RESUMO
AIMS: The aims of this study were (i) to develop a protocol for the entrapment of anaerobic (hyper)thermophilic marine micro-organisms; (ii) to test the use of the chosen polymers in a range of physical and chemical conditions and (iii) to validate the method with batch cultures. METHODS AND RESULTS: The best conditions for immobilization were obtained at 80°C with gellan and xanthan gums. After 5-week incubation, beads showed a good resistance to all tested conditions except those simultaneously including high temperature (100°C), low NaCl (<0â5 mol l(-1) ) and extreme pH (4/8). To confirm the method efficiency, batch cultures with immobilized Thermosipho sp. strain AT1272 and Thermococcus kodakarensis strain KOD1 showed an absence of detrimental effect on cell viability and a good growth within and outside the beads. CONCLUSION: This suggests that entrapment in a gellan-xanthan matrix could be employed for the culture of anaerobic (hyper)thermophilic marine micro-organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: (Hyper)thermophilic marine micro-organisms possess a high biotechnological potential. Generally microbial cells are grown as free-cell cultures. The use of immobilized cells may offer several advantages such as protection against phage attack, high cell biomass and better production rate of desired metabolites.
Assuntos
Bactérias/crescimento & desenvolvimento , Técnicas Microbiológicas/métodos , Polissacarídeos Bacterianos , Thermococcus/crescimento & desenvolvimento , Bactérias/classificação , Técnicas de Cultura Celular por Lotes , Temperatura Alta , Água do Mar/microbiologiaRESUMO
The aim of this work was to study the antifungal properties of durancins isolated from Enterococcus durans A5-11 and of their chemically synthesized fragments. Enterococcus durans A5-11 is a lactic acid bacteria strain isolated from traditional Mongolian airag cheese. This strain inhibits the growth of several fungi including Fusarium culmorum, Penicillium roqueforti and Debaryomyces hansenii. It produces two bacteriocins: durancin A5-11a and durancin A5-11b, which have similar antimicrobial properties. The whole durancins A5-11a and A5-11b, as well as their N- and C-terminal fragments were synthesized, and their antifungal properties were studied. C-terminal fragments of both durancins showed stronger antifungal activities than other tested peptides. Treatment of D. hansenii LMSA2.11.003 strain with 2 mmol l(-1) of the synthetic peptides led to the loss of the membrane integrity and to several changes in the ultra-structure of the yeast cells. Chemically synthesized durancins and their synthetic fragments showed different antimicrobial properties from each other. N-terminal peptides show activities against both bacterial and fungal strains tested. C-terminal peptides have specific activities against tested fungal strain and do not show antibacterial activity. However, the C-terminal fragment enhances the activity of the N-terminal fragment in the whole bacteriocins against bacteria.
Assuntos
Antifúngicos/farmacologia , Bacteriocinas/farmacologia , Debaryomyces/efeitos dos fármacos , Enterococcus , Fungos/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Antibacterianos/química , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Antifúngicos/síntese química , Antifúngicos/química , Antifúngicos/isolamento & purificação , Bacteriocinas/síntese química , Bacteriocinas/química , Bacteriocinas/isolamento & purificação , Queijo/microbiologia , Debaryomyces/ultraestrutura , Enterococcus/isolamento & purificação , Enterococcus/metabolismo , Listeria/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Fragmentos de Peptídeos/síntese química , Fragmentos de Peptídeos/químicaRESUMO
AIMS: To evaluate the survival of Pediococcus acidilactici UL5 and its ability to produce pediocin PA-1 during transit in an artificial gastrointestinal tract (GIT). To investigate the physicochemical and biological stability of purified pediocin PA-1 under GIT conditions. METHODS AND RESULTS: Skim milk culture of Ped. acidilactici UL5 was fed to a dynamic gastrointestinal (GI) model known as TIM-1, comprising four compartments connected by computer-controlled peristaltic valves and simulating the human stomach, duodenum, jejunum and ileum. This strain tolerated a pH of 2·7 in the gastric compartment, while lower pH reduced its viability. Bile salts in the duodenal compartment brought a further 4-log reduction after 180 min of digestion, while high viable counts (up to 5 × 10(7) CFU ml(-1) fermented milk) of Ped. acidilactici were found in both the jejunal and ileal compartments. Pediococcus acidilactici recovered