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Four Gram-positive-staining, microaerophilic, non-spore-forming, fructose-6-phosphate phosphoketolase-positive bacterial strains were isolated from a faecal sample of a 5-year-old ring-tailed lemur (Lemur catta). The strains showed a peculiar morphology, resembling a small coiled snake, a ring shape, or forming a little ‘Y’ shape. The isolated strains appeared identical, and LMC 13T was chosen as a representative strain and characterized further. Strain LMC 13T showed an A3β peptidoglycan type, similar to that found in Bifidobacterium longum . The DNA base composition was 57.2 mol% G+C. Almost-complete 16S rRNA, hsp60, rpoB, dnaJ, dnaG, purF, clpC and rpoC gene sequences were obtained, and phylogenetic relationships were determined. Comparative analysis of 16S rRNA gene sequences showed that strain LMC 13T showed the highest similarity to B. longum subsp. suis ATCC 27533T (96.65 %) and Bifidobacterium saguini DSM 23967T (96.64 %). Strain LMC 13T was located in an actinobacterial cluster and was more closely related to the genus Bifidobacterium than to other genera in the Bifidobacteriaceae . On the basis of these results, strain LMC 13T represents a novel species within the genus Bifidobacterium , for which the name Bifidobacterium lemurum sp. nov. is proposed; the type strain is LMC 13T ( = DSM 28807T = JCM 30168T).

The present research was aimed to investigate the technological potentialities of seven strains of Bifidobacterium aesculapii, a species recently described, in terms of exopolysaccharide (EPS) production and as starter fermentation in soybean milk. The strain production of EPS was firstly evaluated in model system, using different carbon sources. Furthermore, the fermented products obtained by the seven strains of B. aesculapii were tested for their EPS content and strain cell loads, the volatile molecule profiles, the texture features and the overall acceptance. The data showed that all the B. aesculapii strains were able to produce EPS in vitro model in presence of 1.5% and 2% glucose while only four strains were able to produce EPS in presence of lactose 2%. When the strains were employed as fermentation starters in soybean milk, some showed a good growth potential, fermenting the substrate in 14 h and giving rise to fermented products with good firmness and viscosity indexes. Moreover, five strains out seven showed production of EPS (from 5 to 174 μg/mL) in soybean fermented milk.

Introduzione. Come è noto, diversi microrganismi costituiscono un diretto pericolo per la nostra salute in quanto possono infettare l'ospite umano: le malattie infettive causate da batteri e funghi sono ogni anno responsabili di morbilità e mortalità in un numero crescente di individui ospedalizzati e immunocompromessi. Inoltre, negli ultimi anni, la diffusione di ceppi microbici multi-resistenti ai farmaci, (ad es. Staphylococcus aureus, Pseudomonas aeruginosa e Candida spp.), ha provocato infezioni difficili o impossibili da controllare con antimicrobici convenzionali. La necessità di nuove terapie per il trattamento delle infezioni microbiche ha portato i ricercatori a concentrarsi su possibili alternative di tipo naturale. L'idrolato (Id), noto anche come idrosol o acqua aromatica, è la frazione idrofila ottenuta durante il processo di distillazione in corrente di vapore assieme all’olio essenziale (OE), capace di esercitare attività anti-microbiche grazie al contenuto in componenti terpeniche pari a circa l’1% v/v. Sebbene entrambe i prodotti della distillazione abbiano una nota azione antimicrobica, non è mai stata paragonata la loro attività quando estratti dalla stessa pianta. Scopo. Gli scopi di questo lavoro sono stati: (i) quello di paragonare l'attività antimicrobica dell’OE e dell’Id estratti da Monarda citriodora (MC) verso 30 ceppi fungini e batterici coinvolti in infezioni umane, (ii) valutare l’efficacia dell’Id di C. aurantium var. amara (CA) nei confronti di 18 ceppi fungini e batterici potenzialmente responsabili di infezioni cutanee. Materiali e metodi. L’OE e l’Id di MC sono stati ottenuti mediante distillazione in corrente di vapore partendo da piante fiorite coltivate presso l’istituto Scarabelli-Ghini di Imola (fig.1), mentre l’Id-CA era di tipo commerciale. Sono stati eseguiti test di micro-brodo diluizione secondo linee guida internazionali EUCAST per valutare l’efficacia antimicrobica dell’OE-MC (concentrazioni comprese tra 0,0078% e 4%), dell'Id-MC (concentrazioni tra 12,5% e 50%) e dell’Id-CA (concentrazioni tra 3.12% e 50%) vs i ceppi testati. Inoltre, l'OE e gli Id sono stati analizzati in GC-MS per valutarne la composizione terpenica e, nel caso di M. citriodora, paragonare l’efficacia dell’OE a quella dell’Id in funzione del loro contenuto terpenico. Risultati. L'OE-MC e l’Id-CA sono risultati efficaci, a concentrazioni variabili, su tutti i ceppi testati, mentre, l'Id-MC era attivo su tutti i microrganismi tranne che sui ceppi di P. aeruginosa (MIC>4% v/v). Le analisi GC-MS hanno individuato come principali componenti terpeniche il timolo per OE-MC e di Id-MC (19,6% e 66,4% rispettivamente) e il linalolo per Id-CA. Conclusioni. Nonostante sia necessaria una maggiore concentrazione di Id-MC per ottenere lo stesso effetto inibente dell’OE-MC, la quantità di terpeni presenti per unità di volume nell’Id è risultata inferiore rispetto all'EO. La maggiore efficacia dei terpeni contenuti nell’Id è probabilmente dovuta all'ambiente idrofilo che ne promuove una maggiore disponibilità.

