Visualization of probiotics via epifluorescence microscopy and fluorescence in situ hybridization (FISH)

Pasulka, Alexis L.; Howes, Amy L.; Kallet, Julia; VanderKelen, Jennifer; Villars, Clayton

doi:10.1016/j.mimet.2021.106151

Cited by 12 publications

(8 citation statements)

References 31 publications

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“…FISH is on the rise for the detection of probiotic bacteria that are intentionally administered to the GI tract [ 65 ]. In our study, the fluorescently labeled specific probe Lcas467-Cy3 was hybridized with the selected lactose-negative typical LGG-like bacteria.…”

Section: Resultsmentioning

confidence: 99%

Prebiotic Cellulose–Pullulan Matrix as a “Vehicle” for Probiotic Biofilm Delivery to the Host Large Intestine

Savitskaya,

Zhantlessova,

Kistaubayeva

et al. 2023

Polymers

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This study describes the development of a new combined polysaccharide-matrix-based technology for the immobilization of Lactobacillus rhamnosus GG (LGG) bacteria in biofilm form. The new composition allows for delivering the bacteria to the digestive tract in a manner that improves their robustness compared with planktonic cells and released biofilm cells. Granules consisting of a polysaccharide matrix with probiotic biofilms (PMPB) with high cell density (>9 log CFU/g) were obtained by immobilization in the optimized nutrient medium. Successful probiotic loading was confirmed by fluorescence microscopy and scanning electron microscopy. The developed prebiotic polysaccharide matrix significantly enhanced LGG viability under acidic (pH 2.0) and bile salt (0.3%) stress conditions. Enzymatic extract of feces, mimicking colon fluid in terms of cellulase activity, was used to evaluate the intestinal release of probiotics. PMPB granules showed the ability to gradually release a large number of viable LGG cells in the model colon fluid. In vivo, the oral administration of PMPB granules in rats resulted in the successful release of probiotics in the colon environment. The biofilm-forming incubation method of immobilization on a complex polysaccharide matrix tested in this study has shown high efficacy and promising potential for the development of innovative biotechnologies.

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Section: Resultsmentioning

confidence: 99%

Prebiotic Cellulose–Pullulan Matrix as a “Vehicle” for Probiotic Biofilm Delivery to the Host Large Intestine

Savitskaya,

Zhantlessova,

Kistaubayeva

et al. 2023

Polymers

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“…Cultivation methods, such as plate count methods, are the gold standard [47]. The plate count method is simple to perform, but it requires a long incubation time and the selection of appropriate culture media.…”

Section: Cultivation Methodsmentioning

confidence: 99%

“…The most common ones are cultivation methods with the use of appropriate media, as well as the increasingly popular method of flow cytometry. Other methods are also described, such as fluorescence in situ hybridisation (FISH) [47] or nucleic acid abundance methods [48].…”

Section: Determining the Count Of Probiotic Microbes In Productsmentioning

confidence: 99%

Microbiological Testing of Probiotic Preparations

Zawistowska-Rojek

Zaręba

Tyski

2022

IJERPH

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Probiotic microorganisms that are potentially beneficial to the health of the host are commercially available in a great variety of products. Not all microorganism strains present in products have proven beneficial to the health properties. These products include not only foodstuffs but also dietary supplements, food for special medical purposes, medicinal products, as well as cosmetics and medical devices. These products contain from one to a dozen bacterial strains of the same or different species and sometimes also fungal strains. Since the pro-health effects of probiotics depend on a specific strain, the number of its cells in a dose, and the lack of pathogenic microorganisms, it is extremely important to control the quality of probiotics. Depending on the classification of a given product, its form, and its content of microorganisms, the correct determination of the number of microorganisms and their identification is crucial. This article describes the culture-dependent and culture-independent methods for testing the contents of probiotic microorganisms, in addition to biochemical and genetic methods of identification. The microbiological purity requirements for various product categories are also presented. Due to numerous reports on the low quality of probiotic products available on the market, it is important to standardise research methods for this group of products and to increase the frequency of inspections of these products.

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“…They demonstrated both FISH and CARD-FISH techniques effectively detected B. subtilis on plant roots growing in different culture systems. Moreover, Pasulka et al (2021) designed FISH probes to quantitatively measure Bacillus abundance in direct-fed microbial products and crop microbial biostimulants to ensure these probiotics have adequate amounts of bacteria. Still, the potential of FISH to be used for enumerating Bacillus within products has largely been unexplored.…”

Section: Emphasizing Quantification: the Crucial Role Beyond Identifi...mentioning

confidence: 99%

How to identify and quantify the members of the Bacillus genus?

Xu,

Kovács

2024

Environmental Microbiology

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Members of the Bacillus genus are widely distributed throughout natural environments and have been studied for decades among others for their physiology, genetics, ecological functions, and applications. However, despite its prevalence in nature, the characterization and classification of Bacillus remain challenging due to its complex and ever‐evolving taxonomic framework. This review addresses the current state of the Bacillus taxonomic landscape and summarizes the critical points in the development of Bacillus phylogeny. With a clear view of Bacillus phylogeny as a foundation, we subsequently review the methodologies applied in identifying and quantifying Bacillus, while also discussing their respective advantages and disadvantages.

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Visualization of probiotics via epifluorescence microscopy and fluorescence in situ hybridization (FISH)

Cited by 12 publications

References 31 publications

Prebiotic Cellulose–Pullulan Matrix as a “Vehicle” for Probiotic Biofilm Delivery to the Host Large Intestine

Prebiotic Cellulose–Pullulan Matrix as a “Vehicle” for Probiotic Biofilm Delivery to the Host Large Intestine

Microbiological Testing of Probiotic Preparations

How to identify and quantify the members of the Bacillus genus?

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