient and does not display mitochondrial DNA instability. These results provide evidence that only the simultaneous absence of YHM2, ODC1 and ODC2 impairs the export from the mitochondrial matrix of i) 2-oxoglutarate which is necessary for the synthesis of glutamate and ammonium fixation in the cytosol and ii) 2-oxoadipate which is required for lysine biosynthesis in the cytosol. Finally, the data presented allow one to suggest that the yhm2Δodc1Δodc2Δ triple knockout is suitable in complementation studies aimed at assessing the pathogenic potential of human SLC25A21 (ODC) mutations.

Synthetic cystic fibrosis sputum medium diminishes Burkholderia cenocepacia antifungal activity against Aspergillus fumigatus independently of phenylacetic acid production.

Phenylacetic acid (PAA), an intermediate of phenylalanine degradation, is emerging as a signal molecule in microbial interactions with the host. In this work, we explore the presence of phenylalanine and PAA catabolism in 3 microbial pathogens of the cystic fibrosis (CF) lung microbiome: Pseudomonas aeruginosa, Burkholderia cenocepacia, and Aspergillus fumigatus. While in silico analysis of B. cenocepacia J2315 and A. fumigatus Af293 genome sequences showed complete pathways from phenylalanine to PAA, the P. aeruginosa PAO1 genome lacked several coding genes for phenylalanine and PAA catabolic enzymes. High-performance liquid chromatography analysis of supernatants from B. cenocepacia K56-2 detected PAA when grown in Luria-Bertani medium but not in synthetic cystic fibrosis sputum medium (SCFM). However, we were unable to identify PAA production by A. fumigatus or P. aeruginosa in any of the conditions tested. The inhibitory effect of B. cenocepacia on A. fumigatus growth was evaluated using agar plate interaction assays. Inhibition of fungal growth by B. cenocepacia was lessened in SCFM but this effect was not dependent on bacterial production of PAA. In summary, while we demonstrated PAA production by B. cenocepacia, we were not able to link this metabolite with the B. cenocepacia - A. fumigatus microbial interaction in CF nutritional conditions.

Time-to-positivity, type of culture media and oxidase test performed on positive blood culture vials to predict Pseudomonas aeruginosa in patients with Gram-negative bacilli bacteraemia / Tiempo de positividad, tipo de medio de cultivo y prueba de oxidasa realizada en viales de hemocultivo positivos para predecir Pseudomonas aeruginosa en pacientes con bacteriemia por bacilos gramnegativos

Introduction. The aim of this study was to determine the usefulness of oxidase test and time-to-positivity (TTP) in aerobic and anaerobic blood culture vials to detect the presence of Pseudomonas aeruginosa in patients with Gram-negative bacilli (GNB) bacteraemia. Material and methods. TTP was recorded for each aerobic and anaerobic blood culture vial of monomicrobial bacteraemia due to GNB. Oxidase test was performed in a pellet of the centrifuged content of the positive blood culture. An algorithm was developed in order to perform the oxidase test efficiently taking into account TTP and type of vial. Results. A total of 341 episodes of GNB bacteraemia were analysed. Sensitivity, specificity, positive predictive value and negative predictive value of the oxidase test performed on positive vials with GNB to predict P. aeruginosa were 95%, 99%, 91%, and 99%, respectively. When growth was first or exclusively detected in anaerobic vials, P. aeruginosa was never identified hence the performance of the oxidase test could be avoided. When growth was only or first detected in aerobic vials, a TTP≥8h predicted P. aeruginosa in 37% or cases (63 of 169), therefore oxidase test is highly recommended. Conclusions. Oxidase test performed onto positive blood culture vials previously selected by TTP and type of vials is an easy and inexpensive way to predict P. aeruginosa. In most cases, this can lead to optimization of treatment in less than 24 hours (AU)

Introducción. El objetivo del estudio fue determinar la utilidad de la prueba de oxidasa y del tiempo de positividad del hemocultivo (TPH) para detectar la presencia de Pseudomonas aeruginosa en pacientes con bacteriemia por bacilos gramnegativos (BGN). Material y métodos. Se registró el TPH de cada vial aerobio y anaerobio en todos los episodios de bacteriemia monomicrobiana por BGN. La prueba de oxidasa se realizó sobre el contenido centrifugado del hemocultivo positivo. Se diseñó un algoritmo para optimizar la realización de la prueba de oxidasa según el TPH y el tipo de vial. Resultados. Se analizaron 341 episodios de bacteriemia por BGN. La sensibilidad, especificidad, valor predictivo positivo y valor predictivo negativo de la prueba de oxidasa para predecir P. aeruginosa fueron del 95%, 99%, 91% y 99%, respectivamente. Cuando el crecimiento se detectó primero o exclusivamente en viales anaerobios, nunca se identificó P. aeruginosa pudiendo evitar la realización de la prueba de oxidasa. Cuando el crecimiento se detectó antes o exclusivamente en viales aerobios un TPH ≥8h predijo la presencia de P. aeruginosa en el 37% de los casos (63 de 169), por lo que es recomendable la realización de la prueba de oxidasa. Conclusiones. La prueba de oxidasa realizada a viales de hemocultivos positivos previamente seleccionados por el TPH y el tipo de medio es una forma fácil y económica de predecir P. aeruginosa. En la mayoría de los casos, esto puede contribuir a la optimización del tratamiento antibiótico en menos de 24h (AU)

