Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 3.422
Filter
1.
Braz. j. biol ; 83: e250550, 2023. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1345536

ABSTRACT

Abstract Vanillin is the major component which is responsible for flavor and aroma of vanilla extract and is produced by 3 ways: natural extraction from vanilla plant, chemical synthesis and from microbial transformation. Current research was aimed to study bacterial production of vanillin from native natural sources including sewage and soil from industrial areas. The main objective was vanillin bio-production by isolating bacteria from these native sources. Also to adapt methodologies to improve vanillin production by optimized fermentation media and growth conditions. 47 soil and 13 sewage samples were collected from different industrial regions of Lahore, Gujranwala, Faisalabad and Kasur. 67.7% bacterial isolates produced vanillin and 32.3% were non-producers. From these 279 producers, 4 bacterial isolates selected as significant producers were; A3, A4, A7 and A10. These isolates were identified by ribotyping as A3 Pseudomonas fluorescence (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) and A10 Bacillus subtilis (KT962919). Vanillin producers were further tested for improved production of vanillin and were grown in different fermentation media under optimized growth conditions for enhanced production of vanillin. The fermentation media (FM) were; clove oil based, rice bran waste (residues oil) based, wheat bran based and modified isoeugenol based. In FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36, and FM37, the selected 4 bacterial strains produced significant amounts of vanillin. A10 B. subtilis produced maximum amount of vanillin. This strain produced 17.3 g/L vanillin in FM36. Cost of this fermentation medium 36 was 131.5 rupees/L. This fermentation medium was modified isoeugenol based medium with 1% of isoeugenol and 2.5 g/L soybean meal. ech gene was amplified in A3 P. fluorescence using ech specific primers. As vanillin use as flavor has increased tremendously, the bioproduction of vanillin must be focused.


Resumo A vanilina é o principal componente responsável pelo sabor e aroma do extrato de baunilha e é produzida de três formas: extração natural da planta da baunilha, síntese química e transformação microbiana. A pesquisa atual teve como objetivo estudar a produção bacteriana de vanilina a partir de fontes naturais nativas, incluindo esgoto e solo de áreas industriais. O objetivo principal era a bioprodução de vanilina por meio do isolamento de bactérias dessas fontes nativas. Também para adaptar metodologias para melhorar a produção de vanilina por meio de fermentação otimizada e condições de crescimento. Foram coletadas 47 amostras de solo e 13 de esgoto de diferentes regiões industriais de Lahore, Gujranwala, Faisalabad e Kasur; 67,7% dos isolados bacterianos produziram vanilina e 32,3% eram não produtores. Desses 279 produtores, 4 isolados bacterianos selecionados como produtores significativos foram: A3, A4, A7 e A10. Esses isolados foram identificados por ribotipagem como fluorescência A3 Pseudomonas (KF408302), A4 Enterococcus faecium (KT356807), A7 Alcaligenes faecalis (MW422815) e A10 Bacillus subtilis (KT962919). Os produtores de vanilina foram posteriormente testados para produção aprimorada de vanilina e foram cultivados em diferentes meios de fermentação sob condições de crescimento otimizadas para produção aprimorada de vanilina. Os meios de fermentação (FM) foram: à base de óleo de cravo, à base de resíduos de farelo de arroz (resíduos de óleo), à base de farelo de trigo e à base de isoeugenol modificado. Em FM5, FM21, FM22, FM23, FM24, FM30, FM31, FM32, FM34, FM35, FM36 e FM37, as 4 cepas bacterianas selecionadas produziram quantidades significativas de vanilina. A10 B. subtilis produziu quantidade máxima de vanilina. Essa cepa produziu 17,3 g / L de vanilina em FM36. O custo desse meio de fermentação 36 foi de 131,5 rúpias / L. Esse meio de fermentação foi um meio à base de isoeugenol modificado com 1% de isoeugenol e 2,5 g / L de farelo de soja. O gene ech foi amplificado em A3 P. fluorescence usando primers específicos para ech. Como o uso da vanilina como sabor aumentou tremendamente, a bioprodução da vanilina deve ser focada.


Subject(s)
Benzaldehydes/metabolism , Flavoring Agents/metabolism , Bacillus subtilis/metabolism , Industrial Microbiology , Pseudomonas fluorescens/metabolism , Enterococcus faecium/metabolism , Culture Media , Alcaligenes faecalis/metabolism , Fermentation
2.
Braz. j. biol ; 83: e246904, 2023. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1345524

ABSTRACT

Abstract Hyperhydricity is a serious physiological disorder and affects In vitro propagation of many plants and as well of Salvia santolinifolia. The donor material to initiate the in vitro culture was the callus taken from the in vitro shoots produced on Murashig and Skoogs (MS) medium at 4.0 mg/l BA. This callus formed numerous hyperhydric shoots on culturing upon the medium of the same composition. The aim was to systematically evaluate the effect of cytokinins (Benzyladnine (BA) and N6-(-2-isopentenyl) adenine (2iP), culture vessels magnitude, medium solidification, source of nitrogen and calcium chloride for the alleviation of hyperhydricity. In the tissue cultures of S. santolinifolia BA and 2iP induced severe hyperhydricity, when other factors i.e. culture vessels magnitude and a suitable concentration of agar, ammonium nitrate (NH4NO3), potassium nitrate (KNO3) & calcium chloride (CaCl2.2H2O) were not optimized. After 30 days' culture, we observed 83.82% hyperhydric shoots at increased level (1.5 mg/l 2iP) and 81.59% at decreased levels (1.0 mg/l 2iP). On the other hand, hyperhydricity percentage at decreased (0.4%) and at increased (0.8%) levels of agar were 72.37% and 39.08%, respectively. MS medium modification with NH4NO3 (412 mg/l), KNO3 (475 mg/l) and CaCl2.2H2O (880 mg/l) was found the best medium to reduced hyperhydricity (23.6%).


