Man, Rogosa And Sharpe Broth Research Articles

Tofu Whey-Based Media for Probiotic Lactiplantibacillus plantarum D4 as a Halal Starter Culture

In recent years, the development of probiotic-based fermented products with halal status has been a concern. The use of growth medium has relied on de Man Rogosa and Sharpe (MRS) as a relatively expensive standard medium, and its halal status is still uncertain. Extensive research has been carried out to investigate the development of low-cost halal alternative media for the cultivation of probiotic lactic acid bacteria (LAB). This study aimed to develop a probiotic halal and low-cost culture medium using a tofu whey-based medium. This study used three tofu whey-based media – A (tofu whey 100%), B (tofu whey 94.5%, molasses 3%, skim milk 2.5%), C (tofu whey 92.5%, molasses 3%, cheese whey 2.5%, tomato extract 2%), and MRS broth as a standard medium. Bacterial populations, total sugars utilized, total lactic acids produced, low pH (2.0) tolerance, and high bile salt concentration (oxgall 1.5%) were assayed. The highest bacterial population after 48 h of incubation was shown by medium B compared to medium MRS (12.34 ± 0.87 and 11.48 ± 0.3 log CFU/mL). Total sugars utilized by 48.28 ± 2.89, 38.89 ± 6.94, 39.14 ± 4.24, and 76.00 ± 1.41 %; on the other hand, total lactic acids produced by 0.16 ± 0.12, 0.03 ± 0.04, 0.31 ± 0.03, and 2.25 ± 1.48 % in A, B, C, and MRS, respectively. Probiotic tolerance at low pH and the presence of bile salts of Lactiplantibacillus plantarum D4 consistently showed a high survival rate in medium B compared to MRS. Based on these results, the components and proportions used in medium B were suitable for the growth of L. plantarum D4 as a halal probiotic starter candidate.

The Abilities of Different Lactic Acid Bacterial Stains to Deplete Nitrite Level in MRS and M17 Broth

Nitrite is still regarded as essentially undesirable in foods. Therefore, it is necessary to keep its concentration under control for food safety. The current study was conducted to evaluate the ability of two reference lactic acid strains to deplete nitrite. Lactobacillus acidophilus ROO52 and Streptococcus thermophiles YC-180 were examined in De Man Rogosa and Sharpe (MRS) and M17 broth with standard sodium nitrite at 0.10 mg /mL and 1 mL (10-8 cells per mL) for each of the reference strains to determine the ability of these strains to eliminate nitrite during the fermentation period. A control group, MRS and M17 broth and standard nitrite without inoculum were used. The nitrite depletion was determined by a UV-Vis spectrophotometer. After 48 hours of fermentation, there were significant differences between residual nitrite levels of broth inoculated by Streptococcus thermophiles YC-180 compared to control and Lactobacillus acidophilus ROO52. Streptococcus thermophiles YC-180 depleted nitrite levels significantly (P˂0.05) from the initial concentration (0.10 mg/ml) to 0.02 mg/ml with a percentage (45%) after 48 hours compared to control and Lactobacillus acidophilus ROO52, which depleted nitrite levels from the initial concentration of 0.10 mg/ml to 0.04 mg/ml with a percentage (22.5%) after 48 hours. In conclusion, Streptococcus thermophiles YC-180 was more effective at nitrite depletion than Lactobacillus acidophilus ROO52. The reduction of nitrite occurs due to acid production and the enzymatic action of these bacteria.

Polyols Induce the Production of Antifungal Compounds by Lactobacillus plantarum.

Mycotoxins may be present in nuts, coffee, cereals, and grapes, among other products. Increasing concerns about human health and environmental protection have driven the application of biological control techniques that can inhibit fungal contaminants. In this study, the growth inhibition of the ochratoxigenic fungus Aspergillus carbonarius Ac 162 was evaluated using 5 lactic acid bacteria (LAB). The LAB studied were Lactobacillus plantarum MZ801739 (J), Lactobacillus plantarum MZ809351 (31) and Lactobacillus plantarum MZ809350 (34), isolated in the Ivory Coast, and Lactobacillus plantarum MN982928 (3) and Leuconostoc citreum MZ801735 (23), isolated in Mexico. J, 31, 34, 3 and 23 are the internal strain codes from our laboratory. LAB were cultivated in De Man, Rogosa and Sharpe (MRS) broth, and different polyols (glycerol, mannitol, sorbitol, and xylitol) were added to the culture broth to stimulate the production of antifungal compounds. The fungal inhibition studies were performed using the poisoned food technique. The highest inhibition of A. carbonarius growth was obtained by cultivating L. plantarum MZ809351 in the presence of xylitol and glycerol. Under these conditions, 1 L of the L. plantarum MZ809351 cultures were used to identify antifungal compounds. The compounds were concentrated by solid-phase extraction and then characterized by GC-MS. In addition to 9-octadecenoic acid, 3 diketopiperazines or cyclic dipeptides were identified, including cyclo (Leu-Leu), cyclo (Pro-Gly) and cyclo (Val-Phe), which were compounds related to microbial antifungal activities. Xylitol and glycerol induced the production of these antifungal compounds against A. carbonarius Ac 162. On the other hand, adding xylitol and glycerol to the MRS broth reduced the Ochratoxin A (OTA) content to 56.8 and 54.7%, respectively. This study shows the potential for using L. plantarum MZ809351 as a biocontrol agent to prevent the growth of A. carbonarius and reduce the production of OTA in foods.

