Lactobacillus Paracasei Lpc-37 Research Articles

Effects of Probiotics on Inflammatory Biomarkers and Its Associations With Cardiac Autonomic Function in Women With Arterial Hypertension: A Secondary Analysis of a Randomized Clinical Trial.

Preclinical evidence suggests that probiotic administration may exert an anti-inflammatory effect and reduce autonomic dysfunction and blood pressure. This study evaluated the effects of probiotic therapy on inflammatory biomarkers and characterized the correlations between inflammation and cardiac autonomic function in women with arterial hypertension. Women were randomized into probiotics (n = 20) or placebo (n = 20). The probiotic group received 109CFU/day of Lactobacillus (L.) paracasei LPC-37, L. rhamnosus HN001, L. acidophilus NCFM, and Bifidobacterium lactis HN019, and the placebo group received polydextrose. Clinical, electrocardiogram, heart rate variability (HRV) analysis, and cytokine levels were assessed at baseline and after 8weeks. Women who received probiotics for 8weeks had increased serum levels of IL-17A (p = 0.02) and decreased INF-γ (p = 0.02) compared to baseline. Probiotic supplementation increased serum levels of IL-10 compared to the placebo group (p = 0.03). Probiotic or placebo administration did not change serum levels of TNFα and IL-6. Serum levels of IL-2 (p = 0.001, and p = 0.001) and IL-4 (p = 0.001, and p = 0.001) were reduced in women receiving placebo or probiotics, respectively. Correlations between HRV indices and inflammatory variables showed that INF-γ was positively correlated with heart rate (HR) and sympathetic HRV indices and negatively correlated with vagal HRV indices. IL-10 was negatively correlated with HR and sympathetic HRV indices. IL-6 was negatively correlated with parasympathetic HRV indices and positively correlated with SD2/SD1 ratio. Probiotic therapy has a discreet anti-inflammatory effect in hypertensive women, and pro-inflammatory cytokines were negatively correlated with vagal modulation and positively correlated with sympathetic modulation of HRV.The clinical trial was registered in the Brazilian Registry of Clinical Trials (ReBEC) with the identification RBR-9mj2dt.

Adjuvant use of multispecies probiotic in the treatment of peri-implant mucositis: A randomized controlled trial.

This randomized placebo-controlled clinical trial evaluated the effects of multispecies probiotic containing Lactobacillus rhamnosus HN001™, Lactobacillus paracasei Lpc-37®, and Bifidobacterium animalis subsp lactis HN019™ as an adjunct to mechanical debridement (MD) on changes in bleeding on probing (BOP) in edentulous patients with peri-implant mucositis (PiM). Patients were randomly assigned to test (probiotic) or control (placebo) groups. All sites with PiM received MD and topical gel application (probiotic or placebo) at baseline and 12 weeks. After initial MD, patients consumed probiotic or placebo capsules twice a day for 12 weeks. Clinical (modified sulcus bleeding index [mSBI]; modified plaque index [mPI]; probing depth [PD]; and BOP) and immunological parameters were collected at baseline and after 12 and 24 weeks. Data were statistically analysed (p < .05). Thirty-six patients with PiM were recruited. The test group presented higher prevalence (p< .05) of cases of restored peri-implant health at 24 weeks than did the control group (72.2% and 33.3%, respectively). No significant difference was observed between test (n= 18) and control (n= 18) groups for mPI and PD. mSBI %-score 0 was higher in the test group than in the control group at 24 weeks (p< .05). When compared with baseline, both groups presented reduced BOP at 12 and 24 weeks (p< .05). BOP was lower in the test group than in the control group at 12 (mean difference=-14.54%; 95% confidence interval [CI]=-28.87 to 0.22; p= .0163) and 24 (mean difference=-12.56%; 95% CI=-26.51 to 1.37; p= .0090) weeks. At 24 weeks, only the test group presented lower levels of interleukin (IL)-1β, IL-6, IL-8, and tumour necrosis factor (TNF)-α than those at baseline (p< .05). The multispecies probiotic (administered locally and systemically) containing L. rhamnosus HN001™, L. paracasei Lpc-37®, and B. lactis HN019™ as an adjunct to repeated MD promotes additional clinical and immunological benefits in the treatment of PiM in edentulous patients (ClinicalTrials.gov NCT04187222).