Il nostro patrimonio artistico-culturale è estremamente vasto e diversificato. Tra i principali biodeteriogeni dei beni culturali cartacei si annoverano i funghi. Al fine di contrastare il fenomeno di biodeterioramento, recentemente, sono stati sviluppati studi atti a testare l’efficacia di sostanze naturali ad azione biocida, quali oli essenziali (OE) ed idrolati (Id), su biodeteriogeni comunemente isolati da opere d’arte. I trattamenti biocidi naturali ecocompatibili e non tossici per l’operatore, sono indispensabili per la salvaguardia dell’opera, nonché utili per preservare la salute degli operatori dato che alcuni funghi, ad es. quelli presenti negli ambienti chiusi, sono stati associati a molti rischi per la salute. Alcuni sono in grado di indurre, in soggetti defedati, malattie infettive quali le aspergillosi, ma sembra anche siano una delle concause della “Sindrome dell’edificio malato” (Sick Building Syndrome), caratterizzata da un quadro clinico non facilmente riconducibile ad un unico agente causale.

The aim of th epresent study is to describe the bifidobacterial species biodiversity in non-human primates in gut of: marmoset, tamarins, saimiri, saki, gelada, cercopitecae, macaque and lemur.

Bifidobacteria represent one of the main bacterial groups occurring in the gut of primates and other animals and are well known for their positive role on host health. Moreover, bifidobacteriaceae are used in coevolution studies since they are maintained within host lineages across hundreds of thousands of host generations. Therefore, the discovery of bifidobacterial diversity in the gastrointestinal tract together with the isolation and characterization of novel bacterial taxa in different hosts is essential. Until now no evidence has been reported about the occurrence of bifidobacteria in bats which are a crucial part of the ecosystem and are ecologically and economically advantageous to humans especially frugivorous ones. This study aimed at the investigation of bifidobacteria in bats belonging to Rousettus aegyptiacus species which were born in Egypt and hosted in Natura Viva Garda Zoological Park (Bussolengo, Verona, Italy). Faecal samples were serially diluted and plated on selective medium MRS supplemented with 0.05 % (w/v) cysteine, 100 mg/l norfloxacin, 2 mg/l cycloheximide, 50 mg/l mupirocin and 1 ml/l acetic acid. Strains were typed by BOX-PCR. Representative isolates of each cluster were identified sequencing 16S rRNA gene and studied for phylogenetical relationship with other Bifidobacteriaceae. Similarities among profiles were calculated by clustering the Pearson‟s correlation matrix using UPGMA algorithm. A total of 13 bifidobacteria were isolated and two of them belonged to new bifidobacterial species according to 16S rRNA gene analysis. The majority of strains revealed a phylogenetic relationship with bifidobacteria isolated from non-human primates. This evolutive link of bifidobacterial microbiotas between bats and non-human primates will be further investigated studying bifidobacteria characterizing Rousettus aegyptiacus bats from Egypt in order to offer new insights about bats origin.

Irritable Bowel Syndrome (IBS) is a psychosomatic gastrointestinal disorder which aetiology is not well clear, although several factors indicate the involvement of the gut microbiota, remarking the importance of the microbiota-gut-brain axis in IBS pathophysiology. We therefore characterized the gut microbiota in a cohort of 20 IBS subjects and 21 healthy subjects (HS) focusing also on the characterization of the fungal gut microbiota.

The Theropitecus gelada (fig.1), sometimes called the bleeding-heart monkey for their unmistakable red and hairless spot at the center of the chest, useful to indicate the social and reproductive status of the individual, are rocks monkeys living on the stony plateaus of Central Ethiopia, between 1,800 and 4,400 meters high. [1][2] These primates are perfectly suited to running and moving on the ground and find their natural habitat in rocky slopes, especially near rivers and streams. They are diurnal monkeys, and the most important trait of this animals is that they are the only known species of primates to be exclusively herbivores. After a trip of about a thousand kilometers, 6 monkeys of this species are housed in Italy, in the Natura Viva Park of Verona.

