Anti-Obesity Efficacy of Pediococcus acidilactici MNL5 in Canorhabditis elegans Gut Model.

In the present study, thirty two lactic acid bacteria (LAB) were isolated from fermented Indian herbal medicine. In comparison to other strains, MNL5 had stronger bile salt hydrolase (BSH) and cholesterol-lowering properties. Furthermore, it can withstand the extreme conditions found in the GI tract, due to, e.g., pepsin, bile salts, pancreatin, and acids. Pediococcus acidilactici MNL5 was identified as a probiotic candidate after sequencing the 16S rRNA gene. The antibacterial activity of P. acidilactici MNL5 cell-free supernatants (CFS) against Escherichia coli, Staphylococcus aureus, Helicobacter pylori, Bacillus cereus, and Candida albicans was moderate. A Caenorhabditis elegans experiment was also performed to assess the effectiveness of P. acidilactici MNL5 supplementation to increase life span compared to E. coli supplementation (DAF-2 and LIU1 models) (p < 0.05). An immense reduction of the lipid droplets of C. elegans was identified through a fluorescent microscope. The drastic alteration of the expression of fat genes is related to obesity phenotypes. Hence, several paths are evolutionary for C. elegans; the results of our work highlight the nematode as an important model for obesity.

Anti-Tuberculosis Activity of Pediococcus acidilactici Isolated from Young Radish Kimchi against Mycobacterium tuberculosis.

Tuberculosis is a highly contagious disease caused by Mycobacterium tuberculosis. It affects about 10 million people each year and is still one of the leading causes of death worldwide. About 2 to 3 billion people (equivalent to 1 in 3 people in the world) are infected with latent tuberculosis. Moreover, as the number of multidrug-resistant, extensively drug-resistant, and totally drug-resistant strains of M. tuberculosis continues to increase, there is an urgent need to develop new anti-tuberculosis drugs that are different from existing drugs to combat antibiotic-resistant M. tuberculosis. Against this background, we aimed to develop new anti-tuberculosis drugs using probiotics. Here, we report the anti-tuberculosis effect of Pediococcus acidilactici PMC202 isolated from young radish kimchi, a traditional Korean fermented food. Under coculture conditions, PMC202 inhibited the growth of M. tuberculosis. In addition, PMC202 inhibited the growth of drug-sensitive and -resistant M. tuberculosis- infected macrophages at a concentration that did not show cytotoxicity and showed a synergistic effect with isoniazid. In a 2-week, repeated oral administration toxicity study using mice, PMC202 did not cause weight change or specific clinical symptoms. Furthermore, the results of 16S rRNA-based metagenomics analysis confirmed that dysbiosis was not induced in bronchoalveolar lavage fluid after oral administration of PMC202. The anti-tuberculosis effect of PMC202 was found to be related to the reduction of nitric oxide. Our findings indicate that PMC202 could be used as an anti-tuberculosis drug candidate with the potential to replace current chemicalbased drugs. However, more extensive toxicity, mechanism of action, and animal efficacy studies with clinical trials are needed.

Inhibition of acetic acid-induced colitis in rats by new Pediococcus acidilactici strains, vitamin producers recovered from human gut microbiota.

Our aim was to isolate, identify and characterize probiotic bacteria as vitamin producers in particular B2 and B9. 150 human fecal samples were collected and used for isolation of vitamin producers-probiotics. 49 isolates were chosen for screening their genome by PCR for the presence of riboflavin and folic acid genes. As a result, three isolates were selected and their production of the B2 and B9 were confirmed by HPLC. The three isolates were identified on species level by sequencing their 16S rRNA gene which showed 100% identical to strains of Pediococcus acidilactici. Thus, they were named as P. acidilactici WNYM01, P. acidilactici WNYM02, P. acidilactici WNYM03 and submitted to the Genbank database with accession numbers. They met the probiotic criteria by expressing 90-95% survival rate at pH (2.0-9.0) and bile salt up to 2% for 3 h in addition to their antimicrobial activity against gram positive and negative microorganisms. They also showed no hemolytic activity and common pattern for antibiotic susceptibility. Our three strains were tested individually or in mixture in vivo on rat colitis model compared to ulcerative group. The strains were administrated orally to rats in daily dose containing CFU 109 for 14 days then followed by induction of colitis using acetic acid then the oral administration was continued for more four days. The histology results, the anti-inflammatory and anti-oxidative stress biomarkers showed the protective role of the strains compared to the ulcerative group. As a conclusion, we introduce novel three probiotic candidates for pharmaceutical preparations and health applications.

