Multiplex PCR identification and culture-independent quantification of Bacillus licheniformis by qPCR using specific DNA markers

• Type: Article in International Scientific Journal
• Date: 2018

Authors

Almeida, E (Author) Other; Claudia Alexandra dos Reis Serra (Author) FCUP; Albuquerque, P (Author) Other; Guerreiro, I (Author) Other; Oliva Teles, A (Author) FCUP; Enes, P (Author) FCUP; Fernando Tavares (Author) FCUP

Journal

Title: Food Microbiology

Vol. 74

Pages: 1-10

ISSN: 0740-0020

Publisher: Elsevier

Indexing

ISI Web of Knowledge - 4 Citations

Scopus - 4 Citations

Other information

Authenticus ID: P-00N-KJY

DOI: 10.1016/j.fm.2018.02.016

Abstract (EN): Probiotics benefits in fish farming have been usually inferred appraising the effects observed on the host and not through the direct assessment of probiotic dynamics in the host gut microbiota. To overcome this gap, quantitative PCR (qPCR) can be a powerful approach to study the bacterial dynamics in fish gut microbiota. The presented work proposes four B. licheniformis-specific DNA markers and details a qPCR method to track putative probiotics B. licheniformis on fish gut. The four B. licheniformis-specific DNA markers - BL5B (hypothetical protein BL00303), BL8A (serA2), BL13C (rfaB) and BL18A (ligD) - were selected and validated by PCR and multiplex-PCR with 20 B. licheniformis isolates and a broad range of non-target bacteria. To assess the dynamics of B. licheniformis in the digesta of farmed fish, a qPCR was validated using markers BL8A and BL18A and calibration curves obtained for both markers with digesta samples spiked with B. licheniformis cells showed a high correlation (R-2 > 0.99) over 6 log units (CFU/ reaction), and a limit of detection (LOD) as low as 247 CFUs/reaction. Furthermore, the consistent qPCR repeatability and reproducibility underline the specificity and reliability of the qPCR proposed. Ultimately, the possibility to monitor the dynamics of B. licheniformis probiotics in the gut microbiota of farmed fish might be instrumental to optimize best practices in aquaculture.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 10