Recent studies have documented that 29-49% of women has been affected by vulvovaginal candidiasis (VVC) at least once in the lifetime and about 10% of them has history of recurrent VVC. Tea tree oil (TTO), an essential oil extracted by steam distillation from the leaves of Melaleuca alternifolia tree, has been known for many years for its antiseptic properties. Currently, TTO is used in vaginal suppositories (VS) for the treatment of vaginal candidiasis. TTO-VS exhibits an in vitro fungicidal activity towards Candida spp., only slightly affecting some vaginal lactobacilli population isolated from patients with vaginitis (Di Vito et al. 2015). However, preclinical and clinical studies addressing efficacy and safety profile are still few and not exhaustive, especially with regard to the complex vaginal microbial environment. On this basis, we have investigated the efficacy and safety profile of a combined therapy with probiotics and TTO-VS on female volunteers with vaginitisin order to find alternatives to fight the growing antimicrobial resistance to the most common synthetic antifungal remedies.

Many definitions have been proposed for the term “probiotic”. The more widely accepted one is “live microorganisms which, when administered in adequate amounts, confer a health benefit on the host” (FAO/WHO, 2002:1). This definition implies that a health effect must be demonstrated for the probiotic. Recently the economic success and exciting prospects of probiotic products have rapidly increased researches on different species of probiotic (Lactobacillus, Bifidobacterium, Bacillus, etc.). Genomics, comparative and functional genomic investigations have been conducted to gain information about the functionality of probiotics (Pfeiler, 2007:546) even if some insights of probiotic mechanism remain unknown. The following paragraphs will provides some insights in the taxonomy, role in health and disease and safety, aspects related to application on food and regulation about probiotics especially in relation to bifidobacteria and lactobacilli.

Co-evolution between host and gut microbiota is a current hot topic: gut microbe relationships are influenced by host genotype, and gut microbial population differs according to host phylogeny. For example gut microbiota within Great Apes reflects the phylogenetic history of their hosts and hypotheses that close host-species are functionally interchangeable were suggested. The bifidobacterial diversity in several subjects of primates has been studied to better understand the co-evolution with host along the evolutionary time-scale. In Great Apes the human species Bifidobacterium adolescentis was found in Pongo pygmaeus, while B. angulatum and B. dentium were found in Pan troglodytes. The new species B. moukalabense was described in Gorilla gorilla. Ten subjects selected from the two divergent lineages of Strepsirrhini (Lemuridae) and Simiiformes (Callitrichidae) were studied in the present work. Interestingly in Callithrix spp. and in Saguinus spp., together with the five previously described species B. callitrichos, B. stellenboschense, B. reuteri, B. saguini and B. biavati, seventeen new bifidobacterial species were found, showing a strong association with host species. In particular in two adult subjects of Lemur catta and Eulemur macaco (Lemuridae) two novel species were found: B. lemurum and a new one. B. aesculapii and five additional novel species were isolated from 5 baby subjects of the New World Monkey Callithrix jacchus. Nine novel Bifidobacterium spp., together with three bifidobacterial species previously found in both Saguinus midas and Callithrix jacchus, were found in an adult subject of Saguinus oedipus (Callitrichidae).

Dati in letteratura indicano che l’incidenza di candidosi è maggiore nelle donne in gravidanza rispetto alle altre e che il trattamento topico per le candidosi non è spesso sufficiente a decontaminare completamente il microambiente vaginale tanto che, frequentemente, si verificano recidive. Studi in letteratura per la prevalenza delle candidosi vaginali indicano che circa il 13% delle donne non in gravidanza ha recidive per la candidosi. Questa percentuale aumenta nelle gestanti che, pur necessitando di un trattamento preventivo per ridurre il rischio di contagio del neonato durante il parto, non possono essere trattate con i più comuni interventi antimicotici poichè controindicati in gravidanza.

Introduction In the Yaghnob Valley in Tajikistan lives an isolated human population, who still utilize ancient fermentation methods to produce fermented milk as one of their main foods. The area is very isolated, so theinhabitants and their diet have remained protected without influence or contamination from the surroundingworld. Thus, it is interesting to study the microbial biodiversity of their fermented food.The aim of the present work is thephenotypic characterization of yeasts isolated from Yaghnobi fermented milk. Materials and Methods Thirty yeast isolates were obtained from the original Yaghnobi fermented milk and from the sameone reproduced at home for three years.The isolation was performed on M17, MRS, WL,YPD and YPD plus chloramphenicol. Phenotypic characterization were assessed by growth i) on different carbon sources, ii) in presence of ox bile, iii) at high temperatures, iv) in acidic condition and v) in presence of hydrogen peroxide. Also the invasiveness in YPD and hyphae formation was studied. Results The 30 yeast strains, belonging to Kluyveromyces marxianus, Pichia fermentans, Saccharomyces cerevisiae plus one Kazachstania unispora and one Kluyveromyces lactis. revealed different phenotypes within the same species; e.g. invasiveness from 0 to 3 among the isolates of both S. cerevisiae and P. fermentans, the latter species also showed large variations in thermo-tolerance (inhibition from 40°Cto 46°C), and varying tolerance to acidic conditions. Only the strain of K. lactis and about 50%of K .marxianus strains grew at 46°C. All strains of K. marxianus grew at ox bile up to 2%. Of the S. cerevisiae strains, 50% grew in ox bile up to 2%, while two strains were inhibited already at 0.5%. Conclusions The data obtained suggest the presenceof yeasts with interesting probiotic properties in Yaghnobi fermented milk, such as the ability to survive in harsh gastrointestinal environment. The complete probiotic characterization of these yeasts will be investigated further.

