Lactobacillus Casei Subsp Research Articles

Biosynthesis of Copper Oxide Nanoparticles Using Lactobacillus casei Subsp. Casei and its Anticancer and Antibacterial Activities

Background:The present study reveals the synthesis of copper oxide nanoparticles (CuO NPs) by probiotic bacteria (Lactobacillus casei subsp. casei) and demonstrates the cytotoxic effects of these nanoparticles against gram negative and positive bacteria and cancer cell lines.Methods:The CuO NPs are biosynthesized from Lactobacillus casei subsp. casei (L. casei) in an eco-friendly and cost-effective process. These nanoparticles are characterized using Fourier transform infrared spectroscopy (FT-IR), X-ray diffractometer (XRD), field emission scanning electron microscopy (FESEM), energy dispersive X-ray (EDX) and transmittance electron microscope (TEM) analysis. The antibacterial activity is examined by Well-diffusion, minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) assays using Broth microdilution. Anticancer effects of these nanoparticles are evaluated by methyl thiazolyl diphenyl-tetrazolium bromide (MTT) assay and Griess test.Results:Our results confirm the biosynthesis of CuO NPs from L. casei. Antibacterial assays demonstrate that treatment of gram-negative and gram-positive bacteria with CuO NPs inhibits the growth of these bacteria. Furthermore, the cell viability of human cancer cells decreases while treated by nanoparticles. These nanoparticles increase nitric oxide (NO) secretion determined by NO production measurement.Conclusion:These results suggest that CuO NPs may exert antibacterial effects as well as cytotoxic effects on cancer cells by suppressing their growth, increasing the oxidative stress and inducing apoptosis.

Effect of the Alternative Addition of Sodium Acetate and Tween 80 on the Production Curve of Lactic Acid by Lactobacillus Casei Subsp Rhamnosus from date variety Hmira and carob pods syrups

Effect of the Alternative Addition of Sodium Acetate and Tween 80 on the Production Curve of Lactic Acid by Lactobacillus Casei Subsp Rhamnosus from date variety Hmira and carob pods syrups

The use of date waste for lactic acid production by a fed-batch culture using Lactobacillus casei subsp. rhamnosus.

The production of lactic acid from date juice by Lactobacillus caseisubsp. rhamnosus in batch and fed-batch cultures has been investigated. The fed-batch culture system gave better results for lactic acid production and volumetric productivity. The aim of this work is to determine the effects of the feeding rate and the concentration of the feeding medium containing date juice glucose on the cell growth, the consumption of glucose and the lactic acid production by Lactobacillus casei subsp. rhamnosus in fed-batch cultures. For this study, two concentrations of the feeding medium (62 and 100 g/L of date juice glucose) were tested at different feeding rates (18, 22, 33, 75 and 150 mL/h). The highest volumetric productivity (1.3 g/L.h) and lactic acid yield (1.7 g/g) were obtained at a feeding rate of 33 mL/h and a date juice glucose concentration of 62 g/L in the feeding medium. As a result, most of the date juice glucose was completely utilised (residual glucose 1 g/L), and a maximum lactic acid production level (89.2 g/L) was obtained.

Peptidoglycan Hydrolases of Local Lactic Acid Bacteria from Kazakh Traditional Food.

IntroductionPeptidoglycan (PG) is a major component of the cell wall of Gram-positive bacteria and is essential for maintaining the integrity of the bacterial cell and its shape. The bacteria synthesize PG hydrolases, which are capable of cleaving the covalent bonds of PG. They also play an important role in modeling PG, which is required for bacterial growth and division. In an era of increasing antibiotic-resistant pathogens, PG hydrolases that destroy these important structures of the cell wall act as a potential source of new antimicrobials. The aim of this study is to identify the main PG hydrolases of local lactic acid bacteria isolated from traditional foods that enhance probiotic activity of a biological preparation.MethodsLactococcus lactis 17A and Lactococcus garvieae 19A were isolated from the traditional sausage-like meat product called kazy. They were isolated according to standards methods of microbiology. Genetic identification of the isolates were tested by determining the nucleotide sequences of 16S rDNA. The Republican collection of microorganisms took strains of Lactobacillus casei subsp. Rhamnosus 13-P, L. delbrueckii subsp. lactis CG-1 B-RKM 0044 from cheese, Lactobacillus casei subsp. casei B-RKM 0202 from homemade butter. They used the standard technique of renaturating polyacrylamide gel electrophoresis to detect PG hydrolases activity.ResultsAccording to the profiles of PG hydrolase activity on zymograms, the enzymes of Lactococci 17A and 19A in kazy are similar in electrophoretic mobility to major autolysin AcmA, while the lactobacilli of industrial and home-made dairy products have enzymes similar to extracellular proteins p40 and p75, which have probiotic activity.ConclusionsUse of peptidoglycan hydrolases seems to be an interesting approach in the fight against multi-drug resistant strains of bacteria and could be a valuable tool for the treatment of diseases caused by these microorganisms in Kazakhstan.

