Evaluation of the vital viability and their application in fungal spores’ disinfection with flow cytometry

Cao, Ruihua; Wan, Qiqi; Tan, Lili; Xu, Xiangqian; Wu, Gehui; Wang, Jingyi; Xu, Hui; Wen, Gang

doi:10.1016/j.chemosphere.2020.128700

Cited by 31 publications

(9 citation statements)

References 78 publications

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“…In addition, Figure showed that the percentage of spores in the VBNC state obtained from the DRBC culture medium increased after the spores were induced by disinfectant . With increasing disinfection time, the percentage of spores in the VBNC state decreased due to the increased extent of damage, which was similar to the previous study …”

Section: Resultssupporting

confidence: 88%

“…Membrane completeness of Aspergillus spores was observed by flow cytometry (FC) (Accuri C6, BD, USA) . SYBR Green I/Propidium Iodide (SG/PI) was used to stain the Aspergillus spores, where SG can stain all cells but PI stains only membrane-permeabilized cells.…”

Section: Materials and Methodsmentioning

confidence: 99%

“…The primary methods employed for enumerating fungal spores in water include heterotrophic plate counting (HPC), direct microscopic observation, liquid chromatography ergosterol determination, real-time quantitative polymerase chain reaction (qPCR) detection, and flow cytometry (FC). ,− Each method possesses its own strengths and limitations. For instance, the HPC method is cost-effective and straightforward to implement, but 2 days of incubation time are required for fungal growth.…”

Section: Introductionmentioning

confidence: 99%

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Comprehensive Comparisons of Five Kinds of Culture Media for Cultivating Fungal Spores at Different Viable States in Water

Chang,

Xu,

et al. 2023

ACS EST Water

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At present, there is no unified culture medium to quantify the cultivability of fungal spores in water, making comparisons between studies difficult. This study intended to select an appropriate medium to obtain a lower percentage of viable but nonculturable Aspergillus spores at different viable states (freshly prepared, impaired-treated) and fungi in actual water. Five kinds of media ((dichloran rose bengal chloramphenicol (DRBC) agar, potato dextrose agar (PDA), dichloran 18% glycerol (DG-18) agar, rose bengal (RB) agar, and yeast extract dextrose chloramphenicol (YDC) agar) were selected. To obtain the result quickly and colony counting clearly for freshly prepared Aspergillus spores, the optimal incubation time was determined, and the cultivating effectiveness of media was: DRBC > RB > YDC > DG-18 > PDA. The DRBC medium exhibited better performance in quantifying impaired spores treated by ultraviolet or chlorine. Besides, RB medium was optimal for fungal enumeration in actual water. DRBC and RB media exhibited higher potential capacity in cultivating Aspergillus spores at different viable states because the existence of rose bengal restricted the size of the colony and the lower content of glucose limited the rapid growth of colonies. This study provides meaningful information for selecting a suitable medium to quantify fungal spores at different viable states.

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Section: Resultssupporting

confidence: 88%

Section: Materials and Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Comprehensive Comparisons of Five Kinds of Culture Media for Cultivating Fungal Spores at Different Viable States in Water

Chang,

Xu,

et al. 2023

ACS EST Water

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“…This section focuses on the characterization of drinking water quality and wastewater purification [ 74 ] and bioaerosol detection [ 49 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 ]. Although FCM has been widely used for the microbial analysis of aquatic milieus ( Table 2 ), it is less frequently used for the quantitative detection of bioaerosols ( Table 3 ) [ 80 , 83 ].…”

Section: Non-specific State-of-the-art Flow Cytometric Applications For Detection and Monitoringmentioning

confidence: 99%

Potential of Flow Cytometric Approaches for Rapid Microbial Detection and Characterization in the Food Industry—A Review

Zand

Froehling

Schoenher

et al. 2021

Foods

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As microbial contamination is persistent within the food and bioindustries and foodborne infections are still a significant cause of death, the detection, monitoring, and characterization of pathogens and spoilage microorganisms are of great importance. However, the current methods do not meet all relevant criteria. They either show (i) inadequate sensitivity, rapidity, and effectiveness; (ii) a high workload and time requirement; or (iii) difficulties in differentiating between viable and non-viable cells. Flow cytometry (FCM) represents an approach to overcome such limitations. Thus, this comprehensive literature review focuses on the potential of FCM and fluorescence in situ hybridization (FISH) for food and bioindustry applications. First, the principles of FCM and FISH and basic staining methods are discussed, and critical areas for microbial contamination, including abiotic and biotic surfaces, water, and air, are characterized. State-of-the-art non-specific FCM and specific FISH approaches are described, and their limitations are highlighted. One such limitation is the use of toxic and mutagenic fluorochromes and probes. Alternative staining and hybridization approaches are presented, along with other strategies to overcome the current challenges. Further research needs are outlined in order to make FCM and FISH even more suitable monitoring and detection tools for food quality and safety and environmental and clinical approaches.

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“…FCM is an effective method for bacterial cell count and viability determination, which is widely used in medicine, microbiology, on-site environmental detection, and other fields [ 20 , 21 , 22 ]. For example, in medical research, Kalina T and other [ 23 ] people reported the effective use of FCM in primary immunodeficiency, immunophenotype, diagnosis, and functional research; in microbial research, Leonard L and other [ 24 ] people discussed the potential opportunities provided by FCM in detecting the impact of antibiotics on microbial subpopulations; and in environmental testing, some scholars used FCM to evaluate the effect of disinfectants in water on fungal spores and to study their inactivation mechanism [ 25 ]. In recent years, the detection of microorganisms in food by flow cytometry has become more and more extensive.…”

Section: Introductionmentioning

confidence: 99%

Rapid Detection of the Activity of Lacticaseibacillus Casei Zhang by Flow Cytometry

Wang

Dai

et al. 2023

Foods

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Food processing, e.g., freeze-drying, exerts strong pressure on bacteria in the food matrix, decreasing their viability/activity and even forcing them to become viable but unculturable (VBNC), which are often underestimated by traditional plate count. The strict standards of bacterial viability in probiotic products require accurate cell viability/activity enumeration. We developed a staining (5(6)-carboxyfluorescein diacetate succinimide ester, propidium iodide)-based flow cytometry rapid method for detecting the viability/activity of Lacticaseibacillus (Lb.) casei Zhang, a widely used probiotic in the dairy industry in China. We optimized the procedural and instrumental parameters for generating results comparable to that of standard plate counts. This method was also applied to freeze-dried Lb. casei Zhang, yielding 7.7 × 1011 CFU/g, which was non-significantly higher than the results obtained by plate count (6.4 × 1011 CFU/g), possibly due to the detection of VBNC cells in the freeze-dried powder. We anticipated that this method can be used for detecting lactic acid bacteria in other probiotic food/beverages.

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Evaluation of the vital viability and their application in fungal spores’ disinfection with flow cytometry

Cited by 31 publications

References 78 publications

Comprehensive Comparisons of Five Kinds of Culture Media for Cultivating Fungal Spores at Different Viable States in Water

Comprehensive Comparisons of Five Kinds of Culture Media for Cultivating Fungal Spores at Different Viable States in Water

Potential of Flow Cytometric Approaches for Rapid Microbial Detection and Characterization in the Food Industry—A Review

Rapid Detection of the Activity of Lacticaseibacillus Casei Zhang by Flow Cytometry

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