Commercial Probiotic Supplements Research Articles

Çeşitli Kaynaklardan ve Ticari Gıda Takviyelerinden İzole Edilen Bacillus Türlerinin Bazı Probiyotik Özelliklerinin Karşılaştırmalı Değerlendirilmesi

Bacillus species are gram-positive, aerobic, peritrically flagellated and endospore-forming bacteria. They can be found everywhere in the environment, especially in soil (its common habitat), water, dust or in the air. Probiotics, which have beneficial health effects, constitute an important group of Bacillus species. This study aimed to isolate Bacillus from various sources, identify it molecularly and determine its probiotic properties.For this purpose, eight Bacillus subtilis, Bacillus coagulans and Bacillus clausii strains among 58 isolates from fish intestine, soil, ripened cheese and commercial probiotic supplements were identified and their probiotic properties were characterized. Firstly, Bacillus strains were molecularly identified by 16S rRNA PCR analysis. The growth of Bacillus isolates at various temperatures, salt concentrations, and pH levels, as well as tests for esculin hydrolysis, starch hydrolysis, nitrate reduction, and gas generation from glucose, were all investigated to assess the isolates' physiological and biochemical characteristics. In terms of probiotic potential of Bacillus isolates; tolerance of bile salt, cell surface hydrophobicity, auto aggregation, antibiotic susceptibility tests were conducted. In all analyses, strains obtained from food supplements showed high levels of hydrophobicity and auto-aggregation properties, and the highest values following these strains were observed in Bacillus subtilis strains (F1 and S2) isolated from fish intestines and soil, respectively. All strains showed strong growth features in bile salt conditions. It has been determined that antibiotic sensitivity varies depending on the strain. Overall, high sensitivity to tetracycline has been observed. In summary, this study revealed the potential probiotic properties of Bacillus isolates obtained from different sources. The study also compared these probiotic properties with probiotic Bacillus strains isolated from food supplements.

Nanopore Sequencing Discloses Compositional Quality of Commercial Probiotic Feed Supplements

The market for the application of probiotics as a livestock health improvement supplement has increased in recent years. However, most of the available products are quality-controlled using low-resolution techniques and un-curated databases, resulting in misidentification and incorrect product labels. In this work, we deployed two workflows and compared results obtained by full-length 16S rRNA genes (16S) and metagenomic (Meta) data to investigate their reliability for the microbial composition of both liquid and solid forms of animal probiotic products using Oxford Nanopore long-read-only (without short-read). Our result revealed that 16S amplicon data permits to detect the bacterial microbiota even with the low abundance in the samples. Moreover, the 16S approach has the potential to provide species-level resolution for prokaryotes but not for assessing yeast communities. Whereas, Meta data has more power to recover of high-quality metagenome-assembled genomes that enables detailed exploration of both bacterial and yeast populations, as well as antimicrobial resistance genes, and functional genes in the population. Our findings clearly demonstrate that implementing these workflows with long-read-only monitoring could be applied to assessing the quality and safety of probiotic products for animals and evaluating the quality of probiotic products on the market. This would benefit the sustained growth of the livestock probiotic industry.

Safety and the probiotic potential of Bifidobacterium animalis CP-9.

Bifidobacterium animalis CP-9 was a commensal strain isolated from human breast milk. In this study, genetic and 90-day oral toxicity were assessed in rodents for its safety. Ames test as well as in vivo bone marrow micronucleus and spermatocyte chromosomal aberration were surveyed in mice. B. animalis CP-9 exhibited no mutagenic activity in the Ames test at the highest tested dosage (5000µg/plate) with or without metabolic activation. No evidence of in vivo genetic toxicity was observed at the maximum tested dosage of 10g/kg body weight (BW). Furthermore, there was no statistically significant difference of the biochemical and histological parameters in the rats administrated with B. animalis CP-9 at dosages of 0, 0.25, 0.5, or 1.5g/kg BW/day. No indication of concern for pathogenicity was exhibited during evaluation of Bifidobacterium ssp. generally, or B. animalis specifically. It was noted that B. animalis CP-9 was able to survive in gastric acid-like and high bile salt environment, and showed strong adhesion to the intestinal epithelial cells, Caco-2. Intriguingly, B. animalis CP-9 decreased olic acid-induced triglyceral (TG) accumulation in the Caco-2 cells, and viable B. animalis CP-9 had a better bacteriostatic activity compared to another well-documented B. animalis ssp. lactis, BB-12. Based on the present study, B. animalis CP-9 can be a safe probiotic supplement and may improve the health of host. PRACTICAL APPLICATION: Although the health benefits of probiotics are well known, the safety of a probiotic product is acquired particularly for a long-term consumption. We conduct the safety of B. animalis CP-9 isolated from human breast milk, and demonstrate no toxicity concern in vitro and in vivo. Hence, B. animalis CP-9 powder can be used as a commercial and safe probiotic supplement with some health benefits.

Characterization and Viability Prediction of Commercial Probiotic Supplements under Temperature and Concentration Conditioning Factors by NIR Spectroscopy

The quality of probiotics has been associated with bacteria and yeast strains’ contents and their stability against conditioning factors. Near-infrared spectroscopy (NIRS), as a non-destructive, fast, real-time, and cost-effective analytical technique, can provide some advantages over more traditional food quality control methods in quality evaluation. The aim of our study was to evaluate the applicability of NIRS to the characterization and viability prediction of three commercial probiotic food supplement powders containing lactic acid bacteria (LAB) subjected to concentration and temperature conditioning factors. For each probiotic, 3 different concentrations were considered, and besides normal preparation (25 °C, control), samples were subjected to heat treatment at 60 or 90 °C and left to cool down until reaching room temperature prior to further analysis. Overall, after applying chemometrics to the NIR spectra, the obtained principal component analysis-based linear discriminant analysis (PCA-LDA) classification models showed a high accuracy in both recognition and prediction. The temperature has an important impact on the discrimination of samples. According to the concentration, the best models were identified for the 90 °C temperature treatment, reaching 100% average correct classification for recognition and over 90% for prediction. However, the prediction accuracy decreased substantially at lower temperatures. For the 25 °C temperature treatment, the prediction accuracy decreased to nearly 60% for 2 of the 3 probiotics. Moreover, according to the temperature level, both the recognition and prediction accuracies were close to 100%. Additionally, the partial least square regression (PLSR) model achieved respectable values for the prediction of the colony-forming units (log CFU/g) of the probiotic samples, with a determination coefficient for prediction (R2Pr) of 0.82 and root mean square error for prediction (RMSEP) of 0.64. The results of our study show that NIRS is a fast, reliable, and promising alternative to the conventional microbiology technique for the characterization and prediction of the viability of probiotic supplement drink preparations.

The Probiotic Identity Card: A Novel "Probiogenomics" Approach to Investigate Probiotic Supplements.

Probiotic bacteria are widely administered as dietary supplements and incorporated as active ingredients in a variety of functional foods due to their purported health-promoting features. Currently available probiotic products may have issues with regards to their formulation, such as insufficient levels of viable probiotic bacteria, complete lack of probiotic strains that are stated to be present in the product, and the presence of microbial contaminants. To avoid the distribution of such unsuitable or misleading products, we propose here a novel approach named Probiotic Identity Card (PIC), involving a combination of shotgun metagenomic sequencing and bacterial cell enumeration by flow cytometry. PIC was tested on 12 commercial probiotic supplements revealing several inconsistencies in the formulation of five such products based on their stated microbial composition and viability.

Comparative Evaluation of the Antimicrobial and Mucus Induction Properties of Selected Bacillus Strains against Enterotoxigenic Escherichia coli.

Probiotics have been shown to bind to host receptors, which are important for pathogen adhesion and induce the host’s production of defence factors. They can activate the goblet-cell-derived production of mucins, a major component of the mucus layer and a physical barrier participating in limiting the proximity of microorganisms to the epithelial layer. In the last decade, Bacillus spp. strains have gained interest in human and animal health due to their tolerance and stability under gastrointestinal tract conditions. Moreover, Bacillus spp. strains can also produce various antimicrobial peptides that can support their use as commercial probiotic supplements and functional foods. The present study aimed to evaluate and determine the ability of selected Bacillus spp. strains to inhibit the growth of enterotoxigenic Escherichia coli (ETEC) F4 and to reduce binding of ETEC F4 to HT29-16E (mucus-secreting and goblet-like) human intestinal cells. Moreover, mucus production in the HT29 cells in the presence of the Bacillus spp. strains was quantified by ELISA. Bacillus spp. strains (CHCC 15076, CHCC 15516, CHCC 15541, and CHCC 16872) significantly inhibited the growth of ETEC F4. Moreover, the ability of the probiotic Bacillus spp. strains to stimulate mucin release was highly strain dependent. The treatment with Bacillus subtilis CHCC 15541 resulted in a significant increase of both MUC2 and MUC3 in HT29-16E cells. Therefore, this strain could be an up-and-coming candidate for developing commercial probiotic supplements to prevent infections caused by ETEC F4 and, potentially, other pathogens.

Viability and Composition Validation of Commercial Probiotic Products by Selective Culturing Combined with Next-Generation Sequencing.

The consumption of dietary supplements to treat health complications or to improve overall health conditions has become a globally increasing trend that leads to the development of a large number of health-related novel products and expands the associated manufacturing industries around the world. In the current study, we applied selective culturing combined with next-generation sequencing to examine the microbial viability in terms of its culturability on culture medium, composition, and possible contamination in the selected 17 commercial probiotic products sold in the mainland China market. Additionally, the relative abundance of each individual bacterial content was also evaluated by using the generated sequencing reads. The tested probiotic product samples were subjected to Illumina HiSeq-2000 sequencing platform and thoroughly analyzed by the in-house developed bioinformatics pipeline. The comprehensive culturing and sequencing analysis revealed both viability and composition inaccuracy among the several tested probiotic products, however, no contaminant was identified during the analysis. Among the total, five probiotic products (29.41%) were found with an inaccurate or lower colony-forming unit (CFU) counts on culture media while four probiotic products (23.52%) have inaccurately labeled classification. This study provides an ideal qualitative and quantitative assessment approach, which can be used as a diagnostic tool for the accurate assessment of commercial probiotic supplements.

The Spectacular Role of the Human Microbiome in Preventing Post-prandial or Metabolic Endotoxemia (OR23-02-19)

