Characterization of Lactobacillus sake strains associating with production of ropy slime by randomly amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) patterns

Björkroth, Johanna; Ridell, Jouko; Korkeala, Hannu

doi:10.1016/0168-1605(96)00964-6

Cited by 66 publications

(27 citation statements)

References 16 publications

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“…Such experimental variation is well documented for RAPD PCR (11) and limits both the degree of strain discrimination and the establishment of reference patterns that can be used to verify and track specific strains. Under the conditions used in the present study, PFGE was found to discriminate more strains than RAPD PCR and this agrees with the findings presented for a number of other genera (4,6,41). The level of discrimination achieved across the genus agreed well with that previously reported for 67 P. acidilactici and P. pentosaceus strains isolated from human clinical samples that produced 59 distinct macrorestriction patterns following digestion of genomic DNA with either SmaI or NotI (3).…”

Section: Discussionsupporting

confidence: 81%

Genomic Diversity within the Genus Pediococcus as Revealed by Randomly Amplified Polymorphic DNA PCR and Pulsed-Field Gel Electrophoresis

Simpson

Stanton

Fitzgerald

et al. 2002

Appl Environ Microbiol

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The genomic diversity of 33 previously assigned strains from six species within the genus Pediococcus was assessed by randomly amplified polymorphic DNA (RAPD) PCR and pulsed-field-gel electrophoresis (PFGE). The RAPD PCR patterns produced by two separate random primers, termed P1 (ACGCGCCCT) and P2 (ATGTAACGCC), were compared by the Pearson correlation coefficient and the unweighted pair group method with arithmetic averages clustering algorithm. Pattern variations between repeat samples set a strain discrimination threshold of less than 70% similarity. P1 and P2 primers alone and in combination produced 14, 21, and 28 distinct patterns, respectively. When each strain was assigned with a type strain with which it shared the highest level of similarity, both primers grouped 17 of the 27 strains to their proposed species. PFGE following genomic digestion with the restriction enzymes ApaI, NotI, and AscI produced 30, 32, and 28 distinct macrorestriction patterns, respectively. Specific DNA fragments within the NotI and AscI macrorestriction patterns for each strain were observed that allowed 27 of the 33 strains to be assigned to their proposed species. For example, following digestion with AscI, all Pediococcus parvulus strains were characterized by two DNA fragments, one of approximately 220 kb and another between 700 and 800 kb. The exceptions correlated with those observed with both RAPD PCR primers and included three P. damnosus and two P. pentosaceus strains that grew at temperatures regarded as nonpermissive for their proposed species but not for those with which they grouped.

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Section: Discussionsupporting

confidence: 81%

Genomic Diversity within the Genus Pediococcus as Revealed by Randomly Amplified Polymorphic DNA PCR and Pulsed-Field Gel Electrophoresis

Simpson

Stanton

Fitzgerald

et al. 2002

Appl Environ Microbiol

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“…PFGE is considered to offer the highest resolution for strain differentiation of lactic acid bacteria (4,7,22) and was in this study applied to a selection of probiotic and human strains to verify whether members of a given FAFLP cluster represent different strains or may be considered clones of the same parent strain. Because PFGE is a laborious and expensive method, only a limited number of strains were analyzed.…”

Section: Discussionmentioning

confidence: 99%

Intraspecific Genotypic Characterization of Lactobacillus rhamnosus Strains Intended for Probiotic Use and Isolates of Human Origin

