Influence of Zinc Sulphate on the Probiotic Properties of Lactobacillus plantarum CCM 7102

The effects of zinc sulphate on selected properties of L. plantarum CCM 7102 were tested in vitro. The resistance of lactobacilli to higher concentrations of ZnSO4 (up to 5000 mg Zn2+.l−1) in growth media was strain-dependent. Further studies were carried out on the most resistant strain of L. plantarum CCM 7102. While the addition of low concentrations of zinc sulphate into the growth media ( 500 mg Zn2+.l−1 inhibited these properties. The addition of zinc (250 mg Zn2+.l−1) did not affect the resistance to antimicrobials, low pH, and the resistance to bile salt was affected only weakly. Zinc-resistant probiotic Lactobacillus strains are suitable for use in feedstuffs with a higher content of zinc designed for the prevention of post weaning diarrhoea in pigs

The Study of the Probiotic Potential of the Beneficial Bacteria Isolated from Kefir Grains

The aim of this study was to identify beneficial bacteria with probiotic potential from kefir grains. The lactobacilli isolated from kefir grains were characterised as: Lactobacillus plantarum, Lactobacillus paraplantarum, Lactobacillus paracasei, and Lactobacillus kefiri. The strains Lb. plantarum 1Ž, Lb. paraplantarum S10, and Lb. paracasei 2Ž tolerated better the test gastric juice at pH 2 and 2.6 during 120 min of incubation in comparison with the strains Lb. kefiri. On the other hand, the strains Lb. kefiri were resistant to 0.3 % bile acid salts. The Lb. paracasei 2Ž showed the significantly highest survival (P < 0.001) at pH 2 in comparison with all other strains tested and was also able to tolerate 0.3 % concentration of the bile salts. All strains produced medium to strong biofilms on abiotic surfaces and inhibited the growth of selected potential pathogens with varying intensity. All kefir isolates were susceptible to the antibiotics tested and exhibited positive β-galactosidase activity with the exception of Lb. paracasei 2Ž which did not show any activity of undesirable enzymes, such as β-glucosidase and β-glucuronidase. Additional testing and validation of the biological properties and safety of the strain Lb. paracasei 2Ž under in vivo conditions are needed to confirm the prospective use of this strain in practice

Antiparasitic effects of selected isoflavones on flatworms

Medicinal plants have been successfully used in the ethno medicine for a wide range of diseases since ancient times. The research on natural products has allowed the discovery of biologically relevant compounds inspired by plant secondary metabolites, what contributed to the development of many chemotherapeutic drugs. Flavonoids represent a group of therapeutically very effective plant secondary metabolites and selected molecules were shown to exert also antiparasitic activity. This work summarizes the recent knowledge generated within past three decades about potential parasitocidal activities of several flavonoids with different chemical structures, particularly on medically important flatworms such as Schistosoma spp., Fasciola spp., Echinococcus spp., Raillietina spp., and model cestode Mesocestoides vogae. Here we focus on curcumin, genistein, quercetin and silymarin complex of flavonolignans. All of them possess a whole spectrum of biological activities on eukaryotic cells which have multi-therapeutic effects in various diseases. In vitro they can induce profound alterations in the tegumental architecture and its functions as well as their activity can significantly modulate or damage worm´s metabolism directly by interaction with enzymes or signaling molecules in dose-dependent manner. Moreover, they seem to differentially regulate the RNA activity in numbers of worm´s genes. This review suggests that examined flavonoids and their derivates are promising molecules for antiparasitic drug research. Due to lack of toxicity, isoflavons could be used directly for therapy, or as adjuvant therapy for diseases caused by medically important cestodes and trematodes

Effects of Cell Seeding Methods on Chondrogenic Differentiation of Rat Mesenchymal Stem Cells in Polyhydroxybutyrate/Chitosan Scaffolds

The aim of our study was to examine the effects of passive and active cell seeding techniques on in vitro chondrogenic differentiation of mesenchymal stem cells (MSC) isolated from rat bone marrow and seeded on porous biopolymer scaffolds based on polyhydroxybutyrate/chitosan (PCH) blends. This paper is focused on the distribution of the cells on and in the scaffolds, since it influences the uniformity of the created extracellular matrix (ECM), as well as the homogenity of the distribution of chondrogenic markers in vitro which ultimately affects the quality of the newly created tissue after in vivo implantation. The three types of cell-scaffold constructs were examined by: fluorescence microscopy, SEM, histology and quantitative analysis of the glycosaminoglycans after chondrogenic cultivation. The results demonstrated that the active cells seeded via the centrifugation of the cell suspension onto the scaffold guaranteed an even distribution of cells on the bulk of the scaffold and the uniform secretion of the ECM products by the differentiated cells

Concentration-dependent effect of silymarin on concanavalin A-stimulated mouse spleen cells in vitro

Silymarin (SIL), a mixture of phenolic compounds, has a pleiotropic mode of action on various cell types, including immune cells. In this study, we investigated the concentration-dependent effect of SIL on proliferation of concanavalin A (CoA)-stimulated mouse spleen T lymphocytes, their viability, and secretion of IFN-g and IL-4 cytokines ex vivo in relation to gene expressions of transcription factors nuclear factor kappa B and Foxp3. In addition, metabolic activity of T cells was determined as changes in the mitochondrial membrane potential and apoptosis

Differential sensitivity of myeloid and lymphoid cell populations to apoptosis in peritoneal cavity of mice with model larval Mesocestoides vogae infection