Resumo A hiperidricidade é um distúrbio fisiológico sério e afeta a propagação in vitro de muitas plantas e também da Salvia santolinifolia. O material doador para iniciar a cultura in vitro foi o calo retirado dos brotos in vitro produzidos em meio Murashig e Skoogs (MS) a 4,0 mg / l BA. Esse calo formou numerosos rebentos hiperídricos em cultura no meio da mesma composição. O objetivo foi avaliar sistematicamente o efeito das citocininas (Benziladnina (BA) e N6 - (- 2-isopentenil) adenina (2iP), magnitude dos vasos de cultura, solidificação do meio, fonte de nitrogênio e cloreto de cálcio para o alívio da hiperidricidade. culturas de tecidos de S. santolinifolia BA e 2iP induziram hiperidricidade severa, quando outros fatores, como magnitude dos vasos de cultura e uma concentração adequada de ágar, nitrato de amônio (NH4NO3), nitrato de potássio (KNO3) e cloreto de cálcio (CaCl2.2H2O), não foram otimizados. Após 30 dias de cultura, observamos 83,82% de brotos hiperídricos em níveis aumentados (1,5 mg / l 2iP) e 81,59% em níveis reduzidos (1,0 mg / l 2iP). Por outro lado, a porcentagem de hiperidricidade diminuiu (0,4%) e em níveis aumentados (0,8%) de ágar foram 72,37% e 39,08%, respectivamente. A modificação do meio MS com NH4NO3 (412 mg / l), KNO3 (475 mg / l) e CaCl2.2H2O (880 mg / l) foi encontrada melhor hiperidricidade média a reduzida (23,6%).


Subject(s)
Salvia , Plant Shoots , Culture Media
3.
Rev. ADM ; 79(3): 165-176, mayo-jun. 2022. ilus, tab
Article in Spanish | LILACS | ID: biblio-1378976

ABSTRACT

Introducción: El hueso, reservorio de minerales y moléculas orgánicas, es un tejido dinámico que detecta y se adapta a las cargas mecánicas de los órganos y tejidos del cuerpo, el cual mantiene la estructura ósea del esqueleto durante el crecimiento y a través de la vida del ser humano. Las células óseas son sensibles a las cargas mecánicas y microvibra- ciones que recibe el esqueleto. Objetivo: El propósito de este estudio fue realizar una revisión sistemática acerca de los efectos que ejerce la microvibración de alta frecuencia-baja intensidad, en osteocitos cultivados in vitro sobre la síntesis de factores solubles, con el propósito de entender si la microvibración tiene influencia en la aceleración del movimiento dentario. Material y métodos: Se realizó una búsqueda de artículos de revisión de osteocitos y otras células óseas in vitro, a través de la estrategia PICO (Paciente, Intervención, Comparación, Resultado [Outcome]), con el empleo de palabras clave como: «os- teocitos¼, «microvibración¼, «remodelación¼, «osteoclastogénesis¼, «citocinas¼ y «osteoblastos¼. Se estructuró por medio de PRISMA (informe de revisiones sistemáticas y meta-análisis). La captación de datos finales se hizo por medio del método de puntuación de calidad Jadad y Cochrane (modelo de correlación) como herramientas para evaluar el riesgo de sesgo de cada uno de los artículos. Se incluyeron 11 artículos con alta calidad metodológica. Resultados: La mayoría de los experimentos in vitro demostraron que la microvibración tuvo un aumento estadísticamente significativo en la proliferación y dife- renciación de las células madre mesenquimales (MSC), en osteoblastos (MC3T3-E1), en la expresión de proteínas para inducir osteogénesis y en los osteocitos (MLO-Y4). Asimismo, sobrerregularon la expresión de osteoprotegerina (OPG), prostaglandina (PGE2) y óxido nitroso (NO) al alterar y regular los factores solubles como las citocinas, factores de crecimiento y quimiocinas, de las demás células, además de mostrar una disminución en la actividad de los osteoclastos (RAW246.7) en la resorción ósea. Conclusión: La microvibración induce remodelación ósea. Los osteocitos son sensibles a los estímulos mecánicos y producen factores solubles para inducir la remodelación ósea, razón por la cual se emplea la microvibración como una terapia innovadora y prometedora, no invasiva y no farmacológica en la estimulación de la formación ósea de la superficie del hueso (AU)


Subject(s)
Humans , Osteogenesis , Vibration , Bone Remodeling , Osteocytes , Bone Resorption , Analysis of Variance , Cytokines , Culture Media , RANK Ligand
4.
Braz. j. biol ; 82: e242596, 2022. tab, graf
Article in English | LILACS, VETINDEX | ID: biblio-1278487

ABSTRACT

Abstract Hops is a new culture in Brazil. Tissue culture can be an important technique for rapid hop propagation. This paper aims to characterize responses from different genotypes under different growth regulators through the interrelationship of response variables important to hop in vitro growth. Three genotypes were cultivated in six culture media with different combinations of growth regulators, BAP (6-benzylaminopurine), IAA (3-indolacetic acid) and GA3 (gibberellic acid). The means were compared by orthogonal contrasts and the interrelationship of the response variables was performed by path analysis. American genotypes showed favorable root development under the BAP + IAA combination, while the use of IAA improved shoot development. The origin of genotypes was important for defining the best protocol for in vitro cultivation. The path coefficient showed that the variable number of shoots has stronger direct effect on the number of nodal segments. Additionally, in tissue culture assays, the use of a covariable and proper error distribution significantly increased experimental accuracy.


Resumo O lúpulo é uma nova cultura no Brasil. A cultura de tecidos pode ser uma técnica importante para a propagação rápida do lúpulo. Este artigo tem como objetivo caracterizar respostas de diferentes genótipos sob diferentes reguladores de crescimento através da inter-relação de variáveis ​​de resposta importantes para o crescimento in vitro. Três genótipos foram cultivados em seis meios de cultura com diferentes combinações de reguladores de crescimento, BAP (6-benzilaminopurina), AIA (ácido 3-indolacético) e GA3 (ácido giberélico). As médias foram comparadas por contrastes ortogonais e a inter-relação das variáveis ​​de resposta foi realizada por análise de trilha. Os genótipos americanos apresentaram desenvolvimento radicular favorável sob a combinação BAP + AIA, enquanto o uso do AIA melhorou o desenvolvimento da parte aérea. A origem dos genótipos foi importante para definir o melhor protocolo para o cultivo in vitro. O coeficiente de trilha mostrou que a variável número de brotos tem um efeito direto mais forte no número de segmentos nodais. Adicionalmente, em experimentos com cultura de tecidos, o uso de uma covariável e distribuição de erro adequada aumentou significativamente a precisão experimental.


