Bacteriophage ecology in commercial sauerkraut fermentations - PubMed (original) (raw)
Bacteriophage ecology in commercial sauerkraut fermentations
Z Lu et al. Appl Environ Microbiol. 2003 Jun.
Abstract
Knowledge of bacteriophage ecology in vegetable fermentations is essential for developing phage control strategies for consistent and high quality of fermented vegetable products. The ecology of phages infecting lactic acid bacteria (LAB) in commercial sauerkraut fermentations was investigated. Brine samples were taken from four commercial sauerkraut fermentation tanks over a 60- or 100-day period in 2000 and 2001. A total of 171 phage isolates, including at least 26 distinct phages, were obtained. In addition, 28 distinct host strains were isolated and identified as LAB by restriction analysis of the intergenic transcribed spacer region and 16S rRNA sequence analysis. These host strains included Leuconostoc, Weissella, and Lactobacillus species. It was found that there were two phage-host systems in the fermentations corresponding to the population shift from heterofermentative to homofermentative LAB between 3 and 7 days after the start of the fermentations. The data suggested that phages may play an important role in the microbial ecology and succession of LAB species in vegetable fermentations. Eight phage isolates, which were independently obtained two or more times, were further characterized. They belonged to the family Myoviridae or Siphoviridae and showed distinct host ranges and DNA fingerprints. Two of the phage isolates were found to be capable of infecting two Lactobacillus species. The results from this study demonstrated for the first time the complex phage ecology present in commercial sauerkraut fermentations, providing new insights into the bioprocess of vegetable fermentations.
Figures
FIG. 1.
Flow diagram for analyses of a brine sample and isolation of phages and their hosts in a sauerkraut fermentation studied in 2000.
FIG. 2.
Microbial counts in a commercial sauerkraut fermentation.
FIG. 3.
Changes in the concentrations of substrates and products and pH in a commercial sauerkraut fermentation.
FIG. 4.
Restriction profiles of the ITS PCR product obtained from 10 phage hosts. Lane 1, L. mesenteroides 1-A4; lane 2, L. mesenteroides 1-F8; lane 3, L. mesenteroides 3-A4; lane 4, L. mesenteroides 3-B1; lane 5, L. pseudomesenteroides 3-B11; lane 6, L. brevis 7-E1; lane 7, L. plantarum 14-C8; lane 8, L. plantarum 22-D10; lane 9, L. fallax 3-G1; lane 10, Weissella strain 3-H2; lane M, a 100 bp DNA ladder.
FIG. 5.
Electron micrographs of eight phages isolated from commercial sauerkraut fermentations. CsCl-purified phage preparations were negatively stained with 2% uranyl acetate (pH 4.0). (a) φ1-A4; (b) φ1-F8; (c) φ3-A4; (d) φ3-B1; (e) φ3-B11; (f) φ7-E1; (g) φ14-C8; (h) φ22-D10. Magnification, ×50,000.
FIG. 6.
SDS-PAGE analysis of structural proteins from eight phages isolated from a commercial sauerkraut fermentation. Lane M, molecular mass markers; lane 1, φ1-A4; lane 2, φ1-F8; lane 3, φ3-A4; lane 4, φ3-B1; lane 5, φ3-B11; lane 6, φ7-E1; lane 7, φ14-C8; lane 8, φ22-D10.
FIG. 7.
_Hin_dIII restriction digestion analysis of DNAs from eight phages isolated from a commercial sauerkraut fermentation. Lane 1, φ1-A4; lane 2, φ1-F8; lane 3, φ3-A4; lane 4, φ3-B1; lane 5, φ3-B11; lane 6, φ7-E1; lane 7, φ14-C8; lane 8, φ22-D10; lane M, 1-kb DNA ladder.
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