Viability Detection of Foodborne Bacterial Pathogens in Food Environment by PMA-qPCR and by Microscopic Observation

Brauge, Thomas; Midelet, Graziella; Soumet, Christophe

doi:10.1007/978-1-4939-9000-9_9

Cited by 16 publications

(10 citation statements)

References 14 publications

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“…Moreover, Campylobacter strains appear to exhibit different survival characteristics when studied individually though C. coli has emerged as more aerotolerant than C. jejuni (Karki et al, 2018). Important differences between strains as regards residual viable cells have also been reported in biofilms of Listeria monocytogenes cultivated on stainless steel as detected by PMA-qPCR and epifluorescence microscopy indicating a diverse potential of adaptation and survival within the same species (Brauge et al, 2019). The fact that only one strain of C. coli was used in the current study is a recognized limitation regarding the overall survival potential of this species on lamb meat and, therefore, more studies utilizing various strains of this pathogen are needed to provide better insight in this context.…”

Section: Discussionmentioning

confidence: 99%

Method-Dependent Implications in Foodborne Pathogen Quantification: The Case of Campylobacter coli Survival on Meat as Comparatively Assessed by Colony Count and Viability PCR

et al. 2021

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The aim of the present study was to address method-dependent implications during the quantification of viable Campylobacter coli cells on meat over time. Traditional colony counting on selective and non-selective culture media along with an optimized viability real-time PCR utilizing propidium monoazide-quantitative PCR (PMA-qPCR), spheroplast formation and an internal sample process control (ISPC), were comparatively evaluated for monitoring the survival of C. coli on fresh lamb meat during refrigeration storage under normal atmospheric conditions. On day zero of three independent experiments, lamb meat pieces were artificially inoculated with C. coli and then stored under refrigeration for up to 8 days. Three meat samples were tested on different days and the mean counts were determined per quantification method. An overall reduction of the viable C. coli on lamb meat was observed regardless of the applied quantification scheme, but the rate of reduction followed a method-dependent pattern, the highest being observed for colony counting on modified charcoal cefoperazone deoxycholate agar (mCCDA). Univariate ANOVA indicated that the mean counts of viable C. coli using PMA-qPCR were significantly higher compared to Columbia blood agar (CBA) plating (0.32 log10 cell equivalents, p = 0.015) and significantly lower when mCCDA was compared to CBA plating (0.88 log10 CFU, p < 0.001), indicating that selective culture on mCCDA largely underestimated the number of culturable cells during the course of meat storage. PMA-qPCR outperformed the classical colony counting in terms of quantifying both the culturable and viable but non-culturable (VBNC) C. coli cells, which were generated over time on meat and are potentially infectious and equally important from a public health perspective as their culturable counterparts.

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Section: Discussionmentioning

confidence: 99%

Method-Dependent Implications in Foodborne Pathogen Quantification: The Case of Campylobacter coli Survival on Meat as Comparatively Assessed by Colony Count and Viability PCR

et al. 2021

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“…Yet, viability of the detected organisms is questionable as molecular methods rely on the detection of DNA that can persist in the environment long after bacteria have died. To address this issue, previous studies have employed DNA intercalating dyes such as ethidium monoazide (EMA) and propidium monoazide (PMA) to estimate the total number of viable cells in environmental samples [ 5 , 6 , 41 ]. Recently, Cao et al [ 6 ] was able to detect VBNC V. parahaemolyticus in shrimp samples by utilizing PMA dyes [ 6 ].…”

Section: Discussionmentioning

confidence: 99%

Coupled DNA-labeling and sequencing approach enables the detection of viable-but-non-culturable Vibrio spp. in irrigation water sources in the Chesapeake Bay watershed

Malayil

Chattopadhyay

Mongodin

et al. 2021

Environmental Microbiome

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Nontraditional irrigation water sources (e.g., recycled water, brackish water) may harbor human pathogens, including Vibrio spp., that could be present in a viable-but-nonculturable (VBNC) state, stymieing current culture-based detection methods. To overcome this challenge, we coupled 5-bromo-2′-deoxyuridine (BrdU) labeling, enrichment techniques, and 16S rRNA sequencing to identify metabolically-active Vibrio spp. in nontraditional irrigation water (recycled water, pond water, non-tidal freshwater, and tidal brackish water). Our coupled BrdU-labeling and sequencing approach revealed the presence of metabolically-active Vibrio spp. at all sampling sites. Whereas, the culture-based method only detected vibrios at three of the four sites. We observed the presence of V. cholerae, V. vulnificus, and V. parahaemolyticus using both methods, while V. aesturianus and V. shilonii were detected only through our labeling/sequencing approach. Multiple other pathogens of concern to human health were also identified through our labeling/sequencing approach including P. shigelloides, B. cereus and E. cloacae. Most importantly, 16S rRNA sequencing of BrdU-labeled samples resulted in Vibrio spp. detection even when our culture-based methods resulted in negative detection. This suggests that our novel approach can effectively detect metabolically-active Vibrio spp. that may have been present in a VBNC state, refining our understanding of the prevalence of vibrios in nontraditional irrigation waters.

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“…Pathogenic bacteria can cause the epidemic of infectious diseases in livestock and poultry by polluting food and water sources, oral infection in humans may lead to intestinal infections (Chai et al, 2019). Foodborne pathogens are the main cause of foodborne diseases and an important source of food safety problems (Brauge et al, 2019). The common foodborne pathogens are Salmonella , Listeria monocytogenes , Escherichia coli O157:H7, Staphylococcus aureus and so on (Zhang et al, 2020).…”

Section: Introductionmentioning

confidence: 99%

The application and research progress of bacteriophages in food safety

Wang

Zhao

2022

Journal of Applied Microbiology

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The abuse of antibiotics and the emergence of drug-resistant bacteria aggravate the problem of food safety. Finding safe and efficient antibiotic substitutes is an inevitable demand for ensuring the safety of animal-derived food. Bacteriophages are a kind of virus that can infect bacteria, fungi or actinomycetes. They have advantages of simple structure, strong specificity and nontoxic side effects for the human body.Bacteriophages can not only differentiate live cells from dead ones but also detect bacteria in a viable but nonculturable state. These characteristics make bacteriophages more and more widely used in the food industry. This paper describes the concept and characteristics of bacteriophages, and introduces the application of bacteriophages in preharvest production, food processing, storage and sales. Several methods of using bacteriophages to detect foodborne pathogens are listed. Finally, the advantages and limitations of bacteriophages in the food industry are summarized, and the application prospect of bacteriophages in the food industry is discussed.

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Viability Detection of Foodborne Bacterial Pathogens in Food Environment by PMA-qPCR and by Microscopic Observation

Cited by 16 publications

References 14 publications

Method-Dependent Implications in Foodborne Pathogen Quantification: The Case of Campylobacter coli Survival on Meat as Comparatively Assessed by Colony Count and Viability PCR

Method-Dependent Implications in Foodborne Pathogen Quantification: The Case of Campylobacter coli Survival on Meat as Comparatively Assessed by Colony Count and Viability PCR

Coupled DNA-labeling and sequencing approach enables the detection of viable-but-non-culturable Vibrio spp. in irrigation water sources in the Chesapeake Bay watershed

The application and research progress of bacteriophages in food safety

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