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RAPD and REA for Characterization of Lactobacillus plantarum strains, and Enterococcus spp. with Special Reference to Enterococcus spp. from Meat and Humans

Quednau, Mikael LU (1998)
Abstract
Restriction Endonuclease Analysis (REA) was used to characterize strains of Lactobacillus plantarum, Lb. pentosus, and Lb. reuteri. This method was shown to be a powerful tool for separation at the species and strain level. RAPD analysis of raw cell abstracts was used to characterize strains of Lb. plantarum. This method was shown to be useful for separation of most strains, and the results were obtained within one working day.



The species distribution and antibiotic resistance profile of 277 Enterococcus strains from Swedish and Danish retailed pork and chicken were investigated. Enterococcus faecium was the most common species in Swedish and Danish chicken, followed by E. faecalis. E. faecalis was the dominating... (More)
Restriction Endonuclease Analysis (REA) was used to characterize strains of Lactobacillus plantarum, Lb. pentosus, and Lb. reuteri. This method was shown to be a powerful tool for separation at the species and strain level. RAPD analysis of raw cell abstracts was used to characterize strains of Lb. plantarum. This method was shown to be useful for separation of most strains, and the results were obtained within one working day.



The species distribution and antibiotic resistance profile of 277 Enterococcus strains from Swedish and Danish retailed pork and chicken were investigated. Enterococcus faecium was the most common species in Swedish and Danish chicken, followed by E. faecalis. E. faecalis was the dominating species in Danish and Swedish pork. The number of unidentified Enterococcus isolates, was highest in Swedish pork, and lowest in Swedish chicken.



Fourteen percent of the Enterococcus isolates from Danish pork were resistant to one or more of the tested antibiotics. The values for Danish chicken, Swedish pork, and Swedish chicken were 55%, 9%, and 73%, respectively. The most common antibiotic resistance in Danish pork and Swedish chicken was tetracycline. Erythromycin resistance was the most common in Danish chicken. In the Swedish pork, only five resistant isolates were found, resistant to erythromycin, norfloxacin, penicillin V, tetracycline, and trimethoprim/sulfa, respectively. Vancomycin resistant strains, harboring the vanA gene, were found only in Danish meat, and with one exception, only in chicken. One vancomycin resistant isolate from Danish chicken was identified as E. durans, one as E. hirae, one as belonging to the E. faecium group (E. faecium, E. durans, E. hirae, and E. mundtii), and the isolate from Danish pork could not be identified to the species level. The rest were identified as E. faecium.



The use of RAPD analysis, partial 16S rDNA sequence analysis including the variable regions V4 and V9, and Enterococcus specific 16S rDNA hybridization analysis in the identification and classification of enterococci from clinical isolates was compared. 16S rDNA sequencing was more reproducible than RAPD, whereas RAPD was easier to perform, and had a somewhat higher discriminatory power.



RAPD was used to identify clinically relevant species of Enterococcus. It was shown to be a useful way to rapidly identify the two clinically most important species of Enterococcus, i.e. E. faecalis and E. faecium, and species specific profiles were also obtained for other Enterococcus species.



REA was used to characterize Enterococcus faecium strains from chicken, pork, and humans, together with probiotic E. faecium strains. Except for two probiotic strains, all tested strains were clearly separated using REA.



Enterococcus faecium strains cluster according to their source of isolation, indicating that strains from animals do not spread to humans, and that different strains are adapted to their respective source. The number of isolates included in the study was rather limited, however, so the existence of such a spread cannot be ruled out. (Less)
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author
supervisor
opponent
  • Prof Dellaglio, Franco, University of Verona, Italy
organization
publishing date
type
Thesis
publication status
published
subject
keywords
Food and drink technology, pork, chicken, antibiotic resistance, Lactobacillus plantarum, Enterococcus, REA, RAPD, Livsmedelsteknik
pages
98 pages
publisher
Göran Molin, Dept of Food Technology, Chemical Center, Lund University
defense location
Room A, Chemical Center, LTH
defense date
1998-03-13 10:15:00
external identifiers
  • other:ISRN: LUTKDH/TKL0-1033/1-98
language
English
LU publication?
yes
additional info
Article: Johansson, M-L., Quednau, M., Ahrné, S., and Molin, G. (1995) Classification of Lactobacillus plantarum by restriction endonuclease analysis (REA) of total chromosomal DNA, using conventional agarose gel electrophoresis, Int J Syst Bact 45(4), 670-675. Article: Johansson, M-L., Quednau, M., Ahrné, S. and Molin, G. (1995) Randomly amplified polymorphic DNA (RAPD) for rapid typing of Lactobacillus plantarum strains, Lett Appl Microbiol 21(3), 155-159. Article: Quednau, M., Ahrné, S., and Molin, G. Antibiotic resistant strains of Enterococcus isolated from Swedish and Danish retailed chicken and pork, J Appl Bact (in press) Article: Monstein, H-J., Quednau, M., Samuelsson, A., Ahrné, S., Isaksson, B., and Jonasson, J. Division of the genus Enterococcus into species groups using PCR-based molecular typing methods, Microbiology (in press) Article: Quednau, M., Ahrné, S., and Molin, G. Identification of clinically important species of Enterococcus within one day using Randomly Amplified Polymorphic DNA (RAPD), Current Microbiology (in press) Article: Quednau, M., Ahrné, S., and Molin, G. The use of restriction endonuclease analysis (REA) for separation of Enterococcus faecium at the strain level. (submitted)
id
e29e4068-3c72-41c4-a253-81ffee513c3e (old id 38445)
date added to LUP
2016-04-04 11:43:31
date last changed
2018-11-21 21:06:47
@phdthesis{e29e4068-3c72-41c4-a253-81ffee513c3e,
  abstract     = {{Restriction Endonuclease Analysis (REA) was used to characterize strains of Lactobacillus plantarum, Lb. pentosus, and Lb. reuteri. This method was shown to be a powerful tool for separation at the species and strain level. RAPD analysis of raw cell abstracts was used to characterize strains of Lb. plantarum. This method was shown to be useful for separation of most strains, and the results were obtained within one working day.<br/><br>
<br/><br>
The species distribution and antibiotic resistance profile of 277 Enterococcus strains from Swedish and Danish retailed pork and chicken were investigated. Enterococcus faecium was the most common species in Swedish and Danish chicken, followed by E. faecalis. E. faecalis was the dominating species in Danish and Swedish pork. The number of unidentified Enterococcus isolates, was highest in Swedish pork, and lowest in Swedish chicken.<br/><br>
<br/><br>
Fourteen percent of the Enterococcus isolates from Danish pork were resistant to one or more of the tested antibiotics. The values for Danish chicken, Swedish pork, and Swedish chicken were 55%, 9%, and 73%, respectively. The most common antibiotic resistance in Danish pork and Swedish chicken was tetracycline. Erythromycin resistance was the most common in Danish chicken. In the Swedish pork, only five resistant isolates were found, resistant to erythromycin, norfloxacin, penicillin V, tetracycline, and trimethoprim/sulfa, respectively. Vancomycin resistant strains, harboring the vanA gene, were found only in Danish meat, and with one exception, only in chicken. One vancomycin resistant isolate from Danish chicken was identified as E. durans, one as E. hirae, one as belonging to the E. faecium group (E. faecium, E. durans, E. hirae, and E. mundtii), and the isolate from Danish pork could not be identified to the species level. The rest were identified as E. faecium.<br/><br>
<br/><br>
The use of RAPD analysis, partial 16S rDNA sequence analysis including the variable regions V4 and V9, and Enterococcus specific 16S rDNA hybridization analysis in the identification and classification of enterococci from clinical isolates was compared. 16S rDNA sequencing was more reproducible than RAPD, whereas RAPD was easier to perform, and had a somewhat higher discriminatory power.<br/><br>
<br/><br>
RAPD was used to identify clinically relevant species of Enterococcus. It was shown to be a useful way to rapidly identify the two clinically most important species of Enterococcus, i.e. E. faecalis and E. faecium, and species specific profiles were also obtained for other Enterococcus species.<br/><br>
<br/><br>
REA was used to characterize Enterococcus faecium strains from chicken, pork, and humans, together with probiotic E. faecium strains. Except for two probiotic strains, all tested strains were clearly separated using REA.<br/><br>
<br/><br>
Enterococcus faecium strains cluster according to their source of isolation, indicating that strains from animals do not spread to humans, and that different strains are adapted to their respective source. The number of isolates included in the study was rather limited, however, so the existence of such a spread cannot be ruled out.}},
  author       = {{Quednau, Mikael}},
  keywords     = {{Food and drink technology; pork; chicken; antibiotic resistance; Lactobacillus plantarum; Enterococcus; REA; RAPD; Livsmedelsteknik}},
  language     = {{eng}},
  publisher    = {{Göran Molin, Dept of Food Technology, Chemical Center, Lund University}},
  school       = {{Lund University}},
  title        = {{RAPD and REA for Characterization of Lactobacillus plantarum strains, and Enterococcus spp. with Special Reference to Enterococcus spp. from Meat and Humans}},
  year         = {{1998}},
}