The aim of this work is to evaluate the in vitro microbicidal activity of vaginal suppositories (VS) containing tea tree oil (TTO-VS) towards Candida spp. and vaginal probiotics. A total of 20 Candida spp. strains, taken from patients with vaginitis and from an established type collection, including reference strains, were analysed by using the CLSI microdilution method. To study the action of VS towards the beneficial vaginal microbiota, the sensitivity of Bifidobacterium animalis subsp. lactis (DSM 10140) and Lactobacillus spp. (Lactobacillus casei R-215 and Lactobacillus acidophilus R-52) was tested. Both TTO-VS and TTO showed fungicidal activity against all strains of Candida spp. whereas placebo-VS or the Aloe gel used as controls were ineffective. The study of fractional fungicidal concentrations (FFC) showed synergistic interaction with the association between Amphotericin B and TTO (0.25 to 0.08 µg/ml, respectively) against Candida albicans. Instead, the probiotics were only affected by TTO concentration ≥ 4% v/v, while, at concentrations

Ninety-two bifidobacterial strains were obtained from the faeces of 5 baby common marmosets, three known species Bifidobacterium aesculapii, Bifidobacterium callithricos and Bifidobacterium reuteri and 4 novel putative bifidobacterial species were retrieved. The occurrence of bifidobacteria in non-human primate babies is described for the first time.

Candidosis is the most important cause of fungal infections in humans. The yeast Candida albicans can form biofilms, and it is known that microbial biofilms play an important role in human diseases being very difficult to treat. The prolonged treatment with drugs has often resulted in failure and resistance. Due to the emergence of multidrug resistance, alternatives to conventional antimicrobial therapy are needed. The synergistic or additive effect produced by the combination between essential oils and antimicrobial drugs was referred as a strategy for counteracting microbial development (Wagner and Ulrich-Merzenich, 2009). In this work a possible synergistic anti-Candida effect of of Melaleuca alternifolia (tea tree oil, TTO) oils of two different batches (TTO_1 and TTO_2) and amphotericin B towards Candida albicans has been studied with the aim of finding minimum effective dose that maintain antifungal activity but minimize amphotericin side effects and down the onset of microbial resistance. First the minimum inhibitory concentrations (MIC) of the individual oils was determined by using the microdilution method following CLSI guidelines. The checkerboard assay was carried out as described by Lorian (2005) with slight modifications in order to evaluate the synergy between TTOs and amphotericin B (AppliChem GmbH, Germany) against C. albicans ATCC 48274: every possible combinations of seven serial two fold dilution of TTO (from 0,06% v/v to 4% v/v) and antibiotic (from 0,125μg/ml to 4 μg/ml) were tested. All experiments were conducted in triplicate. The components of the TTO oils have been previously characterized by chromatographic analyses. The results obtained showed the occurrence of pronounced fungicidal synergism [FFC (fractional fungicidal concentration) = 0.32 for TTO_1 and FFC= 0.28 for TTO_2] in which a low concentration of TTO (0.06% v/v) enhances the action of amphotericin B at a lower dose (0.125 μg/ml) compared to amphotericin B used alone (1μg/ml) against C. albicans. The in vitro data of the present study suggest the possibility to reduce the effective dose of antibiotic for treatment in the clinical setting decreasing the adverse effects of antibiotics. Further studies are needed to evaluate the toxicology of single components and their combination and to investigate the efficacy of this combination in animal models. Wagner H., Ulrich-Merzenich G. (2009) Synergy research: approaching a new generation of phytopharmaceuticals. Phytomedicine. 16:97-110. Lorian, V. (2005). Antibiotics in laboratory medicine 5th ed, Lippincott Williams & Wilkins, ISBN 978-078-1749-83-1, Philadelphia, USA

Searching for exopolysaccharides (EPS)-producing bacteria could be interested in the exploration of new probiotic strains. EPS have beneficial effects on health and play crucial roles in adhesion mechanisms, control of pathogens, maintain survival/viability of microorganism during technology food process/storage, and contribute to rheological properties of fermented foods. The ability to produce EPS by some bifidobacteria strains is demonstrated. Bifidobacteria, commonly inhabitants of the gastrointestinal tract of humans and animals, are used in foods as probiotic. Although, EPS biosynthesis in bifidobacteria is not known, hypothesis could be proposed based on the functional analysis of few genes and on sequences homology studies (P. Ruas-Madiedo, Salazar, and de los Reyes-Gavilán 2009). The priming glycosyltransferase, p-gtf, is the key enzyme involved in the catalyses of the first step of EPS-units biosynthesis. The role plays by p-gtf genes was confirmed altering or interrupting EPS production through inactivation of these genes (Low et al. 1998). In spite of the low amino acid, the intraspecies homology of a single p-gtf is high for the presence of conserved domains involved in the lipophilic carriers interaction ( Ruas-Madiedo et al. 2007).

