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PubMed1968

Bloater Formation by Gas-forming Lactic Acid Bacteria in Cucumber Fermentations.

The formation of "bloaters" (hollow stock) in cucumbers brined for salt-stock purposes at 5 to 10% salt has been associated with gaseous fermentation caused chiefly by yeasts. Recently, serious early bloater damage, not attributable to yeasts, has been observed in commercial-scale experiments on control of bloaters in overnight dill pickles brined in 50-gal barrels at 3.0 to 4.5% salt. Growth of fermentative species of yeasts was effectively controlled by the addition of 0.025, 0.05, and 0.1% sorbic acid or its sodium salt. In contrast to this, the fermenting brines showed extremely high populations of acid-forming bacteria, identified as Lactobacillus plantarum, L. brevis, and Pediococcus cerevisiae. The gas-forming species (i.e., L. brevis) constituted a high proportion of the total populations. Representative isolates from 36 barrels of overnight dill pickles were tested for their ability to produce bloaters in 1-quart jars of pasteurized cucumbers equilibrated at 4 to 5% salt, 0.25% lactic acid, and pH 4.0. Bloaters, identical with those made by yeast cultures, were produced in all jars inoculated with L. brevis. No bloaters were produced by L. plantarum and P. cerevisiae. These results suggest that the control of bloater damage in cucumber fermentations, particularly at low salt concentrations, may necessitate inhibition of gas-forming lactic acid bacteria.

Journal Article↗
PubMed2000

Ability of dairy strains of lactic acid bacteria to bind aflatoxin M1 in a food model.

Aflatoxin M1 (AFM1) is a highly toxic compound found in milk. Its occurrence poses a threat to the health of consumers, especially young children, and leads to economic losses due to contaminated milk. The problem is global but more severe in developing countries. Consequently, there is a great demand for novel strategies to prevent the contamination and adverse effects of AFM1. To develop a safe and practical decontamination method, a preliminary study was carried out with specific lactic acid bacteria strains that were tested for their ability to remove AFM1 from liquid media. All strains, whether viable or heat-killed, could reduce the AFM1 content of a liquid medium. Two most effective strains were also tested using contaminated skim and full cream milk. The results indicate that specific lactic acid bacteria used in dairy products can offer novel means of decontaminating aflatoxin M1 from milk.

Aflatoxin M1↗
PubMed1982

Effect of lactic acid concentration on growth on meat of Gram-negative psychrotrophs from a meatworks.

The inhibitory effect of the lactic acid in meat on gram-negative psychrotrophs appears to be due mainly to the decrease in pH, not to action of the undissociated acid. Species of Pseudomonas were essentially unaffected by the pH of normal meat. Other gram-negative psychrotrophs isolated from a meatworks included a large number of strains which would not grow on meat of normal pH at chill temperatures. Raising either the pH or the incubation temperature allowed many of the pH-sensitive strains to initiate growth. However, the growth rates of pH-insensitive strains were not affected by increasing the pH, and there were no significant differences in the composition of the spoilage floras which developed on chilled meat of normal and high pH.

Bacteria↗
PubMed2006

Characterization of lactic acid bacteria isolated from the one humped camel milk produced in Morocco.

One hundred and twenty (120) strains of lactic acid bacteria (LAB) were enumerated and isolated from raw dromedary milk in Morocco using various cultured media. Strains isolated were characterized by phenotypic, physiological and biochemical properties. Results showed that high counts of LAB were found. Presumptive lactobacilli counts ranged from 2.5x10(2) to 6x10(7)cfu/ml, presumptive lactococci levels varied from 5x10(2) to 6x10(7)cfu/ml, presumptive streptococci counts varied from 4.2x10(2) to 8x10(7)cfu/ml, presumptive leuconostoc levels ranged from 5.4x10(2) to 5.4x10(7)cfu/ml. Results showed also that Lactobacillus and Lactococcus were the predominant genera with 37.5% and 25.8%, respectively. The dominated species found were Lactococcus lactis subsp. lactis (17.5%), Lactobacillus helveticus (10%), Streptococcus salivarius subsp. thermophilus (9.20%), Lactobacillus casei subsp. casei (5.80%) and Lactobacillus plantarum (5%). This is the first report on the characterization of LAB strains isolated from the one humped camel milk produced in Morocco.