Subject(s)
Plant Growth Regulators , Brazil , Plant Shoots/genetics , Culture Media , Genotype
5.
Chinese Journal of Biotechnology ; (12): 1322-1338, 2022.
Article in Chinese | WPRIM | ID: wpr-927783

ABSTRACT

Aerobic methane oxidizing bacteria (methanotrophs) can use methane as carbon source and energy source, eliminating 10%-20% of global methane. Methanotrophs can also effectively synthesize valuable methane-derived products. This article introduced the methane oxidizing mechanism of methanotrophs, and summarized the practical application and research hotspots of methanotrophs in the field of methane emission reduction in the landfill, ventilation air methane mitigation in coal mines, valuable chemicals biosynthesis, as well as oil and gas reservoir exploration. Main factors influencing the pollutant removal and the biosynthesis efficiency in various applications were also discussed. Based on the study of large-scale cultivation of methanotrophs, some measures to benefit the application and promotion of aerobic methane oxidizing biotechnology were proposed. This includes investigating the effect of intermediate metabolites on methanotrophs activity and population structure, and exploiting economical and efficient alternative culture media and culture techniques.


Subject(s)
Biotechnology , Carbon , Culture Media/chemistry , Methane/metabolism , Methylococcaceae/metabolism , Oxidation-Reduction
6.
Chinese Journal of Biotechnology ; (12): 796-806, 2022.
Article in Chinese | WPRIM | ID: wpr-927745

ABSTRACT

Ergothioneine (ERG) is a natural antioxidant that has been widely used in the fields of food, medicine and cosmetics. Compared with traditional plant extraction and chemical synthesis approaches, microbial synthesis of ergothioneine has many advantages, such as the short production cycle and low cost, and thus has attracted intensive attention. In order to engineer an ergothioneine high-yielding Escherichia coli strain, the ergothioneine synthesis gene cluster egtABCDE from Mycobacterium smegmatis and egt1 from Schizosaccharomyces pombe were introduced into E. coli BL21(DE3) to generate a strain E1-A1 harboring the ergothioneine biosynthesis pathway. As a result, (95.58±3.2) mg/L ergothioneine was produced in flask cultures. To further increase ergothioneine yield, the relevant enzymes for biosynthesis of histidine, methionine, and cysteine, the three precursor amino acids of ergothioneine, were overexpressed. Individual overexpression of serAT410STOP and thrA resulted in an ergothioneine titer of (134.83±4.22) mg/L and (130.26±3.34) mg/L, respectively, while co-overexpression of serAT410STOP and thrA increased the production of ergothioneine to (144.97±5.40) mg/L. Eventually, by adopting a fed-batch fermentation strategy in 3 L fermenter, the optimized strain E1-A1-thrA-serA* produced 548.75 mg/L and 710.53 mg/L ergothioneine in glucose inorganic salt medium and rich medium, respectively.


Subject(s)
Culture Media , Ergothioneine/metabolism , Escherichia coli/metabolism , Fermentation , Histidine/metabolism , Metabolic Engineering
7.
Chinese Journal of Biotechnology ; (12): 760-771, 2022.
Article in Chinese | WPRIM | ID: wpr-927742

ABSTRACT

Fatty acids (FA) are widely used as feed stocks for the production of cosmetics, personal hygiene products, lubricants and biofuels. Ogataea polymorpha is considered as an ideal chassis for bio-manufacturing, due to its outstanding characteristics such as methylotroph, thermal-tolerance and wide substrate spectrum. In this study, we harnessed O. polymorpha for overproduction of fatty acids by engineering its fatty acid metabolism and optimizing the fermentation process. The engineered strain produced 1.86 g/L FAs under the optimized shake-flask conditions (37℃, pH 6.4, a C/N ratio of 120 and an OD600 of seed culture of 6-8). The fed-batch fermentation process was further optimized by using a dissolved oxygen (DO) control strategy. The C/N ratio of initial medium was 17.5, and the glucose medium with a C/N ratio of 120 was fed when the DO was higher than 30%. This operation resulted in a titer of 18.0 g/L FA, indicating the potential of using O. polymorpha as an efficient cell factory for the production of FA.


Subject(s)
Culture Media , Fatty Acids , Fermentation , Metabolic Engineering , Saccharomycetales/metabolism
8.
Rev. ADM ; 78(6): 339-345, nov.-dic. 2021. tab, graf
Article in Spanish | LILACS | ID: biblio-1354635

ABSTRACT

En la práctica clínica, los odontólogos se encuentran expuestos al riesgo de infecciones, que se transmiten a través de instrumentos contaminados con exudados. Instrumentos en contacto con el personal deben estar esterilizados o sometidos a un proceso de desinfección. Se realizó un estudio transversal-prospectivo a 30 pacientes, de los que se tomaron tres muestras con espejos estériles, pasando por fondo de saco, carrillos y lengua, después las muestras se desinfectaron, se realizó el hisopado de cada espejo y se incubó en agar tripticaseína-soya (TSA) 24 horas a 37 oC. Pasadas 24 horas se realizaron diluciones en tubos Eppendorf, y se sembraron en cajas de Petri con agar sangre, se incubaron por 48 horas a 37 oC; se contabilizaron las unidades formadoras de colonias (UFC) y registraron para su análisis. Al obtener los resultados se encontró que ID 213 tuvo mayor reducción con una media = 62.5 en comparación con Zeta 1 Ultra, media = 89.23, y control, media = 164.50, de igual manera se observó una diferencia en reducción de UFC/mL entre ID 213 con respecto a Zeta 1 Ultra con significancia de 0.012. Ambos desinfectantes resultaron efectivos, pero se estableció que ID 213 utilizando la tina ultrasónica resulta más efectivo en la reducción de UFC, que Zeta 1 Ultra (AU)