Bifidobacteria are an important commensal group of the gut microbiota of animals. They belong to key bacterial groups playing symbiotic and probiotic role promoting health activities for the host. The distribution of bifidobacterial species in non human primates is currently poorly investigated. The occurrence of bifidobacterial species in animals is characterized by a peculiar trend where there is a cluster of species typical for non primates animals and a different cluster typical of man (human primates). Recently 5 new bifidobacterial species have been isolate from non-human primates such as new world monkeys (common marmoset, Callithrix jacchus L., and red-handed tamarin, Saguinus midas L.). Due to the novelty of these species they cannot be assigned to any bifidobacterial species cluster. Studies currently under development in our lab show that bifidobacterial isolates from Orangutan and Chimpanzee belong to human primates bifidobacteria cluster: this is in agreement with the strict evolutionary link between hominoids monkey such as Orangutan and Chimpanzee and humans. The aim of the present study is to investigate the biodiversity of bifidobacterial species in primates more distant to human from an evolutionary point of view such as common marmoset, New World monkey, in order to establish where does the bifurcation between the two bifidobacterial cluster can occur. Fecal samples were collected from 5 infants of common marmoset. Colony counts and isolation of bifidobacteria were performed in mTPY (modified TPY) as describe by Rada & Petr (2000). The isolates were tested for the activity of fructose-6-phosphate phosphoketolase (F6PPK) according to Biavati and Mattarelli (2012). Rep-PCR using primer BOXAR1 (Masco et al., 2009) was conducted and the isolates were grouped using R software (Ishii et al., 2009). For the species identification, the 16S rRNA gene amplification and sequencing of one representative strain from each obtained group were performed. Bifidobacteria were isolated from all sampled animals. The faecal Bifidobacterium spp. counts ranged from 9.15 to 9.48 log10 CFU/g in infant common marmosets.. The analysis of BOX profiles reveled richness of species which, basing on the rep-PCR analysis, formed 29 groups. Sequencing analysis of 16S rRNA gene are currently on going and partial results revealed that one of the most representative strains is a novel species: it is not closely related to known Bifidobacterium species as its high sequencing similarity shared 96% to B. scardovii while values

Folates – the natural chemically reduced forms of folic acid (vitamin B9) - are cofactors in essential metabolic pathways such as DNA synthesis and methylation pathways. Humans cannot synthesize folate and depend on intake both from the diet (green vegetables, cereals, rice, milk, fermented milk products, etc.) and from indigenous folate synthesizing bacteria of the intestinal microbiota. Low folate levels increase the risk for neural tube defects and may increase the risk for e.g. certain cancer forms, cardiovascular disease and Alzheimer’s. Screening for folate production of the bifidobacteria isolates from human adult and infant (1-6 month old) was performed. Strains typical of infants, such as Bifidobacterium longum subsp. infantis and B. breve, and of adults ( B. adolescentis) were selected for characterization. The aim of the present work was to investigate bifidobacteria from human host of different age with different feeding habits in order to establish a possible correlation between diet and the folate production. Folate is present in many different forms in humans. The detectable forms studied in the present work are 5-CH3-H4, H4 and total folate content. Bifidobacteria strains were cultivated in folate free synthetic media. Validated HPLC method was used to analyze deconjugated folates extracted from bacterial biomass.

En el marco del desarrollo de nuevos productos lácteos funcionales, yogur y queso untable, que aporten beneficios al consumidor, como ser un eficiente vehículo de microorganismos probióticos (MP), surge la necesidad de evaluar el comportamiento de diversas cepas frente a factores de estrés que puedan afectarlas al ser incorporadas a la matriz alimenticia durante la elaboración de la misma, almacenamiento y/o consumo del producto. Se seleccionaron cepas comerciales: Lactobacillus acidophilus LA5, L. rhamnosus LGG, L. rhamnosus SP1*, Bifidobacterium animalis subsp. lactis BB12; y otras provenientes de la Colección de Bifidobacterias Scardovi, de la Universidad de Bologna, Italia (BUSCOB): L. acidophilus M384*, B. animalis subsp. lactis MB29 (DSM 10140T)*, B. longum subsp. longum B1835* y B. longum subsp. longum RE11 (ATCC 15707T). Sobre cultivos puros se realizaron ensayos de supervivencia en condiciones gastrointestinales simuladas (CGI): bajo pH (supervivencia luego de 0, 15 y 90 min a pH 2,5 con pepsina 3 g/L) y presencia de sales biliares (supervivencia luego de 0, 60 y 210 min a pH 8 con bilis 2%). Los recuentos fueron realizados en condiciones óptimas para cada especie. Se probó también la resistencia a antibióticos (tetraciclina, cefuroxima, kanamicina, cloranfenicol, vancomicina, ampicilina, estreptomicina y eritromicina), evaluando la mínima concentración inhibitoria. Además, se seleccionó 1 cepa de cada especie (*) para ser incorporadas en la fermentación de las matrices lácteas, junto a los cultivos iniciadores, y se evaluó su supervivencia frente a las CGI (bajo pH y bilis, a 90 y 210 min respectivamente) a los 5 días de producidas. En los ensayos de supervivencia, se observó que LA5, MB29 y BB12 mostraron muy buena resistencia a bajo pH, disminuyendo como máximo un orden logarítmico (OL) luego de 90 min. En cambio, frente a bilis todas las cepas mostraron buena resistencia, incluso luego de 210 min, con 1 OL de disminución como máximo, en el caso de B1835. MB29 y BB12, fueron las cepas que menor resistencia a antibióticos mostraron, siendo resistentes a 2 de 8 antibióticos ensayados. En los ensayos sobre los productos, se observó que MB29 es aquella que mejor resistencia mostró frente a las CGI, manteniendo el valor inicial, siendo el recuento siempre superior a 108 UFC/g. Además, desde el punto de vista sensorial, ambos productos adicionados con MB29 resultaron de textura y sabores agradables, concluyéndose que dicha cepa posee muy buena aptitud para ser incorporada a los nuevos productos.

