Genotypic and phenotypic characterization of lactic acid bacteria isolated from Italian ryegrass silage

Abstract

Twenty-three lactic acid bacteria (LAB) isolated from three cultivars (Akiaoba, Nagahahikari and Tachiwase) of Italian ryegrass (Lolium multiflorum Lam.) silage were precisely characterized by a combination of phenotypic tests, genotypic 16S ribosomal DNA sequencing and rapid PCR-based analyses, focusing on their useful phenotypes for silage preparation as inoculants. We successfully identified both at the species and subspecies levels: phenotypically novel Lactococcus lactis subsp. lactis, Lactobacillus brevis, Lactobacillus coryniformis subsp. torquens, Lactobacillus curvatus, Lactobacillus plantarum subsp. plantarum, Lactobacillus sakei subsp. carnosus, Leuconostoc mesenteroides subsp. dextranicum and Pediococcus parvulus. This is the first report to elucidate the presence of Lactobacillus coryniformis ssp. torquens and Leuconostoc mesenteroides subsp. dextranicum in Italian ryegrass silages. Physiological and biochemical tests revealed that phenotypic characteristics are different among the different strains of the same species and subspecies, and that the isolates show unique and diverse phenotypes related to fermentation factors, such as available carbohydrates, optimal growth pH and temperature. These results suggest that, for various well-preserved silage preparations, the isolates may be useful in producing novel inoculants corresponding to their optimally climatic and ecological niches.

Identification and characterization of lactic acid bacteria isolated from mixed pasture of timothy and orchardgrass, and its badly preserved silage

“…1999a). We also conducted a rapid‐PCR method without DNA preparation to amplify the 16S rDNA (Tohno et al . 2011).…”

Update of the list of QPS‐recommended biological agents intentionally added to food or feed as notified to EFSA 4: suitability of taxonomic units notified to EFSA until March 2016

“…In addition, some strains are histamine producers (Landete et al, 2005). P. parvulus is, however, frequently found in fermented foods (Mesas et al, 2011;Abriouel et al, 2012;Ba gder Elmacı et al, 2015) and feed, such as silage (Tohno et al, 2012), and thus there is frequent exposure to it without any signs of pathogenicity for humans or animals. Similarly, it appears that EPS-producing strains are less susceptible to simulated and mice digestive conditions, while lowering the cholesterol levels (Immerstrand et al, 2010;Lindstr € om et al, 2013) reason why they have been proposed as potential probiotics but they were never used as such nor intentionally added to any food.…”

Paenibacillus silagei sp. nov. isolated from corn silage

“…The samples were transferred into sterile homogenisation bags, and silage homogenates were prepared as previously described (Tohno et al, 2012). Strains were isolated from the homogenates (pH 3.7) after incubation on M17 broth (Difco) agar plates for 14 days at 4 C under anaerobic For the initial taxonomic classification of LOOC204 T , the 16S rRNA gene sequence was determined according to methods described previously (Tohno et al, 2014(Tohno et al, , 2015.…”