The use of a quantitative real-time polymerase chain reaction assay for identification and enumeration of Lactobacillus buchneri in silage
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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Schmidt et al. (2008) and Mari et al. (2008) reported greater LAB numbers in corn silages treated with L. buchneri and confirmed greater L. buchneri numbers by quantitative real-time PCR.
“Schmidt et al. (2008) and Mari et al. (2008) also reported greater numbers of LAB in corn silages treated with L. buchneri than in untreated corn silage and further confirmed that treated silages had markedly greater numbers of L. buchneri (as determined by quantitative real-time PCR).”
From The effect of Lactobacillus buchneri 40788 or Lactobacillus plantarum MTD-1 on the fermentation and aerobic stability of corn silages ensiled at two dry matter contents (2009) · cites this paper as Schmidt et al., 2008
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- Authors
- Schmidt, R. J., Emara, M. G., & Kung Jr, L.
- Affiliations
- University of Delaware (US)
- Venue
- Journal of Applied Microbiology
- Publisher
- Oxford University Press (OUP)
- 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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