Animals↗
PubMed1994

Production of histamine and tyramine by lactic acid bacteria isolated from vacuum-packed sugar-salted fish.

The incidence of histamine- or tyramine-producing lactic acid bacteria was examined in several products of vacuum-packed sugar-salted fish (salmon, halibut, mackerel). No histamine-producing isolates were observed, whereas the majority of tyramine-producing isolates were identified as Carnobacterium spp. These organisms were shown to be important members of the microbial flora during storage of vacuum-packed sugar-salted salmon at 5 degrees C. The amount of tyramine produced was reduced by lowering the temperature from 9 degrees C to 4 degrees C for all of five strains of carnobacteria or lactobacilli. The majority of tyramine was produced during the exponential growth phase for Carnobacterium piscicola N 5 and Lactobacillus viridescens N 69. The ability of these bacteria to produce tyramine may be used as an index of microbial quality/acceptability of stored vacuum-packed sugar-salted fish.

Fish Products↗
PubMed2005

Antimicrobial susceptibility of lactic acid bacteria isolated from a cheese environment.

In the production of the Spanish traditional blue-veined Cabrales cheese, lactic acid bacteria strains free of antibiotic resistance that have a transferrable capacity are necessary as components of a specific starter. To select for these bacteria, the minimum inhibitory concentration (MIC) of 12 antibiotics and 2 mixtures (containing beta-lactamase inhibitor and penicillin) were determined by microbroth and agar dilution techniques in 146 strains belonging to the genera Lactococcus, Enterococcus, Lactobacillus, and Leuconostoc. The antibiotic-resistance profiles of Lactococcus and Enterococcus species were different from those of Lactobacillus and Leuconostoc, but clear genus- or species-associated patterns were not observed. Cefoxitin and metronidazole were not effective against bacteria of these genera. The MICs of beta-lactam antibiotics for lactobacilli and leuconostoc isolates were higher than those for lactococci and enterococci, but no strain was clinically resistant. All lactobacilli and leuconostoc isolates were resistant to high levels of vancomycin, a type of resistance not seen among the tested members of the genera Lactococcus and Enterococcus. The majority of the observed resistance appeared to be either intrinsic or nonspecific, although some strains of Lactococcus lactis, Enterococcus spp., and Lactobacillus spp. were resistant to antibiotics, such as chloramphenicol, erythromycin, clindamycin, or tetracycline.

Anti-Bacterial Agents↗
PubMed2006

Human neutrophils reaction to the biodegraded nano-hydroxyapatite/collagen and nano-hydroxyapatite/collagen/poly(L-lactic acid) composites.

The impact of biodegraded nano-hydroxyapatite/collagen (nHAC) composite and nano-hydroxyapatite/collagen/poly(L-lactic acid) (nHAC/PLA) scaffold composite on neutrophils reaction was evaluated in vitro. Neutrophils were separated from human peripheral blood of healthy subjects. The nHAC and nHAC/PLA materials were immersed in the D-Hanks' Balanced Salt Solution (D-HBSS) for 1 day, 7 days and 2, 4, 8 weeks (37 degrees C) as testing solution, which mixed with the neutrophils for 1 h. Both of the nHAC and nHAC/PLA materials were shown the same cell survival rate as blank control, but the lactate dehydrogenase (LDH) and tumor necrosis factor alpha (TNF-alpha) released from the neutrophils were increased significantly after the 2 weeks in nHAC sample. The possible reason relied on the high concentration of calcium due to the quick biodegradation of the nHAC material. Before 2 weeks, the LDH value of nHAC/PLA is higher than that of nHAC sample that corresponded to the initial PLA degradation in vitro. This study provided the biocompatibility test of neutrophils other than common methods, such as osteoblastic cells for biomimetic materials. Moreover, it demonstrated the calcium concentration stimulating effect for cytokine release from neutrophils.