Randomised clinical trial: mesalazine and/or probiotics in maintaining remission of symptomatic uncomplicated diverticular disease - a double-blind, randomised, placebo-controlled study

Placebo-controlled studies in maintaining remission of symptomatic uncomplicated diverticular disease (SUDD) of the colon are lacking. To assess the effectiveness of mesalazine and/or probiotics in maintaining remission in SUDD. A multicentre, double-blind, placebo-controlled study was conducted. Two hundred and ten patients were randomly enrolled in a double-blind fashion in four groups: Group M (active mesalazine 1.6 g/day plus Lactobacillus casei subsp. DG placebo), Group L (active Lactobacillus casei subsp. DG 24 billion/day plus mesalazine placebo), Group LM (active Lactobacillus casei subsp. DG 24 billion/day plus active mesalazine), Group P (Lactobacillus casei subsp. DG placebo plus mesalazine placebo). Patients received treatment for 10 days/month for 12 months. Recurrence of SUDD was defined as the reappearance of abdominal pain during follow-up, scored as ≥5 (0: best; 10: worst) for at least 24 consecutive hours. Recurrence of SUDD occurred in no (0%) patient in group LM, in 7 (13.7%) patients in group M, in 8 (14.5%) patients in group L and in 23 (46.0%) patients in group P (LM group vs. M group, P = 0.015; LM group vs. L group, P = 0.011; LM group vs. P group, P = 0.000; M group vs. P group, P = 0.000; L group vs. P group, P = 0.000). Acute diverticulitis occurred in six group P cases and in one group L case (P = 0.003). Both cyclic mesalazine and Lactobacillus casei subsp. DG treatments, particularly when given in combination, appear to be better than placebo for maintaining remission of symptomatic uncomplicated diverticular disease. (ClinicalTrials.gov: NCT01534754).

Effect of the Essential Oils from Parsley and Fennel Seeds on the Growth of Lactobacillus Casei Subsp Rhamnosus

In this paper the key odorants effects of parsley ( Petroselinum crispum Hoffm) and fennel ( Foeniculum vulgare ) in the growth of Lactobacillus casei subsp rhamnosus , were studied. Parsley and fennel essential oils were obtained by hydrodistillation with 0.14% yield from parsley and 1% from fennel. All experiments were carried out in a 2L jar fermenter with an initial volume of 1.5L at 42°C, and the constant, adjusted to 200 rpm. Lactobacilli suspensions were incubated at 42°C for 12hr at 200 rpm and then a 10% (v/v) of the bacterial suspension was added to the cultures. The culture pH was maintained at 6.25 by automatic addition of 25% (w/w) NH 4 OH. For that purpose, batch cultures on MRS medium containing 15 g.L -1 of glucose were realized: the first fermentation is considered as a control, the second is characterized by the addition of 20 μL of the parsley essential oil in the growth exponential phase and the third contains additionally 20 μL of the fennel essential oil. The results show that Lactobacillus casei subsp rhamnosus can produce up to 10.96 g.L -1 and 13.78 gL -1 of lactic acid on the control fermentation and on the second fermentation, respectively. The addition of the fennel essential oil provokes an inhibition of the Lactobacillus casei subsp rhamnosus growth which is translated by the obtained weak lactic acid concentration of 5.14 g.L -1 .

Lactobacillus casei Abundance Is Associated with Profound Shifts in the Infant Gut Microbiome

Colonization of the infant gut by microorganisms over the first year of life is crucial for development of a balanced immune response. Early alterations in the gastrointestinal microbiota of neonates has been linked with subsequent development of asthma and atopy in older children. Here we describe high-resolution culture-independent analysis of stool samples from 6-month old infants fed daily supplements of Lactobacillus casei subsp. Rhamnosus (LGG) or placebo in a double-blind, randomized Trial of Infant Probiotic Supplementation (TIPS). Bacterial community composition was examined using a high-density microarray, the 16S rRNA PhyloChip, and the microbial assemblages of infants with either high or low LGG abundance were compared. Communities with high abundance of LGG exhibited promotion of phylogenetically clustered taxa including a number of other known probiotic species, and were significantly more even in their distribution of community members. Ecologically, these aspects are characteristic of communities that are more resistant to perturbation and outgrowth of pathogens. PhyloChip analysis also permitted identification of taxa negatively correlated with LGG abundance that have previously been associated with atopy, as well as those positively correlated that may prove useful alternative targets for investigation as alternative probiotic species. From these findings we hypothesize that a key mechanism for the protective effect of LGG supplementation on subsequent development of allergic disease is through promotion of a stable, even, and functionally redundant infant gastrointestinal community.