Antibacterial Activity of Bacteriocin from Pediococcus pentosaceus Strain 2397 and Application as Biopreservative for Fishballs

Meatballs are one of the most popular processed meats in various countries. The meats commonly used to make meatballs are beef, chicken, and fish. The purposes of this study were to evaluate the nitrogen, carbon, and Tween sources on the growth and antimicrobial activity of bacteriocin of P. pentosaceus strain 2397 and to assess the quality of fish balls preserved with bacteriocin from P. pentosaceus strain 2397 during initial storage at freezing temperature in –18 °C. The experimental design used in this study was a factorial, completely randomized design. There were two factors, i.e. bacteriocin concentrations (0, 0.15, 0.30, 0.45, and 0.60%) and storage time (0, 3, 6, and 9 d). Data obtained were statistically analyzed using analysis of variance (ANOVA) and continued with Duncan multiple range test (DMRT) at 5% level. Meanwhile, data on microbiological counts were tabulated and analyzed descriptively. The data showed that the addition of various nitrogen, carbon, or Tween sources did not significantly (p > 0.05) affect the growth of P. pentosaceus strain 2397. The addition of peptone, beef extract or ammonium sulfate, lactose, or Tween 80 into the de Man, Rogosa, and Sharpe broth (MRSB) significantly (p < 0.05) increased the bacteriocin antimicrobial activity of strain 2397 against E. coli. Using 0.60% bacteriocin produced fish balls according to the quality standard of fish balls and maintained the quality of fish balls during initial storage for 9 d at freezing temperatures.

Antituberculosis agents of Lactobacillus plantarum and Pediococcus acidilactici Lactic Acid Bacteria in Breast milk isolates against Mycobacterium tuberculosis

Tuberculosis (TB) is one of the infectious diseases that have become a major problem in Indonesia. This disease is caused by the Mycobacterium tuberculosis bacteria. The bacteria are commonly treated with antibiotics. However, the use of irrelevant antibiotics is the most influential factor of antibiotic resistance. Therefore, natural ingredients are required as novel antibiotic agents, some of which are sourced from lactic acid bacteria. This study aims at investigating the antituberculosis activities of Pediococcus acidilactici and Lactobacillus plantarum isolated from breast milk against M. tuberculosis. Pediococcus acidilactici and Lactobacillus plantarum breast milk isolates were rejuvenated with MRS (De Man, Rogosa and Sharpe) broth media, and the supernatant was then neutralized to pH 7.0 using a pH meter by adding 1N NaOH solution. Antituberculosis activity test was performed with Lowensten Jensen media to investigate the growth of M. tuberculosis. The results of this study showed that Pediococcus acidilactici had antituberculosis activity at 48 hours at concentrations of 80% and 90%, while the secondary metabolites of L. plantarum had antituberculosis activity at 24 hours and 48 hours at concentrations of 60%, 70%, 80%, 90%. Therefore, this study concludes that P. acidilactici and L. plantarum bacteria have the potential to be developed as antituberculosis agents.

Co-Culture Strategy of Lactobacillus kefiranofaciens HL1 for Developing Functional Fermented Milk.