Biocompatible Materials↗
PubMed2001

Development and characterization of CyA-loaded poly(lactic acid)-poly(ethylene glycol)PEG micro- and nanoparticles. Comparison with conventional PLA particulate carriers.

Cyclosporin A (CyA) loaded poly(lactic acid)-poly(ethylene glycol) (PLA-PEG) micro- and nanoparticles have been developed using an emulsion-solvent evaporation method. Physico-chemical properties, peptide loading content and in vitro release profiles of these novel CyA carriers were compared with those corresponding to conventional PLA micro- and nanoparticles. Results obtained confirm the previously described disposition of PEG chains on the surface of the PLA-PEG formulations. In addition, they revealed the presence of CyA molecules on the surface of both PLA and PLA-PEG systems. Further determination of the surface chemical composition by electron spectroscopy for chemical analysis (ESCA) allowed us to quantify the amount of CyA in the nanospheres' top layers, this amount being higher for nanoparticles than for microparticles, and higher for the PLA systems than for those based on PLA-PEG. In vitro release experiments revealed that PLA-PEG particles provided a more adequate control of CyA release than conventional PLA micro- and nanoparticles. Physico-chemical characterization of the systems during the release studies showed that the developed PLA and PLA-PEG micro- and nanoparticles were not degraded, which suggest a diffusion-mediated release mechanism. Furthermore, we have hypothesized that the hydrophilic outer shell of PEG provides a stationary layer for the diffusion of CyA.

Cyclosporine↗
PubMed2001

Characterization of lactic acid bacteria strains on the basis of neutral volatile compounds produced in whey.

AIMS: Seventy-eight strains of lactic acid bacteria belonging to five genera and showing six different phenotype combinations of Lac (lactose fermentation), Prt (proteolytic activity) and Cit (citrate degradation) characters were investigated for their main flavouring properties with the aim to detect variability among and within the groups. METHODS AND RESULTS: High resolution gas chromatography-mass spectrometry analysis of neutral volatile compounds produced in whey showed that, considering both neo-formation compounds and substances quantified in the whey cultures at different concentrations in comparison to the extract from sterile whey, the groups of lactococci, enterococci, thermophilic streptococci and mesophilic lactobacilli produced a higher number of volatiles than thermophilic lactobacilli and leuconostocs. Applying principal component analysis (PCA) to the results, enterococci, mesophilic lactobacilli and thermophilic streptococci showed a broad diversity, while lactococci included rather similar strains as well as strains with special flavouring properties. Applying PCA to thermophilic streptococci and enterococci, to lactococci and enterococci, to lactococci and thermophilic streptococci, or to mesophilic and thermophilic lactobacilli, the strains gathered consistently with their systematic position. CONCLUSION: The study evidenced strains producing some volatile compounds responsible for food flavouring. Flavouring properties were variable among the systematic groups and in some cases different within the same bacterial group. SIGNIFICANCE AND IMPACT OF THE STUDY: The potential of the findings is discussed with reference to the development of flavouring adjuncts for the dairy industry.

Cluster Analysis↗
PubMed2006

Broken or dislodged poly-L-lactic acid bioabsorbable tacks in patients after SLAP lesion surgery.

