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Lactobacilli were isolated from chicken gastrointestinal tract and examined for their potentially probiotic properties towards their inhibitory activity against poultry enteropathogenic bacteria. Biochemical tests, ITS-PCR and cell wall protein analysis were used to characterize the Lactobacillus isolates. The identification of isolated Lactobacillus strains based on phenotypic properties was not always satisfactory. ITS-PCR together with protein profile were found to be helpful in strain identification. Lactobacilli were tested for the inhibitory activity against selected strains of poultry enteropathogenic bacteria (Salmonella Enteritidis, Escherichia coli and Clostridium perfringens). Examined supernatants from Lactobacillus broth cultures demonstrated major antimicrobial activity against C. perfringens. Lower antimicrobial activity were observed against E. coli and Salmonella Enteritidis. The strongest inhibition effect were obtained using supernatant of Lactobacillus acidophilus strain 3D. Results received from this study confirmed that identification of Lactobacillus spp. is often tedious. Some isolates, which are in vitro antagonistic against enteropathogenic bacteria may be considered as potential candidates for poultry probiotics, especially in controlling necrotic enteritis caused by C. perfringens.
Background. Enteroccocci occur and may compete well in fermented sausages and Enterococcus faecium represents that species of the lactic acid bacteria which can be found in the fermented sausages. The representatives of this species can produce bacteriocins with predominant anti-listerial effect. Therefore, the effect of enterocin (Ent) 4231 produced by Enterococcus faecium CCM 4231 strain with probiotic properties was tested in a dry fermented salami Puchov (Slovak product) experimentally inoculated with L. innocua Li1 strain (107 cfu/ml). Material and methods. The bulk salami mixture was prepared in the pilot plant and 2.5 kg for each of three trials were transferred to the laboratory for the experiments. Three independent trials were conducted, each comprising then five salami samples (0.500 g). Trial A (reference control) involved only untreated salami mixture. Trial B represented salami mixture inoculated with Listeria innocua Li1 (107 cfu/ml). For trial C, Ent 4231 possessing activity 6400 AU/ml was added into the salami mixture inoculated with L. innocua Li1 (LilEnt). The mixtures were stuffed into collagen casings and the fiat shape salamis were transferred back to the pilot plant and treated according to conditions typical for this product and stored for 4 weeks. Results. The initial number of L. innocua Li1 in the inoculated salami mixture was 104 cfu/ml. Aft er Ent 4231 addition, the count of Listeria detected in the salami samples inoculated with Li1 and treated with Ent 4231 was 3.64 ±0.14 cfu/ml; difference 0.40 logarithmic cycles was noted between Li samples and Li /Ent samples. On day 2, the difference 1.86 log cycles was noted between Li1 and IM Ent samples. Although, in weeks 3 and 4, slight increase in Li1 cells was determined in Li salamis, the difference in the detection of Li1 cells in Li salamis and Li/Ent samples was even higher than that immedially after Ent addition (difference 2.30; 2.48 log cycles). Bacteriocin activity itself was not recovered from Li/Ent salamis. The pH of the all salamis was almost at the same level. Water activity and water content were not influenced. Conclusion. Addition of Ent 4231 during processing of salami Puchov experimentally inoculated with L. innocua Li1 has lead to decrease of Li1 cell growth, although the bacteriocin activity of Ent itself was not possible to detect in salami samples. The pH value, water activity, as well as sensory character of the final products were not negatively influenced.
The probiotic potential of 3 yeasts strains of Saccharomyces cerevisiae isolated from kefirs and feces was investigated and compared with 3 isolates from medicines and 2 collection strains (ATCC) of Saccharomyces cerevisiae var. houlardii. Genetic identification of yeasts based on karyotypes indicated their affiliation to Saccharomyces spp. although chromosomal polymorphism was observed. Concerning probiotic characteristics survival in simulated gastric and intestinal environment were examined. The survival of all tested yeasts in medium of pH 2.5 was comparable and equaled 86.8-97.1% after 8 hours of incubation at 37°C. The fecal isolate, probiotic and collection yeasts showed also high resistance to pH 1.5 and their survival was 85.3-92.1%, whereas for kefir strains it amounted to 33.1 and 38.9%. All yeasts tested demonstrated high resistance to synthetic bile salts as well. In the presence of 0.1 % sodium etiolate and sodium deoxycholate the reduction of cell number by only 1 log unit after 4 hours of incubation at 37°C was observed. However, 1.0% addition of ox bile did not affect their viability. In simulated gastric and intestinal environment survival of fecal, probiotic and collection strains was 86.3-93.7% after 4 hours of incubation in media with addition of 3 g/l pepsin and 1 g/l pancreatin. Kefir isolates were more sensitive to these conditions and a further 10% reduction of cell number in relation to probiotic yeasts was observed. The tested strains, except for kefir isolates, were able to grow at 37°C. All the tested strains survived in sufficient number to create the possibility of proper action in the human body, although fecal, probiotic and collection strains tolerated the conditions of the human gastrointestinal tract better than food-borne yeasts.
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