Two-weeks repeated-dose oral toxicity study of Pediococcus acidilactici J9 in a mice model.

BACKGROUND: Helicobacter pylori (H. pylori) is an important pathogen that causes chronic gastritis and peptic ulcer, and is related to the development of gastric carcinoma. Several chemicals, including antibiotics, have been used to eradicate H.pylori. However, more studies are yet requred to accomplish a sufficient therapy. Pediococcus acidilactici (P. acidilactici) J9 were studied for inhibition of binding of H.pylori binding to human gastric cell lines. This study was performed in order to investigate the repeated-dose toxicity of P. acidilactici J9 in male and female mice. RESULTS: C57BL/6 male and female Mus musculus were divided into four groups (n = 10 in each group). P. acidilactici J9 was administered daily by oral injection of vehicle control at dosage levels to a low-dose group (500 mg/kg/day), middle-dose group (1000 mg/kg/day), and high-dose group (2000 mg/kg/day) for 2 weeks. After 14 days of exposure, the blood biochemistry and hematology were investigated, along with a histopathology exam. There were no bacterial-related deaths or abnormal clinical signs in either gender of mouse. The data was observed during the period in terms of body weight, food intake, and water consumption. Also, no alterations in organ weights upon administration of P. acidilactici J9 alone were observed. The adhesion and growth of H. pylori were inhibited by a 24 h treatment of H. pylori and P. acidilactici J9 on adenocarcinoma gastric (AGS) cells, which are gastric cancer cells. Compared to the control group (AGS cell and H. pylori), the number of H. pylori analyzed by FACS significantly (p < 0.01) decreased after incubation of AGS cell with P. acidilactici J9 for 24 h. CONCLUSIONS: These results suggest that the oral application of P. acidilactici J9, up to a dosage level of 2000 mg/kg/day, causes no adverse effects in both male and female mice. P. acidilactici J9 inhibits the adhesion of H.pylori to AGS cancer cells. When used as probiotics, P. acidilactici J9 may help decrease the occurrence of gastritis and reduce the risk of H.pylori infection with promising safety issues.

Pediococcus acidilactici intake decreases the clinical severity of atopic dermatitis along with increasing mucin production and improving the gut microbiome in Nc/Nga mice.

Atopic dermatitis (AD) is a chronic inflammatory skin disease that is associated with intestinal microflora. Since specific probiotics may have better efficacy for AD, we determined the efficacy of Pediococcus acidilactici SRCM102024 (PA) for treating AD in HaCaT cells and NC/Nga mice and explored the mechanism of action. AD-like pathology was induced in HaCaT cells and the dorsal skin of Nc/Nga mice by local exposure to 2,4-dinitrochlorobenzene (DNCB). In AD-lesion induced mice, PA in low-, medium- and high-dosages (5 × 10E6, 5 × 10E7 and 5 × 10E8 CFU/kg bw, respectively) and dexamethasone (3 mg/kg bw, positive-control) were orally administered for 5 weeks. The clinical AD severity, serum immunoglobulin E (IgE) and TNF-α, gene expressions of interleukin (IL)-4, IL-13, and TNF-α and gut microflora were measured. PA treatment (100-300 CFU/mL) dose-dependently increased cell survival in DNCB-induced HACAT cells. PA reduced the relative mRNA expression of PAR-2, TNF-α, IL-4 and IL-13 in the cells. In dorsal skin of Nc/Nga mice applied with DNCB, PA dose-dependently attenuated erythema, hemorrhage, edema, excoriation, dryness and scratching behavior and PA-H improved the clinical symptoms similar to the positive-control. PA-M and PA-H treatment significantly prevented the disturbance of the dorsal skin tissues and decreased the inflammatory cellular infiltrate of mast cells, compared to the control. PA dose-dependently reduced serum IgE and TNF-α concentrations and the mRNA expression of TNF-α, IL-4, and IL-13 in dorsal skin. In gut microflora, relative counts of Lactobacillales, Butyricicoccus and Ruminococcus were decreased in the AD-control compared to the positive-control and the PA-M and PA-H prevented their decrease. However, the positive-control increased serum AST and ALT activities, indicating liver damage as an adverse effect. In conclusion, oral treatment of PA (human equivalent 1 × 10E9-1 × 10E10) relieved the AD symptoms by dose-dependently preventing over-activation of the immune response. Oral PA intake may be a safe and effective alternative therapy for AD.

