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PubMed2003

Reclassification of Lactobacillus maltaromicus (Miller et al. 1974) DSM 20342(T) and DSM 20344 and Carnobacterium piscicola (Collins et al. 1987) DSM 20730(T) and DSM 20722 as Carnobacterium maltaromaticum comb. nov.

Phenotypic and genotypic characterizations of Lactobacillus maltaromicus strains DSM 20342(T) and DSM 20344 provided evidence for the reclassification of this species in the genus Carnobacterium. Moreover, phenotypic and genotypic comparisons made between L. maltaromicus and Carnobacterium piscicola highlighted that these two species should be considered synonyms. For these reasons, the species Carnobacterium maltaromaticum comb. nov. (type strain DSM 20342(T) = ATCC 27865(T) = CCUG 30142(T) = CIP 103135(T) = JCM 1154(T) = LMG 6903(T) = NRRL B-14852(T)) is proposed to accommodate L. maltaromicus and C. piscicola.

Bacterial Typing Techniques↗
PubMed2003

Marinilactibacillus psychrotolerans gen. nov., sp. nov., a halophilic and alkaliphilic marine lactic acid bacterium isolated from marine organisms in temperate and subtropical areas of Japan.

A novel marine lactic acid rod bacterium has been described for eight strains isolated from living and decomposing marine organisms collected from temperate and subtropical areas of Japan. The isolates were Gram-positive, catalase-negative, non-sporulating and motile with peritrichous flagella. They were slightly halophilic, highly halotolerant and alkaliphilic; the optimum NaCl concentration for growth was 2.0-3.75% (w/v) with a range from 0 to 17.0-20.5% (depending on the strain); the optimum pH was between 8.0 and 9.5 with a range from 6.0 to 10.0. They were psychrotolerant, growing well at -1.8 degrees C with a maximum at 40-45 degrees C and the optimum at 37-40 degrees C. Lactate yields were 87-100% per consumed glucose; the residual products were formate, acetate and ethanol with a molar ratio of approximately 2 : 1 : 1. The product composition was markedly affected by the pH of fermentation medium; at higher pH, the yield of lactate decreased (60-65% at pH 9.0) and that of other products increased conversely. The cell-wall peptidoglycan type was type A4beta, Orn-D-Glu, whereas that of the genus Alkalibacterium, the phylogenetically closest lactic acid bacterium, was type A4beta, Orn-D-Asp. The major cellular fatty acids were C16 : 0, C16 : 1delta9, C18 : 0 and C18 : 1delta9 (oleic acid). The G + C content of the DNA was 34.6-36.2 mol%. The eight isolates were phenotypically homogeneous and formed a single genomic species. The 16S rRNA gene sequence analysis indicated that the isolates constituted an independent phylogenetic lineage within the radiation of lactic acid bacteria with 96.2% similarity to the genus Alkalibacterium. The secondary structure and the nucleotide sequence of the V6 region of the 16S rRNA were characteristic of the organism among other related lactic acid genera. On the bases of phenotypic and phylogenetic distinctness, the organism was proposed to belong to a new genus and species, Marinilactibacillus psychrotolerans gen. nov., sp. nov. The type strain, M13-2(T) (G + C = 36.2 mol%), has been deposited in the IAM, NBRC, NCIMB and NRIC culture collections as IAM 14980(T), NBRC 100002(T), NCIMB 13873(T) and NRIC 0510(T), respectively.

Animals↗
PubMed1985

Classification of some lactic acid bacteria from vacuum-packed meats by direct probe mass spectrometry.

The technique of direct probe mass spectrometry (DPMS) has been applied to the classification of 40 strains of lactic acid bacteria from refrigerated vacuum-packed beef, pork, lamb and bacon. Relationships between strains were examined by multi-variate statistical techniques using sets of ions selected for reproducibility and sample discrimination. Five groups were distinguished which corresponded closely to those detected in a previous numerical taxonomic study. Two groups contained all 12 representatives of a cluster of unidentifiable non-aciduric streptobacteria whose sub-division is supported by other taxonomic evidence. All twenty-one strains from a cluster of aciduric streptobacteria provisionally identified with Lactobacillus sake were contained in two further groups. The sub-division of these aciduric strains revealed by DPMS has not been verified by other techniques and requires further investigation. The fifth group contained Leuconostoc strains. The study demonstrates the value of DPMS in confirming and clarifying classification schemes obtained by conventional methods.

