Characterization of <i>Carnobacterium</i> species by pyrolysis mass spectrometry

-During the last fifteen years, the Lactobacillus genus has evolved and contains to date more than 80 species. They are present in raw milk and dairy products such as cheeses, yoghurts and fermented milks. Quality assurance programmes associated with research, development, production and validation of the health or technological benefits of these bacteria require their relevant isolation, counting and identification. This review presents the different selective media to isolate lactobacilli, and the numerous different available tools to characterise lactobacilli at genus, species or strain level using either culture-dependent methods: phenotypical, molecular or global methods, or using new culture-independent advanced molecular methods. Enzymes used for PFGE, hybridisation probes and PCR-based method primers are listed in seven tables. In conclusion, the main advantages and disadvantages associated with these techniques are presented.Lactobacillus / media / PFGE restriction enzyme / probe / primer / cheese / dairy product Résumé -Isolement, caractérisation et identification des lactobacilles des produits laitiers. Durant les quinze dernières années, le genre Lactobacillus a subi de nombreux remaniements et compte actuellement plus de 80 espèces. Les lactobacilles sont présents dans le lait cru, les produits laitiers tels que les fromages, les yaourts et les laits fermentés. Pour s'inscrire dans une démarche qualité visant au développement de lactobacilles à effet santé ou d'intérêt technologique, l'isolement, le comptage et la caractérisation parfaite de ces bactéries sont nécessaires. Cette analyse bibliographique présente les différents milieux sélectifs pour isoler les lactobacilles, ainsi que les outils disponibles à ce jour pour caractériser les lactobacilles au niveau du genre, de l'espèce ou de la souche, aussi bien par des méthodes culture dépendantes : phénotypique, moléculaire, globale, que par les nouvelles méthodes se réalisant à partir d'échantillons bruts. Les enzymes utilisées en PFGE, les sondes d'hybridation et les oligonucléotides utilisés pour les différentes techniques de PCR sont répertoriés en sept tableaux. En conclusion, les avantages et inconvénients de ces techniques sont présentés.Lactobacillus / milieu sélectif / enzyme PFGE / sonde moléculaire / amorce PCR / fromage / produit laitier

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“…Another global technique, pyrolysismass spectrometry [125], is also very promising but has not yet been applied to lactobacilli.…”

Section: Global Techniquesmentioning

confidence: 99%

Article

Coeuret

Dubernet

Bernardeau

et al. 2003

Lait

153

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“…In addition, duplicated strains were recovered in the same cluster highlighting the good reproducibility of FT-IR. It is encouraging to note that some of the clusters correspond to groupings recognised in the numerical phenetic work of Lai and Manchester [13] and the PyMS work of Manchester et al [18]. All of the strains in cluster 2, except C. piscicola 1230 and 213, and all the strains in cluster 4 were recovered into the cluster-group which was defined as the species C. piscicola Collins et al 1987 in the numerical phenetic study of Lai and Manchester [13].…”

Section: Resultsmentioning

confidence: 72%

“…Cluster 5 contained strains (NCFB 2763 T , 327, 185, 506, 544, ANP7C33, MT4) which were assigned to a corresponding pyrogroup in the work of Mancehster et al [18], as did some of the strains recovered into cluster 3 (C. divergens 554, MT49, MT51, MT53). There is little correlation between the findings from DNA:DNA relatedness data [3,22].…”

Section: Resultsmentioning

confidence: 99%

“…Methods used in the classification of carnobacteria include physiological and biochemical tests [3,5,13], 16S rDNA sequence analysis [30], nucleic acid hybridisation [2,14,22], and pyrolysis mass spectrometry [18]. However, although these methods are helping to provide a clearer picture of carnobacterial systematics not all are suitable for use in large scale studies as they can be very labour intensive and time consuming.…”

Section: Introductionmentioning

confidence: 99%

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Whole-organism Fingerprinting of the Genus Carnobacterium using Fourier Transform Infrared Spectroscopy (FT-IR)