In clinical practice, dentists are exposed to the risk of infections, which are transmitted through instruments contaminated with exudates. Instruments in contact with personnel must be sterilized or subjected to a disinfection process. A cross-sectional-prospective study was carried out in 30 patients. From which three samples were taken with sterile mirrors, passing through cul-de-sac, cheeks and tongue, later the samples were disinfected with disinfectants, each mirror was swabbed and incubated in TSA 24 hours at 37 oC. After 24 hours, dilutions were made in Eppendorf tubes, and they were seeded in Petri dishes with blood agar, they were incubated 48 hours at 37 oC; CFUs were accounted for and recorded for analysis. When obtaining the results, it was found that ID 213 had a greater reduction with mean = 62.5 compared to Zeta 1 Ultra mean = 89.23 and control mean = 164.50, in the same way a difference in reduction of CFU/mL was observed between ID 213 with respect to Zeta 1 Ultra with significance of 0.012. Both disinfectants were effective but it was established that ID 213 using the ultrasonic tub is more effective in reducing CFU, than Zeta 1 Ultra (AU)


Subject(s)
Humans , Male , Female , Ultrasonics , Infection Control, Dental , Disinfectants , Effectiveness , Colony Count, Microbial , Cross-Sectional Studies , Prospective Studies , Culture Media , Mexico , Military Dentistry
9.
Rev. colomb. biotecnol ; 23(2): 36-40, jul.-dic. 2021. tab
Article in Spanish | LILACS | ID: biblio-1360962

ABSTRACT

RESUMEN La cepa mexicana CP-145 de Ganoderma lucidum debido a la importancia medicinal que ha presentado últimamente, la presente investigación tuvo como objetivo evaluar el efecto de la temperatura y medio de cultivo sobre el crecimiento micelial óptimo en diferentes rangos de pH. Los tratamientos correspondieron en la utilización del medio de cultivo papa dextrosa agar (PDA) y extracto de malta agar (EMA), con dos niveles de temperatura (25 y 28 °C) y seis rangos de pH (4.0, 4.5, 5.0, 5.5, 6.0 y 6.5). El diseño experimental fue completamente al azar con medidas repetidas a través del tiempo, analizados con el paquete REPEATED MEASURE y el efecto tiempo con PROC MIXED de SAS. Como resultado se obtuvieron que el efecto de la temperatura y medios de cultivo en los diferentes rangos de pH, presentaron diferencias significativas (P ≤ 0.05). El crecimiento micelial óptimo de la cepa mexicana de G. lucidum fue en el medio de cultivo EMA en los rangos de pH de 4.0 y 4.5 con 8.3 y 8.2 cm respectivamente. De igual forma, en los rangos de pH 4.0 y 4.5 se obtuvieron los crecimientos miceliales óptimos a temperatura de 25 °C con 8.1 y 8.0 cm respectivamente. El cual concluyó esta investigación que el crecimiento micelial óptimo de la cepa mexicana fueron a pH 4.0 y 4.5, temperatura de 25 °C y medio de cultivo EMA.


ABSTRACT The Mexican strain CP-145 of Ganoderma lucidum due to the medicinal importance it has presented lately, the present investigation had as objective to evaluate the effect of temperature and culture medium on the optimal mycelial growth in different pH ranges. The treatments corresponded to the use of potato dextrose agar (PDA) and malt extract agar (EMA), with two temperature levels (25 and 28 °C) and six pH ranges (4.0, 4.5, 5.0, 5.5, 5.5, 6.0 and 6.5). The experimental design was completely randomised with repeated measures over time, analysed with the REPEATED MEASURE package and the time effect with PROC MIXED of SAS. As a result, the effect of temperature and culture media in the different pH ranges showed significant differences (P ≤ 0.05). The optimal mycelial growth of the Mexican strain of G. lucidum was in the EMA culture medium in the pH ranges of 4.0 and 4.5 with 8.3 and 8.2 cm respectively. Similarly, in the pH ranges 4.0 and 4.5 the optimum mycelial growth was obtained at 25 °C with 8.1 and 8.0 cm respectively. This research concluded that the optimal mycelial growth of the Mexican strain was at pH 4.0 and 4.5, temperature of 25 °C and EMA culture medium.


Subject(s)
Culture Media , In Vitro Techniques
10.
Medisan ; 25(3)2021. tab, ilus
Article in Spanish | LILACS, CUMED | ID: biblio-1287303

ABSTRACT

Introducción: La sangre ovina constituye un suplemento esencial en la elaboración de medios de cultivo, dado que aporta factores nutricionales indispensables para el crecimiento y la recuperación de diversos microorganismos. Objetivo: Evaluar comparativamente el efecto de la sangre ovina, tanto citratada como desfibrinada, en medios de cultivo de base agar en cuanto al crecimiento bacteriano y la producción de hemólisis de cepas de referencia de diferentes bacterias patógenas, así como la recuperación o el aislamiento de microorganismos de muestras clínicas. Métodos: Se realizó un estudio observacional, descriptivo y transversal en 6 laboratorios de microbiología de la ciudad de Santiago de Cuba durante el año 2017, en los cuales se empleó sangre de ovinos de la raza pelibuey, para la elaboración de medios de cultivo. A cada laboratorio se le suministró tanto sangre citratada como desfibrinada y se le entregó una encuesta para valorar los resultados. Resultados: Existió un mejor crecimiento y aislamiento bacteriano en el medio suplementado con sangre desfibrinada, a pesar de que el rendimiento o los resultados en el caso de la sangre citratada resultaron satisfactorios. Conclusiones: Se confirmó la pertinencia del uso de la sangre desfibrinada como suplemento de enriquecimiento nutritivo en medios de cultivo; no obstante, quedó demostrada la utilidad de la sangre citratada en la labor de rutina del laboratorio de microbiología clínica.


Introduction: Sheep blood is an essential supplement in the elaboration of culture media, as it provides important nutritional factors for the growth and recovery of different organisms. Objective: To evaluate comparatively the effect of sheep citrated and defibrinated sheep blood in culture media with agar base as for the bacterial growth and the production of hemolysis in reference strains from different pathogen bacteria, as well as the recovery or isolation of microorganisms from clinical samples. Method: An observational, descriptive and cross-sectional was carried out in 6 microbiology laboratories in Santiago de Cuba city during 2017, in which male sheeps blood from the pelibuey breed for elaborating culture media. Each laboratory received either citrated blood or defibrinated and a survey was delivered to evaluate the results. Results: There was a better growing and bacterial isolation in the media supplemented with defibrinated blood, although yielding or results were favorable with citrated blood. Conclusions: The pertinence of the use of defibrinated blood as a supplement of nutritive enrichment in culture media was confirmed; however, the use of citrated blood was demonstrated in the routine work of the clinical microbiology laboratory.