Improvement of the productivity of ecumicin, a novel anti-tuberculosis agent, from new Nonomuraea sp. MJM5123.

Ecumicin is a novel anti-tuberculosis agent produced by Nonomuraea sp. MJM5123 as a new strain of actinomycetes. First, in order to increase the cell mass of Nonomuraea sp. MJM5123, we optimized the culture conditions with regard to carbon and nitrogen sources. The cell mass of Nonomuraea sp. MJM5123 increased by approximately twofold when glucose and soybean flour were used as carbon and nitrogen sources, respectively. For maximum production of ecumicin, we optimized the culture conditions by adding amino acids as building blocks for ecumicin, by adding vegetable oils and by controlling the temperature and pH. Ecumicin production was two times higher with the addition of valine as the building blocks for ecumicin compared with the production in the absence of valine. Interestingly, with the addition of 1% corn oil, the production of ecumicin increased by 4.6-fold compared with the production in the absence of corn oil. Finally, by controlling the pH and temperature, we established an optimized culture condition in which Nonomuraea sp. MJM5123 produced 576 mg ecumicin per litre of medium, which is about 50 times higher than in the control medium at 30 °C and pH 7.0.

[THE ACTUAL APPROACHES TO PROBLEM OF IMPORT SUBSTITUTION IN TH FIELD OF PRODUCTION GROWTH MEDIUM].

The import substitution becomes one of strategic tasks of Russian economy as a result of imposition of economic sanctions on part of the USA, EU countries, Japan and number of other states. The development of structure and technology of production of national import substituted growth mediums permits satisfying needs of laboratory service of Russia inactive storage and to secure appropriate response to occurring challenges and new biological menaces and support bio-security of state at proper level. The presented data concerning substantiation of nomenclature of growth mediums and transport system permit satisfying in fullness the needs of clinical and sanitary microbiology in growth mediums of national production and to give up of import deliveries without decreasing of quality of microbiological studies.

In vitro culture of previously uncultured oral bacterial phylotypes.

Around a third of oral bacteria cannot be grown using conventional bacteriological culture media. Community profiling targeting 16S rRNA and shotgun metagenomics methods have proved valuable in revealing the complexity of the oral bacterial community. Studies investigating the role of oral bacteria in health and disease require phenotypic characterizations that are possible only with live cultures. The aim of this study was to develop novel culture media and use an in vitro biofilm model to culture previously uncultured oral bacteria. Subgingival plaque samples collected from subjects with periodontitis were cultured on complex mucin-containing agar plates supplemented with proteose peptone (PPA), beef extract (BEA), or Gelysate (GA) as well as on fastidious anaerobe agar plus 5% horse blood (FAA). In vitro biofilms inoculated with the subgingival plaque samples and proteose peptone broth (PPB) as the growth medium were established using the Calgary biofilm device. Specific PCR primers were designed and validated for the previously uncultivated oral taxa Bacteroidetes bacteria HOT 365 and HOT 281, Lachnospiraceae bacteria HOT 100 and HOT 500, and Clostridiales bacterium HOT 093. All agar media were able to support the growth of 10 reference strains of oral bacteria. One previously uncultivated phylotype, Actinomyces sp. HOT 525, was cultivated on FAA. Of 93 previously uncultivated phylotypes found in the inocula, 26 were detected in in vitro-cultivated biofilms. Lachnospiraceae bacterium HOT 500 was successfully cultured from biofilm material harvested from PPA plates in coculture with Parvimonas micra or Veillonella dispar/parvula after colony hybridization-directed enrichment. The establishment of in vitro biofilms from oral inocula enables the cultivation of previously uncultured oral bacteria and provides source material for isolation in coculture.

Biodegradation of pesticide triclosan by A. versicolor in simulated wastewater and semi-synthetic media.

