Identification and clustering of dairy propionibacteria by RAPD-PCR and CGE-REA methods

Rossl, F.; Torriani, Sandra; Dellaglio, Franco

doi:10.1111/j.1365-2672.1998.tb05259.x

Cited by 50 publications

(42 citation statements)

References 17 publications

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“…(27); P2, 5Ј ATGTAACGCC 3Ј (26); P3, 5Ј CTGCGGCAT 3Ј (11); P4, 5Ј CCGCAGCGTT 3Ј (11); P5, 5Ј TGCTCTGCCC 3Ј (46); P6 5Ј GTCCACACGG 3Ј (46); P7, 5Ј AGCAGCGTGG 3Ј (30); P8, 5Ј CGTACAGGCT 3Ј; P9, 5Ј TCACCGTCGC 3Ј; and P10, 5Ј ACTGGCTCCG 3Ј. Each reaction mixture contained each 2Ј-deoxynucleoside 5Ј-triphosphate at a concentration of 200 M, 1 M primer, 1.5 mM MgCl 2 , 1.25 U of Taq DNA polymerase (Life Technologies), 2.5 l of PCR buffer, 25 ng of DNA, and enough sterile bidistilled water to bring the volume to 25 l. The PCR program comprised 45 cycles of denaturation for 1 min at 94°C, annealing for 1 min at 35°C, and extension for 2 min at 72°C; the cycles were preceded by denaturation at 94°C for 4 min and were followed by extension at 72°C for 5 min (40).…”

Section: Methodsmentioning

confidence: 99%

“…Moreover, the resolving power of this method can be easily enhanced by increasing the number of primers used to randomly amplify the bacterial genome (46). RAPD analysis has been used to estimate the diversity of Lactobacillus strains in the Centre National de Recherches Zootechniques collection (46), to type strains of L. plantarum (31), Lactococcus lactis isolated from raw milk used to produce Camembert (33), and dairy propionibacteria (40), to establish the correct nomenclature and classification of strains of L. casei subsp. casei (10), and to study the population of NSLAB in mature commercial cheese (11).…”

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confidence: 99%

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Characterization of Non-Starter Lactic Acid Bacteria from Italian Ewe Cheeses Based on Phenotypic, Genotypic, and Cell Wall Protein Analyses

Angelis

Corsetti

Tosti

et al. 2001

Appl Environ Microbiol

179

120

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Non-starter lactic acid bacteria (NSLAB) were isolated from 12 Italian ewe cheeses representing six different types of cheese, which in several cases were produced by different manufacturers. A total of 400 presumptive Lactobacillus isolates were obtained, and 123 isolates and 10 type strains were subjected to phenotypic, genetic, and cell wall protein characterization analyses. Phenotypically, the cheese isolates included 32% Lactobacillus plantarum isolates, 15% L. brevis isolates, 12% L. paracasei subsp. paracasei isolates, 9% L. curvatus isolates, 6% L. fermentum isolates, 6% L. casei subsp. casei isolates, 5% L. pentosus isolates, 3% L. casei subsp. pseudoplantarum isolates, and 1% L. rhamnosus isolates. Eleven percent of the isolates were not phenotypically identified. Although a randomly amplified polymorphic DNA (RAPD) analysis based on three primers and clustering by the unweighted pair group method with arithmetic average (UPGMA) was useful for partially differentiating the 10 type strains, it did not provide a species-specific DNA band or a combination of bands which permitted complete separation of all the species considered. In contrast, sodium dodecyl sulfate-polyacrylamide gel electrophoresis cell wall protein profiles clustered by UPGMA were species specific and resolved the NSLAB. The only exceptions were isolates phenotypically identified as L. plantarum and L. pentosus or as L. casei subsp. casei and L. paracasei subsp. paracasei, which were grouped together. Based on protein profiles, Italian ewe cheeses frequently contained four different species and 3 to 16 strains. In general, the cheeses produced from raw ewe milk contained a larger number of more diverse strains than the cheeses produced from pasteurized milk. The same cheese produced in different factories contained different species, as well as strains that belonged to the same species but grouped in different RAPD clusters.Non-starter lactic acid bacteria (NSLAB) usually increase from a low number in fresh curd to dominate the microflora of mature cheese (36). In contrast to starters, NSLAB tolerate the hostile environment of cheese during ripening; this environment typically is characterized by 32 to 39% moisture, 4 to 6% salt in moisture, pH 4.9 to 5.3, 5 to 13°C, and a deficiency of nutrients (15,48). Mesophilic lactobacilli predominate in the NSLAB community, although pediococci and micrococci may also be found (6,11,15). Lactobacillus casei subsp. casei, L. casei subsp. pseudoplantarum, Lactobacillus paracasei subsp. paracasei, and Lactobacillus plantarum are the most frequently isolated taxa, but other facultatively and obligately heterofermentative species of lactobacilli are also found (18, 28). Adventitious mesophilic lactobacilli are usually present because of postpasteurization contamination but may also constitute part of the raw milk microflora and survive pasteurization (48).The role of NSLAB in ripening has not been resolved satisfactorily yet, although inclusion of adjunct cultures of some strains of NSLAB or use of ra...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Characterization of Non-Starter Lactic Acid Bacteria from Italian Ewe Cheeses Based on Phenotypic, Genotypic, and Cell Wall Protein Analyses

Angelis

Corsetti

Tosti

et al. 2001

Appl Environ Microbiol

179

120

View full text Add to dashboard Cite

show abstract

“…Strain typing of Staphylococcus spp. was performed by pulsed-field gel electrophoresis; comparison of Propionibacterium acnes strains was performed by random amplification of polymorphic DNA-PCR (RAPD-PCR) with primer OLP-05 as described by Rossi et al [12,13].…”

Section: Identification Of Bacteria and Strain Typingmentioning

confidence: 99%

Screening of platelet concentrates for bacterial contamination: spectrum of bacteria detected, proportionof transfused units, and clinical follow-up

et al. 2009

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“…Different fingerprinting methods such as SDS-PAGE of whole cell proteins [23], 16s rDNA targeted PCR-RFLP [24], ribotyping [25], 16S-23S ribosomal spacer amplification and restriction [26], Pulsed-Field Gel Electrophoresis [27], Conventional Gel Electrophoresis Restriction Endonuclease Analysis (CGE-REA) and Randomly Amplified Polymorphic DNA-PCR [28] have been used for detection and accurate identification of dairy propionibacteria from various environments like milk, cheese, whey and flour. Genus and species-specific primers targeted to the genes encoding 16S rRNA for PCR-based assays were also designed for detection of dairy propionibacteria [29].…”

Section: Introductionmentioning

confidence: 99%

Dairy Propionibacteria: Less Conventional Probiotics to Improve the Human and Animal Health

Zárate¹

2012

Probiotic in Animals

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Identification and clustering of dairy propionibacteria by RAPD-PCR and CGE-REA methods

Abstract: F. ROSSI

Cited by 50 publications

References 17 publications

Characterization of Non-Starter Lactic Acid Bacteria from Italian Ewe Cheeses Based on Phenotypic, Genotypic, and Cell Wall Protein Analyses

Characterization of Non-Starter Lactic Acid Bacteria from Italian Ewe Cheeses Based on Phenotypic, Genotypic, and Cell Wall Protein Analyses

Screening of platelet concentrates for bacterial contamination: spectrum of bacteria detected, proportionof transfused units, and clinical follow-up

Dairy Propionibacteria: Less Conventional Probiotics to Improve the Human and Animal Health

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