Subject(s)
Sheep , Bacterial Growth , Culture Media , Hemolysis
11.
Chinese Journal of Biotechnology ; (12): 3334-3347, 2021.
Article in Chinese | WPRIM | ID: wpr-921429

ABSTRACT

Cordycepin is the key active component of medicinal fungus Cordyceps militaris, and it shows multiple functional activities such as anti-tumor and anti-virus. Cordycepin was conventionally produced by liquid fermentation of C. militaris, but the long production cycle and the low productivity constrained its development and application. In this study, two key genes for cordycepin biosynthesis (ScCNS1 and ScCNS2) were introduced into Saccharomyces cerevisiae S288C, producing 67.32 mg/L cordycepin at 240 h. Analysis of gene expression profiles indicated that ZWF1, PRS4, ADE4, ScCNS1 and ScCNS2 which encode enzymes involved in pentose phosphate pathway, purine metabolism and cordycepin biosynthesis pathway, were significantly up-regulated in the late phage of fermentation. Optimization of fermentation medium determined that 50 g/L initial glucose followed by feeding, supplemented with 5 mmol/L Cu²⁺ and 1.0 g/L adenine were the best condition. Fed-batch fermentation using the engineered yeast in a 5 L stirred fermenter produced 137.27 mg/L cordycepin at 144 h, with a productivity up to 0.95 mg/(L·h) reached, which was 240% higher than that of the control.


Subject(s)
Cordyceps , Culture Media , Deoxyadenosines , Saccharomyces cerevisiae/genetics
12.
Chinese Journal of Biotechnology ; (12): 2543-2553, 2021.
Article in Chinese | WPRIM | ID: wpr-887820

ABSTRACT

We designed and fabricated a novel high throughput brain-on-chip with three dimensional structure with the aim to simulate the in vivo three-dimensional growth environment for brain tissues. The chip consists of a porous filter and 3D brain cell particles, and is loaded into a conventional 96-well plate for use. The filter and the particle molds were fabricated by using computer modeling, 3D printing of positive mold and agarose-PDMS double reversal mold. The 3D cell particles were made by pouring and solidifying a suspension of mouse embryonic brain cells with sodium alginate into a cell particle mold, and then cutting the resulting hydrogel into pieces. The loaded brain-on-chip was used to determine the neurotoxicity of pesticides. The cell particles were exposed to 0, 10, 30, 50, 100 and 200 µmol/L of chlorpyrifos or imidacloprid, separated conveniently from the medium by removing the porous filter after cultivation. Subsequently, cell proliferation, acetylcholinesterase activity and lactate dehydrogenase release were determined for toxicity evaluation. The embryonic brain cells were able to grow and proliferate normally in the hydrogel particles loaded into the filter in a 96-well plate. Pesticide neurotoxicity test showed that both chlorpyrifos and imidacloprid presented dose-dependent inhibition on cell growth and proliferation. Moreover, the pesticides showed inhibition on acetylcholinesterase activity and increase release of lactate dehydrogenase. However, the effect of imidacloprid was significantly weaker than that of chlorpyrifos. In conclusion, a novel brain-on-chip was developed in this study, which can be used to efficiently assess the drug neurotoxicity, pharmacodynamics, and disease mechanism by combining with a microtiterplate reader.


Subject(s)
Animals , Brain , Chlorpyrifos/toxicity , Culture Media , Mice , Oligonucleotide Array Sequence Analysis , Pesticides/toxicity
13.
Braz. j. med. biol. res ; 54(11): e11191, 2021. tab, graf
Article in English | LILACS | ID: biblio-1285664

ABSTRACT

The present study focused on the scenario of confirmed cases of SARS-CoV-2 infection in the state of Minas Gerais (MG), Brazil, from March 2020 to March 2021. We evaluated the evolution of COVID-19 prevalence and death in one municipality from each of the 14 health macro-regions of MG state. Socio-demographic characteristics and variables related to the municipalities were analyzed. The raw dataset used in this study was freely sourced from the website Brasil.io. From the raw dataset, two time series were extracted: the cumulative confirmed cases of COVID-19 and cumulative death counts, and they were compared to the state data using a nowcasting approach. In order to make time series comparisons possible, all data was normalized per 100,000 inhabitants. When analyzing in light of colored wave code interventions initiated in August 2020 in MG, for the majority of the municipalities, there was an absence of clear influence on prevalence and deaths. The national holidays in the first semester of 2020 had a small impact on the COVID-19 prevalence of the municipalities, but the holidays in the second semester of 2020 and beginning of 2021 caused important impacts on COVID-19 prevalence. The low number of ICU beds in some municipalities contributed to the higher number of deaths. The analysis showed here is expected to contribute to the improvement of decision making of the MG government, as it opened a huge possibility to have the total macro-regions and state data analyzed.


Subject(s)
Humans , COVID-19 , Brazil/epidemiology , Cities/epidemiology , Culture Media , SARS-CoV-2
14.
Rev. Fundac. Juan Jose Carraro ; 24(44): 40-47, 2021. ilus, tab
Article in Spanish | LILACS | ID: biblio-1223492

ABSTRACT

Las enfermedades del periodonto tienen una etiopatogenia compleja y puede considerarse multifactorial. El factor etiológico esencial en la patología inflamatoria periodontal es la biopelícula dental y cuando el desequilibrio entre el huésped y los microorganismos cambia la complejidad de la flora. Ciertas bacterias como Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, Fusobacterium nucleatum, Tannerrella forsythia, Campylobacter rectus, Eikenella corrodens y Treponema spp., han sido comúnmente relacionadas con la periodontitis crónica y son consideradas como indicadores de riesgo para la progresión de dicha enfermedad. El objetivo de este trabajo fue establecer la prevalencia de Prevotella spp y Porphyromona spp en los distintos estadios de periodontitis crónicas. Material y métodos: Se estudiaron 48 pacientes sistémicamente saludables con diagnóstico de periodontitis crónica. Se completó el consentimiento informado, se realizó historia clínica y examen periodontal. El estado periodontal se clasificó en distintos grados de severidad: leve, moderada y severa. Se tomaron muestras de dos sitios con mayor profundidad de sondaje con conos de papel absorbente estériles y se transportaron en un medio prerreducido. Para el aislamiento de Prevotella spp se utilizó agar Brucella más sangre ovina al 5%, hemina, vitamina K al que se agregaron vancomicina y kanamicina; Porphyromonas sp se aisló en el mismo medio con el agregado de bacitracina y colistina. Se sembraron 10 µl de muestra entera y las placas fueron incubadas en jarras de anaerobiosis por 5 a 7 días a 37ºC. Resultados: los distintos grados de periodontitis correspondieron a un 17% periodontits leve, 57% moderada y 26% severa. En el total de pacientes se determinó la presencia de Prevotella spp en el 54% de los casos y un 12,5% de Porphyromona spp. Conclusión: De los pacientes estudiados con periodontits crónica, un 52% correspondió al sexo masculino, un 57% de los casos correspondieron a periodontitis moderada. Se aisló Prevotella sp en todos los estadios de periodontitis crónica y Porphyromonas sp sólo en periodontitis severas (AU)