Food Microbiology↗
PubMed1991

Establishment in the piglet gut of lactobacilli capable of degrading mixed-linked beta-glucans.

The establishment in piglets of lactobacilli with ability to degrade mixed-linked (1----3), (1----4) beta-D-glucans was studied in faeces from 15 animals. The piglets had free access to creep feed with an estimated content of 2% beta-D-glucans from 5 days of age. On days 3 and 35, ca log 8 cfu/g of beta-glucan-degrading lactobacilli were found in a majority of the samples. On days 7, 14 and 21 such bacteria were only found occasionally. During establishment of the microflora in the neonate, the faecal lactobacilli of the piglet seemed related to those of the sow. Later, the metabolic activity of the lactobacilli in piglet faeces showed a connection to the composition of the diet. The possible relation of these bacteria to occurrence of beta-glucanases attributed to be endogenous in the pig is discussed.

Analysis of Variance↗
PubMed1992

Use of the polymerase chain reaction and oligonucleotide probes for the rapid detection and identification of Carnobacterium species from meat.

The polymerase chain reaction (PCR) was used selectively to amplify specific rDNA sequences of Carnobacterium divergens, C. mobile, C. piscicola and C. gallinarum in purified DNA extracts, crude cell lysates and food samples. The PCR products were visualized by agarose gel electrophoresis and identified, at species level, by hybridization reactions with three specific oligonucleotide probes for C. divergens, C. mobile and C. piscicola/C. gallinarum designed from 16S rRNA sequence data. The PCR was sufficiently sensitive to amplify DNA from a single bacterium to detectable levels after 30 cycles of amplification. Both radioactive (32P) and non-radioactive alkaline phosphatase labelled probes was able to detect the PCR products. Detection was highly specific and the probes did not hybridize with DNA samples from any other of the bacterial species tested. These methods enabled the rapid and specific detection and identification of carnobacteria from pure cultures and samples of meat.

Base Sequence↗
PubMed1992

Isolation and properties of a bacteriocin-producing Carnobacterium piscicola isolated from fish.

A facultative psychrotrophic lactic acid bacterium isolated from fresh fish was identified as Carnobacterium piscicola on the basis of carbohydrate utilization, G + C content and 16S RNA analysis. Its bacteriocin, designated carnocin UI49, is produced during the mid-exponential phase of growth at temperatures between 15 degrees C and 34 degrees C. Carnocin UI49 is active against a large number of closely-related lactic acid bacteria including carnobacteria, lactobacilli, pediococci and lactococci. Furthermore, the bacteriocin has a bactericidic mode of action which results in lysis of sensitive cells. Maximum bactericidal activity is observed at 34 degrees C with a decrease in activity down to 15 degrees C where it is completely abolished.

Animals↗
PubMed2004

Simultaneous detection of Carnobacterium and Leuconostoc in meat products by multiplex PCR.

AIMS: To develop a multiplex PCR approach for simultaneous detection of Leuconostoc and Carnobacterium and its validation in meat products. METHODS AND RESULTS: Two multiplex PCR assays were developed using newly designed 16S rDNA-directed primers adapted to the current taxonomic situation of genera Leuconostoc and Carnobacterium that allow: (i) simultaneous detection of both genera, and members of the nonmotile species of genus Carnobacterium and (ii) identification in a single assay of the nonmotile species C. divergens, C. maltaromicum and C. gallinarum. Sensitivity values of 10(3) and 10(4) CFU g(-1) were determined for multiplex PCR detection of Carnobacterium and Leuconostoc, respectively, following artificially inoculated meat trials. In addition, both multiplex PCR assays were validated in 14 naturally contaminated samples covering nine types of meat products. Results obtained by colony identification were confirmed by PCR detection. CONCLUSIONS: The methods described in this study provide a rapid and reliable tool for PCR detection of Carnobacterium and Leuconostoc, in meat products, and for colony identification. SIGNIFICANCE AND IMPACT OF THE STUDY: This multiplex PCR approach will help in the analysis of the spoilage microbiota of refrigerated vacuum-packaged meat product in order to determine the appropriate preservation method.

Animals↗
PubMed2006

Isolation and characterization of lactic acid bacteria from suan-tsai (fermented mustard), a traditional fermented food in Taiwan.