from all four compartments was able to produce pediocin at the same level as unstressed cells. The activity of the purified pediocin in the gastric compartment was slightly reduced after 90 min of gastric digestion, while no detectable activity was found in the duodenal, jejunal and ileal compartments during 5 h of digestion. HPLC analysis showed partial degradation of the pediocin peptide in the duodenal compartment and massive breakdown in the jejunal and ileal compartments. CONCLUSIONS: Pediococcus acidilactici UL5 showed high resistance to GIT conditions, and its ability to produce pediocin was not affected, suggesting its potential as a probiotic candidate. The physicochemical and biological stability of pediocin was significantly poor under GIT conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: Pediococcus acidilactici UL5 appears to be a potential probiotic candidate because its capacity to produce pediocin PA-1 is not affected by the GI conditions as well as the strain shows an acceptable survival rate. Meanwhile, purified pediocin PA-1 losses activity during GIT transit; microcapsules could be used to deliver it to the target site.
Assuntos
Bacteriocinas/química , Pediococcus/metabolismo , Trato Gastrointestinal Superior/microbiologia , Animais , Bacteriocinas/isolamento & purificação , Ácidos e Sais Biliares/química , Humanos , Concentração de Íons de Hidrogênio , Viabilidade Microbiana , Leite/microbiologia , Pediocinas , Pediococcus/crescimento & desenvolvimento , Trato Gastrointestinal Superior/químicaRESUMO
AIMS: To detect and enumerate bifidobacteria in faeces with a new quantitative multiplex real-time PCR (qPCR) method and to compare the results obtained with fluorescence in situ hybridization (FISH) methods. METHODS AND RESULTS: A multiplex qPCR assay was developed, which enabled the enumeration of Bifidobacterium spp. by targeting the bifidobacterial xylulose-5-phosphate/fructose-6-phosphate phosphoketolase gene (xfp) and total bacteria using universal Eub-primers targeting 16S rRNA gene from the domain bacteria. The qPCR assay showed high sensitivity and specificity and a low detection limit of about 2.5 x 10(3) bifidobacterial cells per gram of faeces. The qPCR results were compared with FISH combined with microscopy or flow cytometry (FCM). No statistical differences among bifidobacterial counts averages measured in adult faeces with the three methods were observed. Total bacterial count averages were higher with the FISH method coupled with microscopic analyses compared to FISH with FCM, whereas total cell numbers estimated by qPCR were intermediate between the two FISH methods. CONCLUSIONS: The new qPCR assay was shown to be sensitive, rapid and accurate for enumerating bifidobacteria in faeces. SIGNIFICANCE AND IMPACT OF THE STUDY: This method is a valuable alternative for other molecular methods for detecting faecal bifidobacteria, especially when their counts are below the detection limit of the FISH methods.
Assuntos
Aldeído Liases/genética , Proteínas de Bactérias/genética , Bifidobacterium/genética , Fezes/microbiologia , Hibridização in Situ Fluorescente/métodos , Reação em Cadeia da Polimerase/métodos , Adulto , Bifidobacterium/enzimologia , Feminino , Citometria de Fluxo/métodos , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Sensibilidade e EspecificidadeRESUMO
AIMS: To evaluate and optimize the use of denaturing high-performance liquid chromatography (DHPLC) for yeasts identification in red smear cheese surfaces. METHODS AND RESULTS: The resolution of DHPLC was first evaluated and optimized using a mixture of PCR amplicons of the internal transcribed spacer 2 (ITS2) region of 19 yeast reference strains representing 18 species that are common in the cheese microbiota. Sixteen of the 18 yeast species could be resolved by combining runs at temperatures of 57.5 and 59 degrees C. Then, DHPLC was used to investigate the yeast microbiota of pasteurized Maroilles, Munster and Livarot cheese surfaces by comparing their peak profiles with our reference yeast database and by collecting/sequencing of peak fractions. Debaryomyces hansenii and Geotrichum candidum for Munster and Maroilles cheeses, and Candida catenulata, Candida intermedia and G. candidum for Livarot cheese were identified using the reference database and collecting/sequencing of peak fractions. CONCLUSIONS: DHPLC technique was found to have good resolution properties and to be useful for investigating the yeast microbiota of red smear cheese surfaces. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first time that DHPLC is applied to study the yeast microbiota of red smear cheese surfaces.