Targeting the Gut Microbiota in Coronavirus Disease 2019: Hype or Hope?

Targeting the Gut Microbiota in Coronavirus Disease 2019: Hype or Hope?

Do Probiotics During In-Hospital Antibiotic Treatment Prevent Colonization of Gut Microbiota With Multi-Drug-Resistant Bacteria? A Randomized Placebo-Controlled Trial Comparing Saccharomyces to a Mixture of Lactobacillus, Bifidobacterium, and Saccharomyces.

Objective: Most infections with Enterobacteriaceae producing AmpC β-lactamase (AmpC)-, extended-spectrum β-lactamase (ESBL)-, and carbapenemase-producing bacteria, vancomycin-resistant Enterococcus as well as naturally resistant non-fermenting bacteria such as Pseudomonas aeruginosa, are related to a prior colonization of the gut microbiota. The objective of this study was to determine whether treatment with probiotics during an antibiotic treatment could prevent the colonization of the gut microbiota with multi-drug resistant bacteria.Method: In total, 120 patients treated for 10 days with amoxicillin-clavulanate antibiotics were included in a randomized, placebo-controlled, double-blinded trial, comparing the effects of a 30 days treatment with placebo Saccharomyces boulardii CNCM I-745® and a probiotic mixture containing Saccharomyces boulardii, Lactobacillus acidophilus NCFM, Lactobacillus paracasei Lpc-37, Bifidobacterium lactis Bl-04, and Bifidobacterium lactis Bi-07 (Bactiol duo®). Study treatment was initiated within 48 h of the antibiotic being initiated. Most of the patients included were elderly with a mean age of 78 years old with multiple comorbidities. Stools were collected at the time of inclusion in the trial, at the end of the antibiotic treatment, and the end of the study treatment. These were cultured on selective antibiotic media.Results: Treatment with the probiotic mixture led to a significant decline in colonization with Pseudomonas after antibiotic treatment from 25 to 8.3% (p = 0.041). Colonization with AmpC-producing enterobacteria was transiently increased after the antibiotic treatment (p = 0.027) and declined after the probiotic intervention (p= 0.041). No significant changes were observed in the placebo and Saccharomyces groups. Up to 2 years after the trial, no infection with ESBL-producing bacteria was observed in the probiotic mixture group.Conclusion: The association of Saccharomyces boulardii with specific strains of Lactobacillus and Bifidobacterium influences antibiotic treatment by counteracting the colonization of the colon microbiota with antibiotic-resistant pathogens.

Portable CRISPR-Cas9N System for Flexible Genome Engineering in Lactobacillus acidophilus, Lactobacillus gasseri, and Lactobacillus paracasei.