AIMS: To isolate, characterize, and identify lactic acid bacteria (LAB) in suan-tsai (fermented mustard), a traditional fermented food in Taiwan. METHODS AND RESULTS: Suan-tsai samples were collected at five time points from a fixed fermenting bucket. Fifty cultures were isolated from suan-tsai samples, and isolates were divided into classes by phenotype and then into groups by restriction-fragment length polymorphism analysis and sequencing of 16S ribosomal DNA. Phenotypic and biochemical characteristics identified two different bacterial groups (A and B), and the results showed that Pediococcus pentosaceus was the most abundant LAB during the initial fermentation time. However, the more NaCl-tolerant species Tetragenococcus halophilus took the place of P. pentosaceus and became the most abundant LAB later. All isolates were grown in de Man, Rogosa, and Sharpe (MRS) broth containing 6% NaCl, but T. halophilus could grow only in MRS broth containing 10% NaCl. CONCLUSIONS: These results suggest that the LAB P. pentosaceus and T. halophilus play roles in the fermentation of suan-tsai. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report describing the distribution and varieties of LAB that exist in the suan-tsai fermentation process.

Colony Count, Microbial↗
PubMed2006

Degradation of free and sulfur-dioxide-bound acetaldehyde by malolactic lactic acid bacteria in white wine.

AIMS: Acetaldehyde is the major carbonyl compound formed during winemaking and has implications for sensory and colour qualities of wines as well as for the use of the wine preservative SO(2). The current work investigated the degradation of acetaldehyde and SO(2)-bound acetaldehyde by two commercial Oenococcus oeni starters in white wine. METHODS AND RESULTS: Wines were produced by alcoholic fermentation with commercial yeast and adjusted to pH 3.3 and 3.6. While acetaldehyde was degraded rapidly and concurrently with malic acid at both pH values, SO(2)-bound acetaldehyde caused sluggish bacterial growth. Strain differences were small. CONCLUSIONS: Efficient degradation of acetaldehyde can be achieved by commercial starters of O. oeni. According to the results, the degradation of acetaldehyde could not be separated from malolactic conversion by oenococci. While this may be desirable in white winemaking, it may be necessary to delay malolactic fermentation (MLF) in order to allow for colour development in red wines. SO(2)-bound acetaldehyde itself maybe responsible for the sluggish or stuck MLF, and thus bound SO(2) should be considered next to free SO(2) in order to evaluate malolactic fermentability. SIGNIFICANCE AND IMPACT OF THE STUDY: The current study provides new results regarding the metabolism of acetaldehyde and SO(2)-bound acetaldehyde during the MLF in white wine. The information is of significance to the wine industry and may contribute to reducing the concentration of wine preservative SO(2).

Acetaldehyde↗
PubMed2006

Purification and characterization of two bacteriocins produced by lactic acid bacteria isolated from Mongolian airag.

AIMS: The aim of this study was to isolate and identify bacteriocin-producing lactic acid bacteria (LAB) issued from Mongolian airag (traditional fermented mare's milk), and to purify and characterize bacteriocins produced by these LAB. METHODS AND RESULTS: Identification of the bacteria (Enterococcus durans) was carried out on the basis of its morphological, biochemical characteristics and carbohydrate fermentation profile and by API50CH kit and 16S rDNA analyses. The pH-neutral cell-free supernatant of this bacterium inhibited the growth of several Lactobacillus spp. and food-borne pathogens including Escherichia coli, Staphylococcus aureus and Listeria innocua. The antimicrobial agent (enterocin A5-11) was heat stable and was not sensitive to acid and alkaline conditions (pH 2-10), but was sensitive to several proteolytic enzymes. Its inhibitory activity was completely eliminated after treatment with proteinase K and alpha-chymotrypsin. The activity was however not completely inactivated by other proteases including trypsin and pepsin. Three-step purification procedure with high recovery yields was developed to separate two bacteriocins. The applied procedure allowed the recovery of 16% and 64% of enterocins A5-11A and A5-11B, respectively, present in the culture supernatant with purity higher than 99%. SDS-PAGE analyses revealed that enterocin A5-11 has a molecular mass of 5000 Da and mass spectrometry analyses demonstrates molecular masses of 5206 and 5218 Da for fractions A and B, respectively. Amino acid analyses of both enterocins indicated significant quantitative difference in their contents in threonine, alanine, isoleucine and leucine. Their N-termini were blocked hampering straightforward Edman degradation. CONCLUSIONS: Bacteriocins A5-11A and B from Ent. durans belong to the class II of bacteriocins. SIGNIFICANCE AND IMPACT OF THE STUDY: Judging from molecular masses, amino acid composition and spectrum of activities, bacteriocins A5-11A and B from Ent. durans show high degree of similarity with enterocins L50A and L50B isolated from Enterococcus faecium (Cintas et al. 1998, 2000) and with enterocin I produced by Ent. faecium 6T1a, a strain originally isolated from a Spanish-style green olive fermentation (Floriano et al. 1998).

