RESUMEN
In this work, six common stainless steel grades were compared with respect to ennoblement characteristics, corrosion performance and tendency to biofouling in brackish sea water in a pilot-scale cooling water circuit. Two tests were performed, each employing three test materials, until differences between the materials were detected. Open circuit potential (OCP) was measured continuously in situ. Potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) measurements were conducted before and after the tests. Exposed specimens were further subjected to examinations by scanning electron microscopy (SEM) and energy-dispersive spectroscopy (EDS), and the biofouling was studied using epifluorescence microscopy, quantitative polymerase chain reaction (qPCR) and high-throughput sequencing (HTP sequencing). The results revealed dissimilarities between the stainless steel grades in corrosion behaviour and biofouling tendency. The test material that differed from the most of the other studied alloys was grade EN 1.4162. It experienced fastest and most efficient ennoblement of OCP, its passive area shrank to the greatest extent and the cathodic reaction was accelerated to a significant degree by the development of biofilm. Furthermore, microbiological analyses revealed that bacterial community on EN 1.4162 was dominated by Actinobacteria, whereas on the other five test materials Proteobacteria was the main bacterial phylum.
Asunto(s)
Incrustaciones Biológicas , Acero Inoxidable/química , Actinobacteria/aislamiento & purificación , Corrosión , Espectroscopía Dieléctrica , Proteobacteria/aislamiento & purificación , Agua de Mar/química , Agua de Mar/microbiologíaRESUMEN
The food industry, including the meat industry, is currently looking for natural preservatives to prevent the growth of harmful microbes in foods. The potential of plant-derived antimicrobial extracts to increase the shelf life and to delay the microbiological spoilage of marinated broiler chicken cuts in modified atmosphere packages during cold storage was investigated in this study. We evaluated the impact of aqueous ethanolic extracts of Finnish sea buckthorn berries and lingonberries and supercritical CO2-extracted herbal extracts from an antimicrobial blend and oregano leaves on the shelf life of broiler meat. The commercial antimicrobial blend extract and the oregano extract inhibited the growth of lactic acid bacteria (LAB) and Brochothrix thermosphacta in the marinated samples. The antimicrobial blend extract also reduced the growth of psychrotrophic aerobic bacteria, whereas the sea buckthorn and lingonberry extracts did not. Only minor antimicrobial activity against Enterobacteriaceae by all the extracts was observed. Plate count analysis, denaturing gradient gel electrophoresis, and quantitative real-time PCR indicated that LAB, which are the major spoilage group in marinated modified atmosphere-packaged poultry products, were not significantly affected by the berry extracts studied. During this shelf-life study, LAB isolates of Lactobacillus and Leuconostoc were identified in the marinated samples. Antimicrobial blends and oregano leaf extracts can act as antimicrobial agents in marinade blends, although tailoring of the dose is needed because of their strong taste. Further studies for exploiting synergistic effects of plant extracts could contribute to the development of potential and more effective antimicrobial blends. Studies are needed in meat matrices and in product applications to demonstrate the efficacy of these compounds.
Asunto(s)
Antiinfecciosos/farmacología , Embalaje de Alimentos/métodos , Conservación de Alimentos/métodos , Productos de la Carne/microbiología , Animales , Atmósfera , Pollos , Recuento de Colonia Microbiana , Finlandia , Contaminación de Alimentos/análisis , Manipulación de Alimentos/métodos , Microbiología de Alimentos , CarneRESUMEN
PURPOSE: Study how the dietary intake affects the fecal microbiota of a group of obese individuals after a 6-week very low-energy diet (VLED) and thereafter during a follow-up period of 5, 8, and 12 months. Additionally, we compared two different methods, fluorescent in situ hybridization (FISH) and real-time PCR (qPCR), for the quantification of fecal samples. METHODS: Sixteen subjects participated in a 12-month dietary intervention which consisted of a VLED high in protein and low in carbohydrates followed by a personalized diet plan, combined with exercise and lifestyle counseling. Fecal samples were analyzed using qPCR, FISH, and denaturing gradient gel electrophoresis. RESULTS: The VLED affected the fecal microbiota, in particular bifidobacteria that decreased approximately two logs compared with the baseline numbers. The change in numbers of the bacterial groups studied followed the dietary intake and not the weight variations during the 12-month intervention. Methanogens were detected in 56% of the participants at every sampling point, regardless of the dietary intake. Moreover, although absolute numbers of comparable bacterial groups were similar between FISH and qPCR measurements, relative proportions were higher according to FISH results. CONCLUSIONS: Changes in the fecal microbial numbers of obese individuals were primarily affected by the dietary intake rather than weight changes.