Diverse Lactobacillus strains are widely used as probiotic cultures in the dairy and dietary supplement industries, and specific strains, such as Lactobacillus acidophilus NCFM, have been engineered for the development of biotherapeutics. To expand the Lactobacillus manipulation toolbox with enhanced efficiency and ease, we present here a CRISPR (clustered regularly interspaced palindromic repeats)-SpyCas9D10A nickase (Cas9N)-based system for programmable engineering of L. acidophilus NCFM, a model probiotic bacterium. Successful single-plasmid delivery system was achieved with the engineered pLbCas9N vector harboring cas9N under the regulation of a Lactobacillus promoter and a cloning region for a customized single guide RNA (sgRNA) and editing template. The functionality of the pLbCas9N system was validated in NCFM with targeted chromosomal deletions ranging between 300 bp and 1.9 kb at various loci (rafE, lacS, and ltaS), yielding 35 to 100% mutant recovery rates. Genome analysis of the mutants confirmed precision and specificity of the pLbCas9N system. To showcase the versatility of this system, we also inserted an mCherry fluorescent-protein gene downstream of the pgm gene to create a polycistronic transcript. The pLbCas9N system was further deployed in other species to generate a concurrent single-base substitution and gene deletion in Lactobacillus gasseri ATCC 33323 and an in-frame gene deletion in Lactobacillus paracasei Lpc-37, highlighting the portability of the system in phylogenetically distant Lactobacillus species, where its targeting activity was not interfered with by endogenous CRISPR-Cas systems. Collectively, these editing outcomes illustrate the robustness and versatility of the pLbCas9N system for genome manipulations in diverse lactobacilli and open new avenues for the engineering of health-promoting lactic acid bacteria.IMPORTANCE This work describes the development of a lactobacillus CRISPR-based editing system for genome manipulations in three Lactobacillus species belonging to the lactic acid bacteria (LAB), which are commonly known for their long history of use in food fermentations and as indigenous members of healthy microbiotas and for their emerging roles in human and animal commercial health-promoting applications. We exploited the established CRISPR-SpyCas9 nickase for flexible and precise genome editing applications in Lactobacillus acidophilus and further demonstrated the efficacy of this universal system in two distantly related Lactobacillus species. This versatile Cas9-based system facilitates genome engineering compared to conventional gene replacement systems and represents a valuable gene editing modality in species that do not possess native CRISPR-Cas systems. Overall, this portable tool contributes to expanding the genome editing toolbox of LAB for studying their health-promoting mechanisms and engineering of these beneficial microbes as next-generation vaccines and designer probiotics.

Technological Characterisation of Probiotic Lactic Acid Bacteria as Starter Cultures for Dry Fermented Sausages.

The objective of this study was to investigate probiotic microorganisms for use as starter cultures in dry fermented sausages production. A total of eight strains were studied evaluating technological and safety characteristics including the ability to grow, lactic acid production, gas formation, catalase activity, nitrate reductase activity, proteolytic activity, lipolytic activity, hydrogen peroxide production, salt tolerance, performance at low temperatures, decarboxylation of amino acids and antimicrobial activity against pathogens associated with the product. Lactobacillus rhamnosus R0011, L. rhamnosus Lr-32, Lactobacillus paracasei Lpc-37, Lactobacillus casei Shirota and Enterococcus faecium MXVK29 were good candidates for use as fermented sausages starters cultures because they showed the best technological and safety properties since they did not demonstrate amino acid decarboxylation but showed antimicrobial activity against Listeria monocytogenes, Escherichia coli, Salmonella Dublin and Staphylococcus aureus. L. rhamnosus Lr-32 was the strain best tolerating the levels of salt, nitrate and low pH during the simulated stages of fermentation and ripening of sausage. The strain was thus the most promising of the tested probiotics as sausage starter culture. The findings warrant studies in a meat matrix, such as that of raw-cured sausage, to evaluate the effects of L. rhamnosus Lr-32 under actual conditions.

Fecal Recovery of Probiotics Administered as a Multi-Strain Formulation during Antibiotic Treatment.

The present study aimed to investigate whether probiotic recovery is affected when consumed together with antibiotics. Fecal samples were collected from an earlier antibiotic associated diarrhea, randomized, placebo-controlled study with a product consisting of a combination of Lactobacillus acidophilus NCFM, Lactobacillus paracasei Lpc-37, and Bifidobacterium lactis Bi-07, B. lactis Bl-04 at equal numbers and at a total dose of 1010 CFU. Fecal samples were collected during the screening visit (T0), i.e., at the time of antibiotic prescription, and then on the last day of the antibiotic treatment (T1) as well as seven days after the subject had stopped taking the antibiotic treatment (T2) and at two weeks after completing antibiotic treatment and one week after probiotic/placebo consumption stopped (T3). Samples were analyzed for the presence of the four administered strains. The study was registered at clinicaltrials.gov as NCT01596829. Detection levels of all four strains were significantly increased from T0 to T1 and returned to baseline level from T2 to T3. There were also significantly more subjects with detectable levels of L. paracasei Lpc-37, B. lactis Bi-07, and B. lactis Bl-04 at T1 and T2 compared to T0 and T3, and compared to placebo. Each of the four strains could be detected in the feces of patients apparently unaffected by the simultaneous consumption of antibiotics.

