Simultaneous construction of PCR-DGGE-based predictive models of <i>Listeria monocytogenes</i>
 and <i>Vibrio parahaemolyticus</i>
 on cooked shrimps

Liao, Chao; Peng, Zhiyuan; Li, J.B.; Cui, Xiaowen; Zhang, Z.H.; Malakar, Pradeep K.; Zhang, W.J.; Pan, Yingjie; Zhao, Yong

doi:10.1111/lam.12376

Cited by 10 publications

(12 citation statements)

References 22 publications

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“…A f provides the accuracy of the model, which reflects how close the predicted values are to the observed values, while B f indicates the mean difference between observed and predicted value ). The range of 1.0≤ Bf≤Af≤1.1 was defined as a satisfactory limit in this study (Ye et al 2013;Liao et al 2015).…”

Section: Determination Of Behavior Of Two Pathogens In Raw Shrimp Stomentioning

confidence: 99%

Quantifying viable Vibrio parahaemolyticus and Listeria monocytogenes simultaneously in raw shrimp

Zhang

Liu

Lou

et al. 2015

Appl Microbiol Biotechnol

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A novel TaqMan-based multiplex real-time PCR method combined with propidium monoazide (PMA) treatment was firstly developed for the simultaneous quantification of viable Vibrio parahaemolyticus and Listeria monocytogenes in raw shrimp. The optimization of PMA concentration showed that 100 μM was considered optimal to effectively inhibit 10(8) CFU/mL dead cells of both bacteria. The high specificity of this method was confirmed on tests using 96 target and non-target strains. The optimized assay could detect as low as 10(1)-10(2) CFU/g of each strain on the artificially contaminated shrimp, and its amplification efficiencies were up to 100 and 106 % for V. parahaemolyticus and L. monocytogenes, respectively. Furthermore, this assay has been successfully applied to describe the behavior of these two pathogens in raw shrimps stored at 4 °C. In conclusion, this PMA TaqMan-based multiplex real-time PCR technique, where the whole procedure takes less than 5 h, provides an effective and rapid tool for monitoring contamination of viable V. parahaemolyticus and L. monocytogenes in seafood, improving seafood safety and protecting public health.

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Section: Determination Of Behavior Of Two Pathogens In Raw Shrimp Stomentioning

confidence: 99%

Quantifying viable Vibrio parahaemolyticus and Listeria monocytogenes simultaneously in raw shrimp

Zhang

Liu

Lou

et al. 2015

Appl Microbiol Biotechnol

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“…For instance, TCBS agar is widely used to enumerate V. parahaemolyticus colonies and to monitor their growth and survival without enrichment (7). These methods do not count cells in VBNC and stressed states (32) and may underestimate bacterial levels (4,13). In addition, the presence of background bacteria may negatively impact the enumeration efficiency of the plating media (17).…”

Section: Discussionmentioning

confidence: 99%

“…One-way ANOVA was applied using OriginPro 8.0 software (OriginLab Corporation, Northampton, MA, USA) to compare the different predictive models. P Ͻ 0.05 was regarded as representative of a statistically significant difference between models (4,13,29).…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, predictive traditional models (TMs) based on plate counting methods may underestimate the populations of bacteria (15,16). With the development of molecular quantitative methods, predictive models based on real-time PCR and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) methods have been reported to describe the survival and growth of Listeria monocytogenes and V. parahaemolyticus in food matrixes (13,17). However, all of these molecular methods are based on DNA, which can be extracted from both live and dead bacteria (13,18,19).…”

mentioning

confidence: 99%

“…culture the bacteria in a viable but a nonculturable (VBNC) or stressed state (13,14). VBNC pathogenic bacteria are considered a threat to public health and food safety because they retain their viability and ability to express their virulence (15).…”

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confidence: 99%

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Establishment and Validation of RNA-Based Predictive Models for Understanding Survival of Vibrio parahaemolyticus in Oysters Stored at Low Temperatures

Liao

Yong

Wang

2017

Appl Environ Microbiol

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This study developed RNA-based predictive models describing the survival of Vibrio parahaemolyticus in Eastern oysters (Crassostrea virginica) during storage at 0, 4, and 10°C. Postharvested oysters were inoculated with a cocktail of five V. parahaemolyticus strains and were then stored at 0, 4, and 10°C for 21 or 11 days. A real-time reverse transcription-PCR (RT-PCR) assay targeting expression of the tlh gene was used to evaluate the number of surviving V. parahaemolyticus cells, which was then used to establish primary molecular models (MMs). Before construction of the MMs, consistent expression levels of the tlh gene at 0, 4, and 10°C were confirmed, and this gene was used to monitor the survival of the total V. parahaemolyticus cells. In addition, the tdh and trh genes were used for monitoring the survival of virulent V. parahaemolyticus. Traditional models (TMs) were built based on data collected using a plate counting method. From the MMs, V. parahaemolyticus populations had decreased 0.493, 0.362, and 0.238 log 10 CFU/g by the end of storage at 0, 4, and 10°C, respectively. Rates of reduction of V. parahaemolyticus shown in the TMs were 2.109, 1.579, and 0.894 log 10 CFU/g for storage at 0, 4, and 10°C, respectively. Bacterial inactivation rates (IRs) estimated with the TMs (Ϫ0.245, Ϫ0.152, and Ϫ0.121 log 10 CFU/day, respectively) were higher than those estimated with the MMs (Ϫ0.134, Ϫ0.0887, and Ϫ0.0732 log 10 CFU/day, respectively) for storage at 0, 4, and 10°C. Higher viable V. parahaemolyticus numbers were predicted using the MMs than using the TMs. On the basis of this study, RNA-based predictive MMs are the more accurate and reliable models and can prevent false-negative results compared to TMs.IMPORTANCE One important method for validating postharvest techniques and for monitoring the behavior of V. parahaemolyticus is to establish predictive models. Unfortunately, previous predictive models established based on plate counting methods or on DNA-based PCR can underestimate or overestimate the number of surviving cells. This study developed and validated RNA-based molecular predictive models to describe the survival of V. parahaemolyticus in oysters during lowtemperature storage (0, 4, and 10°C). The RNA-based predictive models show the advantage of being able to count all of the culturable, nonculturable, and stressed cells. By using primers targeting the tlh gene and pathogenesis-associated genes (tdh and trh), real-time RT-PCR can evaluate the total surviving V. parahaemolyticus population as well as differentiate the pathogenic ones from the total population. Reliable and accurate predictive models are very important for conducting risk assessment and management of pathogens in food.

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Reduction of infection risk mediated by co‐culturing Vibrio parahaemolyticus and Listeria monocytogenes in refrigerated cooked shrimp

Niu

Xiao

et al. 2018

J Sci Food Agric

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Simultaneous construction of PCR-DGGE-based predictive models of Listeria monocytogenes and Vibrio parahaemolyticus on cooked shrimps

Cited by 10 publications

References 22 publications

Quantifying viable Vibrio parahaemolyticus and Listeria monocytogenes simultaneously in raw shrimp

Quantifying viable Vibrio parahaemolyticus and Listeria monocytogenes simultaneously in raw shrimp

Establishment and Validation of RNA-Based Predictive Models for Understanding Survival of Vibrio parahaemolyticus in Oysters Stored at Low Temperatures

Reduction of infection risk mediated by co‐culturing Vibrio parahaemolyticus and Listeria monocytogenes in refrigerated cooked shrimp

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