Abstract Objectives To determine if 30-d of oral spore-based probiotic supplementation could reduce dietary endotoxemia. Methods Apparently healthy men and women (n = 75) were screened for post-prandial dietary endotoxemia. Subjects whose serum endotoxin concentration increased by at least 5-fold from pre-meal levels at 5-h post-prandial were considered “responders” and were randomized to receive either placebo (rice flour) or a commercial spore-based probiotic supplement [Bacillus indicus (HU36), Bacillus subtilis (HU58), Bacillus coagulans, and Bacillus licheniformis, and Bacillus clausii] for 30-d. The dietary endotoxemia test was repeated at the conclusion of the supplementation period. Dietary endotoxin (LAL) and triglycerides (enzymatic) were measured using an automated chemistry analyzer. Serum disease risk biomarkers were measured using bead-based multiplex assays (Luminex and Milliplex) as secondary, exploratory measures. Results Data were statistically analyzed using repeated measures ANOVA and a P < 0.05. We found that spore-based probiotic supplementation was associated with a 42% reduction in endotoxin (12.9 ± 3.5 vs 6.1 ± 2.6, P = 0.011) and 24% reduction in triglyceride (212 ± 28 vs 138 ± 12, P = 0.004) in the post-prandial period Placebo subjects presented with a 36% increase in endotoxin (10.3 ± 3.4 vs 15.4 ± 4.1, P = 0.011) and 5% decrease in triglycerides (191 ± 24 vs 186 ± 28, P = 0.004) over the same post-prandial period. We also found that spore-based probiotic supplementation was associated with significant post-prandial reductions in IL-12p70 (24.3 ± 2.2 vs 21.5 ± 1.7, P = 0.017) and IL-1β (1.9 ± 0.2 vs 1.6 ± 0.1, P = 0.020). Compared to placebo post supplementation, probiotic subject had less ghrelin (6.8 ± 0.4 vs 8.3 ± 1.1, P = 0.017) compared to placebo subjects. Conclusions 30 days of supplementation with spore-based probiotics showed a 60% reduction in biomarkers of leaky gut when compared to the placebo group. These findings suggest that probiotics may play an integral role in reducing chronic low-grade inflammation that leads to chronic disease and autoimmunity. Funding Sources The present study was funded in part by a competitive research grant from Microbiome Labs, LLC (Glenview, IL) to the University of North Texas. Supporting Tables, Images and/or Graphs

Use of protective culture to control the growth of Listeria monocytogenes and Salmonella typhimurium in ready-to-eat cook-chill products

This study explored the antagonistic activity of Lactobacillus rhamnosus GG (LGG) (Culturelle®), a commercial probiotic supplement, on foodborne pathogens and its heat sensitivity in soup products. Staphylococcus aureus, Escherichia coli O157:H7, Escherichia coli O104:H4, L. monocytogenes and S. Typhimurium were selected to assess the inhibitory effect of LGG on them and analyze their sensitivities towards LGG. Cream of potato (CP) and vegetable beef (VB) soups were selected as food matrix to determine thermal resistance of LGG between 55 and 62.5 °C. The effectiveness of LGG as a protective culture against the growth of Listeria monocytogenes and Salmonella Typhimurium was evaluated at conditions simulating the commercial application of cook-chill processing in CP soup and stored at 5, 10 and 15 °C for 21 days. Overall, LGG showed inhibitory effect against all pathogens tested on spot agar test. E. coli O157:H7 was the most sensitive, whereas S. Typhimurium was the least sensitive to the inhibitory effect of LGG. D-values obtained in CP soup exceeded those obtained in VB soup at the temperatures tested. D55-62.5 values of LGG ranged between 0.7-24.2 min and 0.5–15 min in CP and VG soup, respectively. The z-value for CP and VB soup was around 5 °C. LGG co-inoculated with either target bacterium led to a reduction in S. Typhimurium or L. monocytogenes to undetectable levels. The probiotic population increased about 1-log unit at 15 °C. At 10 and 5 °C, it was maintained throughout the experiment at around 108 CFU g−1. Results demonstrated the potential of LGG as protective culture in controlling S. Typhimurium and L. monocytogenes growth in cream of potato soup.

Effects of commercial dietary prebiotic and probiotic supplements on growth, innate immune responses, and intestinal microbiota and histology of Totoaba macdonaldi

