Development and Use of tuf Gene–Based Primers for the Multiplex PCR Detection of Lactobacillus acidophilus, Lactobacillus casei Group, Lactobacillus delbrueckii, and Bifidobacterium longum in Commercial Dairy Products

Sheu, Sen-Je; Hwang, Wen-Zhe; Chen, Hsin-Chih; Chiang, Yu-Cheng; Tsen, Hau‐Yang

doi:10.4315/0362-028x-72.1.93

Cited by 27 publications

(11 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…With advances in PCR amplification technology and the use of fluorescent (Matsuki et al 2004;Kramer et al 2009). Recent studies have established the applicability of this technology to other fermented dairy products such as yoghurt (Fasoli et al 2003;Rademaker et al 2006;Sieuwerts et al 2008;Garcia-Cayuela et al 2009;Sheu et al 2009), and its applicability to cheese has been demonstrated. Nucleic acid extraction directly from a food matrix was previously limited by high carbohydrate, protein and lipid contents, but methods are now available to extract high-quality DNA for direct use in identification strategies.…”

mentioning

confidence: 99%

“…Challenges in direct nucleic acid extraction from cheese were previously circumvented by prior microbial cell separation from the cheese matrix for nucleic acid extraction (Rudi et al 2005;Fernandez et al 2006;Monnet et al 2006;Fukushima et al 2007) or using large sample quantities (≥10 g cheese) (Monnet et al 2006(Monnet et al , 2008Sohier et al 2012). Else, the methods only provided higher detection limits (≥1000 CFU g À1 ) (Baruzzi et al 2005;Rantsiou et al 2008;Sheu et al 2009) or are incapable of distinguishing added probiotic lactobacilli from NSLAB lactobacilli (Gardiner et al 1998). Such approaches cannot determine changes of species levels in long-term aged cheeses.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Probiotic bacteria survive in Cheddar cheese and modify populations of other lactic acid bacteria

et al. 2014

View full text Add to dashboard Cite

Aims: Starter lactic acid bacteria in Cheddar cheese face physico-chemical stresses during manufacture and ageing that alter their abilities to survive and to interact with other bacterial populations. Nonstarter bacteria are derived from milk handling, cheese equipment and human contact during manufacture. Probiotic bacteria are added to foods for human health benefits that also encounter physiological stresses and microbial competition that may mitigate their survival during ageing. We added probiotic Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus paracasei and Bifidobacterium animalis subsp. lactis to full-fat, reduced-fat and low-fat Cheddar cheeses, aiming to study their survival over 270 days of ageing and to determine the role of the cheese matrix in their survival. Methods and Results: Probiotic and other lactic acid bacterial populations were enumerated by quantitative PCR using primers specifically targeting the different bacterial genera or species of interest. Bifidobacteria were initially added at 10 6 CFU g À1 cheese and survived variably in the different cheeses over the 270-day ageing process. Probiotic lactobacilli that were added at 10 7 CFU g À1 cheese and incident nonstarter lactobacilli (initially at 10 8 CFU g À1 cheese) increased by 10-to 100-fold over 270 days. Viable bacterial populations were differentiated using propidium monoazide followed by species-specific qPCR assays, which demonstrated that the starter and probiotic microbes survived over ageing, independent of cheese type. Addition of probiotic bacteria, at levels 100-fold below that of starter bacteria, modified starter and nonstarter bacterial levels. Conclusions: We demonstrated that starter lactococci, nonstarter lactobacilli and probiotic bacteria are capable of surviving throughout the cheesemaking and ageing process, indicating that delivery via hard cheeses is possible. Probiotic addition at lower levels may also alter starter and nonstarter bacterial survival. Significance and Impact of the Study: We applied qPCR to study multispecies survival and viability and distinctly enumerated bacterial species in commercial-scale Cheddar cheese manufacture.

show abstract

mentioning

confidence: 99%

mentioning

confidence: 99%

Probiotic bacteria survive in Cheddar cheese and modify populations of other lactic acid bacteria

et al. 2014

View full text Add to dashboard Cite

show abstract

“…identification is a valid example of highly related bacterial species that require such approach that provides the polyphasic analysis of two distinct phylogenetic markers: the 16S rDNA gene and the tuf gene encoding for the elongation factor Tu (EF-Tu). [16][17][18]33 Seven products tested in this study showed levels of viable counts similar to or slightly higher than the label claim until 3 months to deadline. The viable bacteria, recognized in two vaginal products without indication, were acceptable.…”

Section: Discussionmentioning

confidence: 96%

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

Blandino

Fazio

Petronio

et al. 2015

Int J Immunopathol Pharmacol

View full text Add to dashboard Cite

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.

show abstract

“…The last 10 years have witnessed a large number of research articles and reviews describing PCR-based applications for probiotics and microbes involved in fermentation processes. Arisen from comparative genomics analyses, these methods aim at specifically quantifying microbial populations (Friedrich and Lenke, 2006; Masco et al, 2007; Randazzo et al, 2009; Sheu et al, 2009, 2010; Abdulamir et al, 2010; Reimann et al, 2010; Sohier et al, 2012), at assessing the role of microbes in dairy products processes (Collado et al, 2006; Ben Amor et al, 2007; Randazzo et al, 2009; Masoud et al, 2011), or at allowing simultaneous identification of dairy and probiotic bacteria containing multiple strains (Wong and Medrano, 2005; Sul et al, 2007; Senan et al, 2008; Smith and Osborn, 2009). …”

Section: The Popular Pcr and Pcr-based Methods: Ready For Routine Anamentioning

confidence: 99%

Evolution of microbiological analytical methods for dairy industry needs

et al. 2014

View full text Add to dashboard Cite

Traditionally, culture-based methods have been used to enumerate microbial populations in dairy products. Recent developments in molecular methods now enable faster and more sensitive analyses than classical microbiology procedures. These molecular tools allow a detailed characterization of cell physiological states and bacterial fitness and thus, offer new perspectives to integration of microbial physiology monitoring to improve industrial processes. This review summarizes the methods described to enumerate and characterize physiological states of technological microbiota in dairy products, and discusses the current deficiencies in relation to the industry’s needs. Recent studies show that Polymerase chain reaction-based methods can successfully be applied to quantify fermenting microbes and probiotics in dairy products. Flow cytometry and omics technologies also show interesting analytical potentialities. However, they still suffer from a lack of validation and standardization for quality control analyses, as reflected by the absence of performance studies and official international standards.

show abstract

Development and Use of tuf Gene–Based Primers for the Multiplex PCR Detection of Lactobacillus acidophilus, Lactobacillus casei Group, Lactobacillus delbrueckii, and Bifidobacterium longum in Commercial Dairy Products

Cited by 27 publications

References 28 publications

Probiotic bacteria survive in Cheddar cheese and modify populations of other lactic acid bacteria

Probiotic bacteria survive in Cheddar cheese and modify populations of other lactic acid bacteria

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

Evolution of microbiological analytical methods for dairy industry needs

Contact Info

Product

Resources

About