Development of a strain-specific assay for detection of viable Lactobacillus sp. HOFG1 after application to cattle feed.
A strain-specific assay was developed for the detection of viable Lactobacillus on cattle feed. The DNA sequences of the 16S rRNA gene and four different 16S/23S rRNA intergenic spacer regions (ISR) from Lactobacillus sp. HOFG1 were determined. Based on these sequences, a strain-specific primer was designed for the amplification of one of the ISRs. When combined with a Lactobacillus group primer, the polymerase chain reaction (PCR) assay detected only Lactobacillus sp. HOFG1 and not other closely related L. animalis or L. murinus strains. The feed assay uses a combination of enrichment culturing and PCR to detect and enumerate viable Lactobacillus sp. HOFG1 after its application onto cattle feed. The high degree of primer specificity and use of selective culturing allows for the detection of viable Lactobacillus which is useful in tracking bacteria applied to complex feed mixtures that contain a high background of endogenous bacteria.
Flint
Joseph F
JF
Department of Microbiology, Cornell University, Wing Hall Ithaca, NY 14853, United States.
Angert
Esther R
ER
D000821:Animal Feed / Q000382:microbiology*; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D015169:Colony Count, Microbial / Q000662:veterinary; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field / Q000662:veterinary; D007778:Lactobacillus / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D019936:Probiotics / Q000032:analysis*; D016415:Sequence Alignment
D017931:DNA Primers; D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer