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Identification of thermo tolerant lactic acid bacteria isolated from silage prepared in the hot and humid climate of Southwestern Japan.

Doi K, Nishizaki Y, Kimura H, Kitahara M, Fujino Y, Ohmomo S, Ohshima T, Ogata S - Springerplus (2013)

Bottom Line: Nearly all of the isolates exhibited high thermotolerance and rapid growth.Using the aforementioned techniques, the isolated strains were identified as L. plantarum, L. rhamonsus, L. rapi, Pediococcus pentosaceus and P. lolii.Our findings also showed that there is greater diversity among thermophilic LABs in silage prepared in a hot and humid environment.

View Article: PubMed Central - PubMed

Affiliation: Microbial Genetic Division, Institute of Genetic Resources, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581 Japan.

ABSTRACT
To develop high-quality silage starters adapted to hot and humid weather, 12 LAB isolates from silage produced in Kyushu and Okinawa, Japan were characterized based on their morphological features, growth curves and sugar utilization. In addition, the nucleotide sequences of the V2-V3 region of their 16S rRNA genes and the 16S-23S rRNA intergenic spacer (ITS) regions were determined. The isolates were also subjected to RAPD-PCR analysis, DNA-DNA hybridization, G+C content analysis and immuno-identification using species-specific monoclonal antibodies and SDS-PAGE profiling. Nearly all of the isolates exhibited high thermotolerance and rapid growth. Combining ITS sequence analysis, RAPD-PCR and immuno-identification enabled rapid and accurate identification of closely related LAB strains that other methods failed to appropriately differentiate; for example, L. plantarum was distinguished from L. pentosus, and L. casei was distinguished from L. rhamonsus. Using the aforementioned techniques, the isolated strains were identified as L. plantarum, L. rhamonsus, L. rapi, Pediococcus pentosaceus and P. lolii. Our findings also showed that there is greater diversity among thermophilic LABs in silage prepared in a hot and humid environment.

No MeSH data available.


Protein profiles of representative LAB isolates and reference strains. The left-hand side of the figure shows the mean correlation coefficients expressed as percentages (r×100) and represented as a dendrogram calculated through UPGMA analysis of the selected strains. The right-hand side of the figure shows the delineation of the 5 clusters.
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Fig5: Protein profiles of representative LAB isolates and reference strains. The left-hand side of the figure shows the mean correlation coefficients expressed as percentages (r×100) and represented as a dendrogram calculated through UPGMA analysis of the selected strains. The right-hand side of the figure shows the delineation of the 5 clusters.

Mentions: Whole-cell protein fingerprinting of the 12 isolates and 5 reference strains yielded distinctly different band patterns (Figure 5), with an intragel reproducibility of approximately 90%. Numerical analysis of the SDS-PAGE patterns obtained with the isolated strains resolved 5 clusters (A-E), emerging at a similarity level of 60%, and confirmed the phenotypic characterization of the isolates. Cluster A includes L. plantarum and L. pentosus, while cluster C includes L. casei and L. rhamnosus.Figure 5


Identification of thermo tolerant lactic acid bacteria isolated from silage prepared in the hot and humid climate of Southwestern Japan.

Doi K, Nishizaki Y, Kimura H, Kitahara M, Fujino Y, Ohmomo S, Ohshima T, Ogata S - Springerplus (2013)

Protein profiles of representative LAB isolates and reference strains. The left-hand side of the figure shows the mean correlation coefficients expressed as percentages (r×100) and represented as a dendrogram calculated through UPGMA analysis of the selected strains. The right-hand side of the figure shows the delineation of the 5 clusters.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3795204&req=5

Fig5: Protein profiles of representative LAB isolates and reference strains. The left-hand side of the figure shows the mean correlation coefficients expressed as percentages (r×100) and represented as a dendrogram calculated through UPGMA analysis of the selected strains. The right-hand side of the figure shows the delineation of the 5 clusters.
Mentions: Whole-cell protein fingerprinting of the 12 isolates and 5 reference strains yielded distinctly different band patterns (Figure 5), with an intragel reproducibility of approximately 90%. Numerical analysis of the SDS-PAGE patterns obtained with the isolated strains resolved 5 clusters (A-E), emerging at a similarity level of 60%, and confirmed the phenotypic characterization of the isolates. Cluster A includes L. plantarum and L. pentosus, while cluster C includes L. casei and L. rhamnosus.Figure 5

Bottom Line: Nearly all of the isolates exhibited high thermotolerance and rapid growth.Using the aforementioned techniques, the isolated strains were identified as L. plantarum, L. rhamonsus, L. rapi, Pediococcus pentosaceus and P. lolii.Our findings also showed that there is greater diversity among thermophilic LABs in silage prepared in a hot and humid environment.

View Article: PubMed Central - PubMed

Affiliation: Microbial Genetic Division, Institute of Genetic Resources, Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581 Japan.

ABSTRACT
To develop high-quality silage starters adapted to hot and humid weather, 12 LAB isolates from silage produced in Kyushu and Okinawa, Japan were characterized based on their morphological features, growth curves and sugar utilization. In addition, the nucleotide sequences of the V2-V3 region of their 16S rRNA genes and the 16S-23S rRNA intergenic spacer (ITS) regions were determined. The isolates were also subjected to RAPD-PCR analysis, DNA-DNA hybridization, G+C content analysis and immuno-identification using species-specific monoclonal antibodies and SDS-PAGE profiling. Nearly all of the isolates exhibited high thermotolerance and rapid growth. Combining ITS sequence analysis, RAPD-PCR and immuno-identification enabled rapid and accurate identification of closely related LAB strains that other methods failed to appropriately differentiate; for example, L. plantarum was distinguished from L. pentosus, and L. casei was distinguished from L. rhamonsus. Using the aforementioned techniques, the isolated strains were identified as L. plantarum, L. rhamonsus, L. rapi, Pediococcus pentosaceus and P. lolii. Our findings also showed that there is greater diversity among thermophilic LABs in silage prepared in a hot and humid environment.

No MeSH data available.