Prebiotics and probiotics constitute a different approach to prevent diseases in cultured fish and shellfish and have shown a number of beneficial impacts, including improved growth, survival, immunological responses, increased area of intestinal absorption, and enhanced general well-being of the host. The dietary supplementation of prebiotics and/or probiotics has not been evaluated in Tototaba macdonaldi, thus, the objective of this study was to evaluate its growth, innate immune responses, and transient intestinal microbiota and histology after being fed commercial prebiotic and probiotic supplements singularly and in combination. Four iso-nitrogenous (43% crude protein) and iso-lipidic (13% crude fat) diets were formulated to contain either 0 (basal diet) or 2% of the yeast-based prebiotic GroBiotic®-A or the Bacillus-containing probiotic Aquablend® at 11 × 109 cell forming units (CFU)/kg, or both supplements, in a 2 × 2 factorial arrangement, feeding fish with an initial mean weight of 215.6 g for 109 days. No statistically significant differences were observed by supplementing either the probiotic, the prebiotic, or both for survival or any of the growth parameters evaluated at the end of the experiment. Lysozyme activity in plasma of fish fed the basal diet was significantly higher (P = 0.0031) than that of fish fed the prebiotic. Histometric analysis of the gastrointestinal tract revealed that fold length in the proximal section was significantly larger (P = 0.0045) in fish fed the diet containing the probiotic (435.6 μm), compared to those of fish fed the basal diet (382.3 μm). No differences were detected for the composition of the transient microbial populations from the gastrointestinal tract (GIT) contents of T. macdonaldi, as evaluated by denaturing gradient gel electrophoresis analysis. However, significant shifts in the overall transient microbial community and diversity were found using massive sequencing of the V3-V4 16S rRNA GENE. Our results showed different bacterial clustering patterns between the basal diet and the diets with additives (prebiotic, probiotic, pre + pro). Furthermore, the genus Bacillus appeared enriched only in the diet combining both supplements. To our knowledge, this is the first report of transient microbial community analysis from intestinal contents of totoaba using the 16S microbiome sequencing. Initial weight of totoaba, or the length of the trial, could partly explain the absence of growth differences among treatments. From these results, it was evident that studies with probiotics and prebiotics fed to sturdy fish like totoaba will benefit from including a challenge experiment in order to assess their potential role in preventing infections.

Bacillus As Potential Probiotics: Status, Concerns, and Future Perspectives.

Spore-forming bacilli are being explored for the production and preservation of food for many centuries. The inherent ability of production of large number of secretory proteins, enzymes, antimicrobial compounds, vitamins, and carotenoids specifies the importance of bacilli in food chain. Additionally, Bacillus spp. are gaining interest in human health related functional food research coupled with their enhanced tolerance and survivability under hostile environment of gastrointestinal tract. Besides, bacilli are more stable during processing and storage of food and pharmaceutical preparations, making them more suitable candidate for health promoting formulations. Further, Bacillus strains also possess biotherapeutic potential which is connected with their ability to interact with the internal milieu of the host by producing variety of antimicrobial peptides and small extracellular effector molecules. Nonetheless, with proposed scientific evidences, commercial probiotic supplements, and functional foods comprising of Bacillus spp. had not gained much credential in general population, since the debate over probiotic vs pathogen tag of Bacillus in the research and production terrains is confusing consumers. Hence, it’s important to clearly understand the phenotypic and genotypic characteristics of selective beneficial Bacillus spp. and their substantiation with those having GRAS status, to reach a consensus over the same. This review highlights the probiotic candidature of spore forming Bacillus spp. and presents an overview of the proposed health benefits, including application in food and pharmaceutical industry. Moreover, the growing need to evaluate the safety of individual Bacillus strains as well as species on a case by case basis and necessity of more profound analysis for the selection and identification of Bacillus probiotic candidates are also taken into consideration.

Oral spore-based probiotic supplementation was associated with reduced incidence of post-prandial dietary endotoxin, triglycerides, and disease risk biomarkers.

