A comparison of fluorescent stains for the assessment of viability and metabolic activity of lactic acid bacteria

Zotta, Teresa; Guidone, Angela; Tremonte, Patrizio; Parente, Eugenio; Ricciardi, Annamaria

doi:10.1007/s11274-011-0889-x

Cited by 41 publications

(28 citation statements)

References 31 publications

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“…This sensitivity to change in the incubation temperature was characterized by a loss of viability between 98% and 100% or about 100% at 54 ° C or 75 ° C, respectively. Significant effect of temperature on the viability of LAB used as starter cultures has already been reported [10,11]. However, the effect of incubation temperature was less evident in our study.…”

Section: Resultscontrasting

confidence: 47%

Assessment of the Potential of Lactic Acid Bacteria as Dried Starter Culture for Cereal Fermentation

Soro-Yao¹,

Aka²,

Thonart³

et al. 2014

TOBIOTJ

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and 0.61 h to 1.33 h, respectively. We observed that the strains were sensitive to a change in water activity (a w 0.32 final), storage and incubation temperatures. The Strain LF obtained a higher cell concentration and viability, as well as a lower g compared to those obtained by the other strains. The kinetic growth parameters (µ m , g) together with viability after stress treatments could be used for the screening of dried lactic acid starter cultures.

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Section: Resultscontrasting

confidence: 47%

Assessment of the Potential of Lactic Acid Bacteria as Dried Starter Culture for Cereal Fermentation

Soro-Yao¹,

Aka²,

Thonart³

et al. 2014

TOBIOTJ

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“…Therefore its application for routine analyses is difficult to implement. The use of other fluorescent labeling techniques, sometimes combined with FISH, is also described (Moreno et al, 2006; Zotta et al, 2012). Fluorescent labeling is always associated with microscopy technologies.…”

Section: Microscopy Flow Cytometry and Other Fluorescent Labeling-bmentioning

confidence: 99%

“…It allows characterizing individual cells within a cell population, and to discriminate between various VBNC and physiological states (Sheehan et al, 2005; Lahtinen et al, 2006; Papadimitriou et al, 2006, 2007; Quiros et al, 2007; Sunny-Roberts and Knorr, 2008; El Arbi et al, 2011; Zotta et al, 2012), which is a major advantage over culture-based methods. The excellent review from Díaz et al (2010) describes the principle of FC, a range of fluorescent probes associated with various cellular functions and applications to industrial microbial bioprocesses including dairy industry applications.…”

Section: Microscopy Flow Cytometry and Other Fluorescent Labeling-bmentioning

confidence: 99%

“…Nowadays, FC appears as the most promising labeling technology (Tracy et al, 2010; Davey, 2011) and many publications describe its use in food microbiology. A large choice of dyes is available to target cell components (nucleic acids, proteins, lipids), intracellular pH, membrane integrity, intracellular ions, viability markers, and membrane energization (Maecker et al, 2004; Sheehan et al, 2005; Papadimitriou et al, 2006; Berney et al, 2007; Quiros et al, 2007; Comas-Riu and Rius, 2009; Díaz et al, 2010; Doherty et al, 2010; El Arbi et al, 2011; Zotta et al, 2012). A fine characterization of the cellular status is possible by combining several dyes (Chen et al, 2011).…”

Section: Microscopy Flow Cytometry and Other Fluorescent Labeling-bmentioning

confidence: 99%

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Evolution of microbiological analytical methods for dairy industry needs

et al. 2014

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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.

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“…The use of automated quantification of fluorescent microscope and double fluorescent dyes (PI and DAPI) may increase the sensitivity of the microscopic method (Rieger et al, 2010;Branco et al, 2012;Zotta et al, 2012), however, identification of hookworm ova may not be possible due to the similar morphology of some other helminth ova (Traub et al, 2007). …”

Section: Vital Stain Methodsmentioning

confidence: 99%

Detection of viable hookworm ova from wastewater and sludge

Gyawali¹

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Infectious helminths are a worldwide major public health problem. In terms of morbidity, approximately 3 × 10 9 people are infected by soil-transmitted helminths (STHs) influencing rates of malnutrition and failure to thrive. All of this increases the global disease burden (GDB) by up to 5.9 × 10 6 Daily Adjusted Life Years (DALYs). Helminth infections are more often seen as a major public health issue for developing countries however, concerns have been expressed in many of the developed nations because of the increased use of treated wastewater and sludge with no clear way of knowing the infection potential that can be attributed from such water.Guidelines have been established to minimise the potential public health risk associated with wastewater and sludge reuse. For example, the WHO guideline specified ≤ 1 ova (Ascaris lumbricoides) per L liquid (wastewater) or 4 g dry solid (sludge) for unrestricted use. Various wastewater treatment processes have been recommended to remove the helminths ova from wastewater depending upon its reuse. However, detection methods used to quantify viable helminths ova from wastewater has limitations. Therefore there is always a potential public health risk associated with reuse of wastewater and sludge.In this research, a real-time PCR method was developed. The new PCR method is rapid and specific. The method was found to be able to detect less than one ovum in one litre treated wastewater and approximately four ova in one litre raw wastewater and ~ 4 g sludge.The real-time PCR method was modified to a quantitative PCR (qPCR) method and further used to quantify hookworm ova from wastewater and sludge. The qPCR had estimated an average of 1.1, 8.6 and 67.3 ova for treated wastewater that was seeded with 1, 10 and 100 ova, respectively. The gene copy numbers obtained for 1, 10 and 100 ova by qPCR varied significantly (P < 0.05) within the tested samples indicating that absolute quantification of ova may not be accurate. Despite the difficulty quantifying accurate numbers of hookworm ova, the lower limit of quantification (LLOQ) of the qPCR method was 30 gene copies. This was a lot less than the gene copies produced by one ovum. Therefore, the qPCR method has potential to use for complying with wastewater guidelines.. iiAlthough the overall aim of this research was to develop a sensitive and specific method for quantitative detection of viable hookworm ova from wastewater, the importance of recovering the ova from wastewater and sludge samples for accurate detection was identified. While determining appropriate recovery rates was not an aim of this research, a suitable method that could be used to standardise research outcomes for further study was established. Therefore, ova recovery rate by different rapid methods was evaluated for further experiments. The result indicated that the ova recovery rate was higher for the treated wastewater (0.2 -50%) than the raw wastewater (0.3 -35%) and sludge (0.02 -4.7%) samples. A significant difference (P < 0.05) was observed between...

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A comparison of fluorescent stains for the assessment of viability and metabolic activity of lactic acid bacteria

Cited by 41 publications

References 31 publications

Assessment of the Potential of Lactic Acid Bacteria as Dried Starter Culture for Cereal Fermentation

Assessment of the Potential of Lactic Acid Bacteria as Dried Starter Culture for Cereal Fermentation

Evolution of microbiological analytical methods for dairy industry needs

Detection of viable hookworm ova from wastewater and sludge

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