PURPOSE: To document the complication of bioabsorbable poly-L-lactic acid (PLLA) tack breakage and repair failure after arthroscopic stabilization of SLAP lesions. METHODS: Six patients (average age, 38 years) were referred to the senior author because of persistent postoperative pain and disability after arthroscopic stabilization of an unstable SLAP lesion was performed with bioabsorbable PLLA tacks. A total of eight tacks were used in the six patients, and the time from index procedure to evaluation at our facility averaged 9.5 months (range, 8 to 12 months). Three patients experienced an identifiable reinjury an average of 4 months after the index procedure was performed. Magnetic resonance imaging (MRI) revealed a broken or dislodged tack in all cases. All patients were taken to the operating room for arthroscopic evaluation of the biceps anchor, tack fragment removal, and restabilization of the SLAP lesion as needed. RESULTS: At repeat arthroscopy, all shoulders were found to have minimal healing of the SLAP lesion with failure of the bioabsorbable tack. Chondral injury was noted in 2 cases. In all cases, revision SLAP repair was performed with the use of arthroscopic suture repair. At an average follow-up of 14 months (range, 8 to 17 months), all patients reported alleviation of symptoms. Five patients had returned to work with no or minimal symptoms. One patient experienced significant improvement in function and range of motion but did not return to work. CONCLUSIONS: Bioabsorbable PLLA tack breakage or dislodgement is a potential postoperative complication after SLAP lesions are arthroscopically stabilized. This event may result in postoperative reports of pain or mechanical symptoms. Surgeons who use these devices must be aware of the potential complications when they evaluate postoperative patients with persistent complaints. MRI may be helpful in identifying tack fragments. Subsequent arthroscopy to evaluate biceps anchor healing, remove tack fragments, and restabilize the labral lesion may be indicated. LEVEL OF EVIDENCE: Level IV, case series.

Absorbable Implants↗
PubMed2002

Growth studies of potentially probiotic lactic acid bacteria in cereal-based substrates.

AIMS: The overall growth kinetics of four potentially probiotic strains (Lactobacillus fermentum, Lact. reuteri, Lact. acidophilus and Lact. plantarum) cultured in malt, barley and wheat media were investigated. The objectives were to identify the main factors influencing the growth and metabolic activity of each strain in association with the cereal substrate. METHODS AND RESULTS: All fermentations were performed without pH control. A logistic-type equation, which included a growth inhibition term, was used to describe the experimental data. In the malt medium, all strains attained high maximum cell populations (8.10-10.11 log10 cfu ml(-1), depending on the strain), probably due to the availability of maltose, sucrose, glucose, fructose (approx. 15 g l(-1) total fermentable sugars) and free amino nitrogen (approx. 80 mg l(-1)). The consumption of sugars during the exponential phase (10-12 h) resulted in the accumulation of lactic acid (1.06-1.99 g l(-1)) and acetic acid (0.29-0.59 g l(-1)), which progressively decreased the pH of the medium. Each strain demonstrated a specific preference for one or more sugars. Since small amounts of sugars were consumed by the end of the exponential phase (17-43%), the decisive growth-limiting factor was probably the pH, which at that time ranged between 3.40 and 3.77 for all of the strains. Analysis of the metabolic products confirmed the heterofermentative or homofermentative nature of the strains used, except in the case of Lact. acidophilus which demonstrated a shift towards the heterofermentative pathway. All strains produced acetic acid during the exponential phase, which could be attributed to the presence of oxygen. Lactobacillus plantarum, Lact. reuteri and Lact. fermentum continued to consume the remaining sugars and accumulate metabolic products in the medium, probably due to energy requirements for cell viability, while Lact. acidophilus entered directly into the decline phase. In the barley and wheat media all strains, especially Lact. acidophilus and Lact. reuteri, attained lower maximum cell populations (7.20-9.43 log10 cfu ml(-1)) than in the malt medium. This could be attributed to the low sugar content (3-4 g l(-1) total fermentable sugar for each medium) and the low free amino nitrogen concentration (15.3-26.6 mg l(-1)). In all fermentations, the microbial growth ceased at pH values (3.73-4.88, depending on the strain) lower than those observed for malt fermentations, which suggests that substrate deficiency in sugars and free amino nitrogen contributed to growth limitation. CONCLUSIONS: The malt medium supported the growth of all strains more than barley and wheat media due to its chemical composition, while Lact. plantarum and Lact. fermentum appeared to be less fastidious and more resistant to acidic conditions than Lact. acidophilus and Lact. reuteri. SIGNIFICANCE AND IMPACT OF THE STUDY: Cereals are suitable substrates for the growth of potentially probiotic lactic acid bacteria.

Culture Media↗
PubMed1996

PCR methods for identification and specific detection of probiotic lactic acid bacteria.