AIMTo determine if 30-d of oral spore-based probiotic supplementation could reduce dietary endotoxemia.METHODSApparently healthy men and women (n = 75) were screened for post-prandial dietary endotoxemia. Subjects whose serum endotoxin concentration increased by at least 5-fold from pre-meal levels at 5-h post-prandial were considered “responders” and were randomized to receive either placebo (rice flour) or a commercial spore-based probiotic supplement [Bacillus indicus (HU36), Bacillus subtilis (HU58), Bacillus coagulans, and Bacillus licheniformis, and Bacillus clausii] for 30-d. The dietary endotoxemia test was repeated at the conclusion of the supplementation period. Dietary endotoxin (LAL) and triglycerides (enzymatic) were measured using an automated chemistry analyzer. Serum disease risk biomarkers were measured using bead-based multiplex assays (Luminex and Milliplex) as secondary, exploratory measures.RESULTSData were statistically analyzed using repeated measures ANOVA and a P < 0.05. We found that spore-based probiotic supplementation was associated with a 42% reduction in endotoxin (12.9 ± 3.5 vs 6.1 ± 2.6, P = 0.011) and 24% reduction in triglyceride (212 ± 28 vs 138 ± 12, P = 0.004) in the post-prandial period Placebo subjects presented with a 36% increase in endotoxin (10.3 ± 3.4 vs 15.4 ± 4.1, P = 0.011) and 5% decrease in triglycerides (191 ± 24 vs 186 ± 28, P = 0.004) over the same post-prandial period. We also found that spore-based probiotic supplementation was associated with significant post-prandial reductions in IL-12p70 (24.3 ± 2.2 vs 21.5 ± 1.7, P = 0.017) and IL-1β (1.9 ± 0.2 vs 1.6 ± 0.1, P = 0.020). Compared to placebo post supplementation, probiotic subject had less ghrelin (6.8 ± 0.4 vs 8.3 ± 1.1, P = 0.017) compared to placebo subjects.CONCLUSIONThe key findings of the present study is that oral spore-based probiotic supplementation reduced symptoms indicative of “leaky gut syndrome”.

Viability, Acid and Bile Tolerance of Spray Dried Probiotic Bacteria and Some Commercial Probiotic Supplement Products Kept at Room Temperature.

Production of probiotic food supplements that are shelf-stable at room temperature has been developed for consumer's convenience, but information on the stability in acid and bile environment is still scarce. Viability and acid and bile tolerance of microencapsulated Bifidobacterium spp. and Lactobacillus acidophilus and 4 commercial probiotic supplements were evaluated. Bifidobacterium and L. acidophilus were encapsulated with casein-based emulsion using spray drying. Water activity (aw ) of the microspheres containing Bifidobacterium or L. acidophilus (SD GM product) was adjusted to 0.07 followed by storage at 25 °C for 10 wk. Encapsulated Bifidobacterium spp. and Lactobacillus acidophilus and 4 commercial probiotic supplement products (AL, GH, RE, and BM) were tested. Since commercial probiotic products contained mixed bacteria, selective media MRS-LP (containing L-cysteine and Na-propionate) and MRS-clindamycin agar were used to grow Bifidobacterium spp. or L. acidophilus, respectively, and to inhibit the growth of other strains. The results showed that aw had a strong negative correlation with the viability of dehydrated probiotics of the 6 products. Viable counts of Bifidobacterium spp. and L. acidophilus of SD GM, AL, and GH were between 8.3 and 9.2 log CFU/g, whereas that of BM and RE were between 6.7 and 7.3 log CFU/g. Bifidobacterium in SD GM, in AL, and in GH products and L. acidophilus in SD GM, in AL, and in BM products demonstrated high tolerance to acid. Most of dehydrated probiotic bacteria were able to survive in bile environment except L. acidophilus in RE product. Exposure to gastric juice influenced bacterial survivability in subsequent bile environment.