Triclosan is known as an antimicrobial agent, a powerful bacteriostat and an important pesticide. In this paper biodegradation of triclosan by Aspergillus versicolor was investigated. Effects of simulated wastewater and semi-synthetic media on fungal triclosan degradation process were detected. HPLC analysis showed that fungal triclosan biodegradation yield was 71.91% at about 7.5 mg/L concentration in semi-synthetic medium and was 37.47% in simulated wastewater. Fungus could be able to tolerate the highest triclosan concentration (15.69 mg/L). The biodegradation yield was 29.81% and qm was 2.22 mg/g at this concentration. Some of the parameters, such as pH, culture media, increasing triclosan and biomass concentrations were optimized in order to achieve the effective triclosan biodegradation process. The highest triclosan biodegradation yields of all microorganisms were achieved by A. versicolor.

Galactose-functionalized polyHIPE scaffolds for use in routine three dimensional culture of mammalian hepatocytes.

Three-dimensional (3D) cell culture is regarded as a more physiologically relevant method of growing cells in the laboratory compared to traditional monolayer cultures. Recently, the application of polystyrene-based scaffolds produced using polyHIPE technology (porous polymers derived from high internal phase emulsions) for routine 3D cell culture applications has generated very promising results in terms of improved replication of native cellular function in the laboratory. These materials, which are now available as commercial scaffolds, are superior to many other 3D cell substrates due to their high porosity, controllable morphology, and suitable mechanical strength. However, until now there have been no reports describing the surface-modification of these materials for enhanced cell adhesion and function. This study, therefore, describes the surface functionalization of these materials with galactose, a carbohydrate known to specifically bind to hepatocytes via the asialoglycoprotein receptor (ASGPR), to further improve hepatocyte adhesion and function when growing on the scaffold. We first modify a typical polystyrene-based polyHIPE to produce a cell culture scaffold carrying pendent activated-ester functionality. This was achieved via the incorporation of pentafluorophenyl acrylate (PFPA) into the initial styrene (STY) emulsion, which upon polymerization formed a polyHIPE with a porosity of 92% and an average void diameter of 33 µm. Histological analysis showed that this polyHIPE was a suitable 3D scaffold for hepatocyte cell culture. Galactose-functionalized scaffolds were then prepared by attaching 2'-aminoethyl-ß-D-galactopyranoside to this PFPA functionalized polyHIPE via displacement of the labile pentafluorophenyl group, to yield scaffolds with approximately ca. 7-9% surface carbohydrate. Experiments with primary rat hepatocytes showed that cellular albumin synthesis was greatly enhanced during the initial adhesion/settlement period of cells on the galactose-functionalized material, suggesting that the surface carbohydrates are accessible and selective to cells entering the scaffold. This porous polymer scaffold could, therefore, have important application as a 3D scaffold that offers enhanced hepatocyte adhesion and functionality.

Development of an industrial medium and a novel fed-batch strategy for high-level expression of recombinant ß-mananase by Pichia pastoris.

An industrial medium, Corn Steep Liquor Powder Dextrose (CSD medium) was developed for constitutive expression of recombinant ß-mananase by Pichia pastoris. The ß-mananase activity (513 U/mL) with CSD medium was 1.64- and 2.5-fold higher than with YPD and BSM in shaken flasks. The ß-mananase productivity with CSD medium was 61.0 U/mL h, which was 1.7- and 2.5-fold higher than with YPD and BSM in a 5-L fermenter based on a novel fed-batch strategy combining the real-time exponential feed mode with the DO-stat feed mode. The ß-mananase activity, dry cell weight and the recombinant enzyme reached up to 5132 U/mL, 110.0 g/L and 4.50 g/L after 50 h cultivation in a 50-L fermenter. The high efficient expression of recombinant ß-mananase by P. pastoris indicated that CSD medium and the novel fed-batch strategy have great potential for the production of recombinant ß-mananase in industrial fermentation.

Biosynthesis of polyhydroxyalkanaotes by a novel facultatively anaerobic Vibrio sp. under marine conditions.