Aquaculture is one of the fastest growing sector of food production in the world. Despite the encouraging trends, several constraints have had a negative impact on the growth of aquaculture. Diseases are the primary limiting factor in large scale production facilities, where aquatic animals are exposed to stressful conditions. Bacterial diseases are responsible for heavy mortality in both wild and cultured fish and result in serious economic lossess. Prevention and control of diseases have led during recent decades to a substantial increase in the use of veterinary medicines. However, the utility of anti-microbial agents as a preventive measure has been questioned, given the extensive documentation on the development and spread of antimicrobial resistance genes among bacteria. Several mechanisms can be responsible for the acquirement of antibiotic resistances, the main common of which are chromosomal mutation or acquisition of plasmids. Chromosomal mutations cannot be transferred to other bacteria but plasmids can transfer resistance rapidly. Several bacterial pathogens can develop plasmid-mediated resistances: plasmids carrying genes for resistance to antibiotics have been found in marine Vibrio species and they could be laterally exchanged. The concept of biological disease control using probiotics has received widespread attention during the last decade. When looking at probiotics intended for an aquatic usage, it is important to take into account the complex relationship that an aquatic organism has with its direct environment, compared to terrestrial animals. Gram-negative facultative anaerobic bacteria are dominant in fish and shellfish digestive tract, but the intestinal microbiota of aquatic animals may change very rapidly with the ingestion of microorganisms coming from water and food. This is probably the reason why a large number of probiotics developed in aquaculture are bacteria directly originating from the aquatic environment. However, more ‘traditional’ bacterial or yeast species marketed for animal nutrition (Lactobacillus spp., Pediococcus spp., Bacillus spp., and S. cerevisiae) are also used. They can target fish eggs and larvae, fish juveniles and adults, crustaceans, bivalve molluscs and also live food such as rotifers, artemia, or unicellular algae (Kesarcodi-Watson et al., 2008). Growth-promoting effects, through better feed utilisation and digestion, as well as biological control of pathogen colonisation are the most important expected benefits of probiotic applications. Disease outbreaks caused by Vibrio spp. or Aeromonas spp. have been recognised as the most significant constraint on aquaculture production, particularly in the shrimp subsector, where vibriosis is currently one of the main diseases identified. Whereas in vitro, antagonism to pathogens has been clearly demonstrated for a wide range of probiotic strains, in vivo evidence of efficacy is still very scarce. Most probiotics proposed as biological control agents in aquaculture belong to the lactic acid bacteria (Lactobacillus spp. and Carnobacterium spp.), to the genus Vibrio (V. alginolyticus), to the genus Bacillus, or to the genus Pseudomonas, although other genera or species have also been mentioned (Aeromonas and Flavobacterium). Few multinational pharmaceutical companies have introduced commercial preparations into the market as probiotics feed/food supplement in various commercial names as Aqualact, Spilac, Protexin (Verschuere et al., 2000). This chapter will give some examples on how probiotics can be applied to reduce the incidence of diseases usually affecting aquatic animals, including fish, crustaceans and mollusks. Moreover, a brief consideration on the recent approach of prebiotic administration will also be provided.