Use of Lactobacillus casei Subspecies Rhamnosus GG and Gastrointestinal Colonization by Candida Species in Preterm Neonates

Candida species increasingly cause morbidity and mortality in the premature infant in neonatal intensive care units, and the gut reservoir is the site from which dissemination most frequently starts in such patients. Specific antifungal prophylaxis is the most suitable strategy with which to limit the severity and the frequent neurodevelopmental impairment in survivors that is associated with neonatal invasive fungal infections. Recent interest has focused on the use of probiotics for the treatment of several diseases in neonatal patients. Pilot studies have implicated these organisms in necrotizing enterocolitis, sepsis, and urinary tract infections. Other applications of probiotic therapy in preterm neonates may also include the prevention of fungal colonization and of Candida-related disorders. Probiotics could provide an innovative and less invasive approach because they modify the bowel flora by colonizing the gastrointestinal tract. Basic research has shown that in mice models, the Lactobacillus casei subsp Rhamnosus GG (LGG) is effective in preventing Candida gut colonization and systemic dissemination. A pilot, randomized, double-blind, placebo-controlled trial in human preterm neonates has demonstrated that LGG administered in the first month of life significantly reduces enteric Candida colonization. The present article summarizes the state of the art about probiotics and Candida-related diseases in the preterm neonate and emphasizes the need for further investigations to determine unequivocally the possible role of LGG in the prevention and management of the fungal diseases in preterm neonates.

Inhibition of Salmonella-induced IL-8 synthesis and expression of Hsp70 in enterocyte-like Caco-2 cells after exposure to non-starter lactobacilli

Oral administration of lactobacilli as probiotics is gaining importance in the treatment of intestinal inflammations. We investigated the effect of non-starter lactobacilli Lactobacillus casei subsp casei 2756, Lactobacillus curvatus 2775, and Lactobacillus plantarum 2142 as well as their spent culture supernatants (SCS) on Salmonella enteritidis 857 growth, interleukin (IL)-8 and heat shock protein 70 (Hsp70) synthesis in undifferentiated crypt-like and differentiated villus-like Caco-2 cells. The cells were infected with graded numbers of non-starter lactobacilli or S. enteritidis 857 for 1 h and allowed to recover for 24 h or exposed to 200 bacteria/cell for 1 h and allowed to recover for different periods of time. In another experiment S. enteritidis 857 was first pre-treated with SCS-lactobacilli for 1 h before infecting the cells. The levels of IL-8 and Hsp70 were assessed using sandwich ELISA and immunostaining of Western blots, respectively. The effect of SCS-lactobacilli on S. enteritidis 857 growth was evaluated by agar plate diffusion test. The non-starter lactobacilli induced a significant increase in the levels of both IL-8 and Hsp70. However, compared with the S. enteritidis 857 induced IL-8 synthesis, the levels of IL-8 induced by the lactobacilli at any equivalent bacterial number were far lower. After exposure of Caco-2 cells to S. enteritidis 857 pre-treated with SCS-lactobacilli, it appeared that their SCS inhibited the S. enteritidis 857 growth and IL-8 synthesis and in addition induced the expression of Hsp70. The differences in response of crypt- and villus-like Caco-2 cells are merely a reflection of their differentiation status. Our data suggest that the beneficial effect of non-starter lactobacilli to the intestinal inflammations might be associated with a decrease of the IL-8 levels. This effect could be mediated, at least in part, by the bacteria themselves or via a secreted antimicrobial product(s) either directly against the pathogens or indirectly through the synthesis of Hsp70.

Oral immunoadjuvant activity of a new synbiotic Lactobacillus casei subsp casei in conjunction with dextran in BALB/c mice

In the present study, Lactobacillus casei subsp casei strains JCM 1134 T (Lcc) and JCM 8129 showed the ability to use dextran, whereas other tested species of Lactobacillus, Bifidobacterium, and intestinal bacteria tested did not. The number of live Lactobacillus species increased significantly in dextran-fed BALB/c mice 30 days after oral administration of Lcc. Further, Lcc was detected specifically using a polymerase chain reaction assay. When the BALB/c mice were orally given Lcc and its specific substrate dextran together with bovine serum albumin (BSA), a greater enhanced production of serum anti-BSA immunoglobulin G was induced in comparison with those given BSA alone. These results clearly showed that a new synbiotic, probiotic Lcc and its prebiotic dextran in combination, exhibited immunoadjuvant activity in BALB/c mice.