Our previous studies indicated that Lactobacillus kefiranofaciens HL1, isolated from kefir grain, has strong antioxidant activities and anti-aging effects. However, this strain is difficult to use in isolation when manufacturing fermented products due to poor viability in milk. Thus, the purpose of this study was to apply a co-culture strategy to develop a novel probiotic fermented milk rich in L. kefiranofaciens HL1. Each of four selected starter cultures was co-cultured with kefir strain HL1 in different media to evaluate their effects on microbial activity and availability of milk fermentation. The results of a colony size test on de Man, Rogosa and Sharpe (MRS) agar agar, microbial viability, and acidification performance in MRS broth and skimmed milk suggested that Lactococcus lactis subsp. cremoris APL15 is a suitable candidate for co-culturing with HL1. We then co-cultured HL1 and APL15 in skimmed milk and report remarkable improvement in fermentation ability and no negative impact on the viability of strain HL1 or textural and rheological properties of the milk. Through a co-culture strategy, we have improved the viability of kefir strain HL1 in fermented skimmed milk products and successfully developed a novel milk product with a unique flavor and sufficient probiotics.

Production of Phenyllactic Acid from Porphyra Residues by Lactic Acid Bacterial Fermentation

The concept of algae biorefinery is attracting attention because of the abundant valuable compounds within algal biomass. Phenyllactic acid (PhLA), a broad-spectrum antimicrobial organic acid that can be produced by lactic acid bacteria (LAB), is considered a potential biopreservative. In this study, a cascading biorefinery approach was developed to harvest PhLA from Porphyra residues by LAB fermentation. LAB strains were cultivated in de Man, Rogosa and Sharpe (MRS) broth to screen their ability to produce PhLA. As the strains of Lactobacillus plantarum KP3 and L. plantarum KP4 produced higher concentrations of PhLA at 0.09 mg/mL, these two strains were employed for fermentation. After phycobiliprotein extraction, the Porphyra residues, ultrafiltration eluate, phenylalanine (Phe) and yeast extract with a volume of 20 mL were inoculated with LAB strain KP3 and fermented at 37 °C for 120 h. The PhLA content of the fermented broth was 1.86 mg. To optimize the biorefinery process, the ultrafiltration eluate was replaced by commercial cellulase. Up to 4.58 mg of PhLA, which was 2.5 times greater than that produced from KP3 cultured in MRS broth, could be harvested. Taken together, the findings provide an optimized process for LAB fermentation, which is shown to be a feasible algae biorefinery approach to obtaining PhLA from Porphyra residues.

Effect of Five Commercial Probiotic Formulations on Candida Albicans Growth: In Vitro Study.

The objective was to evaluate the antagonistic effect of Lactobacillus and Bifidobacterium recovered from five commercial probiotics on the growth of C. albicans. The Lactobacillus and Bifidobacterium strains of five commercial probiotics were recovered and grown: Probio Hp+®, ProBiseis®, Lactipan®, Liolactil®, and Lacteol Fort®; 50 mg of each was hydrated and grown in Lactobacilli MRS (De Man, Rogosa and Sharpe) broth and incubated at 37°C with stirring (120 RPM) for 24 hours. Serial dilutions of 10-1 to 10-7 were made and viability was verified and quantified. For the antagonism tests, a suspension/inoculum of Lactobacillus strains recovered from each commercial preparation (4-30 × 109) and C. albicans ATCC 90028 (1.5-8 × 108) was prepared in MRS broth and incubated for 48 hours at 36°C, then plated on Dextrose Sabouraud Agar with Chloramphenicol and Rogosa Agar and the colony-forming units (CFU) were quantified. Additionally, viability was evaluated using the LIVE/DEAD® Yeast and Bacterial Viability kit. The probiotic that produced the highest acidity of the medium was Lactipan®, followed by Probiseis® and Liolactil®, while Probio Hp+® showed the least change. Probiseis® was determined to have the highest growth of probiotic bacteria and the highest inhibition on C. albicans, followed by Lactipan®; Liolactil® and ProbioHp+® showed the least effect. In fluorescence tests, ProBiseis® showed the best effect, followed by Liolactil® and Lactipan®; Probio Hp+® had less of an effect. Two commercial products (ProBiseis and Lactipan) whose formulations have L. acidophilus, L. casei, L. rhamnosus, L. plantarum, B. infantis, and S. thermophilus have a greater inhibitory effect on C. albicans ATCC 90028.

Development of a Low-Cost and High-Efficiency Culture Medium for Bacteriocin Lac-B23 Production by Lactobacillus plantarum J23.