Bifidobacteria and lactobacilli due to their health promoting activity are commonly used as probiotic in dairy products for human consumption. Their suggested effective concentration is up to 106CFU/g of product. The correct identification and the monitoring of viable probiotics in the final product is an indispensable prerequisite for applications in industrial processes and in safety and quality control applications. The aim of this study was the optimization of routine laboratory methods for the enumeration and identification of probiotic species commercialized in dairy products. Nine fermented milk products of different marketed brands, purchased in Italian supermarkets, were tested. In these products, with or without starter cultures, Bifidobacterium animalis subsp.lactis and/or L. acidophilus, L. casei, L. johnsonii, L. paracasei, L. rhamnosus have been declared in the product label. The shelflife for all the products was about 20 days. Bifidobacteria were counted in TPY-mupirocin medium while lactobacilli were all counted using MRS-bile both in aerobiosis and anaerobiosis; in anaerobiosis MRS-pH 5.4 for L. acidophilus, L. johnsonii and L. paracasei, and MRS-vancomycin for L. casei and L. rhamnosus were also used. The obtained isolates have been catalogued basing on cell morphology and identified by species-specific PCR and ERIC-PCR. TPY-mupirocin has been confirmed as an optimal selective media for bifidobacteria; for L. rhamnosus and L. paracasei all the media tested were suitable while for L. acidophilus and L. johnsonii MRS-pH5.4 gave better results. All the probiotic dairy products contained the recommended levels of 106–107 CFU/g probiotic bacteria except one brand with 105 CFU/g. Morphology cannot help in distinguishing between the species L. casei, L. paracasei and L. johnsonii while L. acidophilus and L. rhamnosus show peculiar morphology; B. animalis subsp.lactis show the typical "bone" morphology. The present work confirm that ERIC-PCR is a valuable method to rapidly discriminate between lactobacilli closely related species while species-specific PCR does not discriminate between L. casei and L. paracasei. All the products declaring the presence of L. casei in the label actually contain L. paracasei.

Folates are water-soluble B-group vitamin that are cofactors in many metabolic pathways such as DNA synthesis and methylation pathways. Humans cannot synthesize folate and its low status has been implicated in a wide variety of disorders. Naturally occurring food folates occur in abundance in leafy green vegetables, yeast extracts, citrus fruit, liver and kidney. Folate could derived, other than from diet, from indigenous bacteria of the microbiota which posses the ability of de novo folate production. Intestinal bacteria such as bifidobacteria can produce this cofactor by themselves from simple precursors; howewer some bifidobacteria are auxotrophic and have a strict growth requirement for folic acid. The study of folate metabolism in bifidobacteria isolated from different animals, such as omnivorous, herbivorous and carnivorous, could advances the knowledge of host-interaction in relation to endogenous folate production for possible microbiota modulation. We aimed to investigate the production of folate by bifidobacteria isolated from animals differing in diet in order to assess the potential indigenous contribute of folate availability provided by the microbiota.We aimed to investigate the production of folate by bifidobacteria isolated from animals differing in diet in order to assess the potential indigenous contribute of folate availability provided by the microbiota.

Study protocol; Journal Article; Multicenter Study; Randomized Controlled Trial; Research Support, Non-U.S. Gov't; BACKGROUND Uncomplicated chronic rachialgia is a highly prevalent complaint, and one for which therapeutic results are contradictory. The aim of the present study is to evaluate the effectiveness and safety of treatment with auriculopressure, in the primary healthcare sector, carried out by trained healthcare professionals via a 30-hour course. METHODS/DESIGN The design consists of a multi-centre randomized controlled trial, with placebo, with two parallel groups, and including an economic evaluation. Patients with chronic uncomplicated rachialgia, whose GP is considering referral for auriculopressure sensory stimulation, are eligible for inclusion. Sampling will be by consecutive selection, and randomised allocation to one of the two study arms will be determined using a centralised method, following a 1:1 plan (true auriculopressure; placebo auriculopressure). The implants (true and placebo) will be replaced once weekly, and the treatment will have a duration of 8 weeks. The primary outcome measure will be the change in pain intensity, measured on a visual analogue scale (VAS) of 100 mm, at 9 weeks after beginning the treatment. A follow up study will be performed at 6 months after beginning treatment. An assessment will also be made of the changes measured in the Spanish version of the McGill Pain Questionnaire, of the changes in the Lattinen test, and of the changes in quality of life (SF-12). Also planned is an analysis of cost-effectiveness and also, if necessary, a cost-benefit analysis. DISCUSSION This study will contribute to developing evidence on the use of auriculotherapy using Semen vaccariae [wang bu liu xing] for the treatment of uncomplicated chronic rachialgia. TRIAL REGISTRATION Current Controlled Trials ISRCTN01897462. Yes

Background Non cephalic presentation in childbirth involves various risks to both the mother and the foetus. The incidence in Spain is 3.8% of all full-term pregnancies. The most common technique used to end the gestation in cases of non cephalic presentation is that of caesarian section, and although it provokes a lower rate of morbi-mortality than does vaginal delivery in such situations, there remains the possibility of traumatic injury to the foetal head and neck, while maternal morbidity is also increased. The application of heat (moxibustion) to an acupuncture point, in order to correct non cephalic presentation, has been practised in China since ancient times, but as yet there is insufficient evidence of its real effectiveness. Methods/Design The experimental design consists of a multi-centre randomised controlled trial with three parallel arms, used to compare real moxibustion, sham moxibustion and the natural course of events, among pregnant women with a non cephalic presentation and a gestational duration of 33–35 weeks (estimated by echography). The participants in the trial will be blinded to both interventions. The results obtained will be analyzed by professionals, blinded with respect to the allocation to the different types of intervention. In addition, we intend to carry out a economic analysis. Discussion This trial will contribute to the development of evidence concerning moxibustion in the correction of non cephalic presentations. The primary outcome variable is the proportion of cephalic presentations at term. As secondary outcomes, we will evaluate the proportion of cephalic presentations at week 38 of gestation, determined by echography, together with the safety of the technique, the specificity of moxibustion and the control of the blinding process. This study has been funded by the Health Ministry of the Andalusian Regional Government. Trial registration Current Controlled Trials ISRCTN10634508.