Periodontal diseases have a complex etiopathogenesis and can be considered multifactorial. The essential etiological factor in periodontal inflammatory pathology is the dental biofilm and when the imbalance between the host and the microorganisms changes the complexity of the flora. Certain bacteria such as Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Prevotella loescheii, Fusobacterium nucleatum, Tannerrella forsythia, Campylobacter rectus, Eikenella corrodens and Treponema spp., Have been commonly related to chronic periodontitis and are considered as risk indicators for the progression of said disease. The objective of this work was to establish the prevalence of Prevotella spp and Porphyromonas spp in the different stages of chronic periodontitis. Forty eight systemically healthy patients diagnosed with chronic periodontitis were studied. Informed consent was completed, a medical history and periodontal examination was carried out. The periodontal state was classified into different degrees of severity: mild, moderate and severe. Samples were taken from two sites with greater depth of probing with sterile absorbent paper cones and transported in a prereduced medium. For the isolation of Prevotella spp, Brucella agar plus 5% sheep blood, hemin, vitamin K to which vancomycin and kanamycin were added. For Porphyromonas spp, the same medium was used and bacitracin and colistin were added. 10 µl of the whole sample was seeded and the plates were incubated in anaerobic jars for 5 to 7 days at 37 ° C. Different degrees of periodontitis corresponded to 17% mild periodontitis, 57% moderate and 26% severe. In the total number of patients, the presence of Prevotella spp was determined in 54% of the cases and 12.5% of Porphyromona spp. Of the patients studied with chronic periodontitis, 52% corresponded to the male sex, 57% of the cases corresponded to moderate periodontitis. Prevotella spp was isolated in all stages of chronic periodontitis and Porphyromonas sp only in severe periodontitis (AU)


Subject(s)
Humans , Male , Female , Adult , Middle Aged , Porphyromonas gingivalis/isolation & purification , Prevotella intermedia/isolation & purification , Chronic Periodontitis/microbiology , Colony Count, Microbial , Risk Factors , Culture Media , Dental Plaque/microbiology , Age and Sex Distribution
15.
Article in English | LILACS, BBO | ID: biblio-1180856

ABSTRACT

ABSTRACT Objective: To compare Transforming growth factor-β1 (TGF-β1) expression in various L-PRF concentrations on the hDPSC differentiation process. Material and Methods: hDPSC cell cultures were subjected to serum starvation by reducing FBS levels in the hDPSC culture media. Lysate PRF was obtained from the PRF gel, which was then incubated at 4°C for 24 h. The supernatant was dried, transferred to a 2-ml Eppendorf tube, and stored at −20°C. The evaluation of TGF-β1 expression in 1%, 5%, 10%, and 25% L-PRF samples and 10% FBS (control) during the process of hDPSC differentiation was quantified using an ELISA reader on day 7. The expression of TGF-β1 was subjected to a one-way ANOVA test, followed by Bonferroni's post hoc test with significant values (p<0.05). Results: Significant differences were noted in TGF-β1 expression between 1%, 5%, 10%, and 25% L-PRF and the control group (10% FBS). The highest TGF-β1 expression occurred with 25% L-PRF (0.645 ± 0.048), followed by 10% L-PRF (0.461 ± 0.035), 10% FBS (0.374 ± 0.013), 5% L-PRF (0.275 ± 0.045), and the lowest expression was with 1% L-PRF (0.160 ± 0.045). Conclusion: The best result of TGF-B1 expression in hDPSC differentiation was in the 25% L-PRF group.


Subject(s)
Humans , Cell Culture Techniques , Culture Media/analysis , Dental Pulp , Platelet-Rich Fibrin/microbiology , Enzyme-Linked Immunosorbent Assay , Transforming Growth Factors , Cell Differentiation/immunology , Analysis of Variance , Indonesia
16.
Rev. Fac. Odontol. (B.Aires) ; 36(84): 21-26, 2021. tab
Article in Spanish | LILACS | ID: biblio-1363852

ABSTRACT

La terapia endodóntica tiene como uno de sus objetivos lograr la completa desinfección del sistema de conductos radiculares. Por esto, se deben seleccionar sustancias irrigantes que tengan la capacidad de eliminar todo el contenido de dicho sistema. La acción antimicrobiana es una de las características más importantes a tener en cuenta en la elección. El hipoclorito de sodio (NaOCl) tiene capacidad bactericida sobre muchos de los microorganismos de la flora endodóntica. El Enterococcus faecalis es una bacteria altamente resistente a antibacterianos que sobrevive en condiciones extremas. El ácido hipocloroso (HOCl) es una molécula derivada del NaOCl que ha demostrado tener alto poder bactericida sobre cepas patogénicas bucales. El objetivo de este trabajo fue evaluar y comparar la efectividad antimicrobiana in vitro del NaOCl 2.5% y el HOCl al 5% frente a Enterococcus faecalis. Una suspensión de Enterococcus faecalis (ATCC29212), de turbidez 0.5 en escala de McFarland, fue inoculada en varios tubos de ensayo, los cuales contenían cada antimicrobiano. Se dejaron actuar durante 1, 5 y 10 minutos para luego neutralizarlos e inclubarlos a 37º C en condiciones de capnofilia durante 48 hs. Todo el procedimiento se realizó por quintuplicado. Los resultados se midieron mediante recuento de UFC/ml. No se evidenció presencia de Enterococcus faecalis en las placas que contenían la solución de NaOCl al 2.5% como tampoco en aquellas que contenían HOCl al 5%. In vitro, el HOCl y el NaOCl en las concentraciones probadas, eliminaron completamente las cepas de Enterococcus faecalis (AU)