At present, De Man, Rogosa and Sharpe (MRS) broth is the medium of choice for promoting bacteriocin production. However, this medium is expensive and not applicable for large-scale production. Therefore, a low-cost and high-efficiency culture medium for bacteriocin Lac-B23 production by Lactobacillus plantarum J23 was developed. First, the effects of the composition of MRS broth on bacteriocin Lac-B23 production and bacterial growth were researched by a one variable at a time approach. Then, a Plackett-Burman design was used to screen significant components for production. Finally, the steepest ascent and central composite designs were used to obtain an optimum medium. The final composition of the modified MRS was much simpler than MRS broth, and the modified MRS contained only glucose, yeast extract, dipotassium phosphate, manganese sulfate monohydrate, Tween 80 and sodium acetate anhydrous. The highest bacteriocin Lac-B23 production reached 2560 activity units (AU)/mL in the modified MRS, which is nine times higher than that in MRS broth (280 AU/mL). Meanwhile, the cost per liter of the modified MRS (8.56 Ren Min Bi (RMB)/L) is 34.70% the cost of MRS broth (13.11 RMB/L), and the cost per arbitrary units of bacteriocin Lac-B23 in the modified MRS is approximately fourteen times more convenient (3.34 RMB/106 AU) than in the MRS broth (46.82 RMB/106 AU).

Effect of Isothiocyanates on the Activity of <i>Lactobacillus plantarum</i> Exposed to Irradiation

Two isothiocyanates, i.e., sulforaphane (SFA) and sulforaphene (SFE), are suggested to be used as an alternative chemopreventive diet. This study was focused on the effect of SFA and SFE on Lactobacillus plantarum, which has been subjected to the irradiation (2-50 Gy). The cultures grown in De Man, Rogosa and Sharpe (MRS) and Tryptone Soya Broth (TSB) were compared in terms of bacteria physiological activity under tested conditions. Broth composition notably influenced the bacteria growth kinetic parameters, as well as culture response to the oxidative stress. Activity of L. plantarum cells after irradiation was evaluated by their dehydrogenase (DHA) and quinone-reductase (QR) activities. The enzyme activity was quantified in living cells. Bacterial cultures obtained in MRS and TSB broth, demonstrated contrasting characteristics in their enzyme activities. The MRS-grown culture did not show any QR activity, whereas the TSB-grown cells showed a non-linear response towards gamma-irradiation with a maximum inhibition being at 10 Gy. Addition of SFA or SFE in concentration of 1 µg/mL to the cultures before irradiation exposure recovered the QR activity from 23% in a non-amended variant up to 102% and 121%, respectively, taking the non-irradiated non-amended variant as 100%.

The Synergistic Effect of Fiber Blends on Lactobacillus rhamnosus

The combination of prebiotics with probiotic microorganisms has been shown to improve gastrointestinal health. Previous studies have demonstrated the beneficial effect of psyllium supplementation on the growth of probiotic bacteria. However, little is known about the synergistic effect of fiber blends. Thus, the main objective of this study was to evaluate the effect of the combination of different fibers with psyllium on the growth of Lactobacillus rhamnosus.Lactobacillus rhamnosus (ATCC® 7469™), a Gram‐positive bacterium with demonstrated probiotic activity, was the bacterial species used in this study. The bacterial inoculum was prepared by growing the bacteria in MRS (De Man, Rogosa and Sharpe) broth for 24 hours under microaerophilic condition (5% CO2) at 37°C, yielding approximately 108 CFU/mL. The Lactobacillus culture media, MRS broth, was supplemented with different combinations of fibers to evaluate their effect on bacterial growth. Three different fibers, psyllium, inulin, and fructooligosaccharides (FOS), were used in the study. To test the combined effect, the fibers were combined with each other at a concentration of 0.5% each and then altogether. The inoculum was grown in an un‐supplemented broth following the same conditions as the fiber‐supplemented broths, to be used as a negative control. Positive controls of psyllium supplemented at 0.5%, 1.0%, and 1.5% were included to test against different treatment concentrations. Each inoculated broth supplemented with each fiber blend were incubated in duplicate. Colony forming units (CFU) were measured on plates in duplicate. The CFU data was converted to a base‐10 algorithm before statistical analysis was performed. All data were analyzed using the general linear model procedure for analysis of variance (ANOVA) and Pearson correlation analysis.The presence of fiber in the growth media, regardless of concentration, improved bacterial growth significantly when compared to the control containing 0% of fiber (9.11 vs 8.57 log CFU/mL, respectively, p<0.005). Bacterial growth increased as the concentration of psyllium increased (r=0.8, p<0.01). This result agrees with earlier studies presented by our team. Treatments containing psyllium supplementation demonstrated significantly better growth than treatments without psyllium (9.14 vs 8.80 log CFU/mL, respectively, p<0.001). The psyllium supplementation at 0.5% concentration, in all blends, exhibited similar growth with no significant differences between treatments. Treatment supplemented with psyllium at 1.5% was similar to the treatment containing a combination of all fibers (psyllium, inulin, and FOS at 0.5% each) (9.22 vs 9.07 log CFU/mL, respectively, p>0.05).In conclusion, no synergistic effect on the growth of Lactobacillus rhamnosus was observed from the blending of fibers. However, the addition of prebiotic fibers to the growth media caused an increase in probiotic bacteria growth. More research is needed to understand if combinations of prebiotics can produce synergistic effects at higher concentrations and with different fiber blends.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The Effect of Cinnamon Essential Oil on Lactobacillus species

