Characterization of homofermentative lactobacilli isolated from kefir grains: potential use as probiotic

2008 ◽  
Vol 75 (2) ◽  
pp. 211-217 ◽  
Author(s):  
Marina A Golowczyc ◽  
Maria J Gugliada ◽  
Axel Hollmann ◽  
Lucrecia Delfederico ◽  
Graciela L Garrote ◽  
...  
Keyword(s):  
Restriction Analysis ◽  
Bacterial Species ◽  
Genotypic Diversity ◽  
23S Rrna ◽  
Pcr Analysis ◽  
High Survival ◽  
Probiotic Properties ◽  
Probiotic Potential ◽  
Rapd Pcr ◽  
Kefir Grains

Considering that several health promoting properties are associated with kefir consumption and a reliable probiotic product requires a complete identification of the bacterial species, the present work evaluates several proved markers of probiotic potential of eleven isolates of homofermentative lactobacilli isolated from kefir grains and molecular identification and genotypic diversity. Using restriction analysis of amplified ribosomal DNA (ARDRA) and analysis of the 16S–23S rRNA internal spacer region we confirmed that all homofermentative lactobacilli belong to the speciesLactobacillus plantarum. RAPD-PCR analysis allowed the discrimination of lactobacilli in five clusters. All isolates exhibited high resistance to bile salt. High survival after one hour of exposure to pH 2·5 was observed inLb. plantarumCIDCA 8313, 83210, 8327 and 8338. All isolates were hydrophilic and non autoaggregative. Isolate CIDCA 8337 showed the highest percentage of adhesion among strains. All tested lactobacilli had strong inhibitory power againstSalmonella typhimuriumandEscherichia coli. Seven out of eleven isolates showed inhibition againstSal. entericaand five isolates were effective againstSal. gallinarum. Only CIDCA 8323 and CIDCA 8327 were able to inhibitSal. sonnei. We did not find any correlation between the five clusters based on RAPD-PCR and the probiotic properties, suggesting that these isolates have unique characteristics.

Molecular identification and typing of lactobacilli isolated from kefir grains

2005 ◽  
Vol 73 (1) ◽  
pp. 20-27 ◽  
Author(s):  
Lucrecia Delfederico ◽  
Axel Hollmann ◽  
Mariano Martínez ◽  
N. Gabriel Iglesias ◽  
Graciela De Antoni ◽  
...  
Keyword(s):  
Restriction Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Restriction Enzymes ◽  
23S Rrna ◽  
Pcr Analysis ◽  
Rrna Gene ◽  
Bacterial Isolates ◽  
Good Tool ◽  
Rapd Pcr ◽  
Kefir Grains

Seventeen heterofermentative lactobacilli isolated from kefir grains were characterized by molecular methods. Bacterial isolates were identified by amplification of 16S rRNA gene and analysis by Amplified Ribosomal DNA Restriction Analysis (ARDRA), using the restriction enzymes Hae III, Dde I, and Hinf I. ARDRA analysis of lactobacilli isolates showed, for each enzyme used, a same banding pattern between the heterofermentative lactobacilli and the reference strains Lactobacillus kefir JCM 5818 and Lb. kefir ATCC 8007. Other reference lactobacilli and one homofermentative isolate showed differences in at least one of these patterns. The 16S-23S rRNA spacer region was also used to discriminate the bacterial isolates at the species level. The data obtained from the analysis of spacer region confirmed that sequencing of this genome region is a good tool for a reliable identification of members of Lb. kefir species. Genotyping of isolates was performed by Random Amplified Polymorphic DNA (RAPD-PCR) analysis using M13, Coc, ERIC-2 and 1254 primers. Patterns obtained allowed the differentiation of isolates in three groups. The three clusters showed by RAPD-PCR analysis could be correlated with at least three different strains of Lb. kefir species in the group of heterofermentative lactobacilli isolates obtained from Argentinian kefir grains.

scholarly journals Application of A Novel Potential Probiotic Lactobacillus paracasei Strain Isolated from Kefir Grains in the Production of Feta-Type Cheese