Dose-Response Recovery of Probiotic Strains in Simulated Gastro-Intestinal Passage

The probiotic definition stipulates “adequate amounts”. Here, we investigated the metabolic output and recovery rate of probiotic strains using a simulated upper gastro-intestinal passage and colonic fermentation. Two different doses, 7 × 109 colony forming units (CFU) and 7 × 1010 CFU, of a probiotic mixture (Bifidobacterium lactis Bl-04, Lactobacillus acidophilus La-14, Lactobacillus paracasei Lpc-37, and Lactobacillus plantarum Lp-115) or placebo were tested. The four strains were quantified by qPCR and the metabolites analyzed by gas chromatography. There was a dose-response in the detection of all four strains. There was a slightly larger increase between the two doses for L. paracasei Lpc-37 as compared with the other strains; this may suggest a greater robustness of this strain. Compared with the placebo, the high dose simulations generated more propionic acid and a higher total of short chain fatty acids (SCFA). Higher doses of a species are required to reach measurable increases above the baseline level of this species.

Effects of probiotic therapy on cardio-metabolic parameters and autonomic modulation in hypertensive women: a randomized, triple-blind, placebo-controlled trial.

We assessed the effects of probiotic therapy for 8 weeks on cardiometabolic variables and autonomic function in women medically diagnosed with arterial hypertension. Forty women with arterial hypertension, 20-50 years, were assigned to two groups in this randomized, triple-blind, placebo-controlled clinical trial. Patients in the probiotic group received a daily sachet containing Lactobacillus para casei LPC-37, Lactobacillus rhamnosus HN001, Lactobacillus acidophilus NCFM, and Bifidobacterium lactis HN019 (109 CFU of each strain) for 8 weeks. Patients in the placebo group received identical sachets with polydextrose (1 g day-1, for 8 weeks). Anthropometric, BP, electrocardiogram, biochemical measurements, fecal microbiota composition, and glucose hydrogen breath test were assessed at baseline and after 8 weeks intervention. Anthropometric variables (weight, BMI, and waist circumference) were similar between the two groups (p > 0.05). Probiotic supplementation significantly reduced fasting glucose (change -10.3 mg dL-1, p < 0.05) and cholesterol levels (change -23.6 mg dL-1, p < 0.05), and increased the HDL-cholesterol (change 6.5 mg dL-1, p < 0.05) compared with the baseline condition. Probiotic supplementation lowered, although without statistical significance, systolic BP by about 5 mmHg and diastolic BP by about 2 mmHg in hypertensive women. Lastly, probiotic administration reduced the low frequency (LF) oscillation and LF/high frequency (HF) ratio (p < 0.05) in the frequency domain of heart rate variability, suggesting an improvement in autonomic modulation. Probiotic therapy for 8 weeks reduced fasting glucose levels, and improved the lipid profile and autonomic modulation in hypertensive women.

Modulation of Anti-Microbial Resistant &amp;lt;i&amp;gt;Salmonella heidelberg&amp;lt;/i&amp;gt; Using Synbiotics (Probiotics and Prebiotics) in Two &amp;lt;i&amp;gt;In-Vitro&amp;lt;/i&amp;gt; Assays (Cross-Streaking and Agar Wells Diffusion)