Evaluation of β-Galactosidase Activity of Commercial Probiotic Supplements Available in the Local Market in Greensboro, North Carolina

The objective of this study was to evaluate the ?-galactosidase activity of commercial probiotic supplements available in the market. Fifteen different supplements (15) were included in the study. Two capsules of each supplement were transferred into 10 ml fresh, sterile MRS broth and incubated at 37 °C for 10-12 h to allow bacterial cells to recover from any environmental stressors. Aliquots from each supplement (100µl) were then inoculated into batches of 10ml TPY broth with lactose (induced) or glucose (un-induced) and incubated for 14 hr. at 37°C. After bacterial growth reached the mid log phase (optical density 0.7-0.9; 610 nm), the ?-gal activity was determined using o-nitrophenyl-?-D-galactoside (ONPG) as a substrate. The activity of ?-gal in the uninduced group ranged between 0 and 600 Miller units, whereas activity in the induced group ranged from 0 to 1300Miller units. These findings indicate that ?-gal activity in the tested supplements varies. This would mean that commercially available probiotic supplements exhibit different degrees of activity, and consumers should be aware of this variability when choosing probiotic supplements that purportedly confer health benefits.

Effect of diet supplementation with a commercial probiotic containingPediococcus acidilactici(Lindner, 1887) on the expression of five quality markers in zebrafish (Danio rerio(Hamilton, 1822)) testis

The effects of a commercial probiotic diet supplement (Bactocell®), containing lactic acid bacteria Pediococcus acidilactici were evaluated by identifying five zebrafish (Danio rerio) sperm quality markers. P. acidilactici (106 colony-forming units per gram) was supplied to young zebrafish males (age 8 months) as a feed probiotic additive during a 10 days test period. Five transcripts, previously described as male zebrafish quality markers, were studied: brain-derived neurotrophic factor (bdnf), BCL2-interacting killer (bik), double-sex and mab-3 related transcription factor 1 (dmrt1), follicle-stimulating hormone beta subunit (fshb) and leptin a (lepa). After exposition, males fed on with the supplemented diet with a commercial probiotic containing P. acidilactici presented an over-expression on three fertility markers comparing to the control group: lepa, dmrt1 and bdnf. These results suggest that P. acidilactici has a potential use as probiotic supplement in zebrafish diet for the improvement of molecular parameters in testicular cells, indicating that probiotic supplementation could affect male reproductive performance.

Short-term daily intake of 6 billion live probiotic cells can be insufficient in healthy adults to modulate the intestinal bifidobacteria and lactobacilli

Most intervention studies on probiotics or prebiotics consist of several-week feeding periods and focus on specific host dysfunctions. However, probiotics are often consumed by healthy people and time can be limited; the efficacy of such dietary intervention, however, has only rarely been considered. In this paper, we report the results of a short-term (1week) study, in which 11 healthy subjects consumed a commercial probiotic food supplement, resulting in a daily intake of about 6 billion viable cells of 10 bacterial species. We measured by quantitative real-time PCR (qPCR) the impact that this product had on faecal Bifidobacterium and Lactobacillus spp. Faecal lactobacilli and bifidobacteria, whose number in the gut is a commonly considered parameter to assess the efficacy of a probiotic/prebiotic intervention, were not significantly affected by the intervention. The only significant increase was observed for intestinal Lactobacillus acidophilus group. However, the significance of this change disappeared after only few days, indicating that it was plausibly due to DNA carryover from the ingestion of a large number of L. acidophilus cells. According to this study, short-term intake of a quite high number of live probiotics can be ineffective in healthy humans on the faecal concentration of bifidobacteria and lactobacilli.

Laboratory Exploration of Survival of Probiotic Cultures Inside the Human Digestive Tract Using In Vitro Models

Scientists often model complex biological phenomena in vitro, mimicking conditions found in living organisms. Understanding the power and limitations of biological models is an important topic in undergraduate science. In this activity, students develop their own in vitro model for testing the survival of bacteria from commercial probiotic supplements. Students work in groups to decide which factors are important for survival of bacteria in a chosen portion of the human digestive tract. Groups of students create their own in vitro models of organs such as stomach and/or intestines. Students expose a probiotic supplement to conditions mimicking the chosen portion of the human digestive tract, and measure the effect of those conditions on the survival of bacteria found in the supplement. Students choose to focus on conditions such as low pH found in stomach or pancreatic enzymes found in the upper intestine. Through this activity, students gain experience with serial dilutions and calculations of colony forming units (CFUs). This project also provides the students with the valuable experience of designing experiments in small groups. Students present their findings in a poster session, which provides a venue for discussing the validity and limitation of various models.