Assuntos
Queijo/microbiologia , Cromatografia Líquida de Alta Pressão/métodos , DNA Fúngico/genética , DNA Fúngico/isolamento & purificação , Micologia/métodos , Leveduras/classificação , Leveduras/genética , Biodiversidade , DNA Intergênico/genética , DNA Intergênico/isolamento & purificação , Análise de Sequência de DNA , Leveduras/isolamento & purificaçãoRESUMO
AIMS: To compare in vitro the inhibitory activity of four bacteriocin-producing Escherichia coli to a well-characterized panel of Salmonella strains, recently isolated from clinical cases in Switzerland. METHODS AND RESULTS: A panel of 68 nontyphoidal Salmonella strains was characterized by pulsed-field gel electrophoresis analysis and susceptibility to antibiotics. The majority of tested strains were genetically different, with 40% resistant to at least one antibiotic. E. coli Mcc24 showed highest in vitro activity against Salmonella (100%, microcin 24), followed by E. coli L1000 (94%, microcin B17), E. coli 53 (49%, colicin H) and E. coli 52 (21%, colicin G) as revealed using a cross-streak activity assay. CONCLUSIONS: Escherichia coli Mcc24, a genetically modified organism producing microcin 24, and E. coli L1000, a natural strain isolated from human faeces carrying the mcb-operon for microcin B17-production, were the most effective strains in inhibiting in vitro both antibiotic resistant and sensitive Salmonella isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: Due to an increasing prevalence of antibiotic resistant Salmonella strains, alternative strategies to fight these foodborne pathogens are needed. E. coli L1000 appears to be a promising candidate in view of developing biotechnological alternatives to antibiotics against Salmonella infections.
Assuntos
Antibacterianos/biossíntese , Antibiose , Bacteriocinas/biossíntese , Escherichia coli/fisiologia , Salmonella/efeitos dos fármacos , Antibacterianos/farmacologia , Técnicas de Tipagem Bacteriana , Bacteriocinas/farmacologia , Análise por Conglomerados , Impressões Digitais de DNA , Eletroforese em Gel de Campo Pulsado , Escherichia coli/isolamento & purificação , Escherichia coli/metabolismo , Genótipo , Humanos , Testes de Sensibilidade Microbiana , Salmonella/classificação , Salmonella/crescimento & desenvolvimento , Salmonella/isolamento & purificação , Infecções por Salmonella/microbiologia , SuíçaRESUMO
Cultivation in a bioreactor of immobilized deep-sea hydrothermal microbial community was tested in order to assess the stability and reactivity of this new system. A community composed of eight hydrothermal strains was entrapped in a polymer matrix that was used to inoculate a continuous culture in a gas-lift bioreactor. The continuous culture was performed for 41 days at successively 60°C, 55°C, 60°C, 85°C and 60°C, at pH 6.5, in anaerobic condition and constant dilution rate. Oxic stress and pH variations were tested at the beginning of the incubation. Despite these detrimental conditions, three strains including two strict anaerobes were maintained in the bioreactor. High cell concentrations (3 × 10(8) cells mL(-1)) and high ATP contents were measured in both liquid fractions and beads. Cloning-sequencing and qPCR revealed that Bacillus sp. dominated at the early stage, and was later replaced by Thermotoga maritima and Thermococcus sp. Acetate, formate and propionate concentrations varied simultaneously in the liquid fractions. These results demonstrate that these immobilized cells were reactive to culture conditions. They were protected inside the beads during the stress period and released in the liquid fraction when conditions were more favorable. This confirms the advantage of immobilization that highlights the resilience capacity of certain hydrothermal microorganisms after a stress period.
