Microbial communities in bees, pollen and honey from Slovakia.

As the honey-bee gastrointestinal tract microflora and pollen are the primary sources for the honey microbial community, the aim of this work was to study and characterize the microbial transit among them. Therefore, an exhaustive microbial analysis of honey, adult honey-bee gastrointestinal tract, and pollen from different Slovakian regions and different seasons, was conducted. Microbial screening revealed that the primary sources of microbial community present in Slovakian honey are pollen and the honey-bees' digestive tract microflora, containing microorganisms normally present in dust, air and flowers. We found that the digestive tract of Slovakian adult honey-bees is highly populated by anaerobic, rather than aerobic bacteria, where coliforms, enterococci, staphylococci, Bacillus sp., Pseudomonas sp., microscopic fungi and yeast were found. Interestingly, statistical differences were found between the microflora of the gastrointestinal tract of summer and winter bees. Pollen revealed the presence of mesophil anaerobic and aerobic microorganisms, coliforms and microscopic fungi. Among these, the most representative genera were Alternaria, Cladosporium and Penicillium . In honey the counts of total anaerobic and total aerobic bacteria, that of coliforms, enterococci, bacilli, microscopic fungi and yeasts were monitored. Most frequently microscopic fungi belonging to genera Penicillium, Cladosporium and Alternaria were found.

Maternal breast-milk and intestinal bifidobacteria guide the compositional development of the Bifidobacterium microbiota in infants at risk of allergic disease.

BACKGROUND: The sources and the impact of maternal bacteria on the initial inoculum of the intestinal microflora of newborn infants remain elusive. OBJECTIVE: To assess the association between maternal breast-milk and fecal bifidobacteria and infants' fecal bifidobacteria. METHODS: Sixty-one mother-infant pairs were included, special emphasis being placed on the maternal allergic status. Bifidobacteria were analysed by a direct PCR method in fecal samples from mothers at 30-35 weeks of gestation and from infants at 1 month of age and from breast-milk samples 1 month post-partum. RESULTS: Fecal Bifidobacterium adolescentis and Bifidobacterium bifidum colonization frequencies and counts among mother-infant pairs correlated significantly (P=0.005 and 0.02 for frequencies, respectively, and P=0.002 and 0.01 for counts, respectively). Only infants of allergic, atopic mothers were colonized with B. adolescentis. Each of the breast-milk samples contained bifidobacteria [median 1.4 x 10(3) bacterial cells/mL; interquartile range (IQR) 48.7-3.8 x 10(3)]. Bifidobacterium longum was the most frequently detected species in breast-milk. Allergic mothers had significantly lower amounts of bifidobacteria in breast-milk compared with non-allergic mothers [median 1.3 x 10(3) bacterial cells/mL (IQR 22.4-3.0 x 10(3)) vs. 5.6 x 10(3) bacterial cells/mL (1.8 x 10(3)-1.8 x 10(4)), respectively, (P=0.004)], and their infants had concurrently lower counts of bifidobacteria in feces [3.9 x 10(8) bacterial cells/g (IQR 6.5 x 10(6)-1.5 x 10(9)) in infants of allergic mothers, vs. 2.5 x 10(9) bacterial cells/g (6.5 x 10(8)-3.2 x 10(10)) in infants of non-allergic mothers, P=0.013]. CONCLUSIONS: Breast-milk contains significant numbers of bifidobacteria and the maternal allergic status further deranges the counts of bifidobacteria in breast-milk. Maternal fecal and breast-milk bifidobacterial counts impacted on the infants' fecal Bifidobacterium levels. Breast-milk bacteria should thus be considered an important source of bacteria in the establishment of infantile intestinal microbiota.

Effect of bile on the lipid composition and surface properties of bifidobacteria.

AIM: The changes produced on the bacterial surface of Bifidobacteria cells when they are grown in bile were compared with those provoked by bile added to bacteria grown in the absence of bile. METHODS AND RESULTS: The adhesive properties, the zeta potential and the lipid composition of Bifidobacterial strains, isolated from human faeces and grown in MRS medium, were determined. Bacteria grown in MRS with bile showed a loss of adherence and autoaggregation in correlation with a decrease in the surface hydrophobicity in comparison to those grown in MRS without bile, concomitant with the absence of two glycolipids, the increase of sugar content and minor changes in fatty acid composition. The surface changes caused by bile shock on bacteria grown in bile-free medium were much less pronounced and, in addition, no effect on the lipid composition was apparent. CONCLUSIONS: The comparison of the results indicates that bile action on surface properties is related to metabolic changes. SIGNIFICANCE AND IMPACT OF THE STUDY: Long-term exposure of bacteria to bile may cause metabolic changes affecting their adhesive properties irreversibly. This may be taken as a criterion to define the probiotic properties of different strains.

Screening of bile resistance and bile precipitation in lactic acid bacteria and bifidobacteria.

A modification of the ecometric method was developed for a rapid screening of bile resistance in lactic acid bacteria and bifidobacteria. Validation of the MEM bile assay (modified ecometric method) was performed comparing the bile resistance index (RIbile) and the bile resistance ratio (R%). Most Bifidobacterium strains assayed were bile sensitive (83.3%), while only 62.1% of lactobacilli showed that behavior. Some bifidobacterial strains (55.6%) showed a crystalline precipitate when grown on solid medium supplemented with 0.5% ox bile. The crystalline structures produced by B. pseudolongum CIDCA 531 were isolated and analyzed by optical and scanning electron microscopy, thin-layer chromatography, melting point, and specific cholesterol reactions. Those studies confirmed the presence of cholesterol in these crystalline structures. On the other hand, none of the lactobacilli and streptococci studied had the ability to produce crystalline precipitates.

Isolation and characterization of Bifidobacterium strains for probiotic formulation.

Twenty-five Bifidobacterium strains isolated from infant feces were identified by sugar fermentation patterns and whole-cell protein analysis. Using gradient SDS-PAGE, six characteristic protein bands of the genus were detected in 40 strains of bifidobacteria but not in lactobacilli. Computerized numerical analysis enabled strains to be grouped in two main clusters. Strains of Bifidobacterium bifidum belong to a well-differentiated cluster that joins the cluster of the remaining species at 0.582 similarity. The predominant species among isolated strains from infant feces were B. bifidum, B. longum, and B. breve. Probiotic and technological indicators such as surface properties, inhibitory capacity, resistance to bile and low pH, and ability to grow under aerobic conditions were studied. Not all desirable characteristics were present in a single strain. In general, adherent and inhibitory strains were not resistant to bile, low pH, and aerobic conditions. Only 10 of 40 strains were resistant to 0.5% bile.