Salmonellosis is the most prevalent bacterial foodborne disease in many countries worldwide. Utilization of probiotics is one of the most accepted ways to reduce Salmonella, especially lactic acid bacteria, as it has proven to reduce the enteric pathogens in monogastric and ruminant livestock animals through different mechanisms such as antimicrobials production, competitive adhesion to the gastrointestinal tract, and immune stimulation. Prebiotics could be utilized solely for health benefits as an alternative to probiotics or in addition to probiotics for a synergistic effect known as synbiotics. The aim of this study was to compare effects of different probiotic strains (Lactobacillus acidophilus (La-14), Lactobacillus paracasei (Lpc-37), Streptococcus thermophiles (St-21), Bifidobacterium bifidum (Bb-06), and Aspergillus niger (ATCC&#17416888TM) and without prebiotics (Mannose; Xylose; Galactooligosaccharides GOS; Inulin; and Dandelion extract) on lowering Salmonella heidelberg CFU in vitro. Different inhibition levels probiotic strains were assessed and compared in the presence and absence of 2.5% prebiotic compounds using cross-streaking and agar well diffusion assays. Recommendations for the growth of selected microorganisms such as temperature and oxygen conditions were taken into consideration. All the analysis was conducted in triplicates. The results showed that all the probiotics strains except S. thermophiles were able to significantly (P < 0.05) inhibit the growth of S. heidelberg in at least one of the assays. The difference in inhibition percentage confirms that probiotic strains have multiple inhibition mechanisms, such as production of antimicrobials, lower pH by producing organic acids (acetic acid, lactic acid, etc.), and inhibition of pathogen’s virulence factor expression, and production of lipopolysaccharide solubilizing compounds.

EFFECT OF SUPPLEMENTATION WITH PROBIOTICS ON COLONIC ANASTOMOSES IN RATS: MORPHOLOGICAL AND TENSIOMETRIC STUDY.

ABSTRACTBackground: The use of probiotics positively modifies the composition and function of intestinal flora, improving the quality of intestinal anastomosis. Aim: To evaluate the impact of probiotic use on intestinal anastomosis of rats. Method: Thirty-six adult male Wistar rats (Rattus norvegicus albinus, Rodentia Mammalia) were used, with body weight ranging from 220-320 g. The animals were housed and acclimated individually in boxes receiving water and ration ad libitum. After initial acclimatization, the control group received perioperative ration ad libitum for 12 days (seven preoperatively and five postoperatively) associated with the maltodextrin formula at a dose of 250 mg/day in isocaloric and isovolumetric form. Likewise, the probiotic group received oral supplementation of probiotics dose of 250 mg/day, associated with isocaloric and isovolumetric diet. The probiotic chosen for this study was composed of strains (doses 1x109 CFU/g)12 Lactobacillus paracasei LPC-37, Bifidobacterium lactis HN0019, Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus NCFM. Probiotics or placebo were administered orally with the aid of a dosimeter spatula. Both groups underwent two colostomies, one in the right colon and the second in rectosigmoid, followed by reanastomosis with eight separate 6-0 mononylon stitches. The sacrifice took place on the fifth day. The parameters evaluated included tensile strength, histology and collagen densitometry. Results: The rate of intestinal fistula for the control and probiotic groups were, respectively, 22.22% and 11.11% (p=0.6581).Perioperative supplementation with probiotics increased collagen deposition of types I and III (p<0.0001), improved maximum traction force and maximum rupture force, p=0.0250 and p=0.0116 respectively, fibrosis area (p<0.0001), and area of the inflammatory infiltrate (p=0.0115). Conclusions: The use of probiotics had a positive impact on the quality of intestinal anastomosis.

Strain specific stress-modulating effects of candidate probiotics: A systematic screening in a mouse model of chronic restraint stress