Assuntos
Reatores Biológicos , Temperatura Alta , Consórcios Microbianos , Água do Mar/microbiologia , Bactérias Anaeróbias/crescimento & desenvolvimento , Técnicas Bacteriológicas , Células Imobilizadas , Meios de Cultura , Concentração de Íons de Hidrogênio , RNA Ribossômico 16SRESUMO
OBJECTIVE: To assess with an isokinetic dynamometer the force and endurance of the spinal flexor and extensor muscles in pre-teens or teens aged 11 to 13 and 14 to 16 years with and without low back pain (LBP). METHOD: The control group and the LBP group were homogeneous in terms of age, weight, height and Body Mass Index (BMI). Assessment was carried out with the isokinetic dynamometer Cybex Norm®. The spinal flexors and extensors were explored concentrically at speeds of 60°, 90° and 120°/sec. The parameters chosen were: maximal moment of force (MMF), mean power (MP), total work (TW), F/E ratios (between the flexors and the extensors for the aforesaid parameters). In the LBP groups, clinical information (pain, extensibility of the spinal and sub-pelvic muscles, sports practice) and sagittal radiological data were all measured. RESULTS: While no significant difference in isokinetic performance was found between asymptomatic and LBP children in the 11-to-13-year-old group, the isokinetic performances of the LBP children were influenced positively by BMI value, number of hours of physical activity and radiologic value of the lumbar lordosis. As regards these pre-teens, assessment with an isokinetic dynamometer does not highlight muscle characteristics that might explain LBP occurrence. As regards the 14-to-16-year-old group, muscle strength has been found to be correlated with age. LBP teens were showed to have weaker extensors and stronger flexors than the healthy teens. It is with regard to this age group that assessment with an isokinetic dynamometer clearly yields interesting results. Since we have yet to standardize our evaluation criteria (working speed, number of trials ), it is difficult to compare our results with those reported in the literature. CONCLUSION: This is a preliminary study involving a relatively low number of patients. That said, given the fact that numerous parameters are connected with the age and height of the subjects, assessment with an isokinetic dynamometer can be constructively carried out from the age of 14. In order to further enhance understanding of this phenomenon, a longitudinal and comparative study of a larger group is needed.
Assuntos
Músculos do Dorso/fisiopatologia , Dor Lombar/fisiopatologia , Força Muscular/fisiologia , Resistência Física/fisiologia , Adolescente , Fatores Etários , Criança , Feminino , Humanos , Masculino , Atividade Motora , Radiografia , Coluna Vertebral/diagnóstico por imagemRESUMO
Antibiotics, of which Fleming has identified the first representative, penicillin, in 1928, allowed dramatical improvement of the treatment of patients presenting with infectious diseases. However, once an antibiotic is used, resistance may develop more or less rapidly in some bacteria. It is thus necessary to develop therapeutic alternatives, such as the use of probiotics, defined by the World Health Organization (WHO) as "micro-organisms which, administered live and in adequate amounts, confer a benefit to the health of the host". The scope of these micro-organisms is broad, concerning many areas including that of infectious diseases, especially respiratory infections. We describe the rational use of probiotics in respiratory tract infections and detail the results of various clinical studies describing the use of probiotics in the management of respiratory infections such as nosocomial or community acquired pneumonia, or on specific grounds such as cystic fibrosis. The results are sometimes contradictory, but the therapeutic potential of probiotics seems promising. Implementing research to understand their mechanisms of action is critical to conduct therapeutic tests based on a specific rational for the strains to be used, the dose, as well as the chosen mode and rhythm of administration.