2018 ◽  
Vol 6 (4) ◽  
pp. 121 ◽  
Author(s):  
Ioanna Mantzourani ◽  
Antonia Terpou ◽  
Athanasios Alexopoulos ◽  
Pelagia Chondrou ◽  
Alex Galanis ◽  
...  
Keyword(s):  
Starter Culture ◽  
Quality Characteristics ◽  
Lactobacillus Paracasei ◽  
Pcr Analysis ◽  
In Vitro Tests ◽  
Probiotic Properties ◽  
The Novel ◽  
Kefir Grains ◽  
Cheese Production

In the present study 38 lactic acid bacteria strains were isolated from kefir grains and were monitored regarding probiotic properties in a series of established in vitro tests, including resistance to low pH, resistance to pepsin and pancreatin, and tolerance to bile salts, as well as susceptibility against common antibiotics. Among them, the strain SP3 displayed potential probiotic properties. Multiplex PCR analysis indicated that the novel strain belongs to the paracasei species. Likewise, the novel strain (Lactobacillus paracasei SP3) was applied as a starter culture for Feta-type cheese production. Feta-type cheese production resulted in significantly higher acidity; lower pH; reduced counts of coliforms, yeasts and fungi; and improved quality characteristics compared with cheese samples produced with no starter culture. Finally, it is highlighted that the application of the novel strain led to Feta-type cheese production with improved overall quality and sensory characteristics.

scholarly journals Genetic Diversity of Pierce's Disease Strains and Other Pathotypes of Xylella fastidiosa

2001 ◽  
Vol 67 (2) ◽  
pp. 895-903 ◽  
Author(s):  
Mavis Hendson ◽  
Alexander H. Purcell ◽  
Deqiao Chen ◽  
Chris Smart ◽  
Magalie Guilhabert ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Xylella Fastidiosa ◽  
Random Amplified Polymorphic Dna ◽  
Consensus Sequence ◽  
23S Rrna ◽  
Pierce's Disease ◽  
Pcr Analysis ◽  
Leaf Scorch ◽  
Pierce’S Disease ◽  
Rapd Pcr

ABSTRACT Strains of Xylella fastidiosa isolated from grape, almond, maple, and oleander were characterized by enterobacterial repetitive intergenic consensus sequence-, repetitive extragenic palindromic element (REP)-, and random amplified polymorphic DNA (RAPD)-PCR; contour-clamped homogeneous electric field (CHEF) gel electrophoresis; plasmid content; and sequencing of the 16S-23S rRNA spacer region. Combining methods gave greater resolution of strain groupings than any single method. Strains isolated from grape with Pierce's disease (PD) from California, Florida, and Georgia showed greater than previously reported genetic variability, including plasmid contents, but formed a cluster based on analysis of RAPD-PCR products,NotI and SpeI genomic DNA fingerprints, and 16S-23S rRNA spacer region sequence. Two groupings of almond leaf scorch (ALS) strains were distinguished by RAPD-PCR and CHEF gel electrophoresis, but some ALS isolates were clustered within the PD group. RAPD-PCR, CHEF gel electrophoresis, and 16S-23S rRNA sequence analysis produced the same groupings of strains, with RAPD-PCR resolving the greatest genetic differences. Oleander strains, phony peach disease (PP), and oak leaf scorch (OLS) strains were distinct from other strains. DNA profiles constructed by REP-PCR analysis were the same or very similar among all grape strains and most almond strains but different among some almond strains and all other strains tested. Eight of 12 ALS strains and 4 of 14 PD strains of X. fastidiosa isolated in California contained plasmids. All oleander strains carried the same-sized plasmid; all OLS strains carried the same-sized plasmid. A plum leaf scald strain contained three plasmids, two of which were the same sizes as those found in PP strains. These findings support a division of X. fastidiosaat the subspecies or pathovar level.

