Carla Orpianesi

Differences in Fecal Microbiota in Different European Study Populations in Relation to Age, Gender, and Country: a Cross-Sectional Study

ABSTRACT A cross-sectional study on intestinal microbiota composition was performed on 230 healthy subjects at four European locations in France, Germany, Italy, and Sweden. The study participants were assigned to two age groups: 20 to 50 years (mean age, 35 years; n = 85) and >60 years (mean age, 75 years; n = 145). A set of 14 group- and species-specific 16S rRNA-targeted oligonucleotide probes was applied to the analysis of fecal samples by fluorescence in situ hybridization coupled with flow cytometry. Marked country-age interactions were observed for the German and Italian study groups. These interactions were inverse for the predominant bacterial groups Eubacterium rectale-Clostridium coccoides and Bacteroides-Prevotella. Differences between European populations were observed for the Bifidobacterium group only. Proportions of bifidobacteria were two- to threefold higher in the Italian study population than in any other study group, and this effect was independent of age. Higher proportions of enterobacteria were found in all elderly volunteers independent of the location. Gender effects were observed for the Bacteroides-Prevotella group, with higher levels in males than in females. In summary, age-related differences in the microbiota makeup were detected but differed between the study populations from the four countries, each showing a characteristic colonization pattern.

Administration of probiotic strain to improve sea bream wellness during development

Two bacterial strains Lactobacillus fructivorans (AS17B), isolated from adult sea bream (Sparus aurata) gut, and Lactobacillus plantarum (906), isolated from human faeces, were administered contemporaneously, during sea bream development using Brachionus plicatilis and/or Artemia salina and dry feed as vectors. The probiotic treatment influenced gut colonization: at 35 days post-hatching (p.h.) L. fructivorans was not present in the gut, but the treatment induced colonization by L. plantarum. At 66 days p.h., L. fructivorans was evident also in the control; moreover, when suitable environmental conditions appeared in the post-metamorphosis gastro-intestinal tract, competition between L. plantarum and L. fructivorans occurred. At 90 days p.h., L. plantarum was displaced by L. fructivorans that became significantly higher with respect to the control. In treated groups, probiotic administration significantly decreased larvae and fry mortality.

EU project Crownalife: functional foods, gut microflora and healthy ageing isolation and identification of Lactobacillus and Bifidobacterium strains from faecal samples of elderly subjects for a possible probiotic use in functional foods

A new EU-funded project called “Crownalife” has been set up both to assess structural and functional alterations of the intestinal microflora with ageing in Europe, and to validate functional foods that promote health by improving the function of the intestinal microflora in the elderly. Species of bifidobacteria and lactobacilli were isolated from faecal samples of healthy elderly Italian subjects and identified for a possible probiotic use in functional foods. Total anaerobic and aerobic, Lactobacillus and Bifidobacterium counts were performed on the faecal samples. Colonies grown on selective media were identified using enzymatic tests. Lactobacillus fermentum and Bifidobacterium longum were the most represented species and could be considered good candidates to be further characterized and then utilized to design appropriate dietary strategies. Some of the identification results related to Bifidobacterium species were validated by using PCR procedure based on 16S rRNA gene sequences species-specific primers showing a good concordance rate with the enzymatic method.

Efficacy of antimicrobial filter treatments on microbial colonization of air panel filters

Aims: To assess the activity of biostatic agents on the microbial colonization of panel filters.

Pilot-scale production and viability analysis of freeze-dried probiotic bacteria using different protective agents.

The functional food industry requires an improvement of probiotic strain stability during storage, especially when they are stored at room temperature. In this study, the viability of freeze-dried Lactobacillus rhamnosus IMC 501(®) and Lactobacillus paracasei IMC 502(®) using different protective agents (i.e., glycerine, mannitol, sorbitol, inulin, dextrin, Crystalean(®)) was determined and compared with semi skimmed milk (SSM) control. No significant differences were observed between the tested protectants and the control (SSM) during storage at refrigerated conditions. During storage at room temperature, only glycerine was found to stabilize viability better than other tested substances.