Antimicrobial properties of cinnamon essential oil solutions (EO) have been identified against pathogenic bacteria, but its effects on commensal microorganisms have not yet been described. In previous studies in our laboratory, cinnamon EO was tested against Salmonella Typhimurium and Escherichia coli utilizing the disc diffusion method. Zones of inhibition were measured, and cinnamon EO at 2 and 4% displayed great effectiveness against these microorganisms. Both microorganisms are Gram negative rods and have the capability to cause disease. Thus, the main objective of this study is to evaluate the effect of cinnamon EO on the commensal Gram positive microorganisms Lactobacillus acidophilus and Lactobacillus rhamnosus.The Lactobacillus acidophilus (ATCC® 4356™) and Lactobacillus rhamnosus (ATCC® 7469™) inoculums were prepared by growing the bacteria in MRS (De Man, Rogosa and Sharpe) broth for 24h under microaerophilic conditions (5% CO2) at 37°C, yielding 6.9 ×108 CFU/ml. One hundred microliters of starting bacterial culture was spread onto MRS agar plates and dried for 15 minutes. Treatments were formed by creating cinnamon EO solutions of 0, 0.5, 1, 2 and 4% using 70% ethyl alcohol as the diluent. Each EO was vortexed and filtered using a 0.2 microliter sterile syringe filter. Twenty microliters of each EO were pipetted into a sterile paper disc and left to dry for 1h. Five paper discs of each treatment containing the EO were placed on standardized locations on each inoculated plate in duplicate. Additionally, three manufactured standard antibiotic paper discs of 5 mcg ciprofloxacin, 10mcg ampicillin, and 10mcg streptomycin were placed on separate plates in duplicate as controls. The zone of inhibition created by each EO or antibiotic was measured using a calibrated digital caliper. All data were analyzed using GLM procedure for ANOVA and Person correlation analysis.The antimicrobial property of the EO observed as the zone of inhibition increased as the concentration of the EO increased for Lactobacillus acidophilus (r=0.86, p<0.0001). EO at 4% showed the greatest antimicrobial effect against Lactobacillus acidophilus compared to 2.0, 1.0, and 0.5% (2.59, 2.11, 0.0 and 0.0mm, respectively; p<0.0001). Lactobacillus rhamnosus showed resistance to EO at all concentrations studied with no zones of inhibition observed. All antibiotics had some effect against Lactobacillus rhamnosus, where ampicillin had the greatest effect compared to ciprofloxacin and streptomycin (6.62, 6.27, and 2.33mm, respectively; p<0.0001). Ampicillin had the greatest effect against Lactobacillus acidophilus compared to streptomycin (9.72 vs. 2.8mm, respectively; p<0.0001). However, L. acidophilus was shown to be resistant to ciprofloxacin with no zone of inhibition formed.In conclusion, Lactobacillus species were generally resistant to cinnamon EO. These Lactobacillus species are commonly used as probiotics and are often adversely affected by antibiotic intake. Using cinnamon essential oil as an antibiotic alternative has shown to be a promising option with limited detrimental effects against the commensal bacteria. Further evaluation of the effects of cinnamon EO on commensal bacteria is needed to further determine the benefits of utilizing a natural alternative.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Antimicrobial Activity of Probiotics against Endodontic Pathogens:- A Preliminary Study

