Identification of indigenous Lactobacillus isolated from artisanal Algerian dairy product by 16S rRNA Gene Sequencing and MALDI-TOF Mass Spectrometry

El-klila, Jben, Rayeb and Lben are artisanal dairy products typical of the North African region. These products are considered a good source of nutrients and a vector par excellence of indigenous lactic acid bacteria with significant technological and therapeutic potential. In this study, fifty-two Lactobacillus isolates from traditional dairy products (El-Klila, Jben and Rayeb) from the Ain Sefra region (southwestern of Algeria) were identified by phenotypic, genotypic and MALDI-TOF mass spectrometry methods. Phenotypic characterization allowed dividing the isolates into two groups: 7.69% of isolates mesophilic and strict heterofermentative and 92.31% isolates mesophilic and optional heterofermentative. The isolates identification by sequencing 16S rDNA was performed using the universal primers w02 and w18. Comparing the sequences of the isolates with the GenBank database (NCBI) revealed a high percentage with five species of Lactobacillus. Of the 52 isolates, thirteen were identified as Lactobacillus paracasei, four isolates were L. brevis, five isolates were L. rhamnosus and ten isolates were L. plantarum. The lasting twenty isolates were related, at the same time, to the plantarum species and the pentosus species. The use of MALDI-TOF mass spectrometry confirmed the identity of 36 strains of Lactobacillus. Of the 20 isolates related to the plantarum / pentosus group by 16S rDNA, 50% were identified as L. plantarum. Finally, the results of the two combined methods allowed us to correctly identify 80.76% of the Lactobacillus isolates at the genus and species level. The analysis by MALDI-TOF SM has proved to be a very reliable, fast (less than an hour) and inexpensive tool for identifying native Lactobacillus species from artisanal dairy products. This method can constitute a major tool for the study of the microbial biodiversity of artisanal food products.

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Evaluation of MALDI-TOF mass spectrometry and MALDI BioTyper in comparison to 16S rDNA sequencing for the identification of bacteria isolated from Arctic sea water

PLoS ONE ◽

10.1371/journal.pone.0181860 ◽

2017 ◽

Vol 12 (7) ◽

pp. e0181860 ◽

Cited By ~ 19

Author(s):

Anna Maria Timperio ◽

Susanna Gorrasi ◽

Lello Zolla ◽

Massimiliano Fenice

Keyword(s):

Mass Spectrometry ◽

16S Rdna ◽

Sea Water ◽

16S Rdna Sequencing ◽

Maldi Tof Mass Spectrometry ◽

Maldi Tof ◽

Rdna Sequencing ◽

Arctic Sea ◽

Maldi Biotyper ◽

Identification Of Bacteria

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Updates on quick identification of acetic acid bacteria with a focus on the 16S–23S rRNA gene internal transcribed spacer and the analysis of cell proteins by MALDI-TOF mass spectrometry

International Journal of Food Microbiology ◽

10.1016/j.ijfoodmicro.2014.12.003 ◽

2015 ◽

Vol 196 ◽

pp. 137-144 ◽

Cited By ~ 19

Author(s):

Janja Trček ◽

François Barja

Keyword(s):

Mass Spectrometry ◽

Acetic Acid ◽

Internal Transcribed Spacer ◽

Acetic Acid Bacteria ◽

23S Rrna ◽

Rrna Gene ◽

Maldi Tof Mass Spectrometry ◽

Maldi Tof ◽

23S Rrna Gene

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Species Diversity of Bifidobacteria in the Intestinal Microbiota Studied Using MALDI-TOF Mass-Spectrometry

Annals of the Russian academy of medical sciences ◽

10.15690/vramn.v70.i4.1409 ◽

2015 ◽

Vol 70 (4) ◽

pp. 435-440 ◽

Cited By ~ 8

Author(s):

A. V. Chaplin ◽

A. G. Brzhozovskii ◽

T. V. Parfenova ◽

L. I. Kafarskaia ◽

N. N. Volodin ◽

...