Lai

Goodacre

Manchester

2004

Systematic and Applied Microbiology

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In the current study, 67 Carnobacterium and related strains were analysed by FT-IR to evaluate the classification of the strains and to determine the potential of FT-IR with multivariate statistical analyses for Carnobacterium systematics. Materials and Methods Strains and cultivationSixty seven strains (Table 1) were subcultured twice on casein-peptone soymeal-peptone agar (CASO broth (Merck), 1.2% (w/v) Lab M No. 1 agar) for 3 days at 25 °C. Ten strains were examined in duplicate to assess the reproducibility of the system. Effect of culture ageType strains of C. divergens (NCFB 2763 T ), C. gallinarum (MT44 T ), C. mobile (NFCB 2765 T ) and C. piscicola (NCFB 2762 T ) were cultivated on CASO agar for 2, 3, 4 and 5 days at 25 °C. Reproducibility over six monthsThe reproducibility of the method was examined using 15 randomly selected Carnobacterium strains (Table 1). The strains were grown on two occasions separated by a period of six months as previously described on CASO agar at 25 °C for 3 days. Whole-organism Fingerprinting of the Genus SummarySixty seven strains of Carnobacterium, atypical Lactobacillus, Enterococcus durans, Lactobacillus maltaromicus and Vagacoccus salmoninarum were examined by Fourier transform infrared (FT-IR) spectroscopy. The effects of culture age and reproducibility over a six month period were also investigated. The results were analysed by multivariate statistics and compared with those from a previous numerical phenetic study, a pyrolysis mass spectrometry (PyMS) study and with investigations which used DNA-DNA and 16S rRNA sequencing homologies. Taxonomic correlations were observed between the FT-IR data and these studies. Culture age was observed to have little effect on the spectra obtained. The reproducibility study indicated that there was correlation between spectra produced on two occasions over the six month period. It was concluded that FTIR is a reliable method for investigating carnobacterial classification, and may have further potential as a rapid method for use in Carnobacterium identification.

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“…While some other reports indicated the cations and cold-hot alternation in the process of transformation causes the cell membrane to transform into a more regular packing structure with a less fluid-like behavior. This solidified membrane may increase the permeability to DNA, which may be due to the complex forming of PHB/Ca 2+ /PPi leading to small pores in the membrane structure (Addison, Chu & Reusch, 2004;Huang & Reusch, 1995;Pavlov et al, 2005). Meanwhile, the pores could be enlarged by adding of other bioorganic compounds, such as 10% ethanol (Sarkar, Choudhuri & Basu, 2002a;Sarkar, Choudhuri & Basu, 2002b;Sharma, Singh & Gill, 2007) and cyclodextrins (Aachmann & Aune, 2009).…”

Section: Discussionmentioning

confidence: 99%

Effects of Sr^2 + on the preparation of Escherchia coli DH5α competent cells and plasmid transformation

Wang

et al. 2020

PeerJ

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Bacterial gene transformation used with Escherichia coli as a desired microorganism is one of the important techniques in genetic engineering. In this study, the preparation of E. coli DH5α competent cells treated with SrCl2 and transformation by heat-shock with pUC19 plasmid was optimized by Response Surface Methodology (RSM). Other five E. coli strains including BL21 (DE3), HB-101, JM109, TOP10 and TG1, three different sizes plasmids (pUC19, pET32a, pPIC9k) were used to verify the protocol, respectively. The transformation mechanism was explored by scanning electron microscope combined with energy dispersive spectrometer (SEM-EDS), atomic absorption spectroscopy (AAS) and Fourier-transform infrared spectroscopy (FT-IR). An equation of regression model was obtained, and the ideal parameters were Sr2 + ions of 90 mM, heat-shock time of 90 s and 9 ng of plasmid. Under this conditions, the transformation efficiency could almost reach to 106 CFU/µg DNA. A small change of the cell surface structure has been observed between E. coli DH5α strain and competent cells by abovementioned spectrum technologies, which implied that a strict regulation mechanism involved in the formation of competent cells and transformation of plasmids. An equation of regression model for the competent cells preparation and plasmid transformation could be applied in gene cloning technology

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Characterization of Carnobacterium species by pyrolysis mass spectrometry

Cited by 14 publications

References 36 publications

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Article

Whole-organism Fingerprinting of the Genus Carnobacterium using Fourier Transform Infrared Spectroscopy (FT-IR)

Effects of Sr^2 + on the preparation of Escherchia coli DH5α competent cells and plasmid transformation

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Characterization of Carnobacterium species by pyrolysis mass spectrometry

Cited by 14 publications

References 36 publications

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Article

Whole-organism Fingerprinting of the Genus Carnobacterium using Fourier Transform Infrared Spectroscopy (FT-IR)

Effects of Sr2 + on the preparation of Escherchia coli DH5α competent cells and plasmid transformation

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Effects of Sr^2 + on the preparation of Escherchia coli DH5α competent cells and plasmid transformation