AbstractIntroduction: The purpose of this study was to evaluate the antibacterial effectiveness of probiotics lactobacilli group and Bifidobacterium against Enterococcus faecalis and Candida albicans in both planktonic stage and biofilm stage. Materials and Methods: Phase 1 of the study was conducted by agar well diffusion method. About 0.5 ml of test pathogen culture was inoculated on 20 ml of molten agar and allowed to solidify. 4–5 circular wells of diameter 8–10 mm were punched in each poured plates and 150 μl of diluted test samples were added to the wells. Phase 2 was deferred antagonism test, wherein purified culture of pathogen strain was streaked at right angle to the original producer growth and incubated at 37°C for 24 h. Zone of inhibition was measured for both the phases. Phase 3 biofilm stage evaluation was conducted by mixing 9 ml of 30% poloxamer 407 and De Man, Rogosa and Sharpe (MRS) broth in a test tube with 500 μl of either pathogen, together with 500 μl of test probiotic strains and incubated (37°C, 48 h), followed by serially diluting the mixture by 1 ml into 9 ml sterile saline till 108 dilutions for evaluation of colony-forming unit/ml counts. Controls were endodontic pathogens in 30% poloxamer with MRS broth and no probiotics. Results: Results were evaluated and statistically analysed using one-way analysis of variance and unpaired t-test. In the planktonic stage, probiotics showed inhibitory activity against endodontic pathogens with valid statistical significance (P < 0.05), while there was no activity by deferred antagonism method. In biofilm stage, all three probiotics showed growth reduction for E. faecalis, while lactobacilli group showed reduction in C. albicans colonies. Conclusion: This preliminary study suggested that probiotics are effective for preventing the growth of endodontic pathogens in vitro. Poloxamer could be utilised as an ideal delivery vehicle for probiotics.

Optimizational production of phenyllactic acid by a Lactobacillus buchneri strain via uniform design with overlay sampling methodology

Abstract A Lactobacillus buchneri GBS3 strain isolated from the traditional Chinese pickles was used for the production of 3-phenyllactic acid (PLA), an important compound with antimicrobial activities against a wide species of gram-positive and gram-negative bacteria and some fungi. The growth performance of this strain in the de Man, Rogosa and Sharpe (MRS) medium, the production of metabolites of valuable organic acids, and the biosynthesis of PLA using this strain as the whole-cell biocatalyst and phenylpyruvic acid (PPA) as the precursor, were investigated experimentally. The uniform design method with overlay sampling was developed for the optimization of the biotransformation conditions. The results showed that although it produced naturally lactic acid with the maximum concentration of 1.84 g·L− 1 and PLA with the concentration of 0.015 g·L− 1 after 66 to 72 h cultivation in MRS broth by fermentation, the present strain displayed an effective utilization ability by transforming PPA to PLA. By the uniform design method with overlay sampling for the design and optimization of transformation conditions, a maximum yield of 10.93 g·L− 1 PLA with the mole conversion ratio of 83.07% from PPA to PLA was achieved under the optimized condition, i.e., 20 g·L− 1 glucose, 270 g·L− 1 cells, 13 g·L− 1 PPA, pH 8.0 and the reaction time of 15 h, indicating that Lactobacillus buchneri GBS3 was an interesting strain for the biosynthesis of PLA via the microbial transformation. The prediction of PLA yield under different conditions was achieved successfully based on the limited information of only a small number of experiments by the uniform design with overlay sampling. Therefore, the present methodology is effective and helpful for the optimization of the biosynthesis processes of PLA.

Effect of Fiber Concentration on the Growth of Lactobacillus rhamnosus

The use of probiotics and prebiotics has become widely accepted as beneficial for intestinal health. In addition to the enhanced intestinal function, probiotics and prebiotics are being recognized for their capacity to modulate the immune system, and their effectiveness and safety in diseases, such as diabetes, cancer and heart disease. The best known probiotic bacteria are those belonging to the genera Lactobacillus and Bifidobacterium. The most common prebiotics are fibers, also known as dietary fibers, which have the potential to stimulate the growth of commensal bacteria residing in the intestinal tract. Gaining insight on how fibers can modulate or enhance the growth of probiotics is of great importance. Therefore, this study aims to evaluate the effect of different fiber sources and different concentrations of these fibers on in vitro bacterial growth. It is hypothesized that higher concentration of fiber will result in higher microbial populations.Lactobacillus rhamnosus, a Gram‐positive bacterium with demonstrated probiotic activity, was the bacterial species used in this study. The bacterial inoculum was prepared by growing the bacteria in MRS (De Man, Rogosa and Sharpe) broth for 24 hours under microaerophilic condition (5% CO2) at 37°C, yielding about 108 CFU/ml. Lactobacillus culture media, MRS broth, was supplemented with different fibers at different concentrations to evaluate the effect of these different nutrients and their concentration on bacterial growth. Three different fibers were used in the study, which were inulin, fructooligosaccharides (FOS), and psyllium. To test the concentration effect, three different concentrations were used, 1%, 2% and 4%.In a previous study, we have proven the beneficial effect of the addition of fiber to the growth media. The present study results agree with those findings. In addition, based on the current results, the hypothesis that higher concentration of the fibers enhances bacterial growth was proven correct for psyllium (8.7, 9.0, 9.3 log CFU/ml for 1, 2, 4% respectively, p&lt;0.05), but not for the other fibers used in this study. Psyllium supplemented at 2% provided a greater bacterial growth than the other fibers (9.0 vs. 8.7 log CFU/ml, p=0.01). The highest bacterial growth observed with psyllium supplementation compared to the other treatments was achieved at 4% supplementation (9.3 vs 8.7 log CFU/ml, p&lt;0.001).In conclusion, Lactobacillus rhamnosus growth can be differently affected by the different fibers. Psyllium resulted in the highest population growth, and its effect increased as concentration increased. Distinct probiotics are influenced by different types of prebiotic at different concentrations, which is correlated to the growth needs of the bacterial cells. Therefore, the present study findings add important information regarding the complex interaction between probiotic bacteria and prebiotics; however, more research is needed to better understand these intricate relationships.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Development of Whey Protein Concentrate-Pectin-Alginate Based Delivery System to Improve Survival of B. longum BL-05 in Simulated Gastrointestinal Conditions.