Amino Acids↗
PubMed2006

Impact of winemaking practices on arginine and citrulline metabolism during and after malolactic fermentation.

AIMS: To study arginine degradation and carcinogenic ethyl carbamate precursor citrulline formation during and after malolactic fermentation (MLF). METHODS AND RESULTS: MLF was induced in white wine with two commercial Oenococcus oeni strains under different winemaking conditions regarding the type of alcoholic fermentation (spontaneous, induced) and the lees management (racked, on lees). Arginine degradation and citrulline formation did not occur during malic acid degradation in any treatment. In five of the six treatments in which arginine degradation took place, it occurred 3 weeks after malic acid depletion and significant amounts of citrulline were formed. Presence of yeast lees in wines led to increased citrulline formation. CONCLUSIONS: This study suggests that arginine metabolism is inhibited in oenococci at low pH values (< 3.5) and that in the postalcoholic fermentation phase, citrulline formation from arginine degradation can be avoided if MLF is induced by pure cultures of O. oeni with inhibition of the bacterial biomass after malic acid depletion. Residual yeast lees in the wine have been identified as a significant risk factor for increased citrulline formation. SIGNIFICANCE AND IMPACT OF THE STUDY: Conclusions drawn from this study allow reducing the risk of carcinogenic ethyl carbamate formation from citrulline excretion by wine lactic acid bacteria.

Arginine↗
PubMed2007

The microbiological quality of hot water-washed broccoli florets and cut green beans.

AIMS: To determine the effect of hot water washing on the microbiological quality of cut broccoli florets and trimmed green beans. METHODS AND RESULTS: Broccoli florets and trimmed beans were washed for 90 s in tap water at either 20 degrees C or 52 degrees C and stored at 7 and 10 degrees C. The numbers of naturally occurring aerobic mesophilic organisms, Pseudomonas spp., Enterobacteriaceae, yeast and moulds and lactobacilli or lactic acid bacteria were enumerated at intervals for up to 2 weeks. The ability of Listeria monocytogenes, Bacillus cereus and Escherichia coli O157:H7 inoculated onto the tissue post heat treatment to survive or grow was also measured to mimic the effect of postprocess contamination. Using a hot wash treatment improved the initial appearance of the vegetables and resulted in a small, but significant, reduction in populations of all groups of endogenous flora measured. The number of yeast and moulds on the vegetables washed at 52 degrees C remained below the levels observed on the 20 degrees C washed vegetables throughout the observation period, but Pseudomonas spp., lactobacilli and Enterobacteriaceae were better able to grow on the hot-washed vegetables such that the counts at the end of storage were greater on hot-washed than ambient-washed vegetables. All three of the pathogens tested were better able to grow on hot-washed broccoli and beans than on equivalent product washed at 20 degrees C. CONCLUSIONS: Hot water washing can be used to control enzymic browning or yeast and moulds growth but it can also allow more rapid and extensive growth by pathogens and spoilage organisms. SIGNIFICANCE AND IMPACT OF THE STUDY: Reduced protection against growth by pathogens means that the hot wash treatment of vegetables should be used with caution and requires careful assessment of risk.

Bacillus cereus↗
PubMed1994

Identification of Carnobacterium spp. and Leuconostoc spp. in meat by genus-specific 16S rRNA probes.

Oligonucleotide probes specific for Carnobacterium and Leuconostoc species were constructed from the variable regions of 16S rRNA obtained from the literature and sequence data bases. The probes were hybridized with crude nucleic acid extract from 32 type strains of lactic acid bacteria (LAB) commonly found on meat. Two of the probes hybridized only to the four Carnobacterium species whereas the other two hybridized only to five of the six Leuconostoc species tested. The probes were also hybridized with nucleic acids from unknown strains of LAB. The identification was consistent with the results of biochemical tests used to characterize the two genera.

Animals↗
PubMed2007

Vanillin production from simple phenols by wine-associated lactic acid bacteria.