BackgroundChanges in the gut microbiota have been implicated in mood and cognition. In rodents, supplementation with certain bacteria have been shown to alleviate adverse effects of stress on gut microbiota composition and behaviour, but little is known of how the performance of different strains compare to each other. We took a systematic approach to test the efficacy of twelve candidate probiotic strains from ten species/sub-species of Bifidobacterium and Lactobacillus on behaviours and neuroendocrine responses of chronically stressed mice. MethodsThe strains were tested in four screening experiments with non-stressed and chronically stressed vehicle groups. The three most efficacious strains were re-tested to validate the results. Mice were administered a daily oral gavage containing either 1 × 109 colony forming units (CFU) of selected candidate probiotic or saline solution for one week prior to and for three weeks during daily chronic restraint stress. Behavioural tests including the elevated plus maze, open field, novel object recognition, and forced swim test were applied during week five. Corticosterone and adrenocorticotropic hormone (ACTH) were analysed to measure the neuroendocrine response to stress. Plasma and tissue samples were collected for biomarker analyses. ResultsOf the twelve candidate probiotics, Lactobacillus paracasei Lpc-37, Lactobacillus plantarum LP12407, Lactobacillus plantarum LP12418 and Lactobacillus plantarum LP12151 prevented stress-associated anxiety and depression-related behaviours from developing compared with chronically stressed vehicle mice. In addition, Lpc-37 improved cognition. ConclusionThis systematic screening indicates species- and strain-dependent effects on behavioural outcomes related to stress and further suggests that strains differ from each other in their effects on potential mechanistic outcomes.

The effect of a probiotic blend on gastrointestinal symptoms in constipated patients: a double blind, randomised, placebo controlled 2-week trial.

Selected strains of lactobacilli and bifidobacteria are known to ameliorate constipation-related symptoms and have previously shown efficacy on digestive health. In this clinical trial, the safety and effectiveness of a probiotic blend containing lactobacilli and bifidobacteria were evaluated in adults with self-reported bloating and functional constipation. Constipation was diagnosed by the Rome III criteria. A total of 156 adults were randomised into this double-blind and placebo-controlled trial. Participants consumed the combination of Lactobacillus acidophilus NCFM (1010 cfu), Lactobacillus paracasei Lpc-37 (2.5×109 cfu), Bifidobacterium animalis subsp. lactis strains Bl-04 (2.5×109 cfu), Bi-07 (2.5×109 cfu) and HN019 (1010 cfu) (n=78), or placebo (microcrystalline cellulose) (n=78) for two weeks. After treatment the following were measured: primary outcome of bloating and secondary outcomes of colonic transit time, bowel movement frequency, stool consistency, other gastrointestinal symptoms (flatulence, abdominal pain, and burbling), constipation-related questionnaires (PAC-SYM and PAC-QoL) and product satisfaction. Faecal recovery of consumed strains was determined. The enrolled population was defined as constipated, however, the initial bloating severity was lower than in previous similar studies. No clinically significant observations related to the safety of the product were reported. Product efficacy was not shown in the primary analysis for bloating nor for the secondary efficacy analyses. The placebo functioned similarly as the probiotic product. In post-hoc analysis, a statistically significant decrease in flatulence in favour of the probiotic group was observed; day 7 (intention-to-treat (ITT): P=0.0313; per-protocol (PP): 0.0253) and on day 14 (ITT: P=0.0116; PP: P=0.0102) as measured by area under the curve (AUC) analysis. The mean AUC of all symptoms decreased in favour of the probiotic group, indicating less digestive discomfort. The study was registered at the ISRCTN registry (ISRCTN41607808).

Influence of polysaccharide as co-encapsulant on powder characteristics, survival and viability of microencapsulated Lactobacillus paracasei Lpc-37 by spray drying

In the present work, five polysaccharides were selected as co-encapsulant of skim milk based matrix and used for the microencapsulation of Lactobacillus paracasei Lpc-37 by spray drying. The morphological characterization, physical properties, encapsulation efficiency, gastrointestinal simulation, and storage stability of microencapsules were examined to evaluate the protective capacity against the adverse conditions of the different wall matrices. Addition of polysaccharides was associated with the enhancement of physical performance of microencapsulated powders, such as the flowability and water activity. Encapsulation matrices containing gum Arabic showed the greatest encapsulation efficiency of 97.1% (maximum survival 7.78 log CFU/g). L. paracasei Lpc-37 encapsulated with the matrices incorporating sodium alginate or sodium carboxymethylcellulose exhibited higher survival (>6.5 log CFU/g) after exposed to simulated gastrointestinal condition and lower viability loss (<0.3 log CFU/g) after 8 weeks of refrigerated storage. The application of polysaccharides, however, showed the tendency of higher hygroscopicity and lower stability of the powders in high temperature and humid environment.