In vitro and in vivo anti-clostridial activity of newly isolated Pediococcus acidilactici SPM138 against Clostridium difficile.

Clostridium difficile infection (CDI) has become a growing health concern, as evident from the increase in the mortality rate among elderly or hospitalized patients. Treatment of CDI is usually based on antibiotics (metronidazole and vancomycin), but it has some limitations, including cost and antibiotic resistance. Probiotics could offer an effective remedy to prevent CDI and could be an auxiliary agent in treatment CDI. Here, the anti-clostridial activity of a newly isolated probiotic strain, Pediococcus acidilactici SPM138 (SPM138) was evaluated. The cultivation of C. difficile (CD) with SPM138, inhibited the growth of CD and significantly reduced CD toxins level. The result of MTT assay showed that, incubation with 25% CD culture supernatant decreased the survival rate of HT-29 cells to less than 20%. However, the survival rate of these cells increased to 98% in the presence of the 25% CD + SPM138 supernatant. The mRNA expression of IL-6, IL-8 and PTGS2 in HT-29 cells decreased by more than 60% upon incubation with CD + SPM138 co-cultures as compared to the levels observed after treatment with CD supernatant only. The concentration of IL-8 also decreased by more than 60% upon treatment with CD + SPM138 co-culture supernatant. In a C. difficile PCR ribotype 027-infected mouse model, the concentration of CD toxin in stool samples of SPM138-fed mice was only 37% of that reported in C. difficile 027-infected group. These findings show that P. acidilactici SPM138 may be a promising probiotic in CDI.

Aggregation, adhesion and efficacy studies of probiotic candidate Pediococcus acidilactici NCDC 252: a strain of dairy origin.

Aggregation and adhesion capability and survival efficacy of candidate probiotic strain Pediococcus acidilactici NCDC 252 under simulated gastric, intestinal and vaginal conditions was studied. The strain exhibited strong autoaggregation phenotype and coaggregation with other Lactic acid bacteria (LAB) and E. coli. The adhesion studies of NCDC 252 to pig's intestinal epithelial cells showed its adhesive ability. Aggregation and adhesiveness were related through cell surface proteins as removal/extraction of surface proteins resulted in altered aggregation and no adhesiveness. Cell surface proteins were analysed by SDS-PAGE and also in silico analysed from its genome. SDS-PAGE analysis of cell surface proteins of NCDC 252 revealed two potential proteins of approximately 74.3 and 53.6 kDa to be involved in host-probiotic interaction. Removal of cell surface proteins by LiCl-treatment (5 mol l-1) resulted in loss of aggregation and adhesiveness. Further survival of NCDC 252 under simulated gastrointestinal and vaginal conditions in terms of high viable counts confirmed its efficacy for its survival under gut and urogenital conditions. These observations suggest that it can be used further in functional foods, nutraceuticals and in combating urogenital infections. As NCDC 252 was able to survive in intestinal conditions, interaction of its cell surface proteins with intestinal mucins was studied in silico by docking. Highest affinity of adhesion was observed for MUC3B. In conclucion, NCDC 252, exhibited aggregation phenotype and adhesion capability. Survivability of NCDC 252 under simulated conditions and its interaction with human mucins confirms its efficacy to be used as probiotic.

Probiotic strains improve high-fat diet-induced hypercholesterolemia through modulating gut microbiota in ways different from atorvastatin.