Bifidobacterium longum BL-05 encapsulated beads were developed by using whey protein concentrate (WPC) and pectin (PE) as encapsulating material through extrusion/ionic gelation technique with the objective to improve survival of probiotics in harsh gastrointestinal conditions. B. longum BL-05 was grown in MRS (de man rogosa and sharpe) broth, centrifuged and mixed with polymeric gel solution. Bead formulations E4 (2.5% WPC + 1.5% PE) and E5 (2% PE) showed the highest value for encapsulation efficiency, size, and textural properties (hardness, cohesiveness, springiness) due to increasing PE concentration. The survivability and viability of free and encapsulated B. longum BL-05 was assessed through their resistance to simulated gastric juice (SGJ), tolerance to bile salt, release profile in simulated intestinal fluid (SIF), and storage stability during 28days at 4°C. The microencapsulation provided protection to B. longum BL-05 and encapsulated cells were exhibited significant (p < 0.05) resistance to SGJ and SIF as compared to free cells. Bead formulations E3 (5.0% WPC + 1.0% PE) and E4 (2.5% WPC + 1.5% PE) exhibited more resistance to SGJ (at pH 2 for 2h) and at 2% bile salt solution but comparatively slow release as compared to other bead formulations. Free cells lost their viability when stored at 4°C after 28days but microencapsulated cells demonstrated promising results during storage and viable cell count was > 107CFU/g. This study revealed that extrusion using WPC and PE as encapsulating material could be considered as one of the novel technologies for protection and effective delivery of probiotics.

Potensi Limbah Cair Industri Tapioka sebagai Media Pertumbuhan Starter Bakteri Asam Laktat Pediococcus pentosaceus E.1222

Fermentation of Lactic Acid Bacteria (LAB) Pediococcus pentosaceus can improve the quality of food and its shelf life. Using commercial LAB specific media, de Man Rogosa and Sharpe (MRS) for growth on industrial scale application is not efficient. Tapioca wastewater (TW) still contains some of the nutrients needed for the growth of P. pentosaceus, but needs the enrichment of carbon sources (5% of glucose) and nitrogen sources (ammonium sulfate). This study aimed to use tapioca industrial wastewater with the addition of glucose and ammonium sulfate as an alternative growth media for lactic acid bacteria P. pentosaceus E.1222. The results showed that glucose and nitrogen had no significant effect on the number of bacterial colonies. The highest substrate efficiency was tapioca wastewater (86.81%), MRS broth (53.73%), and TW with 5% of glucose and 1% of ammonium sulfate (43.53%) respectively. Maximum growth rate (μmaks) was found in TW with 5% of glucose and 1% of ammonium sulfate (0,52 hours-1). Increasing the starter volume until 1000 mL in TW with 5% of glucose and 1% of ammonium sulfate showed a slight decrease in the log number of bacteria from 8,836 (50 mL), 8,401 (500 mL), to 8,063 (1000 mL).

Growth of Bacillus Coagulans Using Molasses as a Nutrient Source

Bacillus coagulans (B. coagulans) can be utilized as microbial inoculant to speed up the bioremediation of wastewater. The effectiveness of the microbial inoculant for treatment is highly selective and relying on the characteristics of the wastewater. A feasible carbon source must be first chosen to pre-culture the microbe prior to the bioremediation. Commercial nutrient broths are efficient to grow the microbial; they are costly for the treatment of large volume of wastewater treatment. This study aims to evaluate the growth rate of B. coagulans using a cheaper nutrient source, i.e. molasses. The commercial Man, Rogosa and Sharpe (MRS) media was used as a benchmark. The growth rate of B. coagulans was conducted at different concentrations of molasses (1 %, 3 % and 5 % (w/v)) and in the MRS broth as benchmark. B. coagulans grown in the MRS has shown a much higher maximum specific growth rate (0.69 h-1) compared to that grown in 1 % (w/v) molasses (0.14 h-1). No growth was observed in the higher concentrations of molasses (3 and 5 % (w/v)). Measuring the colonyforming units of B. coagulans in both the MRS and molasses mediums validated the results. Molasses is a desirable carbon source as it is relatively cheaper and easily available. More studies are needed to improve the maximum specific growth rate of B. coagulans in 1 % (w/v) of molasses.

