[Enterotoxin-producing Bacteroides fragilis strains isolated from horses].
Abstract: Seven Bacteroides fragilis strains were cultured from samples collected from horses. From all the tested strains, as well as from the reference B. fragilis strains: enterotoxigenic NCTC 11925 and nonenterotoxigenic IPL 323 strain, DNA was isolated using Genomic DNA PREP PLUS isolation kit manufactured by A&A Biotechnology (Poland). To detect the enterotoxin (fragilysin) gene, polymerase chain reaction (PCR) was applied, using the following starters: 404 (GAG CCG AAG ACG GTG TAT GTG ATT TGT) and 407 (TGC TCA GCG CCC AGT ATA TGA CCT AGT). DNA obtained from bacterial cells was amplified in a thermocycler (Techne). The temperature profile was as follows: 1 cycle (4 min. 94 degrees C), 40 cycles (1 min. 94 degrees C, 1 min. 52 degrees C, 1 min. 74 degrees C). Amplification products were detected by electrophoresis in agarose gel (1%) with ethidium bromide added. The presence of the fragilysin gene was detected in two strains. Among the strains isolated from horses enterotoxin gene-possessing Bacteroides fragilis strains (ETBF) can be detected.
Publication Date: 2002-01-05 PubMed ID: 11757425
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- English Abstract
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research article is about the isolation of seven strains of Bacteroides fragilis from horses, two of which were found to have the enterotoxin (fragilysin) gene.
Introduction and methodology
- The research is aimed at determining the presence of enterotoxin producing Bacteroides fragilis strains in horses.
- These harmful bacteria can cause intestinal infections that can lead to diarrhea and other intestinal disorders.
- The study started by culturing seven different strains of Bacteroides fragilis from horses.
- The DNA from the cultured strains, including reference strains of B. fragilis (enterotoxigenic NCTC 11925 and nonenterotoxigenic IPL 323), was isolated using a Genomic DNA PREP PLUS isolation kit made by A&A Biotechnology (Poland).
Detection of Enterotoxin (Fragilysin) Gene
- To determine the presence of the enterotoxin (fragilysin) gene in the B. fragilis strains, they used Polymerase Chain Reaction (PCR).
- PCR is a technique used in molecular biology to amplify a single copy or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence.
- The PCR process was initiated with specific primers (starters), and the DNA obtained from the bacterial cells was amplified using a thermocycler.
- The temperature profile used was: 1 cycle (4 minutes at 94 degrees Celsius), 40 cycles (1 minute at 94 degrees Celsius, 1 minute at 52 degrees Celsius, 1 minute at 74 degrees Celsius).
Results and conclusions
- The output of PCR was detected by performing electrophoresis in agarose gel (1%) with ethidium bromide added.
- This process helps visualize DNA fragments based on their size and charge.
- The discovery concludes that the presence of the fragilysin gene, the enterotoxin, was detected in two out of seven of the tested B. fragilis strains.
- Thus, it was concluded that enterotoxin gene-possessing Bacteroides fragilis strains (ETBF) could be found in the strains isolated from horses.
Cite This Article
APA
Obuch-Woszczatyński P, Pituch H, Martirosian G, Silva J, Meisel-Mikołajczyk F, Łuczak M.
(2002).
[Enterotoxin-producing Bacteroides fragilis strains isolated from horses].
Med Dosw Mikrobiol, 53(2), 161-166.
Publication
Researcher Affiliations
- Katedra i Zakład Mikrobiologii Lekarskiej Centrum Biostruktury, Akademia Medyczna w Warszawie.
MeSH Terms
- Animals
- Bacteroides fragilis / genetics
- Bacteroides fragilis / isolation & purification
- Bacteroides fragilis / metabolism
- Bone and Bones / microbiology
- DNA, Bacterial / isolation & purification
- Enterotoxins / biosynthesis
- Horses
- Intestines / microbiology
- Lung / microbiology
- Lymph Nodes / microbiology
- Metalloendopeptidases / analysis
- Tendons / microbiology
Citations
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