Hypercholesterolemia is a major risk factor for cardiovascular disease (CVD). Probiotics are one of the most popular dietary supplements for hypercholesterolemia, but there are questions as to whether there are differences between probiotics and cholesterol-lowering drugs like atorvastatin (ATO) both in effectiveness and in the underlying mechanisms. In this study, the hypocholesterolemia effects of 4 probiotic strains were investigated and compared with ATO, focusing on their impacts on the gut microbiota. A hypercholesterolemia model was established via high-fat diet (HFD) in golden hamsters after which ATO and the 4 probiotics were orally administered individually for 8 weeks. All probiotics were effective, but less than ATO, on body weight, serum parameters (TG, TC, LDL, INS, HbA1c) and expression of inflammatory factors (INF-α, IL-1ß, CRP), with strain JQII-5 being most significant. Besides, these effects were associated with restoration of microbiota dysbiosis induced by HFD. It was worth noting that ATO and probiotics induced different shifts of the gut microbiota in both structure and key phylotypes. Most interestingly, Allobaculum, a HFD-suppressed genus, reported to be involved in alleviating oxidative stress, was enriched by all tested probiotic strains, but not by ATO. Furthermore, Prevotella, also a HFD-suppressed genus, was uniquely reversed by JQII-5. Importantly, most of the alerted genera and reversed genera were found to be correlated with the inflammatory state and serum lipid level. Compared with ATO, the probiotic strains were less effective on body weight, hypercholesterolemia, and inflammation. However, probiotics exert additional favorable effects on the gut microbiota, making them excellent potential complements to cholesterol-lowering drugs like ATO.

Assessment of Pediococcus acidilactici ATCC 8042 as potential Salmonella surrogate for thermal treatments of toasted oats cereal and peanut butter.

The control of Salmonella in low water activity foods poses a challenge for the food industry because of its thermal resistance. The use of surrogate bacteria in a food plant is considered a critical component to validate processing steps. The objective of this study was to evaluate the use of Pediococcus acidilactici ATCC 8042, a generally recognized as safe bacterium (GRAS), as potential surrogate for Salmonella in commercial toasted oats cereal (TOC) and peanut butter. P. acidilactici was compared to a five-serovar cocktail of Salmonella and Enterococcus faecium NRRL-B2354, separately. Cultures were inoculated into TOC and thermal kinetic parameters (δ, ß) were determined at 80, 85, 90, and 95 °C using the Weibull model. In peanut butter, δ and ß parameters were obtained at 63, 68, 73, and 77 °C. In TOC, the δ values (initial decimal reduction time) of P. acidilactici were 63 and 7 min at 80 and 95 °C, respectively, and at all four temperatures they were not significantly different from δ values of E. faecium. The δ value of Salmonella at 80 °C (139 min) was two-fold greater than the other two bacteria's values (p < 0.05). In peanut butter, δ values of P. acidilactici ranged from 31 min at 63 °C to 2.6 min at 77 °C, and at all temperatures they were not significantly different from E. faecium's δ values. In peanut butter, all Salmonella cocktail's δ values were significantly smaller than P. acidilactici's with values of 2 min at 63 °C and 0.4 min at 77 °C. These results indicated that P. acidilactici was as heat tolerant as E. faecium in these food matrices. However, the thermal inactivation kinetic parameters suggested that P. acidilactici can only be considered a Salmonella surrogate in TOC at temperatures above 85 °C. Because of its greater thermal tolerance in peanut butter, P. acidilactici may be used as Salmonella surrogate if an additional safety factor is recommended.

Effects of Lactobacillus plantarum, Pediococcus acidilactici, and Pediococcus pentosaceus on the Growth of Listeria monocytogenes and Salmonella on Alfalfa Sprouts.