Subject(s)
Sodium Hypochlorite/therapeutic use , Enterococcus faecalis/drug effects , Hypochlorous Acid/therapeutic use , Anti-Bacterial Agents/therapeutic use , Root Canal Irrigants/therapeutic use , In Vitro Techniques , Colony Count, Microbial , Culture Media , Dental Pulp Cavity/microbiology
17.
Braz. j. biol ; 80(4): 914-920, Oct.-Dec. 2020. tab, graf
Article in English | LILACS | ID: biblio-1142526

ABSTRACT

Abstract Growth and biological conditions of Messastrum gracile were evaluated to compare the effect of photoautotrophic and mixotrophic cultivation on the increase of biomass production and chemical conditions cultured in macrophyte and commercial culture media. The growth rate (k) of M. gracile was different in the culture media, higher in mixotrophic cultivation for Lemna minor culture medium, whilst to Eichhornia crassipes and NPK culture media were higher in photoautotrophic cultivation. Mean lipid contents in photoautotrophic cultivation were 8.2% biomass dry weight, whereas they reached 19% biomass dry weight in mixotrophic cultivation. Protein contents were below 48% biomass dry weight in photoautotrophic cultivation and 30% biomass dry weight in mixotrophic cultivation. Messastrum gracile cultured in macrophyte culture media (E. crassipes and L. minor) and NPK culture medium provided satisfactory results with regard to lipid and protein contents in mixotrophic and photoautotrophic cultivations, respectively. Lipid and protein contents in alternative media were higher or similar to the CHU12 commercial culture medium.


Resumo O crescimento e as condições biológicas da microalga Messastrum gracile foram avaliados para comparar o efeito do cultivo foto-autotrófico e mixotrófico na produção de biomassa e condições químicas em meios de cultura comercial e de macrófitas. A taxa de crescimento (k) de M. gracile foi diferente entre os meios de cultura, sendo maior no cultivo mixotrófico para o meio Lemna minor, enquanto para os meios Eichhornia crassipes e NPK foram maiores no cultivo foto-autotrófico. Os teores de lipídios no cultivo foto-autotrófico foram de 8,2% da biomassa seca, enquanto que no mixotrófico atingiram 19% da biomassa seca. Os teores de proteína em cultivo foto-autotrófico estiveram abaixo de 48% da biomassa seca e 30% de biomassa seca no cultivo mixotrófico. Messastrum gracile cultivada em meios de cultura de macrófitas (E. crassipes e L. minor) e NPK apresentaram resultados satisfatórios em relação aos teores de lipídeos e proteínas nos cultivos mixotróficos e foto-autotróficos, respectivamente. Os teores lipídicos e proteicos em meios alternativos foram maiores ou semelhantes ao meio comercial CHU12.


Subject(s)
Eichhornia , Microalgae , Chlorophyceae , Biomass , Culture Media
18.
Rev. Asoc. Odontol. Argent ; 108(2): 46-51, mayo-ago. 2020. tab
Article in Spanish | LILACS | ID: biblio-1121108

ABSTRACT

Objetivos: Comparar ex vivo la eficacia del instrumento XP-endo Finisher y del sistema EndoActivator en la reducción/eliminación del biofilm microbiano en conductos radiculares infectados. Materiales y métodos: Se utilizaron 23 premolares inferiores humanos extraídos cuya longitud fue estandarizada en 17 mm. Todos los conductos se prepararon con el sistema WaveOne Gold Medium (#35.06). Los dientes se esterilizaron, se inocularon con Enterococcus faecalis y se separaron en dos grupos experimentales de 10 piezas cada uno. De los 3 dientes remanentes, 1 fue utilizado como control positivo y 2, como controles negativos. En el grupo 1, las soluciones irrigantes se agitaron con XP-endo Finisher. En el grupo 2, se utilizó EndoActivator. Se tomaron muestras antes de la contaminación, luego de esta y después de la agitación de los irrigantes mediante conos de papel estériles. La carga microbiana fue sembrada en agar sangre y los conos se cultivaron en caldo tripteína de soja. La remoción de la carga microbiana se determinó por la presencia o ausencia de turbiedad del medio. Las unidades formadoras de colonias (UFC) remanentes se cuantificaron y los resultados se categorizaron como R1 (≤10 UFC) o R2 (>10 UFC). Los datos fueron analizados mediante la prueba de Fisher. Resultados: No hubo diferencias significativas entre XP-endo Finisher y EndoActivator (P>0,05). El número de usos no influyó sobre la capacidad operativa de ambos instrumentos (AU)


Aim: To compare ex vivo the effectiveness of the XP-endo Finisher and the EndoActivator in biofilm reduction/ removal from infected root canals. Materials and methods: Twenty three extracted human single-rooted lower premolars were selected and standardised to 17 mm in length. All the canals were prepared with WaveOne Gold Medium reciprocating files (#35.06). The teeth were autoclaved and inoculated with Enterococcus faecalis. The infected teeth were then assigned to 2 experimental groups of 10 teeth each according to the final irrigation/agitation protocol. Of the three remaining teeth, one was used as a positive control, and the other two were used as negative controls. In Group 1 the irrigating solutions were agitated with XP-endo Finisher while in Group 2 the EndoActivator was used. All root canals were sampled before and after contamination, and again after irrigant agitation with sterile paper points. The microbial load was spread on blood agar plates and the paper points were cultured in sterile trypticase soy broth. The removal of the microbial load was determined by visual observation of the turbidity of the media and by quantification of the number of colony-forming units (UFC). The results were categorized as R1 (≤10 UFC) or R2 (>10 UFC). Data were analysed by the Fisher's exact test at P<0.05. Results: No significant differences was found between XP-endo Finisher and EndoActivator (P>0.05) regarding their effectiveness in the reduction/removal of the microbial biofilm. The number of uses of both instruments did not affect their operative performance (AU) Conclusion: XPF and EA were both equally effective for microbial biofilm reduction/removal from ex vivo infected root canals (AU)


