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  • Author: R. Nemcová x
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Open access

K. Englerová, R. Nemcová and D. Mudroňová

Abstract

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.

Open access

R. Nemcová, M. Maďar, S. Gancarčíková and J. Pistl

Abstract

FISH (fluorescence in situ hybridization) analysis of the intestinal tract of conventional mice, following 14-day supplementation of feed with host non-specific (porcine) strain L. reuteri L2/6, showed in the presence of complex microbiota, a significant increase in the counts of representatives of the genera Lactobacillus and Bifidobacterium, and a significant decrease in the representatives of the genera Clostridium, Bacteroides and Enterobacteriaceae. At the same time, the supplemented strain stimulated the population of caecal lactobacilli of the species L. reuteri. These results demonstrated that the L. reuteri L2/6 colonised the jejunum, ileum and caecum and modulated the investigated intestinal microbiota.

Open access

M. Fedorová, R. Nemcová, D. Mudroňová, E. Styková, M. Brudňáková and K. Reiffová

Abstract

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).

Open access

D. Sopková, R. Vlčková, Z. Andrejčáková, Z. Hertelyová, S. Gancarčíková and R. Nemcová

Abstract

This study investigated for 14 days post-weaning, the influence of dietary supplementation of synbiotics in the form of probiotic cheeses containing cultures of L. plantarum and L. fermentum and crushed flaxseed (source of ω-3 polyunsaturated fatty acids — PUFAs and fibre) on 36 commercial piglets originating from an infected herd (Coronavirus and E. coli) during the critical period of weaning. We focused on the health and metabolism of PUFAs in this critical period of a piglet’s life. The dietary supplementation positively affected: the overall health state of weaners, reduced diarrhoea by 29 % by 14 days post-weaning and significantly increased the counts of lactic acid bacteria, bifidobacteria and the production of volatile fatty acids. The PUFA concentrations in the m. biceps femoris of the piglets were analysed by gas chromatography. High levels of ω-3 alpha-linolenic acid (ALA) in flaxseed increased significantly the level of ALA, eicosapentaenic acid (EPA) and docosahexaenic acid (DHA) in the pig muscles on days 7 and 14 post-weaning. The levels of ω-6 linolenic acid (LA) were less affected by the diet, but were increased on day 14 post-weaning, while the conversion products of LA, and arachidonic acid (AA), were decreased on days 7 and 14. The increased level of dietary ALA favoured the activity of Δ-6-desaturase for the conversion of ALA to EPA and DHA, at the expense of AA synthesis from LA. The ability of synbiotics to incorporate high levels of DHA in the pig muscles appear prospective for improving the nutritional properties of pork and reducing the occurrence of civilization diseases in consumers of this product of animal origin.

Open access

K. Hlubeňová, D. Mudroňová, R. Nemcová, S. Gancarčíková, M. Maďar and Ľ. Sciranková

Abstract

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.