The germination conditions of sprouted vegetables consisting of relatively high temperatures and humidity, low light, and abundance of nutrients are ideal for pathogen survival and growth. The continual occurrence of outbreaks and recalls associated with sprout vegetables indicate additional measures are needed to improve product safety. The objective of this study was to evaluate the efficacy of a mixture of Lactobacillus plantarum, Pediococcus acidilactici, and Pediococcus pentosaceus (LPP) against Listeria monocytogenes and Salmonella on alfalfa sprouts during 5 days of sprouting at 20°C and its influence on sprout quality. Alfalfa seeds were inoculated with L. monocytogenes or Salmonella (each at 1 and 3 log CFU/g) and LPP (7 log CFU/g). Populations of LPP were maintained at 7.5 to 8.0 log CFU/g throughout sprouting. LPP had a significant effect on the growth of L. monocytogenes and Salmonella ( P < 0.05). After 5 days of sprouting, populations of L. monocytogenes at an initial concentration of 1 and 3 log CFU/g of seeds treated with LPP were approximately 4.5 and 1.0 log CFU/g less than the untreated seeds, respectively. Populations of Salmonella at an initial concentration of 1 and 3 log CFU/g were 1.0 log CFU/g less than the control. LPP did not compromise the yield, seedling length, or pH of the sprouts.

Evaluation of the microbiological safety and sensory quality of a sliced cured-smoked pork product with protective cultures addition and modified atmosphere packaging.

The aim of this study was to evaluate the effect of two protective lactic acid bacteria cultures combined with modified atmosphere packaging on the survival/growth of Listeria innocua 2030c (as a surrogate for Listeria monocytogenes) and on sensory attributes of ready-to-eat 'lombo' over storage time. Sliced 'lombo', a traditional cured-smoked pork loin, was inoculated with L. innocua 2030c, Lactobacillus sakei ST153 (isolated from 'salpicão') and BLC35 culture (with Lactobacillus curvatus, Staphylococcus xylosus and Pediococcus acidilactici; CHR Hansen) as protective cultures. Samples were packed in two modified atmosphere packaging conditions (20% CO2/80% N2 and 40% CO2/60% N2) and stored at 5 ℃ for 124 days. Both cultures led to a reduction of 1-2 log CFU/g of L. innocua 2030c after 12 h; however, at the end of storage only Lb. sakei ST153 maintained this antilisterial effect, which was more evident at 40% CO2/60% N2. The influence of cultures addition and modified atmosphere packaging conditions on the sensory characteristics of the product were not significant. Thus, Lb. sakei ST153 combined with modified atmosphere packaging is a strong candidate to be used in a biopreservation strategy maintaining the traditional sensory quality of cured-smoked pork products and increasing their safety with respect to Listeria spp.

Probiotic supplementation with Lactobacillus plantarum and Pediococcus acidilactici for Helicobacter pylori therapy: A randomized, double-blind, placebo-controlled trial.

OBJECTIVE: To evaluate the safety, tolerability and efficacy of a probiotic supplementation for Helicobacter pylori (H. pylori) eradication therapy. DESIGN: Consecutive adult naive patients with a diagnosis of H. pylori infection who were prescribed eradication therapy according to clinical practice (10-day triple or nonbismuth quadruple concomitant therapy) randomly received probiotics (1 × 109 colony-forming units each strain, Lactobacillus plantarum and Pediococcus acidilactici) or matching placebo. Side effects at the end of the treatment, measured through a modified De Boer Scale, were the primary outcome. Secondary outcomes were compliance with therapy and eradication rates. RESULTS: A total of 209 patients (33% triple therapy, 66% non-bismuth quadruple therapy) were included [placebo (n = 106) or probiotic (n = 103)]. No differences were observed regarding side effects at the end of the treatment between groups (ß -0.023, P 0.738). Female gender (P < 0.001) and quadruple therapy (P 0.007) were independent predictors of side effects. No differences in compliance were observed, regardless of the study group or eradication therapy. Eradication rates were similar between groups [placebo 95% (95% confidence interval (CI), 89% to 98%) vs probiotic 97% (95% CI, 92% to 99%), P 0.721]. There were no relevant differences in cure rates (>90% in all cases) between triple and quadruple concomitant therapy. CONCLUSION: Probiotic supplementation containing Lactobacillus Plantarum and Pediococcus acidilactici to H. pylori treatment neither decreased side effects nor improved compliance with therapy or eradication rates.