Probiotics are defined as "microbes improving animal feed." Three lactic acid bacteria, previously selected as probiotic for pig feeding, were identified by sequencing the variable V1 region of the 16S rDNA after PCR amplification primed in the flanking constant region. A VR region showing strong nucleotide differences between the three probiotic and the reference strains was delimited. Oligonucleotides specific for each strain were designed. A specific assay for probiotic detection was developed, based on a PCR reaction with three primers.

Animal Feed↗
PubMed1997

Identification of a cold shock gene in lactic acid bacteria and the effect of cold shock on cryotolerance.

When Lactic Acid Bacterial cultures were frozen at -20 degrees C for 24 h, the cell viability decreased drastically, but when they were cold shocked at 10 degrees C for 2 h prior to freezing, viability improved significantly for the Lactococcus lactis subsp. lactis strains (25-37%) and Pediococcus pentosaceus PO2 (18%), but not for the Lactococcus lactis subsp. cremoris strains tested or for one strain of Lactobacillus helveticus LB1 and Streptococcus thermophilus TS2. When the period for cold shock was extended to 5 h, the viability increased even further for those strains that displayed cold shock cryotolerance. Use of degenerate PCR primers based on the major cold shock protein (csp) of both Escherichia coli and Bacillus subtilis resulted in PCR products from all strains tested. The PCR product from Lactococcus lactis ssp. lactis M474 was cloned and sequenced, and the deduced amino acid sequence displayed a high sequence similarity to other csp's. Use of PCR primers based on the M474 sequence resulted in PCR products being produced only from the lactococcal strains studied and not from the Lactobacillus helveticus, Streptococcus thermophilus, or Pediococcus pentosaceus strains tested.

Amino Acid Sequence↗
PubMed2003

Pediocin production by recombinant lactic acid bacteria.

Production of the anti-listerial bacteriocin, pediocin, by lactic acid bacteria (LAB) transformed with the cloning vector pPC418 (Ped+, 9.1 kb) was influenced by composition of media and incubation temperature. Maximum pediocin production, tested against Listeria innocua, by electrotransformants of Lactococcus lactis ssp. lactis was measured in tryptone/lactose/yeast extract medium after 24 h growth at 30 degrees C, while incubation at 40 degrees C was optimum for Ped+ transformants of Streptococcus thermophilus and Enterococcus faecalis. The amount of pediocin produced by S. thermophilus in skim milk and cheese whey supplemented with 0.5% yeast extract was estimated as 51,000 units ml(-1) and 25,000 units ml(-1), respectively. Pediocin production remained essentially unchanged in reconstituted skim milk or whey media diluted up to 10-fold. The results demonstrate the capacity of recombinant strains of LAB to produce pediocin in a variety of growth media including skim milk and inexpensive cheese whey-based media, requiring minimum nutritional supplementation.

Animals↗
PubMed2003

Behavior of psychrotrophic lactic acid bacteria isolated from spoiling cooked meat products.

Three kinds of lactic acid bacteria were isolated from spoiling cooked meat products stored below 10 degrees C. They were identified as Leuconostoc mesenteroides subsp. mesenteroides, Lactococcus lactis subsp. lactis, and Leuconostoc citreum. All three strains grew well in MRS broth at 10 degrees C. In particular, L. mesenteroides subsp. mesenteroides and L. citreum grew even at 4 degrees C, and their doubling times were 23.6 and 51.5 h, respectively. On the other hand, although the bacteria were initially below the detection limit (<10 CFU/g) in model cooked meat products, the bacterial counts increased to 10(8) CFU/g at 10 degrees C after 7 to 12 days.

Animals↗
PubMed2001

Isolation, characterization, and influence of native, nonstarter lactic acid bacteria on Cheddar cheese quality.

