scholarly journals Analysis of Serpulina hyodysenteriae strain variation and its molecular epidemiology using pulsed-field gel electrophoresis

1999 ◽  
Vol 123 (1) ◽  
pp. 133-138 ◽  
Author(s):  
R. F. ATYEO ◽  
S. L. OXBERRY ◽  
D. J. HAMPSON
Keyword(s):  
Molecular Epidemiology ◽  
Gel Electrophoresis ◽  
Restriction Endonuclease Analysis ◽  
Pulsed Field Gel Electrophoresis ◽  
Enzyme Electrophoresis ◽  
Swine Dysentery ◽  
Pulsed Field ◽  
Serpulina Hyodysenteriae ◽  
Single Pattern ◽  
Endonuclease Analysis

Pulsed-field gel electrophoresis (PFGE) was applied as a molecular typing tool for the spirochaete Serpulina hyodysenteriae, the agent of swine dysentery. Analysis of a collection of 40 mainly Australian isolates, previously characterized by other methods, divided these into 23 PFGE types. This confirmed that there are many strains of the spirochaete in Australia. PFGE was more discriminatory for strain typing than both multilocus enzyme electrophoresis and serotyping. It had similar discriminatory power to restriction endonuclease analysis, but the results of PFGE were easier to interpret. When applied to 29 isolates collected from 4 farms over periods of up to 8 years, 2 PFGE patterns were found on 3 farms, and a single pattern on the other. In each case a new strain had apparently emerged as a variant of an original parent strain. PFGE was found to be a powerful technique for investigating the molecular epidemiology of swine dysentery outbreaks.

Monitoring lactic acid bacteria strains during ‘Cacioricotta’ cheese production by restriction endonuclease analysis and pulsed-field gel electrophoresis

2001 ◽  
Vol 68 (1) ◽  
pp. 139-144 ◽  
Author(s):  
GIUSEPPE BLAIOTTA ◽  
GIANCARLO MOSCHETTI ◽  
ERMENEGILDA SIMEOLI ◽  
ROSAMARIA ANDOLFI ◽  
FRANCESCO VILLANI ◽  
...  
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Restriction Endonuclease ◽  
Gel Electrophoresis ◽  
Water Buffalo ◽  
Restriction Endonuclease Analysis ◽  
Buffalo Milk ◽  
Pulsed Field Gel Electrophoresis ◽  
Pulsed Field ◽  
Endonuclease Analysis

Lactic acid bacteria (LAB) play an important role in food fermentation, as the products obtained with their aid are characterized by hygienic safety, storage stability and attractive sensory properties. A major aim of the research in this field is the selection of LAB strains that could be used. Hence it is very important to be able to apply a reliable method to distinguish a particular strain specifically and unambiguously, which allows studies of population dynamics of mixed cultures and monitoring starter strains during fermentation (Ramos & Harlander, 1990).Molecular methods are a powerful alternative to the traditional differentiation of bacteria. A highly reproducible method for characterizing and distinguishing closely related strains, is represented by REA-PFGE (restriction endonuclease analysis by pulsed-field gel electrophoresis) performed by infrequently cutting endonucleases. Genetic differentiation of strains in several species of LAB has been successfully performed by this technique (Moschetti et al. 1997; Villani et al. 1997) obtaining very clear and reproducible restriction patterns (Moschetti et al. 1998).In this study selected inoculated strains (lactococci or lactobacilli) were monitored by REA-PFGE during the whole process of water-buffalo ‘cacioricotta’ cheese-making. This product is a typical and traditional cheese in southern Italy produced from cow, goat, ewe or water-buffalo milk. The original technology of this preparation permits the recovery of whey proteins due to the high heat treatment of whole milk employed, allowing interesting yields to be achieved in terms of cheese. The use of starters is not common in traditional technology but low acid protection of the final product suggested the use of LAB as starter (Emaldi et al. 1987).

scholarly journals Comparison of Genomic Methods for Differentiating Strains of Enterococcus faecium: Assessment Using Clinical Epidemiologic Data

1998 ◽  
Vol 36 (11) ◽  
pp. 3327-3331 ◽  
Author(s):  
Connie Savor ◽  
Michael A. Pfaller ◽  
Julie A. Kruszynski ◽  
Richard J. Hollis ◽  
Gary A. Noskin ◽  
...  
Keyword(s):  
Enterococcus Faecium ◽  
Gel Electrophoresis ◽  
Genomic Dna ◽  
Restriction Endonuclease Analysis ◽  
Pulsed Field Gel Electrophoresis ◽  
Agarose Gel Electrophoresis ◽  
Subspecies Level ◽  
Vancomycin Resistant ◽  
Ideal Method ◽  
Endonuclease Analysis

Genomic DNA extracted from 45 vancomycin-resistantEnterococcus faecium (VRE) isolates was cleaved withHindIII and HaeIII and subjected to agarose gel electrophoresis. The ability of this method (restriction endonuclease analysis [REA]) to distinguish strains at the subspecies level was compared with results previously determined by pulsed-field gel electrophoresis (PFGE). Chart reviews were performed to provide a clinical correlation of possible epidemiologic relatedness. A likely clinical association was found for 29 patients as part of two outbreaks. REA found 21 of 21 isolates were the same type in the first outbreak, with PFGE calling 19 strains the same type. In the second outbreak with eight patient isolates, HindIII found six were the same type and two were unique types. HaeIII found three strains were the same type, two strains were a separate type, and three more strains were unique types, while PFGE found three were the same type and five were unique types. No single “ideal” method can be used without clinical epidemiologic investigation, but any of these techniques is helpful in providing focus to infection control practitioners assessing possible outbreaks of nosocomial infection.

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