Marine bacteria have recently attracted attention as potentially useful candidates for the production of practical materials from marine ecosystems, including the oceanic carbon dioxide cycle. The advantages of using marine bacteria for the biosynthesis of poly(hydroxyalkanoate) (PHA), one of the eco-friendly bioplastics, include avoiding contamination with bacteria that lack salt-water resistance, ability to use filtered seawater as a culture medium, and the potential for extracellular production of PHA, all of which would contribute to large-scale industrial production of PHA. A novel marine bacterium, Vibrio sp. strain KN01, was isolated and characterized in PHA productivity using various carbon sources under aerobic and aerobic-anaerobic marine conditions. The PHA contents of all the samples under the aerobic-anaerobic condition, especially when using soybean oil as the sole carbon source, were enhanced by limiting the amount of dissolved oxygen. The PHA accumulated using soybean oil as a sole carbon source under the aerobic-anaerobic condition contained 14% 3-hydroxypropionate (3HP) and 3% 5-hydroxyvalerate (5HV) units in addition to (R)-3-hydroxybutyrate (3HB) units and had a molecular weight of 42 × 10³ g/mol. The present result indicates that the activity of the beta-oxidation pathway under the aerobic-anaerobic condition is reduced due to a reduction in the amount of dissolved oxygen. These findings have potential for use in controlling the biosynthesis of long main-chain PHA by regulating the activity of the beta-oxidation pathway, which also could be regulated by varying the dissolved oxygen concentration.

Efecto de distintos antimicrobianos sobre el crecimiento de Listeria monocytogenes / Effect of different antimicrobial agents on the growth of Listeria monocytogenes

En este estudio se ha evaluado la eficacia de dos ácidos orgánicos contemplados en la lista positiva de aditivos alimentarios, el lactato sódico (E-325) y el diacetato sódico (E-262), sobre el crecimiento de Listeria monocytogenes. Estos aditivos se adicionaron en diferentes concentraciones a un medio de cultivo líquido, determinando el incremento de densidad óptica del medio a 600 nm durante 24 horas a 37 ºC, con respecto al medio sin inocular que se tomó como blanco, realizando la medida cada hora. El incremento de absorbancia se midió con respecto al tiempo, evaluando el crecimiento de la bacteria a través de la interpretación de la tasa máxima de incremento de absorbancia (micro) y el tiempo mínimo requerido para detectar un incremento en la densidad óptica del medio (epsilon). Este último parámetro se puede equiparar al tiempo de latencia o tiempo de adaptación al medio. Así, para el lactato sódico, se observó que ejerce un efecto negativo dosis dependiente sobre el crecimiento de L. monocytogenes, prolongando el tiempo que necesitó la bacteria para adaptarse al medio de cultivo (epsilon), sin afectar a la tasa de crecimiento (micro) una vez que esta comenzó a crecer. El diacetato sódico mostró ser más efectivo que el lactato sódico frente al crecimiento de la bacteria, incrementando el tiempo de adaptación al medio, así como disminuyendo la tasa de crecimiento. Además, el diacetato sódico consiguió inhibir de forma completa el crecimiento de la bacteria a concentraciones iguales o superiores a 0.2%(AU)

The effectiveness of two organic acids included in the positive list for additives, sodium lactate (E-325) and sodium diacetate (E-262), was evaluated against Listeria monocytogenes growth. Different concentrations of these additives were added to the liquid culture medium. The optical density increments at 600 nm was measured for a 24 hours period under 37 0C, using non-inoculated medium as blank. The measurements were taken every hour in sterile 96 wells plates each. After this analysis, a graphical representation of absorbance increment against time was done, extrapolating the maximum absorbance increment rate (micro) and the minimum time required to detect an absorbance increment (epsilon) from the graphic. These two parameters made possible to evaluate the bacterial growth. After the analysis of epsilon and micro for lactate concentrations, a negative effect in bacterial growth was observed, extending epsilon value. Nevertheless, once the bacterial growth started, any effect on micro value was detected. A higher inhibitory effect was observed after the analysis of these parameters for diacetate concentrations, an extension on epsilon value as well as a micro value descent was found. In this way, a total inhibition of growth occurred when diacetate concentration was 0,2% or higher(AU)

Variabilidad en la sensibilidad de tigeciclina frente a Acinetobacter baumannii en diferentes medios de cultivo / Variability in the sensitivity to tigecycline against Acinetobacter baumannii in different culture medium

Introducción. Tigeciclina puede suponer una alternativaterapéutica para el control de A. baumannii multirresistentes, sibien no existe consenso en cuanto a los puntos de corte desensibilidad ni a la variabilidad de su CMI en función del mediode cultivo utilizado para realizar el antibiograma frente a estemicroorganismo. Por ello, nuestro objetivo ha sido verificardicha variabilidad, así como proponer el medio de cultivo quemás se aproxime al método estándar.Métodos. Se seleccionaron 41 cepas de A. baumanniicarbapenem-resistentes. Se analizó la sensibilidad frente atigeciclina en diferentes medios de cultivo: Mueller Hinton agarcomercial de Oxoid (MH-C); Mueller Hinton agar fresco de BDand Co, USA (MH-F) e ISO-sensitest agar en fresco de Oxoid,utilizando la técnica de E-test y disco. Las CMIs se compararonfrente a las obtenidas mediante la técnica estándar demacrodilución.Resultados. La CMI y halos de inhibición medios obtenidosen los diferentes medios de cultivos correspondieron a 9,26mg/L y 15,1 mm de diámetro para MH-C; 1,71 mg/L y 22,7 mmpara MH-F; 2,68 mg/L y 20,8 mm para ISO-sensitest. La CMImedia obtenida mediante el método estándar de dilución fuede 0,47 mg/L (SD=0,21), con rango entre 0,25 a 1 mg/L.Conclusión. En los tres medios de cultivo estudiados, seobservan CMIs superiores al estándar, lo que supone interpretarfalsas resistencias en muchos casos. No obstante, el medio quese aproxima más al de referencia es el MH-F(AU)