The effect of sucrose or starch‐based diet on short‐chain fatty acids and faecal microflora in rats

An investigation was carried out to determine whether variations of dietary carbohydrates could modify the colonic flora in rats. Sprague‐Dawley rats were fed with two equicaloric diets based on the AIN‐76 diet ( American Institute of Nutrition 1977 ) but differing from that diet in content of carbohydrates, i.e. high sucrose (64%) or high corn starch (64%). Feeding was continued for 9 months ad libitum and no variation in weight gain was recorded among the different diets. A prevalence of aerobes, and a significant reduction in the ratio anaerobes/aerobes in the faeces of rats on the high starch diet compared with the high sucrose diet, was observed. The anaerobe genera identified included Actinomyces, Bacteroides, Bifidobacterium, Clostridium, Eubacterium, Lactobacillus and Propionibacterium. Bacteroides was the most prevalent genus in both dietary groups (51·2 and 29·5% in the faeces of rats fed the sucrose and starch diets, respectively). In contrast, clostridia were prevalent in the starch‐fed group (23·8%) and less so in the sucrose diet (11·5%), as propionibacteria were prevalent in faeces of rats fed the starch diet (15·5%), and low in the sucrose diet (3·9%). The remaining genera were scarce in faeces from rats on either diet. Total short‐chain fatty acids (SCFA) were significantly higher in the faeces of animals fed the starch diet compared with those fed the sucrose diet. The relative concentrations of acetic, propionic and butyric acids were not significantly different between the two dietary groups. In conclusion, high starch diet can markedly modify the composition of faecal flora and alter considerably the faecal concentration of SCFAs, compounds which might have a health‐promoting effect.

In vitro evaluation of antimicrobial activity of Lactobacillus rhamnosus IMC 501®, Lactobacillus paracasei IMC 502® and SYNBIO® against pathogens

Probiotic lactobacilli have a great potential to produce antimicrobial compounds that inhibit and control the microbial pathogen growth. The antimicrobial and antifungal activities of two probiotic strains, Lactobacillus rhamnosus IMC 501® and Lactobacillus paracasei IMC 502®, and their 1 : 1 combination, named SYNBIO®, were studied using four different methods.

Evaluation of antipathogenic activity and adherence properties of human Lactobacillus strains for vaginal formulations

To test different Lactobacillus strains for their antipathogenic activity towards Candida strains and their adhesion properties for the preparation of vaginal ovules and douches to be used in vaginal candidiasis prevention.

Effect of the addition of Lactobacillus delbrueckii subsp. delbrueckii on the gut microbiota composition and contribution to the well-being of European sea bass (Dicentrarchus labrax, L.)

The present study aimed to test the effects of probiotic treatment on gut microbiota and the contribution to the well-being of European sea bass (Dicentrarchus labrax, L.). A bacterial strain of Lactobacillus delbrueckii subsp. delbrueckii (AS13B), isolated from adult European sea bass gut, was administered during sea bass development using Brachionus plicatilis and/or Artemia salina as carriers. The effective strain colonization and modulation of the gut microbiota, the mortality and the cortisol levels of sea bass were monitored during early developmental stages. The L. delbrueckii subsp. delbrueckii strain showed good capability to colonize the gut of sea bass larvae, therefore modifying the gut microbiota, and exerting positive effects on the survival of the treated sea bass. L. delbrueckii subsp. delbrueckii as a probiotic dietary supplement represents an advantageous and ecological strategy for improving the health of fish in aquaculture and increasing fish production.

Effect of buckwheat flour and oat bran on growth and cell viability of the probiotic strains Lactobacillus rhamnosus IMC 501®, Lactobacillus paracasei IMC 502® and their combination SYNBIO®, in synbiotic fermented milk.

Fermented foods have a great significance since they provide and preserve large quantities of nutritious foods in a wide diversity of flavors, aromas and texture, which enrich the human diet. Originally fermented milks were developed as a means of preserving nutrients and are the most representatives of the category. The first aim of this study was to screen the effect of buckwheat flour and oat bran as prebiotics on the production of probiotic fiber-enriched fermented milks, by investigating the kinetics of acidification of buckwheat flour- and oat bran-supplemented milk fermented by Lactobacillus rhamnosus IMC 501®, Lactobacillus paracasei IMC 502® and their 1:1 combination named SYNBIO®. The probiotic strains viability, pH and sensory characteristics of the fermented fiber-enriched milk products, stored at 4 °C for 28 days were also monitored. The results showed that supplementation of whole milk with the tested probiotic strains and the two vegetable substrates results in a significant faster lowering of the pH. Also, the stability of L. rhamnosus IMC 501®, L. paracasei IMC 502® and SYNBIO® during storage at 4 °C for 28 days in buckwheat flour- and oat bran-supplemented samples was remarkably enhanced. The second aim of the study was to develop a new synbiotic product using the best combination of probiotics and prebiotics by promoting better growth and survival and be acceptable to the consumers with high concentration of probiotic strain. This new product was used to conduct a human feeding trial to validate the fermented milk as a carrier for transporting bacterial cells into the human gastrointestinal tract. The probiotic strains were recovered from fecal samples in 40 out of 40 volunteers fed for 4 weeks one portion per day of synbiotic fermented milk carrying about 10(9) viable cells.