Asunto(s)
Restricción Calórica , Heces/microbiología , Conducta Alimentaria , Microbiota , Obesidad/dietoterapia , Índice de Masa Corporal , Peso Corporal , Recuento de Colonia Microbiana , ADN Bacteriano/genética , Electroforesis en Gel de Gradiente Desnaturalizante , Ingestión de Energía , Ejercicio Físico , Femenino , Finlandia , Estudios de Seguimiento , Humanos , Hibridación Fluorescente in Situ , Estilo de Vida , Masculino , Obesidad/microbiología , ARN Ribosómico 16S/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Población BlancaRESUMEN
The aim of the study was to evaluate the potential of utilising the information on expression levels of selected stress genes in assessing the quality of probiotic products. For this purpose RT-qPCR methods were developed to study the expression of clpL1 and clpL2 stress genes in Lactobacillus rhamnosus VTT E-97800 (E800) cells after exposure to processing-related stress conditions or to freeze-drying. Heat treatments in laboratory scale were performed with E800 cells incubated at 47 °C or 50 °C for 60 min. Acid treatments were performed both at laboratory and fermenter scale. At laboratory scale E800 cells were inoculated into General Edible Medium (GEM) adjusted to pH 4.0 and pH 3.5 and incubated at 37 °C for 180 min, whereas fermenter-grown cells were exposed to pH 4.0 for 60 min at the end of the fermentation. RNA from fresh cells and freeze-dried powders was reverse transcribed after isolation, quantification and standardisation. clpL1 and clpL2 transcripts were analysed by RT-qPCR with SYBR Green I. clpL1 was induced in L. rhamnosus E800 cells exposed to 50 °C and to a much lesser extent to 47 °C. No induction was observed for clpL2 in E800 cells during either acid or heat treatment, in any of the conditions applied. RNA isolation from freeze-dried powders was unsuccessful although several attempts were made with high quality products. In conclusion, our results suggest that developing quality indicators for probiotic products based on differences in the expression of stress genes is a challenging task for several reasons: at least with some genes (like in the present study with clpL) quite harsh conditions are needed to detect differences in the gene expression; mRNA isolation from freeze-dried powders was unsuccessful which hampers the quality analysis of large proportion of probiotic products; and furthermore RT-qPCR proved to be a too laborious procedure for routine use.
Asunto(s)
Ácidos/farmacología , Proteínas Bacterianas/genética , Endopeptidasa Clp/genética , Regulación Enzimológica de la Expresión Génica , Lacticaseibacillus rhamnosus/química , Lacticaseibacillus rhamnosus/enzimología , Proteínas Bacterianas/metabolismo , Endopeptidasa Clp/metabolismo , Liofilización , Regulación Bacteriana de la Expresión Génica/efectos de los fármacos , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Calor , Concentración de Iones de Hidrógeno , Lacticaseibacillus rhamnosus/efectos de los fármacos , Lacticaseibacillus rhamnosus/genéticaRESUMEN
AIM: To evaluate the influence of doxycycline therapy on the composition and antibiotic susceptibility of intestinal bifidobacteria. METHODS AND RESULTS: Faecal samples were collected from nine subjects receiving doxycycline therapy and ten control subjects, and analysed for bifidobacteria by culturing and PCR-DGGE (denaturing gradient gel electrophoresis). A marked decrease in the diversity (average number of amplicons detected by PCR-DGGE 0.8 in the antibiotic vs 4.3 in the control group) of Bifidobacterium populations was observed during doxycycline therapy. The proportion of a tetracycline-resistant bifidobacterial population was higher in the antibiotic group than in the control group (83%vs 26%). Based on the tet gene PCR, resistance could be associated with the presence of tet(W). In two subjects, strains representing highly similar genetic fingerprints but different tetracycline susceptibilities were detected. A mutation causing lack of functionality in the tet(W) was observed in one of the susceptible strains. CONCLUSIONS: Doxycycline therapy had a drastic effect on the diversity and tetracycline susceptibility of intestinal Bifidobacterium populations. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of broad-spectrum antibiotics increased the pool of tetracycline-resistant commensal bacteria in the intestine. The detection of resistance genes alone is not sufficient for the evaluation of bacterial antibiotic resistance.
Asunto(s)
Antibacterianos/administración & dosificación , Bifidobacterium/fisiología , Doxiciclina/administración & dosificación , Intestinos/microbiología , Administración Oral , Antibacterianos/uso terapéutico , Técnicas Bacteriológicas , Bifidobacterium/efectos de los fármacos , Bifidobacterium/genética , Estudios de Casos y Controles , Doxiciclina/uso terapéutico , Heces/microbiología , Variación Genética/efectos de los fármacos , Humanos , Pruebas de Sensibilidad Microbiana/métodos , Mutación , Reacción en Cadena de la Polimerasa/métodos , Resistencia a la Tetraciclina/genéticaRESUMEN
The aim of this study was to find out whether liquorice-containing starch gel could affect plaque accumulation and its microbial composition. Sixteen healthy volunteers (mean age: 30.4+/-6.9 years) used 6 g of either control [8% acid-hydrolyzed corn starch, 25% maltitol syrup, water (w/w)] or liquorice gel (control + 2.5% liquorice extract), three times a day for 2 weeks. The gels were used in a random order with a 2-week washout period in between. At the end of each fortnight, plaque was allowed to accumulate for 2 days and all available plaque from the right side of the mouth was collected, weighed, and transferred to transport medium. The plaque on the left side was dyed and photographed in a standardized manner. Mutans streptococci, total streptococci, and facultative bacteria were assessed from the plaque using plate culturing. Plaque index (0-5) of incisors and canines on the left side was evaluated from the photographs. The clinical study was preceded by an in vivo acid production test. The acid production from gels containing 2.5-10% liquorice extract was monitored with a microelectrode. The in vivo acid production potential of the maltitol-containing starch gel was about 50% compared to the sucrose control. Liquorice inhibited acid production from the gel. In the clinical study, the weight of plaque after consumption of the liquorice gel did not differ from that of the control gel. No differences were found in the microbial counts nor in the plaque index between the two gels. In addition, the liquorice gel had no effect on the stability of the predominant bacterial populations of the plaque samples of 16 individuals as detected by PCR-denaturing gradient gel electrophoresis. In conclusion, an addition of liquorice extract to starch-containing gel with a low acid production potential had no effect on the plaque formed during a 2-week gel consumption period.