AIMS: The ability of lactic acid bacteria (LAB) to metabolize certain phenolic precursors to vanillin was investigated. METHODS AND RESULTS: Gas chromatography-mass spectrometry (GC-MS) or HPLC was used to evaluate the biosynthesis of vanillin from simple phenolic precursors. LAB were not able to form vanillin from eugenol, isoeugenol or vanillic acid. However Oenococcus oeni or Lactobacillus sp. could convert ferulic acid to vanillin, but in low yield. Only Lactobacillus sp. or Pediococcus sp. strains were able to produce significant quantities of 4-vinylguaiacol from ferulic acid. Moreover, LAB reduced vanillin to the corresponding vanillyl alcohol. CONCLUSIONS: The transformation of phenolic compounds tested by LAB could not explain the concentrations of vanillin observed during LAB growth in contact with wood. SIGNIFICANCE AND IMPACT OF THE STUDY: Important details of the role of LAB in the conversion of phenolic compounds to vanillin have been elucidated. These findings contribute to the understanding of malolactic fermentation in the production of aroma compounds.

Benzaldehydes↗
PubMed2005

Statistical aspects in physical mapping application to the genome of O. oeni strain GM.

This work addresses issues around physical maps, in particular, for circular genomes. The overlapping relationship between two fragments obtained by applying two different restriction enzymes, separately, is classified as nonoverlapping, partial overlapping, and total overlapping. A double partial overlapping can also appear in a particular situation. Taking into account DNA fragment lengths and under the assumption that the left-hand endpoints of the two restriction fragments are independent random variables, each of which with a uniform distribution along a circular genome, we present expressions for prior probabilities of those events. This information is combined with hybridization data via Bayes' theorem, in order to evaluate corresponding posterior probabilities. Additionally, we explore a sensitivity analysis to quantify the effect of length variation in the results.

Algorithms↗
PubMed2000

Effect of Alloiococcus otitidis and three pathogens of otitis media in production of interleukin-12 by human monocyte cell line.

Alloiococcus otitidis is detected in middle ear effusion of otitis media with effusion (OME). Only a limited number of studies are available concerning the immunological profile of A. otitidis. We have studied the ability of A. otitidis and three other representative pathogens of otitis media to stimulate the production of interleukin-12 (IL-12) from a monocytic cell line THP-1. Viable A. otitidis induced the production of IL-12 in THP-1 cells but IL-12 production was reduced if glutaraldehyde-fixed bacteria were used as stimulants. When viable bacteria were physically separated from THP-1 cells during the stimulation period, remarkable reductions of IL-12 secretion were shown after challenge with gram-positive bacteria A. otitidis and S. pneumoniae. When stimulated with soluble extracts of A. otitidis, THP-1 secreted IL-12 in a dose-dependent manner. The subfraction with a molecular mass over 100 kDa showed a strong ability to induce IL-12 production. Our results show that A. otitidis has immunostimulatory capacity with regard to IL-12 production. We also show that soluble antigen(s) of A. otitidis can modulate the immune response in OME.

Antigens, Bacterial↗
PubMed1995

pUCL287 plasmid from Tetragenococcus halophila (Pediococcus halophilus) ATCC 33315 represents a new theta-type replicon family of lactic acid bacteria.

A cryptic plasmid, pUCL287, was isolated from Tetragenococcus halophila (Pediococcus halophilus) ATCC 33315. It had a theta-type mechanism of replication in its natural host. Its minimal replicon, Rep287, was isolated on a 1.6-kb EcoRI fragment. The Rep287 host range included the genera Pediococcus, Enterococcus, Lactobacillus and Leuconostoc but not genus Lactococcus. Plasmids hybridizing to pUCL287 are rare among lactic acid bacteria. As assessed by hybridization, Rep287 is dissimilar to pAM beta 1, pIP501 and pUCL22, representatives of the most common theta-type replicon groups in Gram-positive bacteria. Therefore, pUCL287 appears to represent a new theta-type replicon family from lactic acid bacteria.

Base Sequence↗
PubMed2000

Ethyl carbamate precursor citrulline formation from arginine degradation by malolactic wine lactic acid bacteria.

Major commercially available strains for induction of malolactic fermentation in wine were examined for arginine metabolism in a resting cell system at wine pH with the aim of evaluating their ability to excrete and utilize citrulline, a precursor of carcinogenic ethyl carbamate (urethane). All strains tested excreted citrulline from arginine degradation. Citrulline was stored intracellularly during growth in arginine rich medium and was released upon lysis of the cells. All strains were found to degrade citrulline as a sole amino acid and some of them were able to reutilize previously excreted citrulline.

Arginine↗