[Show abstract][Hide abstract] ABSTRACT: To reduce high-salt waste from cucumber fermentations, low-salt fermentations are under development. These fermentations may
require the use of starter cultures to ensure normal fermentations. Because potential phage infection can cause starter culture
failure, it is important to understand phage ecology in the fermentations. This study investigated the phage ecology in a
commercial cucumber fermentation. Brine samples taken from a fermentation tank over a 90-day period were plated onto deMan-Rogosa-Sharpe
agar plates. A total of 576 lactic acid bacterial isolates were randomly selected to serve as potential hosts for phage isolation.
Filtered brine served as a phage source. Fifty-seven independent phage isolates were obtained, indicating that 10% of the
bacterial isolates were sensitive to phage attack. Phage hosts include Lactobacillus brevis (67% of all hosts), Lactobacillus plantarum (21%), Weissella paramesenteroides, Weissella cibaria, and Pediococcus ethanolidurans. Nearly 50% of phages were isolated on day 14, and the majority of them attacked L. brevis. Some phages had a broad host range and were capable of infecting multiple hosts in two genera. Other phages were species
specific or strain specific. About 30% of phage isolates produced turbid pinpoint plaques or only caused reduced cell growth
on the bacterial lawns. Six phages with distinct host ranges were characterized. The data from this study showed that abundant
and diverse phages were present in the commercial cucumber fermentation, which could cause significant mortality to the lactic
acid bacteria population. Therefore, a phage control strategy may be needed in low-salt cucumber fermentations.
[Show abstract][Hide abstract] ABSTRACT: Vegetable fermentations rely on the proper succession of a variety of lactic acid bacteria (LAB). Leuconostoc mesenteroides initiates fermentation. As fermentation proceeds, L. mesenteroides dies off and other LAB complete the fermentation. Phages infecting L. mesenteroides may significantly influence the die-off of L. mesenteroides. However, no L. mesenteroides phages have been previously genetically characterized. Knowledge of more phage genome sequences may provide new insights into phage genomics, phage evolution, and phage-host interactions. We have determined the complete genome sequence of L. mesenteroides phage Phi1-A4, isolated from an industrial sauerkraut fermentation. The phage possesses a linear, double-stranded DNA genome consisting of 29,508 bp with a G+C content of 36%. Fifty open reading frames (ORFs) were predicted. Putative functions were assigned to 26 ORFs (52%), including 5 ORFs of structural proteins. The phage genome was modularly organized, containing DNA replication, DNA-packaging, head and tail morphogenesis, cell lysis, and DNA regulation/modification modules. In silico analyses showed that Phi1-A4 is a unique lytic phage with a large-scale genome inversion ( approximately 30% of the genome). The genome inversion encompassed the lysis module, part of the structural protein module, and a cos site. The endolysin gene was flanked by two holin genes. The tail morphogenesis module was interspersed with cell lysis genes and other genes with unknown functions. The predicted amino acid sequences of the phage proteins showed little similarity to other phages, but functional analyses showed that Phi1-A4 clusters with several Lactococcus phages. To our knowledge, Phi1-A4 is the first genetically characterized L. mesenteroides phage.
[Show abstract][Hide abstract] ABSTRACT: The extent of glucose and fructose utilization during cucumber juice fermentation was affected differentially by the addition of 10 to 360 mM of selected anions (chloride, nitrate, sulfate, phosphate, acetate, lactate, and citrate) and cations (Na+, K+, NH4+, Ca2+, Mg2+, and Mn2+). Inorganic anions generally suppressed utilization of both sugars, particularly at higher concentrations. Acetate and lactate increased utilization of fructose, but not glucose, while citrate increased utilization of both sugars. Of the cations tested, 10-60 mM Mn+2 significantly (P < 0.05) increased utilization of both sugars, but higher concentrations reduced utilization, as compared with the control. Evidence indicates that brine composition can significantly influence sugar utilization during cucumber fermentation, and may be important in developing controlled fermentation strategies for brined cucumbers.
