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1.
Can J Microbiol ; 57(8): 629-37, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21823898

RESUMO

This work aimed to study the changes over time in the bacterial communities associated with the fluid and food particle fractions of the cow rumen following a change in diet. Four cannulated cows were fed a hay-based diet for 21 days and were then switched to a corn-silage-based diet for 33 days. The bacterial communities were regularly characterized by capillary electrophoresis - single-strand conformation polymorphism (CE-SSCP) and qPCR, and the main ruminal parameters were determined. The dietary change led to slight reductions in the diversity index, bacterial concentration, pH, and NH(3)-N concentration, and to an increase in the redox potential and volatile fatty acid concentrations. CE-SSCP profiles were not significantly affected by the dietary change but did change over time, with frequent fluctuations in both fluid and food particle fractions before and after the dietary change. The food particle fraction had a higher diversity index of bacterial community (+1.2 points, P < 0.001) and slightly more total bacteria than the fluid fraction of the rumen.


Assuntos
Bactérias/crescimento & desenvolvimento , Bovinos/microbiologia , Dieta/veterinária , Digestão/fisiologia , Rúmen/microbiologia , Amônia/análise , Animais , Bactérias/genética , Bovinos/fisiologia , Eletroforese Capilar , Ácidos Graxos Voláteis/análise , Feminino , Concentração de Íons de Hidrogênio , Nitrogênio/análise , Reação em Cadeia da Polimerase , Polimorfismo Conformacional de Fita Simples , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Rúmen/química , Silagem , Fatores de Tempo
2.
J Appl Microbiol ; 107(5): 1642-50, 2009 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-19457023

RESUMO

AIMS: Improved knowledge of the bacterial community of the digestive tract is required to enhance the efficiency of digestion in herbivores. This work aimed to study spatial and temporal variations of the bacterial communities in the bovine digestive tract and their correlation with gut environmental parameters. METHODS AND RESULTS: Rumen content and faeces of five cows were sampled for 3 weeks. In addition, reticulum content was sampled during the third week. Bacterial communities were assessed by studying capillary electrophoresis single-stranded conformation polymorphism (CE-SSCP) profiles of 16S rRNA genes. The bacterial community structure differed between the forestomach and faecal contents. The abundance of several operational taxonomic units changed from week to week. Bacterial community structure of the rumen was correlated to propionic acid and NH(3)-N concentrations. CONCLUSIONS: The bacterial community of the bovine digestive tract varied in space and time. SIGNIFICANCE AND IMPACT OF THE STUDY: The study of the bacterial communities of the digestive tract in herbivores should be widened from the rumen to the large intestine. The amplitude and origin of the temporal variation of the ruminal bacterial community need to be better understood to improve the control of the fermentative activity in herbivores.


Assuntos
Bactérias/genética , Bactérias/isolamento & purificação , Bovinos/microbiologia , Trato Gastrointestinal/microbiologia , RNA Ribossômico 16S/análise , Animais , Derivados de Benzeno/análise , Biodiversidade , DNA Bacteriano/análise , Eletroforese Capilar , Fezes/microbiologia , Reação em Cadeia da Polimerase/métodos , Polimorfismo Conformacional de Fita Simples , Propionatos/análise , Retículo/microbiologia , Rúmen/química , Rúmen/microbiologia
3.
Int J Data Min Bioinform ; 9(4): 417-23, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25757248

RESUMO

Oligonucleotide sets are widely used in molecular biology to target a group of nucleic acid sequences using Polymerase Chain Reaction (PCR)-based technologies. Currently, the global matching efficiency of an oligonucleotide set is considered to be equal to the lower matching efficiency calculated for each oligonucleotide. However, sequences matching the limiting oligonucleotide did not always match the other oligonucleotide of the set, resulting in a biased evaluation of the matching efficiency. The OligoSpecificitySystem program avoid this bias by calculations of the real global matching efficiency of oligonucleotide sets. It can process all kinds of oligonucleotide sets, including the number of oligonucleotides, base pair degeneracy occurrences or mismatch occurrences.


Assuntos
Pareamento Incorreto de Bases , Biologia Computacional/métodos , Oligonucleotídeos/genética , Algoritmos , Bacteroides/genética , Hibridização de Ácido Nucleico , Reação em Cadeia da Polimerase em Tempo Real , Reprodutibilidade dos Testes
4.
Animal ; 5(11): 1761-8, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22440416

RESUMO

This work aimed to study the response of the growing rabbit caecal ecosystem (bacterial community and caecal environmental parameters) after a switch from a control to a low-fibre diet (LFD). A group of 160 rabbits were fed ad libitum a control diet (ADF: 20.4%) from weaning (36 days). At 49 days of age (day 0), 75 rabbits were switched to a LFD group (ADF: 10.7%), whereas 85 others (control group) remained on the control diet, for 39 days. Caecal contents were regularly sampled throughout the trial (60 rabbits per group). The bacterial community structure was characterized using CE-SSCP (capillary electrophoresis single strand conformation polymorphism) and total bacteria were quantified using real-time PCR. Redox potential (Eh), pH, NH(3)-N, volatile fatty acid (VFA) were measured in the caecum to characterize environmental parameters. The reduction of fibre in the diet modified the CE-SSCP profiles (P < 0.001) but not the diversity index (5.6 ± 0.8, ns). The number of 16S rRNA gene copies of total bacteria decreased (P < 0.01) in LFD rabbits compared with controls. In LFD rabbits, the caecal environment was less acid (+0.2 units; P < 0.01), more reductive (-11 mV; P < 0.05) and drier (+3.4 g 100 per g; P < 0.001), with an increase in NH3-N (+77%; P < 0.001) and a decrease in total VFA concentration (-17%; P < 0.001). We found significant correlations between the bacterial community, the quantity of bacteria and the caecal traits of the caecal ecosystem. Indeed, in both groups, the caecal traits barely constrained the total inertia of the CE-SSCP profile set (less than 14%), whereas total bacteria were positively related to total VFA, acetic acid and butyric acid levels, and Eh, and negatively related to pH. All the microbial and environmental modifications had occurred by day 2 and remained stable thereafter. These results suggest that the bacterial community in the growing rabbit caecum is able to adapt quickly after a change to in the dietary fibre supply to reach a new steady-state equilibrium.

5.
Mol Ecol Resour ; 9(5): 1359-63, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21564907

RESUMO

Molecular fingerprint methods are widely used to compare microbial communities in various habitats. The free program StatFingerprints can import, process, and display fingerprint profiles and perform numerous statistical analyses on them, and also estimate diversity indexes. StatFingerprints works with the free program R, providing an environment for statistical computing and graphics. No programming knowledge is required to use StatFingerprints, thanks to its friendly graphical user interface. StatFingerprints is useful for analysing the effect of a controlled factor on the microbial community and for establishing the relationships between the microbial community and the parameters of its environment. Multivariate analyses include ordination, clustering methods and hypothesis-driven tests like 50-50 multivariate analysis of variance, analysis of similarity or similarity percentage procedure and the program offers the possibility of plotting ordinations as a three-dimensional display.

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