Oral probiotic combination of Lactobacillus and Bifidobacterium alters the gastrointestinal microbiota during antibiotic treatment for Clostridium difficile infection.

Perturbations in the gastrointestinal microbiome caused by antibiotics are a major risk factor for Clostridium difficile infection (CDI). Probiotics are often recommended to mitigate CDI symptoms; however, there exists only limited evidence showing probiotic efficacy for CDI. Here, we examined changes to the GI microbiota in a study population where probiotic treatment was associated with significantly reduced duration of CDI diarrhea. Subjects being treated with standard of care antibiotics for a primary episode of CDI were randomized to probiotic treatment or placebo for 4 weeks. Probiotic treatment consisted of a daily multi-strain capsule (Lactobacillus acidophilus NCFM, ATCC 700396; Lactobacillus paracasei Lpc-37, ATCC SD5275; Bifidobacterium lactis Bi-07, ATCC SC5220; Bifidobacterium lactis B1-04, ATCC SD5219) containing 1.7 x 1010 CFUs. Stool was collected and analyzed using 16S rRNA sequencing. Microbiome analysis revealed apparent taxonomic differences between treatments and timepoints. Subjects administered probiotics had reduced Verrucomicrobiaceae at week 8 compared to controls. Bacteroides were significantly reduced between weeks 0 to 4 in probiotic treated subjects. Ruminococcus (family Lachnospiraceae), tended to be more abundant at week 8 than week 4 within the placebo group and at week 8 than week 0 within the probiotic group. Similar to these results, previous studies have associated these taxa with probiotic use and with mitigation of CDI symptoms. Compositional prediction of microbial community function revealed that subjects in the placebo group had microbiomes enriched with the iron complex transport system, while probiotic treated subjects had microbiomes enriched with the antibiotic transport system. Results indicate that probiotic use may impact the microbiome function in the face of a CDI; yet, more sensitive methods with higher resolution are warranted to better elucidate the roles associated with these changes. Continuing studies are needed to better understand probiotic effects on microbiome structure and function and the resulting impacts on CDI.

Safety evaluation of HOWARU® Restore (Lactobacillus acidophilus NCFM, Lactobacillus paracasei Lpc-37, Bifidobacterium animalis subsp. lactis Bl-04 and B. lactis Bi-07) for antibiotic resistance, genomic risk factors, and acute toxicity

Although probiotic lactobacilli and bifidobacteria are generally considered safe by various regulatory agencies, safety properties, such as absence of transferable antibiotic resistance, must still be determined for each strain prior to market introduction as a probiotic. Safety requirements for probiotics vary regionally and evaluation methods are not standardized, therefore methodologies are often adopted from food ingredients or chemicals to assess microbial safety. Four individual probiotic strains, Lactobacillus acidophilus NCFM®, Lactobacillus paracasei Lpc-37®, Bifidobacterium animalis subsp. lactis strains Bl-04®, and Bi-07®, and their combination (HOWARU® Restore) were examined for antibiotic resistance by broth microdilution culture, toxin genes by PCR and genome mining, and acute oral toxicity in rats. Only B. lactis Bl-04 exhibited antibiotic resistance above a regulated threshold due to a tetW gene previously demonstrated to be non-transferable. Genomic mining did not reveal any bacterial toxin genes known to harm mammalian hosts in any of the strains. The rodent studies did not indicate any evidence of acute toxicity following a dose of 1.7–4.1 × 1012 CFU/kg body weight. Considering a 100-fold safety margin, this corresponds to 1.2–2.8 × 1012 CFU for a 70 kg human. Our findings demonstrate a comprehensive approach of in vitro, in silico, and in vivo safety testing for probiotics.