Introduction. The tigecycline may represent a therapeuticalternative for the control of multiresistant A.baumannii, although there is no consensus regarding thecutoff points for sensitivity or variability of MIC as afunction of culture medium used for the antibiogramagainst this microorganism. Therefore, our objective wasto verify this variability, and propose the culture mediumthat comes closest to the standard method.Methods. We selected 41 strains of carbapenem-resistantA. baumannii. We analyzed the sensitivity to tigecyclinein different culture medium: Mueller Hintonagar Oxoid commercial (C-MH), Mueller Hinton freshagar BD and Co., USA (F-MH) and ISO-sensitest freshagar Oxoid, using the E-test and disk. The MICs werecompared against those obtained using the techniquestandard of macrodilution.Results. The mean MIC and inhibition diameters obtainedin the different culture medium corresponded to9.26 mg/L and 15.1 mm in diameter for MH-C, 1.71mg/L and 22.7 mm for MH-F; 2.68 mg/L and 20.8 mmfor ISO-sensitest. Half the MIC obtained by the standardmethod of dilution was 0.47 mg/L (SD = 0.21), with valuesbetween 0.25 and 1 mg/L.Conclusion. In the three growth media studied, MICssuperior to the standard are observed, which is false tointerpret resistance in many cases. However, the mediumthat comes closer more that of reference is the MH-F(AU)

Characteristics of Candida albicans biofilms grown in a synthetic urine medium.

Urinary tract infections (UTIs) are the most common type of nosocomial infection, and Candida albicans is the most frequent organism causing fungal UTIs. Presence of an indwelling urinary catheter represents a significant risk factor for UTIs. Furthermore, these infections are frequently associated with the formation of biofilms on the surface of these catheters. Here, we describe the characterization of C. albicans biofilms formed in vitro using synthetic urine (SU) medium and the frequently used RPMI medium and compare the results. Biofilms of C. albicans strain SC5314 were formed in 96-well microtiter plates and on silicon elastomer pieces using both SU and RPMI media. Biofilm formation was monitored by microscopy and a colorimetric XTT [2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide] reduction assay. As in biofilms grown in RPMI medium, time course studies revealed that biofilm formation using SU medium occurred after an initial adherence phase, followed by growth, proliferation, and maturation. However, microscopy techniques revealed that the architectural complexity of biofilms formed in SU medium was lower than that observed for those formed using RPMI medium. In particular, the level of filamentation of cells within the biofilms formed in SU medium was diminished compared to those in the biofilms grown in RPMI medium. This observation was also corroborated by expression profiling of five filamentation-associated genes using quantitative real-time reverse transcriptase PCR. Sessile C. albicans cells were resistant to fluconazole and amphotericin B, irrespective of the medium used to form the biofilms. However, caspofungin exhibited potent in vitro activity at therapeutic levels against C. albicans biofilms grown in both SU and RPMI media.

Development of an industrial medium for economical 2,3-butanediol production through co-fermentation of glucose and xylose by Klebsiella oxytoca.

An industrial medium containing urea as a sole nitrogen source, low levels of corn steep liquor and mineral salts as nutrition factors to retain high 2,3-butanediol production through co-fermentation of glucose and xylose (2:1, wt/wt) by Klebsiella oxytoca was developed. Urea and corn steep liquor were identified as the most significant factors by the two-level Plackett-Burman design. Steepest ascent experiments were applied to approach the optimal region of the two factors and a central composite design was employed to determine their optimal levels. Under the optimal medium, the yield of 2,3-butanediol plus acetoin relative to glucose and xylose was up to 0.428 g/g, which was 85.6% of theoretical value. The cheap nitrogen source and nutrition factors combining the co-fermentation process using lignocellulose derived glucose and xylose as the carbon source in the developed medium would be a potential solution to improve the economics of microbial 2,3-butanediol production.