Assuntos
Pneumonia Bacteriana/terapia , Probióticos/uso terapêutico , Animais , Infecções Comunitárias Adquiridas/microbiologia , Infecções Comunitárias Adquiridas/terapia , Infecção Hospitalar/microbiologia , Infecção Hospitalar/terapia , Fibrose Cística/complicações , Suscetibilidade a Doenças , Método Duplo-Cego , Humanos , Sistema Imunitário/imunologia , Camundongos , Microbiota , Pneumonia Bacteriana/microbiologia , Pneumonia Associada à Ventilação Mecânica/microbiologia , Pneumonia Associada à Ventilação Mecânica/fisiopatologia , Pneumonia Associada à Ventilação Mecânica/terapia , Probióticos/efeitos adversos , Doença Pulmonar Obstrutiva Crônica/complicações , Percepção de Quorum , Ensaios Clínicos Controlados Aleatórios como Assunto , Sistema Respiratório/microbiologia , Especificidade da EspécieRESUMO
Antifungal lactic acid bacteria (ALAB) biodiversity was evaluated in raw milk from ewe, cow and goat over one year period. Lactic acid bacteria were enumerated using 8 semi-selective media, and systematically screened for their antifungal activity against 4 spoilage fungi commonly encountered in dairy products. Depending on the selective medium, between 0.05% (Elliker agar) and 5.5% (LAMVAB agar) screened colonies showed an antifungal activity. The great majority of these active colonies originated from cow (49%) and goat (43%) milks, whereas only 8% were isolated from ewe milk. Penicillium expansum was the most frequently inhibited fungus with 48.5% of colonies active against P. expansum among the 1235 isolated, followed by Mucor plumbeus with 30.6% of active colonies, Kluyveromyces lactis with only 12.1% of active colonies and Pichia anomala with 8.7% of active colonies. In the tested conditions, 94% of the sequenced active colonies belonged to Lactobacillus. Among them, targeted fungal species differed according to the Lactobacillus group, whose presence largely depended on year period and milk origin. The Lb. casei and Lb. reuteri groups, predominantly recovered in summer/fall, were overrepresented in the population targeting M. plumbeus, whereas isolates from the Lb. plantarum group, predominantly recovered in spring, were overrepresented in the population targeting K. lactis, the ones belonging to the Lb. buchneri group, predominantly recovered in spring, were overrepresented in the population targeting P. anomala. Raw milk, especially cow and goat milks from the summer/fall period appeared to be a productive reservoir for antifungal lactobacilli.
Assuntos
Biodiversidade , Lactobacillus/classificação , Leite/microbiologia , Animais , Antibiose , Bovinos , Contagem de Colônia Microbiana , Feminino , Fungos/classificação , Fungos/isolamento & purificação , Cabras , Kluyveromyces/classificação , Kluyveromyces/isolamento & purificação , Ácido Láctico , Lactobacillus/isolamento & purificação , Estações do Ano , OvinosRESUMO
UNLABELLED: : Thoracic hyperkyphosis is a frequent problem and can impact greatly on patient's quality of life during adolescence. This condition can be idiopathic or secondary to Scheuermann disease, a disease disturbing vertebral growth. To date, there is no sound scientific data available on the management of this condition. Some studies discuss the effects of bracing, however no guidelines, protocols or indication's of treatment for this condition were found. The aim of this paper was to develop and verify the consensus on managing thoracic hyperkyphosis patients treated with braces and/or physiotherapy. METHODS: The Delphi process was utilised in four steps gradually modified according to the results of a set of recommendations: we involved the SOSORT Board twice, then all SOSORT members twice, with a Pre-Meeting Questionnaire (PMQ), and during a Consensus Session at the SOSORT Lyon Meeting with a Meeting Questionnaire (MQ). RESULTS: There was an unanimous agreement on the general efficacy of bracing and physiotherapy for this condition. Most experts suggested the use of 4-5 point bracing systems, however there was some controversy with regards to physiotherapeutic aims and modalities. CONCLUSION: The SOSORT panel of experts suggest the use of rigid braces and physiotherapy to correct thoracic hyperkyphosis during adolescence. The evaluation of specific braces and physiotherapy techniques has been recommended.
