Probiotic Lactobacillus fermentum UCO-979C biofilm formation on AGS and Caco-2 cells and Helicobacter pylori inhibition

The ability of the human isolate Lactobacillus fermentum UCO-979C to form biofilm and synthesize exopolysaccharide on abiotic and biotic models is described. These properties were compared with the well-known Lactobacillus casei Shirota to better understand their anti-Helicobacter pylori probiotic activities. The two strains of lactobacilli synthesized exopolysaccharide as detected by the Dubois method and formed biofilm on abiotic and biotic surfaces visualized by crystal violet staining and scanning electron microscopy. Concomitantly, these strains inhibited H. pylori urease activity by up to 80.4% (strain UCO-979C) and 66.8% (strain Shirota) in gastric adenocarcinoma (AGS) cells, but the two species showed equal levels of inhibition (~84%) in colorectal adenocarcinoma (Caco-2) cells. The results suggest that L. fermentum UCO-979C has probiotic potential against H. pylori infections. However, further analyses are needed to explain the increased activity observed against the pathogen in AGS cells as compared to L. casei Shirota.     

Administration of the probiotic Lactobacillus rhamnosus IMC 501 as a strategy for the control of Vibrio bacteria in the brine shrimp Artemia

The present study aimed to address the capability of the probiotic bacterium Lactobacillus rhamnosus IMC 501^® to survive in seawater and the ability of Artemia metanauplii to incorporate it, as well as to analyse the potential effect of the probiotic as a control agent for potentially pathogenic Vibrionaceae bacteria in Artemia. The results demonstrate the ability of L. rhamnosus IMC 501^® to survive in seawater for up to 30 h. They also advocate their capability to be efficiently incorporated into Artemia metanauplii at concentrations of 10⁴ CFU per Artemia after 30 min of suspension in probiotic solution, thereby promoting a 1-log reduction in Vibrionaceae levels after 3 h. These low levels of Vibrio bacteria were maintained for about 30 min after transfer into clear seawater, a sufficient time for Artemia to be ingested by aquatic organisms. These results contribute to broaden the knowledge on the suitability of probiotics as sustainable alternatives for the prevention/reduction of diseases in aquaculture facilities.     

Protoplast formation and regeneration in <Emphasis Type="Italic">Lactobacillus delbrueckii</Emphasis>

Method for production and regeneration of Lactobacillus delbrueckii protoplasts are described. The protoplasts were obtained by treatment with a mixture of lysozyme and mutanolysin in protoplast buffer at pH 6.5 with different osmotic stabilizers. The protoplasts were regenerated on deMan, Rogosa and Sharpe (MRS) with various osmotic stabilizers. Maximum protoplast formation was obtained in protoplast buffer with sucrose as an osmotic stabilizer using a combination of lysozyme (1 mg/ml) and mutanolysin (10 μg/ml). Maximum protoplast regeneration was obtained on MRS medium with sucrose (0.5 M) as an osmotic stabilizer. The regeneration medium was also applicable to other species of lactobacilli as well. This is, to our knowledge, the first report on protoplast formation and efficient regeneration in case of L. delbrueckii.     

Modulatory effect of Lactobacillus acidophilus KLDS 1.0738 on intestinal short‐chain fatty acids metabolism and GPR41/43 expression in β‐lactoglobulin–sensitized mice

We investigated the correlation between the beneficial effect of Lactobacillus acidophilus on gut microbiota composition, metabolic activities, and reducing cow's milk protein allergy. Mice sensitized with β‐lactoglobulin (β‐Lg) were treated with different doses of L. acidophilus KLDS 1.0738 for 4 weeks, starting 1 week before allergen induction. The results showed that intake of L. acidophilus significantly suppressed the hypersensitivity responses, together with increased fecal microbiota diversity and short‐chain fatty acids (SCFAs) concentration (including propionate, butyrate, isobutyrate, and isovalerate) when compared with the allergic group. Moreover, treatment with L. acidophilus induced the expression of SCFAs receptors, G‐protein–coupled receptors 41 (GPR41) and 43 (GPR43), in the spleen and colon of the allergic mice. Further analysis revealed that the GPR41 and GPR43 messenger RNA expression both positively correlated with the serum concentrations of transforming growth factor‐β and IFN‐γ (p < .05), but negatively with the serum concentrations of IL‐17, IL‐4, and IL‐6 in the L. acidophilus–treated group compared with the allergic group (p < .05). These results suggested that L. acidophilus protected against the development of allergic inflammation by improving the intestinal flora, as well as upregulating SCFAs and their receptors GPR41/43.     

Structural similarity and distribution of small cryptic plasmids of Lactobacillus curvatus and L. sake

Plasmid profiles of strains of Lactobacillus curvatus and L. sake isolated from meat or sauerkraut were analysed to investigate plasmid homology and distribution in relation to the ecology of these organisms in fermenting foods. A hybridisation probe was constructed by cloning of pLc2, a cryptic, 2.6-kbp plasmid from L. curvatus LTH683, into the Escherichia coli plasmid pRV50. In Southern hybridisations with the digoxygenine labeled pLc2 probe, pLc2-related small plasmids were frequently detected in meat-borne strains of L. casei subsp. pseudoplantarum, L. curvatus, L. sake, L. alimentarius, L. farciminis and L. halotolerans and in L. curvatus and L. sake isolated from sauerkraut. Among 27 Lactobacillus type strains originally isolated from habitats other than meat this type of homology was detected only with plasmids of L. buchneri and L. mali. Restriction-enzyme mapping of six small cryptic plasmids from L. curvatus and L. sake revealed strong structural homology but no similarity to previously characterized plasmids of lactobacilli. The presence of a variable region in addition to a conserved one and the occurrence of deletions during cloning of pLc2 suggest that vectors derived from these plasmids are likely to be structurally unstable.     

Distribution of lactobacilli in the porcine gastrointestinal tract

Abstract Characteristics of the lactobacilli colonizing the various regions of the porcine gastrointestinal tract were investigated. The lumenal contents from the gastric, jejunal, cecal and colonic regions, and biopsies from the gastric non-secreting and secreting regions were homogenized and serial dilutions were spread on Rogosa agar and colonies (50 per region and animal) were randomly sampled. The lactobacillus isolates were grouped according to their protein profiles of lysozyme-treated whole cells. Several different groups of lactobacillus could be detected. Specific groups were associated and often unique for the stomach, jejunal, cecal and colonic regions of the gastrointestinal tract. A few groups were uniformly distributed through the tract. Although published studies of lactobacilli colonizing stomach mucosa have been restricted mainly to the non-secreting region, significant levels were also detected on the secreting stomach tissue. Population densities ranged from 2 × 10⁴ to 2 × 10⁶ CFU per cm² on both the secretory and non-secretory regions. Of the isolates which colonized the gastric secreting epithelium, 26% formed smooth colonies and 74% formed rough colonies. In contrast, the non-secreting epithelium was mainly colonized by lactobacilli forming smooth colonies (67%). From this study, one can suggest that different lactobacilli are associated with specific regions of the gastrointestinal tract, and that some specific groups appear to be restricted to defined regions.