A New Method of Producing a Natural Antibacterial Peptide by Encapsulated Probiotics Internalized with Inulin Nanoparticles as Prebiotics.

Synbiotics are a combination of probiotics and prebiotics, which lead to synergistic benefits in host welfare. Probiotics have been used as an alternative to antibiotics. Among the probiotics, Pediococcus acidilactici (PA) has shown excellent antimicrobial activity against Salmonella Gallinarum (SG) as a major poultry pathogen and has improved the production performances of animals. Inulin is widely used as a prebiotic for the improvement of animal health and growth. The main aim of this study is to investigate the effect of the antimicrobial activity of inulin nanoparticles (INs)-internalized PA encapsulated into alginate/chitosan/alginate (ACA) microcapsules (MCs) in future in vivo application. The prepared phthalyl INs (PINs) were characterized by DLS and FE-SEM. The contents of phthal groups in phthalyl inulin were estimated by ¹H-NMR measurement as 25.1 mol.-%. The sizes of the PINs measured by DLS were approximately 203 nm. Internalization into PA was confirmed by confocal microscopy and flow cytometry. Antimicrobial activity of PIN-internalized probiotics encapsulated into ACA MCs was measured by co-culture antimicrobial assays on SG. PIN-internalized probiotics had a higher antimicrobial ability than that of ACA MCs loaded with PA/inulin or PA. Interestingly, when PINs were treated with PA and encapsulated into ACA MCs, as a natural antimicrobial peptide, pediocin was produced much more in the culture medium compared with other groups inulin-loaded ACA MCs and PA-encapsulated into ACA MCs.

Efficacy of species-specific probiotic Pediococcus acidilactici FT28 on blood biochemical profile, carcass traits and physicochemical properties of meat in fattening pigs.

The present study investigates the influence of supplementing Pediococcus acidilactici strain FT28 on serum biochemistry, carcass and physicochemical properties of meat in fattening pigs. A total of 36 piglets (28day) were randomly divided into three groups of four replicates of three animals in each. Each group was fed one of the experimental diet as basal diet alone (Control-T0); basal diet supplemented with Pediococcus acidilactici strain FT28 (swine origin probiotic-T1) and basal diet supplemented with Lactobacillus acidophilus NCDC-15 (dairy origin probiotic-T2). Increased (P<0.05) level of serum total protein and albumin, and decreased (P<0.05) triglyceride were observed with probiotic supplementation either of swine or dairy origin, compared to control. The level globulin and glucose were higher (P<0.05) in T1 group among the dietary groups. Carcass traits remained unaltered except for the weight of ham, kidney and spleen which were higher (P<0.05) in the T1 group. Water holding capacity and TBARS were improved (P<0.05) in the T1 group than control, however comparable to the T2 group. The ether extract, pH and sensory attributes (juiciness and appearance) were improved (P<0.05) in the T1 group among the dietary groups. It is evident from the present study that P. acidilactici FT28 could serve as probiotic for enhancing carcass quality and physicochemical properties of pork without influencing the blood metabolites.

Effect of lactic acid bacteria-treated King grass silage on the performance traits and serum metabolites in New Zealand white rabbits (Oryctolagus cuniculus).

This study examined the effects of Lactobacillus plantarum and Pediococcus acidilactici-treated silage of King grass (Pennisetum purpureophoides) on the productive traits and blood biochemistry of New Zealand white (NZW) rabbits. King grass was ensiled without or with L. plantarum (T1), P. acidilactici (T2) and P. acidilactici + L. plantarum (T3). A total of 72 male NZW rabbits (6 weeks) of similar weight and appearance were housed in groups in metallic cages in a completely randomized design. The performance traits, daily feed intake, body weight gain and feed convention ratio were not affected by the silage treatment. Triglyceride and alanine aminotransferase (ALT) concentration was increased (p < .05) in T2 (0.92 mmol/L) compared to the other treatments. The superoxide dismutase (SOD), phosphorous (P) and magnesium (Mg) increased (p < .05) in T1 (658.01 nmol/ml, 5.1 mg/kg and 2.43 mg/kg, respectively) compared to the control and other experimental groups. Cu and Mn decreased (p < .05), while Zn increased (p < .05) in the treated groups compared to the control. The CP decreased (p < .05) in the treatment groups compared to the control. Neutral detergent fibre (NDF) increased (p < .05) in T2 (54.40%) compared to the other treatments. The inoculation of lactic acid bacteria-treated King grass silage supplementation positively affected rabbit performance and improved blood cholesterol profile, antioxidant status and improve nutrients digestibility.