Preparation and prebiotic potential of pectin oligosaccharides obtained from citrus peel pectin

Pectin oligosaccharide (POS) fractions were obtained by controlled chemical degradation of citrus peel pectin. By adjusting trifluoroacetic acid (TFA) concentration, three oligosaccharides were obtained, of molecular weight (Mw) range 3000–4000 Da, 2000–3000 Da and lower than 2000 Da. Varying hydrogen peroxide (H2O2) concentration and reaction time produced oligosaccharides of 3000–4000 Da and 2000–3000 Da. The relative proportions of acidic monosaccharide increased from 68.58% to 89.93% (TFA) and from 63.74% to 83.26% (H2O2) as the reaction conditions intensified. Prebiotic activity scores were used to quantify POS ability to promote selective growth of specific probiotics. Sample POSH1 from H2O2 degradation showed the highest prebiotic potential with prebiotic activity score 0.41 for Lactobacillus paracasei LPC-37 and 0.92 for Bifidobacterium bifidum ATCC 29521.

The influence of an adjunct culture of Lactobacillus paracasei LPC-37 on the physicochemical properties of Dutch-type cheese during ripening

The main objective of this study was to evaluate the effect of adjunct culture Lactobacillus paracasei LPC-37 on the physicochemical properties of ripening Dutch-type cheeses. Those products were produced under industrial conditions in accordance with the technological regulations. The use of adjunct culture Lactobacillus paracasei LPC-37 in the production of Dutch-type ripening cheeses did not affect the modification of their chemical composition, nor did it change their acidity during ripening. No significant effect of Lactobacillus paracasei LPC-37 culture on the scope of proteolysis and peptidolysis in cheeses products was noted. Although the experimental cheeses were characterized by a higher range and depth of proteolysis than the control group, the differences were not statistically significant.

Simulating colonic survival of probiotics in single-strain products compared to multi-strain products.

ABSTRACT Background: Probiotic formulations can be single- or multi-strain. Commercially, multi-strain preparations have been suggested to have improved functionality over single-strain cultures. Probiotics are often tested as single-strain preparations but may subsequently be commercially formulated as multi-strain products. Objective: The aim of this study was to determine what happens at the site of action, the intestine, with probiotics as single- compared to multi-strain preparations. The human gastrointestinal tract contains a broad mixture of different microbes which may affect the survival of different probiotics in different ways. Design: The current study was performed to evaluate, in an in vitro colon simulation, whether probiotics influence each other’s survival when they are taken as a combination of several strains (HOWARU Restore; Lactobacillus acidophilus NCFM, Lactobacillus paracasei Lpc-37, Bifidobacterium lactis Bl-04 and B. lactis Bi-07) compared to the strains as single preparations. Results: All strains could be detected after the colon simulations and there were no substantial differences in levels of the same strain when comparing single- and multi-strain products. Conclusions: It can be concluded that probiotics do not have an antagonistic effect on each other’s survival when used in a multi-strain product compared to a single-strain product, at least within a microbiota in a simulated colonic environment.

The effect of milk fat substitution with palm fat on lactic acid bacteria counts in cheese-like products

This study evaluated the influence of complete substitution of milk fat with palm fat on the growth of lactic acid bacteria in Gouda-type cheese-like products, as compared with Gouda cheeses of the same type. Cheese-like products and ripened cheeses were produced in industrial plants with or without the addition of probiotic cultures. The analysed products were evaluated during ripening and storage based on the counts of starter lactic acid bacteria (SLAB), non-starter lactic acid bacteria and the viability of Lactobacillus acidophilus NCFM and Lactobacillus paracasei LPC-37 probiotic cultures.The highest SLAB counts were determined in control and experimental Gouda cheeses. SLAB were significantly less abundant in cheese-like products. During ripening, a greater reduction in SLAB counts was observed in control ripened cheeses than in control cheese-like products. Lactococcus sp. counts were lower in experimental products containing L. acidophilus NCFM than in products with L. paracasei LPC-37.The viability of L. acidophilus NCFM was significantly correlated with product type and the time of ripening and storage. The counts of L. acidophilus NCFM were higher in ripened cheeses than in cheese-like products. L. paracasei LPC-37 cultures were characterised by higher viability rates in products than L. acidophilus NCFM.