Subject(s)
Root Canal Irrigants/chemistry , Dental High-Speed Equipment , Biofilms , Dental Instruments , Dental Pulp Cavity/microbiology , In Vitro Techniques , Colony Count, Microbial/methods , Efficacy , Data Interpretation, Statistical , Enterococcus faecalis/isolation & purification , Culture Media
19.
Vaccimonitor (La Habana, Print) ; 29(2)mayo.-ago. 2020. graf
Article in Spanish | LILACS, CUMED | ID: biblio-1127511

ABSTRACT

Streptococcus pneumoniae es un patógeno oportunista que puede causar infecciones como otitis media, neumonía, sepsis y meningitis. Sin embargo, existen muchas limitaciones para el cultivo en zaranda de este microorganismo en los laboratorios de microbiología. Por esta razón, se realizó un estudio de la consistencia del cultivo en zaranda de Streptoccoccus pneumoniae 19A a escala de 40 L. Para esto se desarrolló previamente la curva de crecimiento patrón hasta las 5 h. Desde el pre-inóculo se inocularon 6 frascos de 100 mL, de los cuales se realizó la inoculación en botellones de 1 L y 5 L. En todos los casos, se determinó la pureza a partir de la tinción de Gram y el crecimiento bacteriano se monitoreó por el método de conteo de viables y la densidad óptica cada 1 h. Además, se evaluó el rendimiento del cultivo a partir de la cantidad de biomasa obtenida por peso húmedo. Cada proceso se llevó a cabo en condiciones iguales por triplicado. En los tres procesos se obtuvieron curvas de crecimiento similares, tanto por densidad óptica como por conteo de viables, alcanzando una viabilidad máxima de 109 UFC/mL en la última escala. Además, se obtuvieron rendimientos de biomasa de 11,62; 11,92 y 11,60 g/L, respectivamente. Estos resultados demuestran que la metodología utilizada ofrece una consistencia de este proceso, a pesar del alto volumen de cultivo en zaranda, lo cual no afectó la calidad de la biomasa, demostrado por la viabilidad final(AU)


Streptococcus pneumoniae is an opportunistic pathogen that can cause infections including otitis media, pneumonia, sepsis and meningitis. However, there are many limitations for the cultivation in shaker of this microorganism in microbiology laboratories. For this reason, a study of the consistency of the culture in shaker of Streptoccoccus pneumoniae 19A was carried out at a scale of 40 L. The pattern growth curve was made until 5 h, under our laboratory conditions and culture medium. From the pre-inoculum 6 bottles of 100 mL were inoculated, from which the scaling was accomplished to bottles of 1 L and 5 L. In all cases the purity was determined by Gram staining and the bacterial growth by viable counting method and the optical density were monitored every 1 h. In addition, the yield was evaluated from the determination of the amount of biomass obtained by wet weight. Each process has been made in equal conditions in triplicate. In the three processes, similar growth curves were obtained both by optical density and by viable counts, reaching a maximum viability of 109 CFU/mL on the last scale. In addition, biomass yields of 11.62, 11.92 and 11.60 g/L were obtained, respectively. These results demonstrate that the methodology used offers a high process consistency, despite the high volume of culture in rotational shaker, and did not affect the quality of the biomass, which could be demonstrated by the viable count(AU)


Subject(s)
Pneumococcal Infections , Biomass , Culture Media
20.
Medisan ; 24(2)mar.-abr. 2020. tab
Article in Spanish | LILACS, CUMED | ID: biblio-1098393

ABSTRACT

Introducción: Las enfermedades transmitidas por alimentos se producen por ingestión de un alimento, incluido el agua, que puede estar contaminado por diversos agentes. Objetivo: Caracterizar los agentes bacterianos aislados en brotes de enfermedades transmitidas por alimentos. Métodos: Se realizó un estudio observacional, descriptivo y transversal de 100,0 % de los brotes de enfermedades transmitidas por alimentos en la provincia de Santiago de Cuba, desde enero de 2018 hasta diciembre de 2019, para lo cual se seleccionaron muestras de alimentos y heces fecales. La caracterización de las bacterias aisladas se basó en los resultados del crecimiento y otras pruebas bioquímicas-metabólicas. Se utilizaron resultados del aislamiento y confirmación de los agentes identificados en cada uno de los brotes a partir de las muestras antes citadas. Entre las variables analizadas figuraron: número de brotes, muestras de alimentos, de heces fecales y resultados de pruebas bioquímicas y metabólicas. Resultados: Se obtuvo un aislamiento de agentes bacterianos en 100,0 % de las muestras de alimentos. Hubo una mayor frecuencia de bacterias Gram negativas (82,0 %) y la menor correspondió a microorganismos Gram positivos (18,0 %). La Salmonella D fue el microorganismo más frecuente. Conclusiones: Este resultado representa un instrumento para el diagnóstico etiológico de los brotes de enfermedades transmitidas por alimentos en Santiago de Cuba.


Introduction: Diseases transmitted by foods are produced due to ingestion of a food, including water that can be contaminated by diverse agents. Objective: To characterize the bacterial agents isolated in diseases outbreaks transmitted by foods. Methods: An observational, descriptive and cross-sectional study of 100.0 % of the diseases outbreaks transmitted by foods in Santiago de Cuba, from January, 2018 to December, 2019 was carried out, for which samples of foods and stools were selected. The characterization of the isolated bacterias was based on the results of growth and other biochemical-metabolic tests. Results of the isolation and confirmation of agents identified in each one of the outbreaks from the samples mentioned above were used. Among the analyzed variables we can mention: number of outbreaks, samples of foods, samples of stools and results of biochemical and metabolic tests. Results: An isolation of bacterial agents was obtained in 100.0 % of foods samples. There was a higher frequency of Gram negative bacterias (82.0 %) and the lower corresponded to Gram positive microorganisms (18.0 %). Salmonella D was the most frequent microorganism. Conclusions: This result represents an instrument for the etiological diagnosis of diseases outbreaks transmitted by foods in Santiago de Cuba.


Subject(s)
Bacteria , Culture Media , Foodborne Diseases/epidemiology , Disease Outbreaks
SELECTION OF CITATIONS
SEARCH DETAIL