Vancanneyt¹,

Huys

Lefebvre³

et al. 2006

Appl Environ Microbiol

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A set of 118 strains of the species Lactobacillus rhamnosus was collected, including probiotic strains, research strains with potential probiotic properties, food starter cultures, and human isolates. The majority of the strains were collected from companies, hospitals, or culture collections or were obtained after contacting authors who reported clinical case studies in the literature. The present work aimed to reveal the genotypic relationships between strains of these diverse sources. All strains were initially investigated using fluorescent amplified fragment length polymorphism (FAFLP) with three different primer combinations. Numerical analysis of FAFLP data allowed (i) confirmation of the identification of all strains as members of L. rhamnosus and (ii) delineation of seven stable intraspecific FAFLP clusters. Most of these clusters contained both (potentially) probiotic strains and isolates of human origin. For each of the clusters, strains of different sources were selected for pulsed-field gel electrophoresis (PFGE) of macrorestriction fragments obtained with the enzymes NotI and AscI. Analysis of PFGE data indicated that (i) some (potentially) probiotic strains were indistinguishable from other probiotic strains, suggesting that several companies may use duplicate cultures of the same probiotic strain, and (ii) in a number of cases human isolates from sterile body sites were indistinguishable from a particular probiotic strain, suggesting that some of these isolates may be reisolations of commercial strains.Lactobacillus species are members of the commensal microflora in the oral cavity and the gastrointestinal and genitourinary system in humans and animals and are usually considered nonpathogenic (1). Selected strains of some Lactobacillus species are being used as probiotics and are claimed to influence the health of the host in a beneficial manner (10). To a very limited extent, strains belonging to the same Lactobacillus species as those used in probiotic applications have also been isolated from clinical specimens and have been associated with rare cases of endocarditis, meningitis, deep abscesses, and bacteremia (1, 5, 11). It should be noted that in these types of infections underlying disease or immunosuppression is a common feature and that infection in previously healthy humans is extremely rare.Among the lactobacilli, strains of Lactobacillus rhamnosus are commonly used as probiotics including the GG (GorbachGoldin) strain, which is one of the most widely used and welldocumented probiotics (13, 25). On the other hand, L. rhamnosus has also been reported as one of the most common Lactobacillus taxa in human clinical cases (5). Harty et al. (8,9) identified possible virulence factors in L. rhamnosus endocarditis strains compared to laboratory or oral isolates. Furthermore, L. rhamnosus strains have been shown to produce enzymes enabling the breakdown of glycoproteins and synthesis or lysis of fibrin clots (18).Aggregate epidemiologic studies have not found a link between the number of clinica...

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“…Of these methods, PFGE has been shown to be highly effective in epidemiological studies (Hudson et al 2001;Murase et al 1995;Seo et al 2006). A number of researchers have suggested that this genotyping method could provide a reproducible subdivision of Lactobacillus and that it is in fact the most discriminatory method (Björkroth et al 1996;Tynkkynen et al 1999;Vandamme et al 1996).…”

Section: Introductionmentioning

confidence: 97%

PFGE genotyping and antibiotic resistance of Lactobacillus distributed strains in the fermented dairy products

Guo

Wang

et al. 2011

Ann Microbiol

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The aim of this study was to analyze the antibiotic resistance and pulsed-field gel electrophoresis (PFGE) genotypes of Lactobacillus strains present in fermented dairy products available on the Chinese market and to analyze the correlation between these. A total of 33 Lactobacillus strains identified in fermented dairy products were tested for resistance to 16 antibiotics using the broth microdilution method and also analyzed by PFGE. Almost all of the strains were multidrug resistant and showed high resistance rates to fosfomycin (97.0%), chloramphenicol (87.9%), vancomycin (87.9%), ceftriaxone (81.8%), imipenem (66.7%), trimethoprim/sulfamethoxazone (54.6%), tetracycline (33.3%), cefotaxime (21.2%), gentamicin (18.2%), erythromycin (12.1%), gatifloxacin (12.1%), amoxicillin/clavulanic acid (12.1%), rifampin (9.1%), and clindamycin (9.1%). Based on the PFGE results, the 33 strains were subtyped into 17 pulsotypes (PFPs, pulsed-field profiles), of which six were represented by more than one strain each. In conclusion, the Lactobacillus strains identified here in fermented dairy products available on the Chinese market showed a high rate of antibiotic resistance. Lactobacillus strains were genotyped well by PFGE; the resistance patterns of the strains among the same PFP were not necessarily similar to each other.

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Characterization of Lactobacillus sake strains associating with production of ropy slime by randomly amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) patterns

Cited by 66 publications

References 16 publications

Genomic Diversity within the Genus Pediococcus as Revealed by Randomly Amplified Polymorphic DNA PCR and Pulsed-Field Gel Electrophoresis

Genomic Diversity within the Genus Pediococcus as Revealed by Randomly Amplified Polymorphic DNA PCR and Pulsed-Field Gel Electrophoresis

Intraspecific Genotypic Characterization of Lactobacillus rhamnosus Strains Intended for Probiotic Use and Isolates of Human Origin

PFGE genotyping and antibiotic resistance of Lactobacillus distributed strains in the fermented dairy products

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