To determine whether adventitious nonstarter lactic acid bacteria (NSLAB) might affect cheese flavor and quality, we studied a population of NSLAB present in 30 premium quality Cheddar cheeses (3-mo ripened) produced at a commercial facility in the United States. DNA fingerprinting analysis with a sensitive strategy for arbitrary priming polymerase chain reaction showed that 75 isolates corresponded to at least 18 distinct nonstarter organisms. According to ribotype database comparisons of representatives from the 18 groups, 9 matched Lactobacillus (closest to paracasei species), 8 matched Streptococcus thermophilus, and 1 matched to a Lactococcus species. This finding indicated that among the 75 NSLAB isolates, Lactobacillus made up 64%, S. thermophilus 32%, and Lactococcus 4%. Isolates representing 11 NSLAB groups were characterized for protease, peptidase, and diacetyl production. Based on this phenotypic analysis, two Lactobacillus isolates were evaluated as adjuncts in Cheddar cheese. All of the NSLAB identified from the adjunct cheese at 3 mo by DNA fingerprinting consisted of the adjunct lactobacilli, showing that the adjunct strains predominated throughout the early stages of ripening. The impact of adjunct lactobacilli was evident after 6 mo when free amino acids significantly increased and sensory scores improved in adjunct cheese as compared with a control cheese. The largest impact was found in adjunct cheese containing a blend of both lactobacilli strains. These results show that certain adventitious NSLAB positively contribute to flavor development.

Amino Acids↗
PubMed2005

Invited review: methods for the screening, isolation, and characterization of exopolysaccharides produced by lactic acid bacteria.

The ability to produce exopolysaccharides (EPS) is widespread among lactic acid bacteria (LAB), although the physiological role of these molecules has not been clearly established yet. Some EPS confer on LAB a "ropy" character that can be detected in cultures that form long strands when extended with an inoculation loop. When EPS are produced in situ during milk fermentation they can act as natural biothickeners, giving the product a suitable consistency, improving viscosity, and reducing syneresis. In addition, some of these EPS may have beneficial effects on human health. The increasing demand by consumers of novel dairy products requires a better understanding of the effect of EPS on existing products and, at the same time, the search for new EPS-producing strains with desirable properties. The use of genetically modified organisms capable of producing high levels of EPS or newly designed biopolymers is still very limited. Therefore, exploration of the biodiversity of wild LAB strains from natural ecological environments is currently the most suitable approach to search for the desired EPS-phenotype. The screening of ropy strains and the isolation and characterization of EPS responsible for this characteristic have led to the application over the past years of a wide variety of techniques. This review summarizes the available information on methods and procedures used for research on this topic. The information provided deals with methods for screening of EPS-producing LAB, detection of the ropy phenotype, and the physicochemical and structural characterization of these molecules, including parameters related to their viscosifying properties. To our knowledge, this is the first compilation of methods available for the study of EPS produced by LAB.

Dairy Products↗
PubMed1993

Effect of antioxidative lactic acid bacteria on rats fed a diet deficient in vitamin E.

Lactic acid bacteria, including Bifidobacterium, with antioxidative activity were selected by in vitro screening. The effect of the antioxidative activity was investigated by in vivo experiments using rats that were deficient in vitamin E. In the first stage of screening, 570 strains were examined; intracellular cell-free extracts of 19 strains (16 lactobacilli, 2 streptococci, and 1 lactococci) had antioxidative activity as determined by an assay using rat liver microsomes and thiobarbituric acid. In the second stage of screening, 7 strains of lactobacilli showed over 70% inhibition of oxidation activity. The highest activity was obtained by heterofermentative Lactobacillus sp. SBT 2028. The effect of two strains, Lactobacillus sp. SBT 2028 and Lactobacillus casei ssp. rhamnosus SBT 2257, was evaluated for improvement of the condition of rats fed a diet deficient in vitamin E. Intracellular cell-free extracts of those two strains were also used for in vivo experiments. Hemolysis of red blood cells was inhibited in rats that were administered the extract of Lactobacillus sp. SBT 2028, which proved that the extract improved the vitamin E deficiency status. Antioxidative activity of an extract from L. casei ssp. rhamnosus SBT 2257 determined by hemolysis was relatively weak compared with the activity of Lactobacillus sp. SBT 2028 extract.

Animals↗