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Rapid genus identification of selected lactic acid bacteria isolated from Mugil cephalis and Oreochromis niloticus organs using MALDI-TOF

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Abstract

Lactic acid bacteria (LAB) are traditionally used in food and feed bio-industries as they have health-promoting probiotic effect for their host and currently considered safe for human and animal consumption. Recently, we isolated 177 strains from freshwater fishes of dams of Tunisia exhibiting antimicrobial activities against various food-borne pathogens and generated a laboratory biobank to be characterized. Herein, we investigated whether MALDI-TOF could assist rapid identification of LAB genus. First, a lactic microflora-selective Man Rogosa Sharpe medium was used. Isolates were further screened according to the antimicrobial activity, using well-difusion agar. In total, four major genera have been well identified and species frequency has been estimated (74% Enterococcus, 24% Leuconostoc, 3% Lactococcus, and 2% Vagococcus). Eighteen isolates was further analyzed using MALDI-TOF. Comparative analysis of spectral fingerprints to six referenced MALDI-TOF fingerprints was carried out from the recently developed USC library (www.spectrabank.org). The intra- and inter-specific phyloproteomic relationships among strains were compared to available phylogenetic data based on 16S rDNA genes. This study showed that MALDI-TOF MS is a potent and reliable rapid method for both discrimination and identification of LABs. This highlights the insight of proteomic approach into the screening of food-derived beneficial microorganisms.

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Acknowledgments

The authors would like to thank Dr. Mohamed El Ayeb, Director of Laboratoire des Venins et Molécules Thérapeutiques, Institut Pasteur de Tunis for his constant encouragements. Special thanks are addressed to Mr. Marcos Quintela Baluja (LHICA) for his technical assistance.

Funding

This work was initially co-funded by a grant from the Tunisia-Spain bilateral AECI PROJECT A1/038311/11 and secondly supported by the project BIOVecQ P.S.1.3/08 from the National Institute of Marine Sciences and Technology (INSTM) and the Pasteur Institute of Tunis (IPT), Ministry of Higher Education and Scientific Research and Technology of Information and Communication, Tunisia.

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Authors and Affiliations

  1. Laboratory of Microbiology and Pathology of Aquatic Organisms, Institut National des Sciences et Technologies de la Mer (INSTM), rue 2 mars 1934, 2025, Salammbô, Tunisia

    Rim El-Jeni & Monia El Bour

  2. Laboratory of Venoms and Therapeutic Molecules, Pasteur Institute of Tunisia/University of Tunis El Manar, 13 Place Pasteur, BP-74, 1002, Tunis, Tunisia

    Rim El-Jeni & Balkiss Bouhaouala-Zahar

  3. Laboratory of Food Technology, LHICA, Department of Analytical Chemistry, Nutrition and Food Science, University of Santiago de Compostela, Lugo, Spain

    Karola Böhme, Pilar Calo-Mata & Jorge Barros-Velázquez

  4. Biomedical and oncogenetic genomics laboratory, Pasteur Institute of Tunisia/University of Tunis El Manar, 13 Place Pasteur, BP-74, 1002, Tunis, Tunisia

    Rym Kefi

  5. Medical School of Tunis, University of Tunis El Manar, 15 Rue Djebel Lakhdhar. La Rabta, 1007, Tunis, Tunisia

    Balkiss Bouhaouala-Zahar

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  1. Rim El-Jeni
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  2. Karola Böhme
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  3. Monia El Bour
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  4. Pilar Calo-Mata
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Correspondence to Balkiss Bouhaouala-Zahar.

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El-Jeni, R., Böhme, K., El Bour, M. et al. Rapid genus identification of selected lactic acid bacteria isolated from Mugil cephalis and Oreochromis niloticus organs using MALDI-TOF. Ann Microbiol 69, 1–15 (2019). https://doi.org/10.1007/s13213-018-1357-8

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