RESUMO
AIMS: To evaluate the sensitivity of 21 common intestinal bacteria to six antibiotics and three broad-spectrum bacteriocins (nisins Z and A and pediocin PA-1). METHODS AND RESULTS: Neutralized cell-free culture supernatants containing active bacteriocins, and antibiotics were tested with the agar diffusion test and the disc-diffusion method, respectively. The tested intestinal strains showed high sensitivity to most antibiotics except for streptomycin and oxacillin. Nisins A and Z (8 mug per well) had similar activity spectra and inhibited all Gram-positive intestinal bacteria at different levels (except Streptococcus salivarius), with bifidobacteria (except Bifidobacterium breve and Bif. catenulatum), Collinsella aerofaciens and Eubacterium biforme being the most sensitive strains, but they were not active against Gram-negative bacteria. Surprisingly, none of the tested strains were inhibited by pediocin PA-1 (16 mug per well). CONCLUSION: Pediocin PA-1 which is very active against Listeria spp. and other food pathogens did not inhibit major intestinal species in the human intestine in contrast to both nisins A and Z. SIGNIFICANCE AND IMPACT OF THE STUDY: Our data suggest that pediocin PA-1 has potential to inhibit Listeria within the intestinal microbiota without altering commensal bacteria.
Assuntos
Antibacterianos/farmacologia , Bacteriocinas/farmacologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Nisina/análogos & derivados , Trato Gastrointestinal/microbiologia , Lactobacillus/efeitos dos fármacos , Listeria monocytogenes/efeitos dos fármacos , Nisina/farmacologia , PediocinasRESUMO
Bacteria isolated from infant feces were immobilized in polysaccharide gel beads (2.5% gellan gum, 0.25% xanthan gum) using a two-phase dispersion process. A 52-day continuous culture was carried out in a single-stage chemostat containing precolonized beads and fed with a medium formulated to approximate the composition of infant chyme. Different dilution rates and pH conditions were tested to simulate the proximal (PCS), transverse (TCS), and distal (DCS) colons. Immobilization preserved all nine bacterial groups tested with survival rates between 3 and 56%. After 1 week fermentation, beads were highly colonized with all populations tested (excepted Staphylococcus spp. present in low numbers), which remained stable throughout the 7.5 weeks of fermentation, with variations below 1 log unit. However, free-cell populations in the circulating liquid medium, produced by immobilized cell growth, cell-release activity from gel beads, and free-cell growth, were altered considerably by culture conditions. Compared to the stabilization period, PCS was characterized by a considerable and rapid increase in Bifidobacterium spp. concentrations (7.4 to 9.6 log CFU/mL), whereas Bifidobacterium spp., Lactobacillus spp., and Clostridium spp. concentrations decreased and Staphylococcus spp. and coliforms increased during TCS and DCS. Under pseudo-steady-state conditions, the community structure developed in the chemostat reflected the relative proportions of viable bacterial numbers and metabolites generally encountered in infant feces. This work showed that a complex microbiota such as infant fecal bacteria can be immobilized and used in a continuous in vitro intestinal fermentation model to reproduce the high bacterial concentration and bacterial diversity of the feces inoculum, at least at the genera level, with a high stability during long-term experiment.
Assuntos
Bactérias/metabolismo , Colo/fisiologia , Fezes/microbiologia , Modelos Biológicos , Análise de Variância , Ácidos Carboxílicos/metabolismo , Células Imobilizadas/fisiologia , Cromatografia Líquida de Alta Pressão , Colo/microbiologia , Meios de Cultura , Fermentação/fisiologia , Humanos , Concentração de Íons de Hidrogênio , LactenteRESUMO
Species-specific 16S rRNA-targeted, Cy3 (indocarbocyanine)-labeled oligonucleotide probes were designed and validated to quantify different Eubacterium species in human fecal samples. Probes were directed at Eubacterium barkeri, E. biforme, E. contortum, E. cylindroides (two probes), E. dolichum, E. hadrum, E. lentum, E. limosum, E. moniliforme, and E. ventriosum. The specificity of the probes was tested with the type strains and a range of common intestinal bacteria. With one exception, none of the probes showed cross-hybridization under stringent conditions. The species-specific probes were applied to fecal samples obtained from 12 healthy volunteers. E. biforme, E. cylindroides, E. hadrum, E. lentum, and E. ventriosum could be determined. All other Eubacterium species for which probes had been designed were under the detection limit of 10(7) cells g (dry weight) of feces(-1). The cell counts obtained are essentially in accordance with the literature data, which are based on colony counts. This shows that whole-cell in situ hybridization with species-specific probes is a valuable tool for the enumeration of Eubacterium species in feces.