Evaluation of the Role of Probiotics in Endodontic Treatment: A Preliminary Study.

Aims and Objectives:The principal goal of endodontics is the prevention of periapical infection. Acute and chronic apical periodontitis occur due to the persistence of pathogenic microorganisms such as Enterococcus faecalis and Candida albicans harboring the root canal systems of the teeth. The concept of the use of probiotics in addressing endodontic disease is new and has not been studied adequately. On the basis of the success of probiotics in periodontal treatment, this preliminary work was performed (a) to evaluate the antibacterial efficacy of probiotics against common endodontic pathogens, i.e. E. faecalis and C. albicans, and (b) to evaluate the potential use of probiotic therapy as an additive in endodontic treatment procedures.Materials and Methods:Two commercial probiotics were selected and evaluated based upon the numbers and concentration of organisms. Pathogenic test organisms were C. albicans (ATCC 10231) and E. faecalis (ATCC 29212). Phase 1 of the study was conducted by agar cup method test to evaluate the antibacterial activity of the selected probiotics against E. faecalis and C. albicans by measuring zones of inhibition (ZOI) in mm. Microorganisms from probiotic samples were isolated following manufacturer's instructions. Pathogenic organisms were set to a 0.1 McFarland standard challenge. Circular wells of 8 mm diameter were punched in each of the poured plates. Appropriately diluted test samples were added to the above-punched wells. The volume of the solution added to each well was 100 μl. The plates were incubated in an upright position at 37°C for 24 hours under aerobic conditions. Post incubation, ZOI was measured (mm). Phase 2 was conducted by mixing 9 ml of 30% poloxamer 407 and de Man, Rogosa and Sharpe (MRS) broth in a test tube with 500 μl of either E. faecalis or C. albicans set at an optical density (OD) of 0.252, together with 500 μl of test probiotic strain, set at a respective OD. Samples were then incubated at 37°C for 48 hours, followed by serial dilutions by 1 ml till 108. This was done to calculate colony forming units (CFU)/ml counts. Controls used were endodontic pathogens in 30% poloxamer with MRS broth without any probiotic group.Results:Probiotic groups showed inhibitory activity against E. faecalis by the agar cup method, whereas there was no effect on C. albicans. In the biofilm stage, both the test groups had an antibacterial effect on pathogenic organisms.Conclusion:This study suggests that probiotic organisms of the species Lactobacillus and Bifidobacterium are effective for preventing the growth of E. faecalis and C. albicans in vitro. Because probiotics are available in varied compositions and concentrations, further evaluation for their role in treating endodontic infection is suggested and warranted. In addition, the study suggested that poloxamer 407 could be utilized as an ideal delivery vehicle for probiotics for use as a potential endodontic intracanal medicament.

Enhancement of Nisin Production by Lactococcus lactis subsp. lactis.

Lactococcus lactis subsp lactis BSA (L. lactis BSA) was isolated from a commercial fermented product (BSA Food Ingredients, Montreal, Canada) containing mixed bacteria that are used as starter for food fermentation. In order to increase the bacteriocin production by L. lactis BSA, different fermentation conditions were conducted. They included different volumetric combinations of two culture media (the Man, Rogosa and Sharpe (MRS) broth and skim milk), agitation level (0 and 100rpm) and concentration of commercial nisin (0, 0.15, and 0.30µg/ml) added into culture media as stimulant agent for nisin production. During fermentation, samples were collected and used for antibacterial evaluation against Lactobacillus sakei using agar diffusion assay. Results showed that medium containing 50% MRS broth and 50% skim milk gave better antibacterial activity as compared to other medium formulations. Agitation (100rpm) did not improve nisin production by L. lactis BSA. Adding 0.15µg/ml of nisin into the medium-containing 50% MRS broth and 50% skim milk caused the highest nisin activity of 18,820AU/ml as compared to other medium formulations. This activity was 4 and ~3 times higher than medium containing 100% MRS broth without added nisin (~4700AU/ml) and 100% MRS broth with 0.15µg/ml of added nisin (~6650AU/ml), respectively.