National survey outcomes on commercial probiotic food supplements in Italy

To assess whether the probiotic food supplements, produced and distributed on the Italian market during 2005–2006, complied with the Italian Guidelines on Prebiotics and Probiotics, 72 samples from 29 processing plants were analyzed. The survey included 41 samples from processing plants and 31 samples of the same brand from retailers collected at timed intervals (3, 8 and 13 months). A polyphasic approach based on a suitable analytical collection method (genotypic identification of total bacteria – differential presumptive enumeration – genotypic identification of viable bacteria) was adopted to identify and quantify the microorganisms labelled and recovered from the probiotic supplements examined. Most supplements analyzed (87%) did not conform to the Italian guidelines and the differences were both quantitative and qualitative (number determination, purity, types and viability of microorganisms). Even though most labelled supplements (25 samples) indicated the presence of Bifidobacterium bifidum, this organism was only detected sporadically and always as dead cells. Unexpected results were obtained during our survey due to the absence of viability of Bacillus coagulans spores in some labelled supplements. Besides this, some of these supplements also contained other spore-forming species, identified as B. cereus that are toxin producing. We have also documented a widespread use of misclassified microbial species or species with fictitious names. The main factors involved in the absence of compliance were examined and the poor quality control applied by manufacturers was emphasized.

Use of a commercial probiotic supplement in meat goats1

Sixty-three Boer crossbred goats were used in 5 separate experiments (Exp. 1 to 5) to evaluate the effects of a commercial probiotic supplement on growth performance (Exp. 1 to 4), diet digestibility (Exp. 5), carcass traits (Exp. 3), and fecal bacterial populations (Exp. 4). Goats were either fed a commercially pelleted concentrate diet and supplemented with a commercial probiotic (PRO) that had shown anecdotal positive effects on goat growth and performance according to local goat producers, or they remained as controls. The dose of PRO used was within the labeled dose for sheep for all studies. For Exp. 1, goat BW and feed intake were measured and G:F was calculated every 7 d for 56 d. For Exp. 2 to 4, BW and feed intake were measured and G:F was calculated every 14 d. The first day of supplementation was considered d 0. Carcass traits were also collected at slaughter on d 57 for Exp. 3, and fecal samples were collected every 14 d for microbial culture for Exp. 4. For Exp. 5, which was a digestibility trial that lasted for 10 d, animals were placed in metabolic pens for collection of feces and orts. Growth performance of goats was not affected by probiotic supplementation, with the exception of performance in Exp. 2, in which ADG and G:F were improved (P < 0.03) in PRO goats compared with control goats on d 56 only (treatment x day interaction; P < 0.05), averaging 0.21 +/- 0.02 kg/d for PRO goats and 0.11 +/- 0.02 kg/d for control goats for ADG and 0.17 +/- 0.02 for PRO goats and 0.10 +/- 0.02 for control goats for G:F. Carcass weights and weights of fabricated cuts (shoulder, loin, leg, rack, shank, and total parts) as well as carcass length, leg circumference, loin eye area, and backfat were not influenced by PRO supplementation. Apparent digestibilities of OM, DM, NDF, ADF, CP, and GE (on a DM basis) were similar for the PRO and control treatments. Fecal culture analysis of Escherichia coli and coliforms, Lactobacillus, and Bifidobacterium populations were not influenced by the PRO treatment. Overall, although the PRO treatment affected goat ADG and G:F in Exp. 2, no PRO treatment effects were noted on growth performance for Exp. 1, 3, and 4. Furthermore, the PRO treatment did not affect diet digestibility, carcass traits, or fecal microbial populations in goats. In conclusion, no consistent benefits were noted from supplementing healthy, growing meat goats with PRO.