[Show abstract][Hide abstract] ABSTRACT: Glucose (32 mM) and fructose (36 mM) of cucumber juice were degraded simultaneously by Lactobacillus plantarum, but at different rates and extents. Glucose depletion was slightly more rapid than fructose during the exponential growth phase, but slower thereafter and stopped before exhaustion. In contrast, fructose degradation continued until all naturally present fructose was exhausted. When cucumber juice was supplemented with fructose and/or glucose, the starter culture continued to ferment fructose, but not glucose, resulting in an increase in lactic acid production and a decrease in terminal pH. Fructose utilization was not affected by the presence of glucose, but the presence of fructose reduced glucose utilization.
[Show abstract][Hide abstract] ABSTRACT: The complete genomic sequence of a Lactobacillus plantarum virulent phage PhiJL-1 was determined. The phage possesses a linear, double-stranded, DNA genome consisting of 36,677 bp with a G+C content of 39.36%. A total of 52 possible open reading frames (ORFs) were identified. According to N-terminal amino acid sequencing and bioinformatic analyses, proven or putative functions were assigned to 21 ORFs (41%), including 5 structural protein genes. The PhiJL-1 genome shows functionally related genes clustered together in a genome structure composed of modules for DNA replication, DNA packaging, head and tail morphogenesis, and lysis. This type of modular genomic organization was similar to several other phages infecting lactic acid bacteria. The structural gene maps revealed that the order of the head and tail genes is highly conserved among the genomes of several Siphoviridae phages, allowing the assignment of probable functions to certain uncharacterized ORFs from phage PhiJL-1 and other Siphoviridae phages.
[Show abstract][Hide abstract] ABSTRACT: A virulent Lactobacillus plantarum bacteriophage, phiJL-1, was isolated from a commercial cucumber fermentation. The phage was specific for two related strains of L. plantarum, BI7 and its mutant (deficient in malolactate fermenting ability) MU45, which have been evaluated as starter cultures for controlled cucumber fermentation and as biocontrol microorganisms for minimally processed vegetable products. The phage genome of phiJL-1 was sequenced to reveal a linear, double-stranded DNA (36.7 kbp). Sodium dodecyl sulfate-polyacryamide gel electrophoresis (SDS-PAGE) profiles indicated that phiJL-1 contains six structural proteins (28, 34, 45, 50, 61, and 76 kDa). Electron microscopy revealed that the phage has an isometric head (59 nm in diameter), a long non-contractile tail (182 nm in length and 11 nm in width), and a complex base plate. The phage belongs to the Bradley group B1 or Siphoviridae family. One-step growth kinetics of the phage showed that the latent period was 35 min, the rise period was 40 min, and the average burst size was 22 phage particles/infected cell. Phage particles (90%) adsorbed to the host cells 20 min after infection. Calcium supplementation (up to 30 mM CaCl2) in MRS media did not affect the first cycle of phage adsorption, but promoted rapid phage propagation and cell lysis in the infection cycle subsequent to adsorption. The D values of phiJL-1 at pH 6.5 were estimated to be 2.7 min at 70 degrees C and 0.2 min at 80 degrees C by a thermal inactivation experiment. Knowledge of the properties of L. plantarum bacteriophage phiJL-1 may be important for the development of controlled vegetable fermentations.
International Journal of Food Microbiology 08/2003; 84(2). DOI:10.1016/S0168-1605(03)00111-9 · 3.08 Impact Factor
[Show abstract][Hide abstract] 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.
[Show abstract][Hide abstract] ABSTRACT: : The composition of pickling cucumbers varied with fruit size, which affected buffer capacity, sugar utilization, terminal pH, and texture of the fermented fruit. We found that as cucumber size increased (from less than 27 to 51 mm in dia), malic acid, pH, buffer capacity, and dry matter content decreased, and glucose and fructose contents increased. Fruit firmness and bloater damage were greater in large than in small, fermented, whole cucumbers. Blanching (75 °C, 30 s) had little effect on the fermentation and prevention of bloater formation in finished products, regardless of fruit sizes. It was demonstrated that cucumber juice can serve as a model system for studying the metabolic, but not the physical (texture, bloater damage), consequences of lactic acid bacteria chosen as starter cultures for cucumber fermentation.