The metacestode stage of the tapeworm Mesocestoides vogae (M. vogae) has the ability of asexual growth in the peritoneal cavity of rodents and other intermediate hosts without restriction. Early immunological events have decisive role in the establishment of infection. In the present study we investigated the kinetic of myeloid and lymphoid cell populations and the proportions of cells undergoing apoptosis in peritoneal cavities of mice within the first month after oral infection with M. vogae larvae. Proportions of cell phenotypes and apoptotic cells were examined by flow cytometry and by microscopical analysis of cells following May/Grünwald staining and fluorescent stain Hoechst 33234, respectively. Total numbers of peritoneal cells increased and their distribution changed towards accumulation of myelo-monocytic cell lineage in the account of reduced proportions of lymphoid cells. CD4+ T cell subpopulations were more abundant than CD8+ and their proportions elevated within two weeks post infection (p.i.) which was followed by a significant decline. Expression level of CD11c marker on myelo-monocytic cells revealed phenotype heterogeneity and proportions of cells with low and medium expression elevated from day 14 p.i. along with concurrent very low presence of CD11chigh phenotype. Lymphoid cell population was highly resistant to apoptosis but elevated proportions of myeloid cells were in early/late stage of apoptosis. Apoptosis was detected in a higher number of adherent cells from day 14 p.i. onwards as evidenced by nuclear fluorescent staining. By contrast, cells adherent to larvae, mostly macrophages and eosinophils, did not have fragmented nuclei. Our data demonstrated that apoptosis did not account for diminished population of peritoneal lymphoid cells and substantial proportions of myeloid cells seem to be more susceptible to apoptotic turnover in peritoneal cavity of mice with ongoing M. vogae infection, suggesting their important role in the host-parasite interactions

Exopolysaccharides May Increase Gastrointestinal Stress Tolerance of Lactobacillus reuteri

This study investigated a possible relationship between exopolysaccharides (EPS) production and the resistance to bile salts and low pH in intestinal strains of Lactobacillus reuteri. The strains displayed a mucoid phenotype, when grown in the presence of 10 % sucrose. Scanning electron microscopy (SEM) revealed strands of exopolysaccharide linking neighbouring cells. The strains (except L. reuteri B1/1) produced EPS in the range from 15.80 to 650.70 mg.l−1. The strains were tested for tolerance to bile salts (0.15; 0.3 %) and low pH (1.5—2.0—2.5—3.0). The survival rate, after the treatment with artificial gastric and intestinal juices, was determined by flow cytometric analysis. The strains of L. reuteri that produced 121—650 mg.l−1 of EPS showed a significantly higher tolerance (P < 0.001) to the gastric juice at pH 3 and 2.5, throughout the entire exposure time, in comparison to the strains that produced less than 20 mg.l−1 of EPS. L. reuteri L26, with the highest production of EPS, exhibited the highest survival rate (60 %) at pH 2 after the 120 minutes of in-cubation and was able to tolerate pH 1.5 for 30 minutes. Higher production of EPS significantly (P < 0.001) increased the strains’ tolerance against the intestinal juice in the presence of 0.15 and 0.3 % bile salts and was time dependent. L. reuteri L26 showed the highest tolerance (P < 0.001) against 0.3 % bile salts. This investigation revealed a positive correlation between the EPS production and the resistance of intestinal L. reuteri to the stress conditions of the gastrointestinal tract (GIT)

The Efect of Probiotic Lactobacilli and Alginite on the Cellular Immune Response in Salmonella Infected Mice

Alginite is organic matter rich in humic substances and commonly found in nature, but despite that, the knowledge of its biological effects is limited. In our study we focused on monitoring the effects of alginite alone, as well as its effect as a carrier of probiotic lactobacilli on the cellular immune response in SPF mice after infection with Salmonella Typhimurium. Sixty six conventional SPF female mice of the Balb/c line were divided into 4 groups: 1. infection free negative control (NK) supplied neither alginite nor probiotic lactobacilli in the feed; 2. infection free alginite control (Alg) supplied feed with 10 % alginite; infected control supplied alginite in the feed but no lactobacilli; 3. infectious control (Alg + Sal) - animals infected with salmonella and supplied 10 % alginite in the feed but no lactobacilli;and 4. probiotic group (Lab + Alg + Sal) - animals infected with salmonella and administered 10 % alginite and Lactobacillus reuteri 2/6 in the feed. On day 21 of the experiments, the mice were bled and their mesenteric lymph nodes were taken after their death. The peripheral blood of the mice was analysed for the activity of phagocytes and the percentage of selected lymphocyte subpopulations was determined in the mesenteric lymph nodes and blood. The significantly highest phagocytic activity (FA) was noted in the infected group with alginite (Alg + Sal). The FA was significantly increased in groups Alg and Lab + Alg + Sal in comparison with the NK group. The highest engulfing ability of phagocytes (phagocytic index) was observed in the Lab + Alg + Sal group in comparison with other groups, but also in Alg group in comparison with NK. In the Lab + Alg + Sal group, we observed a significantly higher percentage of B-lymphocytes, CD4+CD8+ and natural killer T cells (NKT), but more significant impact on the numbers of subpopulations of lymphocytes was observed in the mesenteric lymph nodes, with the significantly highest proportions of CD4+CD8+lymphocytes and NK and NKT cells. Our results indicated immunomodulatory properties of alginite and L. reuteri 2/6 in salmonella infected mice, especially at the level of the innate immune system components. This activation of phagocytosis and NK cells can be used in the treatment of various infections