Assuntos
Eubacterium/classificação , Eubacterium/isolamento & purificação , Fezes/microbiologia , RNA Ribossômico 16S/genética , Contagem de Colônia Microbiana , Eubacterium/genética , Eubacterium/crescimento & desenvolvimento , Humanos , Hibridização In Situ , Sondas de Oligonucleotídeos , Reação em Cadeia da Polimerase/métodos , Especificidade da EspécieRESUMO
AIMS: In vitro studies have suggested that fructo-oligosaccharides (FOS) and resistant starch (two fermentable non-digestible carbohydrates) display different fermentation kinetics. This study investigated whether these substrates affect the metabolic activity and bacterial composition of the intestinal microflora differently depending on the caecocolonic segment involved. METHODS AND RESULTS: Eighteen rats were fed a low-fibre diet (Basal) or the same diet containing raw potato starch (RPS) (9%) or short-chain FOS (9%) for 14 days. Changes in wet-content weights, bacterial populations and metabolites were investigated in the caecum, proximal and distal colon and faeces. Both substrates exerted a prebiotic effect compared with the Basal diet. However, FOS increased lactic acid-producing bacteria (LAPB) throughout the caecocolon and in faeces, whereas the effect of RPS was limited to the caecum and proximal colon. As compared with RPS, FOS doubled the pool of caecal fermentation products, while the situation was just the opposite distally. This difference was mainly because of the anatomical distribution of lactate, which accumulated in the caecum with FOS and in the distal colon with RPS. Faeces reflected these impacts only partly, showing the prebiotic effect of FOS and the metabolite increase induced by RPS. CONCLUSIONS: This study demonstrates that FOS and RPS exert complementary caecocolonic effects. SIGNIFICANCE AND IMPACT OF THE STUDY: The RPS and FOS combined ingestion could be beneficial by providing health-promoting effects throughout the caecocolon.
Assuntos
Bactérias Anaeróbias/isolamento & purificação , Intestino Grosso/microbiologia , Oligossacarídeos/farmacologia , Amido/farmacologia , Animais , Bactérias Anaeróbias/efeitos dos fármacos , Bactérias Anaeróbias/metabolismo , Ceco/microbiologia , Colo/microbiologia , Contagem de Colônia Microbiana , Fermentação/efeitos dos fármacos , Ácido Láctico/biossíntese , Masculino , Ratos , Ratos Wistar , Solanum tuberosumRESUMO
Some data suggest that the colonic microflora may adapt to produce more butyrate if given time and the proper substrate. To test this hypothesis, we investigated the effect of prolonged feeding of resistant potato starch on butyrate production. Rats were fed on either a low-fibre diet (basal) or the same diet supplemented with 90 g resistant potato starch/kg (PoS) for 0.5, 2 and 6 months. Short-chain fatty acid (SCFA) concentrations were determined in caecal and colonic contents at the end of each ingestion period. Total SCFA concentration increased over time throughout the caecocolonic tract with PoS, but was not modified with the basal diet. While propionate concentration was unchanged, butyrate concentration was highly increased by PoS at each time period in both the caecum and colon. Moreover, the butyrogenic effect of PoS increased over time, and the amount of butyrate was increased 6-fold in the caecum and proximal colon and 3-fold in the distal colon after 6 months compared with 0.5 months. Accordingly, the ratio butyrate:- total SCFA increased over time throughout the caecocolonic tract (12.6 (SE 2.8) v. 28 (SE 1.8)% in the caecum, 10.5 (SE 1.4) v. 26.8 (SE 0.9)% in the proximal colon, and 7.3 (SE 2.4) v. 23.9 (SE 2.7)% in the distal colon at 0.5 v. 6 months respectively), while the proportion of acetate decreased. Neither the proportion nor the concentration of butyrate was modified over time with the basal diet. Butyrate production was thus promoted by long-term ingestion of PoS, from the caecum towards the distal colon, which suggests that a slow adaptive process occurs within the digestive tract in response to a chronic load of indigestible carbohydrates.