L'analisi di targets associabili ad una sorgente specifica consente di distingure la natura dell'inquinamento fecale, umana o animale di un corpo idrico. Uno studio condotto nell'interland bolognese ha valutato l'impiego di bifidobatteri come Microbial Source Tracking ospite specifico oltre che come strumento di stima dell'età della contaminazione fecale.

The general objectives of the unit are related to the exposed lectures. The first step is give an introductory point of view about the principal factor influencing safety in food chain. This is a first step which conduct the food industry employer to the principal hygienic rules to follow during food handling and food production. That can serve also as introduction to the principal low of European Community in matter of food production and food industry. This module is also a mean to introduce the user to the world of the safety and quality certification (HACCP, UNI EN ISO). As general objective can be recognized an introduction to food quality, with the intent of trying to define it considering the question in each aspect (safety, salubrity, nutrition) and from different point of view (producer and consumers).

The outlines of antibiotic resistance of some probiotic microorganisms were studied. This study was conducted with the double purpose of verifying their ability to survive if they are taken simultaneously with an antibiotic therapy and to increase the selective properties of suitable media for the isolation of samples containing mixed bacterial populations. We isolated from commercial dairy and pharmaceutical products, 34 strains declared as probiotics, belonging to the genera Bifidobacterium and Lactobacillus, and 21 strains of starter culture bacteria. All the microorganisms have been compared by electrophoresis of the soluble proteins for the purpose of identifying them. A Multiplex-PCR with genus- and species-specific primers was used to detect for Bifidobacterium animalis subsp. lactis presence. All bifidobacteria were B. animalis subsp. lactis except one Bifidobacterium longum. Sometimes the identification showed that the used strain was not the one indicated on the label. The lactobacilli were Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus delbrueckii subsp. bulgaricus. The streptococci were all Streptococcus thermophilus. The minimal inhibitory concentration (MIC) of 24 common antibiotic substances has been valued by the broth microdilution method. All tested strains were susceptible to ampicillin, bacitracin, clindamycin, dicloxacillin, erytromycin, novobiocin, penicillin G, rifampicin (MIC(90) ranging from 0.01 to 4 microg/ml); resistant to aztreonam, cycloserin, kanamycin, nalidixic acid, polymyxin B and spectinomycin (MIC(90) ranging from 64 to >1000 microg/ml). The susceptibility to cephalothin, chloramphenicol, gentamicin, lincomycin, metronidazole, neomycin, paromomycin, streptomycin, tetracycline and vancomycin was variable and depending on the species.

La riduzione all’uso di farmaci spinge a cercare nuove strategie alimentari per migliorare accrescimento, salute, risposta immunitaria dei suini in allevamenti da reddito. Probiotici e prebiotici offrono la possibilità di modulare, da soli o in combinazioni nutraceutiche, la microflora gastrointestinale dell’ospite. L’integrazione della dieta con bifidobatteri può stimolare positivamente l’ecosistema intestinale, migliorando velocità di accrescimento e utilizzazione della razione alimentare. Sono state valutate le potenzialità probiotiche di bifidobatteri, le capacità prebiotiche di due diverse concentrazioni di frutto- e galattooligosaccaridi, gli effetti sinbiotici di sei combinazioni nutraceutiche. La verifica della colonizzazione di Bifidobacterium spp. nel contenuto di ceco è stata determinata attraverso il metodo colturale classico ed un protocollo di PCR diretta quantitativa, genere specifica. Si è valutata (64 soggetti) la capacità colonizzante di 12 ceppi selezionati tra le 4 specie B. breve, B. animalis subsp. lactis, B. suis e B. choerinum. Le migliori potenzialità probiotiche appartengono ai ceppi M 354 e RA 18 di B. animalis subsp. lactis ed al ceppo SU 891 di B. choerinum: la somministrazione comporta una riduzione del pH intestinale, incremento del livello di bifidobatteri nel ceco, assenza di diarrea. Si sono testate (64 soggetti), le capacità prebiotiche di diverse concentrazioni (2% e 4% della dieta) di due fruttooligosaccaridi, uno da inulina e uno da barbabietola e di un Galattooligosaccaride da siero di latte. Un effetto significativo sul livello dei bifidobatteri endogeni si osserva per il FOS da barbabietola al 4%. Il ceppo RA 18 di Bifidobacterium animalis subsp. lactis ed il ceppo Su 891 di B. choerinum sono stati testati infine (128 soggetti) combinando quattro diverse dosi di probiotico (0, 107, 109, 1011 ufc/day) con la concentrazione prebiotica di FOS individuata. Il solo ceppo di B. animalis subsp. lactis RA 18 determina un significativo aumento di peso corporeo linearmente con la dose somministrata. Il FOS interagisce sinergicamente con RA 18 incrementando significativamente i bifidobatteri.