Contribution of Lactococcus lactis subsp lactis IFPL 359 and Lactobacillus casei subsp casei IFPL 731 to the Proteolysis of Caprine Curd Slurries

ABSTRACT: Proteolysis was studied in low‐fat caprine curd slurries pre‐incubated (24 h at 30 °C) with an extra dose of rennet, 10.0 or 22.0 μg g‐1 (LR and HR slurries, respectively), and treated at 70 °C for 15 min prior to incubation with sonicated cells of Lactococcus lactis subsp lactis IFPL 359 alone or combined with Lactobacillus casei subsp casei IFPL 731. The addition of lactobacilli to slurries containing lactococci increased the conversion of water‐soluble nitrogen to nonprotein nitrogen and amine nitrogen. Major differences were found for some free amino acids, such as glutamic acid and proline, and hydrolysis of most hydrophobic peptides produced by lactococci, which are associated with bitterness in cheese.

Effect of pasteurization and use of a native starter culture on proteolysis in a ewes’ milk cheese

The effect of different manufacturing methods on the production of a ewes’ milk cheese was studied. Three batches were made: raw milk without added starter, raw milk with added starter, and pasteurized milk with added starter. The starter culture was composed of Lactococcus lactis subsp lactis biovar diacetylactis and Lactobacillus casei subsp casei. Samples were taken from all three cheese batches on 1, 90, and 180 days of ripening for analysis of the soluble nitrogen (SN) and non-protein nitrogen (NPN). SN and NPN levels were the highest in the batch made from the pasteurized milk, which also exhibited the highest level of degradation of the αs and β-caseins. Conversely, that batch had the lowest total free amino acid (FAA) content. This parameter was the highest in the batch made from the raw milk with added starter. The cheeses made from the pasteurized milk presented the highest serine and asparagine contents, but γ-aminobutyric acid levels were negligible.

Combined effects of concentrated thermophilic and mesophilic cultures and conditions of curd acidifications on the manufacture and quality of kasseri cheese

The effect offour process variables on the biological acidification of curd to be usedfor the manufacture of kasseri cheese was examined. The variables were (i) concentrated starter cultures: a thermophilic mixture of Streptococcus thermophilus and Lactobacillus delbrueckii subsp bulgar‐icus compared with a blend of Lactococcus lactis subsp lactis, Lactococcus lactis subsp cremoris and Lactobacillus casei subsp casei, and (ii) acidification temperatures: 30 and 40° C for the thermophilic cultures and 20 and 30°C for the mesophiles. The use of concentrated starter cultures enabled the cheese to be made in one day as compared with three in the traditional system usually using raw milk and no culture. The mesophilic blend combined with a lower scalding temperature (35°C) gave a higher yield than the alternative culture, and the cheeses were higher in total solids, acidity andsalt; the same cheeses were also harder and less elastic. The use of mesophilic cultures and a low acidification temperature (20° C) is recommended.

Flavour development in aseptic cheese curd slurries prepared with single-strain starter bacteria in the presence and absence of adjuncts

Summary - Eight strains of Lactococcus starter bacteria were tested under microbiologically controlled conditions for their ability to pro duce Cheddar cheese flavour, For this purpose, aseptic Cheddar chee se curds were prepared with each of the organisms, and the resulting curds slurried with sterile salt and water, and stored at 30 C for accelerated f1avour development. Distinct though weak Cheddar cheese flavour was produced by the Lactococcus lactis subsp cremoris strain SK Il, but not by strains ASCC457, ASCC50, ASCC 109, ASCC660, ASCC360, or the L lactis subsp lactis strain ASCC381, or the L lactis subsp lactis biovar diacetylactis strain ASCC250. Different starter strains produced different degrees of bitterness, with strains ASCC457, 109 and 660 producing the most bitter slurries. Strains SKII, ASCC50, ASCC360, and ASCC250 produced only slight bitterness in slurries ripened for 15 days, Free fatty acid concentrations in the slurries differed depending on the starter strain used, implying that different starter strains have different lipase activity. Cheese flavour development was slow in the slurries containing only starter bacteria. In contrast, slurries made with the strain SKII in the presence of the adjuncts Lactobacillus helveticus ASCC395, Brevibacterium linens CSCC750 or Lactobacillus casei subsp casei NCDOl51 underwent rapid f1avour development, giving distinct and strong chee se (but not Cheddar) flavours for each adjunct. These differences in flavour, however, cou Id not be explained in terms of differences in the non-sulphur volatile composition of the slurries determined by static headspace analysis.

Differences between Lactobacillus casei subsp. casei 2206 and Citrate-Positive Lactococcus lactis subsp. lactis 3022 in the Characteristics of Diacetyl Production

Lactobacillus casei subsp. casei 2206 exhibited much lower levels of diacetyl reductase activity than CitrLactococcus lactis subsp. lactis 3022 but two-, three-, and more than eightfold-higher levels of diacetyl synthase, lactate dehydrogenase, and NADH oxidase activities, respectively. A requirement for metal ions by the diacetyl synthases in both species was observed. The extracts of strain 2206 but not strain 3022 produced more diacetyl from pyruvate when the reaction for diacetyl synthase was aerated than when it was conducted statically.