Assuntos
Butiratos/metabolismo , Fibras na Dieta/administração & dosagem , Intestino Grosso/metabolismo , Solanum tuberosum , Amido/administração & dosagem , Animais , Ceco/metabolismo , Ceco/microbiologia , Colo/metabolismo , Colo/microbiologia , Contagem de Colônia Microbiana , Ácidos Graxos Voláteis/análise , Fezes/química , Fermentação , Intestino Grosso/microbiologia , Ácido Láctico/análise , Masculino , Ratos , Ratos Wistar , Fatores de TempoRESUMO
While the prebiotic effects of fructo-oligosaccharides (FOS), short-chain polymers of fructose, have been thoroughly described after 2-3 wk of ingestion, effects after intake for several months are unknown. We tested the hypothesis that these effects would differ after ingestion for short and long periods in rats. Rats were fed a basal low-fiber diet (Basal) or the same diet containing 9 g/100 g of FOS for 2, 8 or 27 wk, and cecal contents were collected at the end of each time period. Cecal short-chain fatty acid concentration was higher in rats fed FOS than in those fed Basal, and this effect persisted over time: 83.8 +/- 4.1 vs. 62.4 +/- 6.5 micromol/g at 2 wk and 103.5 +/- 5.8 vs. 73.2 +/- 7.4 micromol/g at 27 wk (P < 0.05). The molar butyrate ratio was higher in rats fed FOS regardless of the time period (14.8 +/- 0.6% vs. 6.7 +/- 1.1% at 27 wk, P < 0.05). Lactate concentration in rats fed FOS was elevated after 2 wk and then decreased: 63.5 +/- 21.6 micromol/g at 2 wk vs. 8.8 +/- 3.3 micromol/g at 8 wk (P < 0.05). After 2 wk, FOS increased the concentrations of total lactic acid-producing bacteria, and Lactobacillus sp. (P < 0.05), without modifying total anaerobes. However, most of these effects were abolished after 8 and 27 wk of FOS consumption. In the long term, the FOS-induced increase in intestinal lactic acid-producing bacteria was lost, but the butyrogenic properties of FOS were maintained.
Assuntos
Bactérias/crescimento & desenvolvimento , Bactérias/metabolismo , Butiratos/metabolismo , Ceco/metabolismo , Ácido Láctico/biossíntese , Oligossacarídeos/farmacologia , Animais , Contagem de Colônia Microbiana , Ácidos Graxos Voláteis/metabolismo , Concentração de Íons de Hidrogênio , Ácido Láctico/metabolismo , Masculino , Oligossacarídeos/administração & dosagem , Ratos , Ratos Wistar , Fatores de TempoRESUMO
Short-chain fatty acids are the main end products of bacterial fermentation of carbohydrates. Their role on the metabolism and biology of colonocytes is now well characterized. However, the functional consequences of their presence on intestinal smooth muscle cells remain poorly studied. We aimed to assess the effect of different short-chain fatty acids on ileal and colonic smooth muscle cells in primary culture and on A7R5 line. Butyrate (above 0.1 mM) inhibited A7R5 cell proliferation, while at low concentration (0.05 to 0.5 mM) butyrate significantly stimulated the proliferation of ileal and colonic myocytes in primary culture. An inhibition was observed at higher concentrations. Collagenous and noncollagenous protein synthesis was stimulated by butyrate. Moreover, butyrate stimulated actin and myosin expression. Thus, butyrate, which is produced by dietary fiber fermentation, may affect intestinal muscles by directly acting at the molecular level on myocytes.