Keyword(s):

Mass Spectrometry ◽

16S Rrna ◽

16S Rrna Gene ◽

Intestinal Microbiota ◽

Rrna Gene ◽

Growing Up ◽

Gene Fragment ◽

Bifidobacterium Adolescentis ◽

Maldi Tof Mass Spectrometry ◽

Maldi Tof

Background: The members of genus Bifidobacterium represent a significant part of intestinal microbiota in adults and predominate in infants. Species repertoire of the intestinal bifidobacteria is known to be subjected to major changes with age; however, many details of this process are still to be elucidated.Objective: Our aim was to study the diversity of intestinal bifidobacteria and changes of their qualitative and quantitative composition characteristics during the process of growing up using MALDI-TOF mass-spectrometric analysis of pure bacterial cultures.Methods: A cross-sectional study of bifidobacteria in the intestinal microbiota was performed in 93 healthy people of the ages from 1 month to 57 years. Strains were identified using Microflex LT MALDI-TOF MS, the confirmation was performed by 16S rRNA gene fragment sequencing.Results: 93% of isolated bifidobacterial strains were successfully identified using MALDI-TOF mass-spectrometry. At least two of the strains from each species were additionally identified by 16S rRNA gene fragment sequencing, in all of the cases the results were the same. It was shown that the total concentration of bifidobacteria decreases with age (p 0.001) as well as the frequency of isolation of Bifidobacterium bifidum (p =0.020) and Bifidobacterium breve (p 0.001), and the frequency of isolation of Bifidobacterium adolescentis, increases (p 0.001), representing the continuous process of transformation of microbiota.Conclusion: The method of MALDI-TOF mass spectrometry demonstrated the ability to perform rapid and reliable identification of bifidobacteria that allowed the study of changes in the quantitative and qualitative characteristics of human microbiota in the process of growing up.

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Francisella philomiragia bacteremia in an immunocompromised patient: a rare case report

Annals of Clinical Microbiology and Antimicrobials ◽

10.1186/s12941-021-00475-2 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Hui Shan Chua ◽

Yih Harng Soh ◽

Shih Keng Loong ◽

Sazaly AbuBakar

Keyword(s):

Mass Spectrometry ◽

16S Rdna ◽

Acute Pulmonary Oedema ◽

Molecular Technique ◽

Type I ◽

16S Rdna Sequencing ◽

Maldi Tof Mass Spectrometry ◽

Prompt Treatment ◽

Maldi Tof ◽

Rdna Sequencing

Abstract Background Francisella philomiragia is a very rare opportunistic pathogen of humans which causes protean diseases such as pneumonia and other systemic infections. Subsequent failure of prompt treatment may result in poor prognosis with mortality among infected patients. Case presentation The present report describes a case of F. philomiragia bacteraemia first reported in Malaysia and Asian in a 60-year-old patient with underlying end-stage renal disease (ESRF) and diabetes mellitus. He presented with Acute Pulmonary Oedema with Non-ST-Elevation Myocardial Infarction (NSTEMI) in our hospital. He was intubated in view of persistent type I respiratory failure and persistent desaturation despite post haemodialysis. Blood investigation indicated the presence of ongoing infection and inflammation. The aerobic blood culture growth of F. philomiragia was identified using the matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (Score value: 2.16) and confirmed by 16S Ribosomal DNA (16S rDNA) sequencing. He was discharged well on day 26 of admission, after completing one week of piperacillin/tazobactam and two weeks of doxycycline. Conclusion Clinical suspicion should be raised if patients with known risk factors are presenting with pneumonia or pulmonary nodules especially as these are the most common manifestations of F. philomiragia infection. Early diagnosis via accurate laboratory identification of the organism through MALDI-TOF mass spectrometry and molecular technique such as 16S rDNA sequencing are vital for prompt treatment that results in better outcomes for the afflicted patients.

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