Genetic Diversity of the Carrion and Jungle Crows as Evidenced by RAPD–PCR Analysis

2003 ◽  
Vol 39 (11) ◽  
pp. 1281-1291 ◽  
Author(s):  
L. N. Spiridonova ◽  
G. N. Chelomina ◽  
A. P. Kryukov
Keyword(s):  
Genetic Diversity ◽  
Pcr Analysis ◽  
Rapd Pcr

scholarly journals First Report of Xylella fastidiosa in Peach in New Mexico

Plant Disease ◽  
2011 ◽  
Vol 95 (7) ◽  
pp. 871-871 ◽  
Author(s):  
J. J. Randall ◽  
J. French ◽  
S. Yao ◽  
S. F. Hanson ◽  
N. P. Goldberg
Keyword(s):  
New Mexico ◽  
Prunus Persica ◽  
Xylella Fastidiosa ◽  
Its Region ◽  
23S Rrna ◽  
Pcr Analysis ◽  
Sequencing Analysis ◽  
First Report ◽  
Dark Green ◽  
Branch Dieback

Xylella fastidiosa is a gram-negative bacterium that causes disease in a wide variety of plants such as grapes, citrus trees, oleanders, and elm and coffee trees. This bacterium is xylem limited and causes disease symptoms such as leaf scorch, stunting of plant growth, branch dieback, and fruit loss. The presence of X. fastidiosa was previously reported in New Mexico where it was found to be infecting chitalpa plants and grapevines (3). In the summer of 2010, peach (Prunus persica (L.) Batsch) trees from two locations in northern New Mexico exhibited leaf deformity and stunting, dark green venation, slight mottling, and branch dieback. Preliminary viral diagnostic screening was performed by Agdia (Elkhart, IN) on one symptomatic tree and it was negative for all viruses tested. Three trees from two different orchards tested positive for X. fastidiosa by ELISA and PCR analysis using X. fastidiosa-specific primer sets HL (1) and RST (2). Bacterial colonies were also cultured from these samples onto periwinkle wilt media. Eight colonies obtained from these three plants tested PCR positive using the X. fastidiosa-specific primers. The 16S ribosomal and 16S-23S rRNA internal transcribed spacer (ITS) region (557 nucleotides) (GenBank Accession No. HQ292776) along with the gyrase region (400 nucleotides) (GenBank Accession No. HQ292777) was amplified from the peach total DNA samples and the bacterial colonies. Sequencing analysis of these regions indicate that the X. fastidiosa found in peach is 100% similar to other X. fastidiosa multiplex isolates including isolates from peach, pecan, sycamore, and plum trees and 99% similar to the X. fastidiosa isolates previously found in New Mexico. Further analysis of the 16S ribosomal and 16S-23S rRNA ITS sequences with maximum likelihood phylogenetic analysis using Paup also groups the peach isolates into the X. fastidiosa multiplex subspecies. The gyrase sequence could not be used to differentiate the peach isolates into a subspecies grouping because of the lack of variability within the sequence. This X. fastidiosa multiplex subspecies could possibly be a threat to the New Mexico pecan industry since pecan infecting X. fastidiosa isolates belong to the same bacterial subspecies. It is not known if X. fastidiosa subspecies multiplex isolates from peach are capable of infecting pecans but they are closely genetically related. It is interesting to note that the isolates from peach are different than previously described X. fastidiosa isolates in New Mexico that were infecting chitalpa and grapes (3). X. fastidiosa has previously been described in peach; the disease is called “phony peach”. The peach trees exhibited stunting and shortened internodes as reported for “phony peach”. They also exhibited slight mottling and branch dieback that may be due to the environment in New Mexico or perhaps they are also exhibiting mineral deficiency symptoms in association with the X. fastidiosa disease. To our knowledge, this is the first report of X. fastidiosa in peach in New Mexico. References: (1) M. H. Francis et al. Eur. J. Plant Pathol. 115:203, 2006. (2) G. V. Minsavage et al. Phytopathology 84:456, 1994. (3) J. J. Randall et al. Appl. Environ. Microbiol. 75:5631, 2009.

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