Evolution of microbiological analytical methods for dairy industry needs

Sohier, Danièle; Pavan, Sonia; Riou, Armelle; Combrisson, Jérôme; Postollec, Florence

doi:10.3389/fmicb.2014.00016

Cited by 54 publications

(46 citation statements)

References 123 publications

(184 reference statements)

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“…In this scenario, the polyphasic taxonomy suggested by Vandamme et al 31 and by other authors, 17,18,32 is the better choice for bacterial identification.…”

Section: Discussionmentioning

confidence: 99%

Labeling quality and molecular characterization studies of products containing Lactobacillus spp. strains.

Blandino

Fazio

Petronio

et al. 2015

Int J Immunopathol Pharmacol

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The objective of the study was to characterize at species level by phenotypic and different molecular methods the strains of Lactobacillus spp. used as constituents of five oral and four vaginal products. Susceptibilities to representative antibiotics were evaluated. In addition, total viable counts at mid and 3 months to deadline of shelf life, in the different formulations and the presence of eventual contaminant microorganisms were investigated. In all oral products the molecular characterization at species level of the strains of Lactobacillus spp. confirmed the strains stated on the label, except for one strain cited on the label as Lactobacillus casei, that our study characterized as Lactobacillus paracasei. In oral products total viable cell content complied with content claimed on the label. In three out four vaginal products (one product claimed “bacillo di Döderlein”), molecular characterization complied with the bacterial name stated on the label. Two vaginal products reported viable counts on the label that were confirmed by our study. The other vaginal products, which did not report bacterial counts on the label, showed a similar decrease of viable counts at different dates to deadline compared to the others. From all the tested products, contaminant microorganisms and acquired resistance to representative antibiotics by the probiotic strains were not detected.

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“…In this scenario, the polyphasic taxonomy suggested by Vandamme et al 31 and by other authors, 17,18,32 is the better choice for bacterial identification.…”

Section: Discussionmentioning

confidence: 99%

Labeling quality and molecular characterization studies of products containing Lactobacillus spp. strains.

Blandino

Fazio

Petronio

et al. 2015

Int J Immunopathol Pharmacol

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show abstract

“…However, RNA manipulation is difficult due to the short half‐life of microbial RNA (Sohier et al . ). Molecular biology protocols require several delicate steps (nucleic acid extraction and amplification) that can be tedious.…”

Section: Introductionmentioning

confidence: 97%

“…The use of flow cytometry in combination with fluorescent staining has increased within the microbiology field over the last 20 years (Comas‐Riu and Rius ; Davis ; Sohier et al . ). Viability staining and flow cytometry (viable flow cytometry) has been extensively used to monitor bacteria physiological state (Joux and Lebaron ; Doherty et al .…”

Section: Introductionmentioning

confidence: 97%

“…Another way to quantify viable bacteria is to employ RNA-targeting techniques (Lahtinen et al 2008). However, RNA manipulation is difficult due to the short halflife of microbial RNA (Sohier et al 2014). Molecular biology protocols require several delicate steps (nucleic acid extraction and amplification) that can be tedious.…”

Section: Introductionmentioning

confidence: 99%

“…Flow cytometry is a technology used for the multiparametric analysis of cells based on light scattering and fluorescence at the singlecell level (Robinson and Roederer 2015). The use of flow cytometry in combination with fluorescent staining has increased within the microbiology field over the last 20 years (Comas-Riu and Rius 2009;Davis 2014;Sohier et al 2014). Viability staining and flow cytometry (viable flow cytometry) has been extensively used to monitor bacteria physiological state (Joux and Lebaron 2000;Doherty et al 2010), survival upon physical or chemical stresses (Amor et al 2002;Rault et al 2007;Chen et al 2012), antibiotic susceptibility (Novo et al 2000) and microbial communities study (Apajalahti et al 2002;Muller and Nebe-von-Caron 2010;Koch et al 2014).…”

Section: Introductionmentioning

confidence: 99%

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Flow cytometry: a versatile technology for specific quantification and viability assessment of micro-organisms in multistrain probiotic products

2018

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Aims: Classical microbiology techniques are the gold standard for probiotic enumeration. However, these techniques are limited by parameters of time, specificity and incapacity to detect viable but nonculturable (VBNC) microorganisms and nonviable cells. The aim of the study was to evaluate flow cytometry as a novel method for the specific quantification of viable and nonviable probiotics in multistrain products. Methods and Results: Custom polyclonal antibodies were produced against five probiotic strains from different species (Bifidobacterium bifidum R0071, Bifidobacterium longum ssp. infantis R0033, Bifidobacterium longum ssp. longum R0175, Lactobacillus helveticus R0052 and Lactobacillus rhamnosus R0011). Evaluation of specificity confirmed that all antibodies were specific at least at the subspecies level. A flow cytometry method combining specific antibodies and viability assessment with SYTO â 24 and propidium iodide was applied to quantify these strains in three commercial products. Analyses were conducted on two flow cytometry instruments by two operators and compared with classical microbiology using selective media. Results indicated that flow cytometry provides higher cell counts than classical microbiology (P < 0Á05) in 73% of cases highlighting the possible presence of VBNC. Equivalent performances (repeatability and reproducibility) were obtained for both methods. Conclusions: This study showed that flow cytometry methods can be applied to probiotic enumeration and viability assessment. Combination with polyclonal antibodies can achieve sufficient specificity to differentiate closely related strains. Significance and Impact of the Study: Flow cytometry provides absolute and specific quantification of viable and nonviable probiotic strains in a very short time (<2 h) compared with classical techniques (>48 h), bringing efficient tools for research and development and quality control.

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Enumeration and Identification of Mixed Probiotic and Lactic Acid Bacteria Starter Cultures

Majhenič¹,

Lorbeg²,

Treven³

2017

Probiotic Dairy Products

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Evolution of microbiological analytical methods for dairy industry needs

Cited by 54 publications

References 123 publications

Labeling quality and molecular characterization studies of products containing Lactobacillus spp. strains.

Labeling quality and molecular characterization studies of products containing Lactobacillus spp. strains.

Flow cytometry: a versatile technology for specific quantification and viability assessment of micro-organisms in multistrain probiotic products

Enumeration and Identification of Mixed Probiotic and Lactic Acid Bacteria Starter Cultures

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