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Strain-specific detection of orally administered canine jejunum-dominated Lactobacillus acidophilus LAB20 in dog faeces by real-time PCR targeted to the novel surface layer protein.

Abstract：

UNLABELLED:Lactobacillus acidophilus LAB20 has potential to be a probiotic strain because it can be present at high numbers in the jejunum of dog. To specifically detect LAB20 from dog faecal samples, a real-time PCR protocol was developed targeting the novel surface (S) layer protein gene of LAB20. The presence of S-layer protein was verified by N-terminal sequencing of the approximately 50-kDa major band from SDS-PAGE gel. The corresponding S-layer gene was amplified by inverse PCR using homology to known S-layers and sequenced. This novel S-layer protein has low sequence similarity to other S-layer proteins in the N-terminal region (32-211 aa, 7-39%). This enabled designing strain-specific PCR primers. The primer set was utilized to study intestinal persistence of LAB20 in dog that was fed with LAB20 fermented milk for 5 days. The results showed that LAB20 can be detected from dog faecal sample after 6 weeks with 10(4·53) DNA copies g(-1) postadministration. It suggested that LAB20 could be a good candidate to study the mechanism behind its persistence and dominance in dog intestine and maybe utilize it as a probiotic for canine. SIGNIFICANCE AND IMPACT OF THE STUDY:A real-time PCR method was developed to detect Lactobacillus acidophilus LAB20, a strain that was previously found dominant in canine gastrointestinal (GI) tract. The quantitative detection was based on targeting to variation region of a novel S-layer protein found in LAB20, allowing to specifically enumerate LAB20 from dog faeces. The results showed that the real-time PCR method was sensitive enough to be used in later intervention studies. Interestingly, LAB20 was found to persist in dog GI tract for 6 weeks. Therefore, LAB20 could be a good candidate to study its colonization and potentially utilize as a canine probiotic.

journal_name

Lett Appl Microbiol

journal_title

Letters in applied microbiology

authors

Tang Y,Saris PE

doi

10.1111/lam.12117

subject

Has Abstract

pub_date

2013-10-01 00:00:00

pages

330-5

issue

eissn

0266-8254

issn

1472-765X

journal_volume

pub_type

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Strain-specific detection of orally administered canine jejunum-dominated Lactobacillus acidophilus LAB20 in dog faeces by real-time PCR targeted to the novel surface layer protein.