Isolation and molecular identification of lactic acid bacteria from King grass and their application to improve the fermentation quality of sweet Sorghum.

The aim of the present study was isolation and molecular identification of lactic acid bacteria from King grass and their application to improve the fermentation quality of sweet Sorghum. Seventy-six strains of LAB were isolated; five strains were selected for Physiological and morphological tests and 16S rRNA sequencing. All five strains grew at different pH 3.5-8.0, different temperature 35, 40, 45, 50 °C and different NaCl concentrations 3, 6.5, 9.5%. Strains HDASK were identified Lactobacillus plantarum and SK3907, SK2A32, SK3A42 and ASKDD Pediococcus acidilactici. Three isolated strains and one commercial strain were added to sweet sorghum. Silage was prepared of four treatments and one control with three replicates as control (SKC, adding 2 ml/kg sterilizing water), L. plantarum commercial bacteria (SKP), L. plantarum (HDASK) isolated from King grass (SKA), P. acidilactici (SK3907) isolated from King grass (SKB) and P. acidilactici (ASKDD) isolated from King grass (SKD). All silage were prepared using polyethylene terephthalate bottles, and incubated at room temperature for different ensiling days. The level of pH, acetic acid, NH3-N, water soluble carbohydrate and butyric acid was significantly (P < 0.05) decreased. Lactic acid, ethanol and propionic acid (PA) was significantly (P < 0.05) increased in treatments compared to control. The dry matter, propionic acid neutral detergent fiber, acid detergent fiber did not significantly (P < 0.05) differ among the treatments but the values were increased and decreased. The number of yeast, mold and LAB were significantly (P < 0.05). It is suggested that the supplementation of LAB could enhanced the fermentation quality of sweet Sorghum silage.

Cholesterol lowering by Pediococcus acidilactici LAB4 and Lactobacillus plantarum LAB12 in adult zebrafish is associated with improved memory and involves an interplay between npc1l1 and abca1.

This study assessed the cholesterol lowering effect of Pediococcus acidilactici LAB4 and Lactobacillus plantarum LAB12 using adult zebrafish. Animals were fed with a high cholesterol diet (HCD) with/without LAB for seven weeks. Serum and liver cholesterol was quantified using colorimetric and dye staining methods. Expressions of npc1l1 and abca1 in the liver and intestine and appa in the brain were quantified using RT-PCR. Serum and liver cholesterol was significantly lowered in LAB4- and LAB12-fed zebrafish (≤64% and ≤71%, respectively), with reduced liver cholesterol deposition. The cholesterol lowering effect was accompanied by down-regulation of npc1l1 in intestines (≤28.7%), up-regulation of abca1 in the liver (≥30.5%) and down-regulation of appa in the brain (≤24.5%). A moderately strong positive Pearson correlation (r = 0.617, p < 0.01) was found between appa and serum cholesterol. LAB-fed zebrafish exhibited improved spatial learning and memory. LAB4 and LAB12 can be potentially used in preventing hypercholesterolaemia and Alzheimer's diseases.

Improved antimicrobial activity of Pediococcus acidilactici against Salmonella Gallinarum by UV mutagenesis and genome shuffling.

Pediococcus acidilactici is a widely used probiotic, and Salmonella enterica serovar Gallinarum (SG) is a significant pathogen in the poultry industry. In this study, we improved the antimicrobial activity of P. acidilactici against SG using UV mutation and genome shuffling (GS). To improve antimicrobial activity against SG, UV mutagenesis was performed against wild-type P. acidilactici (WT), and five mutants showed improved antimicrobial activity. To further improve antimicrobial activity, GS was performed on five UV mutants. Following GS, four mutants showed improved antimicrobial activity compared with the UV mutants and WT. The antimicrobial activity of GS1 was highest among the mutants; however, the activity was reduced when the culture supernatant was treated with proteinase K, suggesting that the improved antimicrobial activity is due to a proteinous substance such as bacteriocin. To validate the activity of GS1 in vivo, we designed multi-species probiotics and performed broiler feeding experiments. Groups consisted of no treatment (NC), avilamycin-treated (PC), probiotic group 1 containing WT (T1), and probiotic group 2 containing GS1 (T2). In broiler feeding experiments, coliform bacteria were significantly reduced in T2 compared with NC, PC, and T1. The cecal microbiota was modulated and pathogenic bacteria were reduced by GS1 oral administration. In this study, GS1 showed improved antimicrobial activity against SG in vitro and reduced pathogenic bacteria in a broiler feeding experiment. These results suggest that GS1 can serve as an efficient probiotic, as an alternative to antibiotics in the poultry industry.

In vitro assessment of Pediococcus acidilactici Kp10 for its potential use in the food industry.

BACKGROUND: Selection of a microbial strain for the incorporation into food products requires in vitro and in vivo evaluations. A bacteriocin-producing lactic acid bacterium (LAB), Pediococcus acidilactici Kp10, isolated from a traditional dried curd was assessed in vitro for its beneficial properties as a potential probiotic and starter culture. The inhibitory spectra of the bacterial strain against different gram-positive and gram-negative bacteria, its cell surface hydrophobicity and resistance to phenol, its haemolytic, amylolytic and proteolytic activities, ability to produce acid and coagulate milk together with its enzymatic characteristics and adhesion property were all evaluated in vitro. RESULTS: P. acidilactici Kp10 was moderately tolerant to phenol and adhere to mammalian epithelial cells (Vero cells and ileal mucosal epithelium). The bacterium also exhibited antimicrobial activity against several gram-positive and gram-negative food-spoilage and food-borne pathogens such as Listeria monocytgenes ATCC 15313, Salmonella enterica ATCC 13311, Shigella sonnei ATCC 9290, Klebsiella oxytoca ATCC 13182, Enterobacter cloaca ATCC 35030 and Streptococcus pyogenes ATCC 12378. The absence of haemolytic activity and proteinase (trypsin) and the presence of a strong peptidase (leucine-arylamidase) and esterase-lipase (C4 and C8) were observed in this LAB strain. P. acidilactici Kp10 also produced acid, coagulated milk and has demonstrated proteolytic and amylolactic activities. CONCLUSION: The properties exhibited by P. acidilactici Kp10 suggested its potential application as probiotic and starter culture in the food industry.

Isolation, Identification and Screening of Potential Probiotic Bacteria in Milk from South African Saanen Goats.

This study aimed to evaluate lactic acid bacteria isolates from Saanen goats' milk for probiotic attributes, thereby determining their potential as direct-fed microbials for goats. Isolates were identified using API 50CH system, 16S rDNA sequencing and matrix-assisted laser desorption ionization-time of flight mass spectrometry. All 17 isolates obtained were identified as Lactobacillus plantarum except one identified as Pediococcus acidilactici. Four isolates identified as L. plantarum (Accession numbers KJ026587.1, KM207826.1, KC83663.1 and KJ958428.1) by at least two of the techniques used and isolate 17 differently identified by all the methods used were selected as representatives and then screened for probiotic properties. These isolates displayed phenotypic probiotic attributes including tolerance to acid and bile salts, ability to adhere to intestines and possession of antagonistic activities against Proteus vulgaris, Staphylococcus aureus, Salmonella typhimurium, Pseudomonas aeruginosa and Escherichia coli. The lactic acid bacteria isolated from Saanen goats' milk showed potential to be used as sustainable probiotics in goats' industry. Successful use of probiotics in animals depends upon availability of appropriate isolates originating from the specific host animal. This study is a positive contribution towards identification of isolates with potential for formulation as direct-fed microbials for South African Saanen goats.