Cited by 31Probiotics and Fermented FoodsRuminant Nutrition and Digestive PhysiologyPlant-Microbe Interactions and Immunity

The use of a quantitative real-time polymerase chain reaction assay for identification and enumeration of Lactobacillus buchneri in silage

Schmidt, R.J., Emara, M.G., Kung Jr, L.·2008·Journal of Applied Microbiology

Abstract

AIMS: To detect and quantify Lactobacillus buchneri in plant samples with the aid of polymerase chain reaction (PCR) methods. METHODS AND RESULTS: DNA from silage samples spiked with different amounts of L. buchneri cells was isolated using a lysozyme/sodium dodecyl sulfate lysis and phenol/chloroform extraction method. The DNA served as a template for PCR amplification with primers specific for the bacterium. The primers were developed by comparison of 16S rDNA sequences from different lactic acid bacteria (LAB) and testing for specificity with 11 different strains of LAB. As few as 100 L. buchneri colony-forming units per gram of silage could be detected. Additionally, the technique was successfully applied to quantify the population of L. buchneri in two cultivars of corn with or without inoculation. CONCLUSIONS: The PCR assay provided a specific and rapid tool for identifying and enumerating L. buchneri in silage samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of microbial inoculants for silage production is a safe and environment friendly practice, but the full potential of such additives can only be achieved with a better understanding of the fate and activity of the microbes involved. The current study describes a methodology to detect and enumerate L. buchneri, a micro-organism used as an inoculant.

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Crossref · 10.1111/j.1365-2672.2008.03834.x

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Authors
Schmidt, R. J., Emara, M. G., & Kung Jr, L.
Affiliations
University of Delaware (US)
Venue
Journal of Applied Microbiology
Published
2008-09
Volume
105
Issue
3
Pages
920-929
ISSN
  • 1364-5072 (ISSN-L)
  • 1365-2672
OpenAlex
W2042414072
Cited by
31
References
38
Citations by year
2026: 1 · 2025: 2 · 2024: 1 · 2022: 3 · 2021: 1 · 2020: 2 · 2018: 3 · 2017: 2 · 2016: 2 · 2015: 3 · 2014: 1 · 2013: 2 · 2012: 1
FWCI
3.74
Type
journal article
Language
en
Open access
closed
Locations
  • Journal of Applied Microbiology (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Agricultural and Biological Sciences › Food Science › Probiotics and Fermented Foods
  • Life Sciences › Agricultural and Biological Sciences › Agronomy and Crop Science › Ruminant Nutrition and Digestive Physiology
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant-Microbe Interactions and Immunity
Keywords
Lactobacillus buchneri, silage, lactic acid bacteria, bacterial enumeration, PCR detection, silage inoculants, corn silage, microbial DNA extraction
MeSH terms
Animal Feed, Zea mays, DNA, Bacterial, Food Microbiology, Lactobacillus, Silage, Colony Count, Microbial, DNA Primers, Reverse Transcriptase Polymerase Chain Reaction

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