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Identification and adhesion profile of Lactobacillus spp. strains isolated from poultry.

ABSTRACT: In the aviculture industry, the use of Lactobacillus spp. as a probiotic has been shown to be frequent and satisfactory, both in improving bird production indexes and in protecting intestine against colonization by pathogenic bacteria. Adhesion is an important characteristic in selecting Lactobacillus probiotic strains since it impedes its immediate elimination to enable its beneficial action in the host. This study aimed to isolate, identify and characterize the in vitro and in vivo adhesion of Lactobacillus strains isolated from birds. The Lactobacillus spp. was identified by PCR and sequencing and the strains and its adhesion evaluated in vitro via BMM cell matrix and in vivo by inoculation in one-day-old birds. Duodenum, jejunum, ileum and cecum were collected one, four, 12 and 24 h after inoculation. The findings demonstrate greater adhesion of strains in the cecum and an important correlation between in vitro and in vivo results. It was concluded that BMM utilization represents an important technique for triage of Lactobacillus for subsequent in vivo evaluation, which was shown to be efficient in identifying bacterial adhesion to the enteric tract.

SUBMITTER: Rocha TS

PROVIDER: S-EPMC4204948 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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Identification and adhesion profile of Lactobacillus spp. strains isolated from poultry.

Rocha Ticiana Silva TS Baptista Ana Angelita Sampaio AA Donato Tais Cremasco TC Milbradt Elisane Lenita EL Okamoto Adriano Sakai AS Andreatti Filho Raphael Lucio RL

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology] 20141009 3

In the aviculture industry, the use of Lactobacillus spp. as a probiotic has been shown to be frequent and satisfactory, both in improving bird production indexes and in protecting intestine against colonization by pathogenic bacteria. Adhesion is an important characteristic in selecting Lactobacillus probiotic strains since it impedes its immediate elimination to enable its beneficial action in the host. This study aimed to isolate, identify and characterize the in vitro and in vivo adhesion of ...[more]

PMID: 25477944

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Project description:The objective of our study is to evaluate the potential use of Amplified 16S Ribosomal DNA Restriction Analysis (16S-ARDRA) and MALDI-TOF mass spectrometry (MS) as methods for species identification of Lactobacillus strains in poultry.A total of 80 Lactobacillus strains isolated from the cloaca of chicken, geese and turkeys were identified to the species level by MALDI-TOF MS (on-plate extraction method) and 16S-ARDRA. The two techniques produced comparable classification results, some of which were additionally confirmed by sequencing of 16S rDNA. MALDI-TOF MS enabled rapid species identification but produced more than one reliable identification result for 16.25 % of examined strains (mainly of the species L. johnsonii). For 30 % of isolates intermediate log(scores) of 1.70-1.99 were obtained, indicating correct genus identification but only presumptive species identification. The 16S-ARDRA protocol was based on digestion of 16S rDNA with the restriction enzymes MseI, HinfI, MboI and AluI. This technique was able to distinguish 17 of the 19 Lactobacillus reference species tested and enabled identification of all 80 wild isolates. L. salivarius dominated among the 15 recognized species, followed by L. johnsonii and L. ingluviei.The MALDI-TOF MS and 16S-ARDRA assays are valuable tools for the identification of avian lactobacilli to the species level. MALDI-TOF MS is a fast, simple and cost-effective technique, and despite generating a high percentage of results with a log(score) <2.00, the on-plate extraction method is characterized by high-performance. For samples for which Biotyper produces more than one reliable result, MALDI-TOF MS must be used in combination with genotypic techniques to achieve unambiguous results. 16S-ARDRA is simple, repetitive method with high power of discrimination, whose sole limitation is its inability to discriminate between species with very high 16S rDNA sequence homology, such as L. casei and L. zeae. The assays can be used for discrimination of Lactobacillus bacteria from different habitats.

Project description:Probiotics are considered an alternative to antibiotics in the prevention and treatment of Salmonella diseases in poultry. However, to use probiotics as proposed above, it is necessary to evaluate their properties in detail and to select the most effective bacterial strains in the application targeted. In this study, probiotic properties of new Lactobacillus sp. strains were investigated and their antimicrobial activity against 125 environmental strains of Salmonella sp. was determined using the agar slab method. Furthermore, their survival in the presence of bile salts and at low pH, antibiotics susceptibility, aggregation and coaggregation ability, adherence to polystyrene and Caco-2 cells, and cytotoxicity were investigated. Each strain tested showed antagonistic activity against at least 96% of the environmental Salmonella sp. strains and thus representing a highly epidemiologically differentiated collection of poultry isolates. In addition, the probiotic properties of new Lactobacillus strains are promising. Therefore, all strains examined showed a high potential for use in poultry against salmonellosis.Probiotics are considered an alternative to antibiotics in the prevention and treatment of Salmonella diseases in poultry. However, to use probiotics as proposed above, it is necessary to evaluate their properties in detail and to select the most effective bacterial strains in the application targeted. In this study, probiotic properties of new Lactobacillus sp. strains were investigated and their antimicrobial activity against 125 environmental strains of Salmonella sp. was determined using the agar slab method. Furthermore, their survival in the presence of bile salts and at low pH, antibiotics susceptibility, aggregation and coaggregation ability, adherence to polystyrene and Caco-2 cells, and cytotoxicity were investigated. Each strain tested showed antagonistic activity against at least 96% of the environmental Salmonella sp. strains and thus representing a highly epidemiologically differentiated collection of poultry isolates. In addition, the probiotic properties of new Lactobacillus strains are promising. Therefore, all strains examined showed a high potential for use in poultry against salmonellosis.

Project description:The objective of this study was to evaluate the functional properties of lactic acid bacteria (LAB) isolated from Tibetan kefir grains. Three Lactobacillus isolates identified as Lactobacillus acidophilus LA15, Lactobacillus plantarum B23 and Lactobacillus kefiri D17 that showed resistance to acid and bile salts were selected for further evaluation of their probiotic properties. The 3 selected strains expressed high in vitro adherence to Caco-2 cells. They were sensitive to gentamicin, erythromycin and chloramphenicol and resistant to vancomycin with MIC values of 26 µg/ml. All 3 strains showed potential bile salt hydrolase (BSH) activity, cholesterol assimilation and cholesterol co-precipitation ability. Additionally, the potential effect of these strains on plasma cholesterol levels was evaluated in Sprague-Dawley (SD) rats. Rats in 4 treatment groups were fed the following experimental diets for 4 weeks: a high-cholesterol diet, a high-cholesterol diet plus LA15, a high-cholesterol diet plus B23 or a high-cholesterol diet plus D17. The total cholesterol, triglyceride and low-density lipoprotein cholesterol levels in the serum were significantly (P<0.05) decreased in the LAB-treated rats compared with rats fed a high-cholesterol diet without LAB supplementation. The high-density lipoprotein cholesterol levels in groups B23 and D17 were significantly (P<0.05) higher than those in the control and LA15 groups. Additionally, both fecal cholesterol and bile acid levels were significantly (P<0.05) increased after LAB administration. Fecal lactobacilli counts were significantly (P<0.05) higher in the LAB treatment groups than in the control groups. Furthermore, the 3 strains were detected in the rat small intestine, colon and feces during the feeding trial. The bacteria levels remained high even after the LAB administration had been stopped for 2 weeks. These results suggest that these strains may be used in the future as probiotic starter cultures for manufacturing novel fermented foods.

Dataset Information

Identification and adhesion profile of Lactobacillus spp. strains isolated from poultry.

Publications

Identification and adhesion profile of Lactobacillus spp. strains isolated from poultry.

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