Luxin Wang

Survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on inoculated almonds and pistachios stored at -19, 4, and 24° C.

The survival of Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes was determined on almonds and pistachios held at typical storage temperatures. Almond kernels and inshell pistachios were inoculated with four- to six-strain cocktails of nalidixic acid-resistant Salmonella, E. coli O157:H7, or L. monocytogenes at 6 log CFU/g and then dried for 72 h. After drying, inoculated nuts were stored at -19, 4, or 24°C for up to 12 months. During the initial drying period after inoculation, levels of all pathogens declined by 1 to -log CFU/g on both almonds and pistachios. During storage, moisture content (4.8%) and water activity (0.4) of the almonds and pistachios were consistent at -19°C; increased slowly to 6% and 0.6, respectively, at 4°C; and fluctuated from 4 to 5% and 0.3 to 0.5 at 24°C, respectively. Every 1 or 2 months, levels of each pathogen were enumerated by plating; samples were enriched when levels fell below the limit of detection. No reduction in population level was observed at -19 or 4°C for either pathogen, with the exception of E. coli O157:H7-inoculated almonds stored at 4°C (decline of 0.09 log CFU/g/month). At 24°C, initial rates of decline were 0.20, 0.60, and 0.71 log CFU/g/month on almonds and 0.15, 0.35, and 0.86 log CFU/g/month on pistachios for Salmonella, E. coli O157:H7, and L. monocytogenes, respectively, but distinct tailing of the survival curves was noted for both E. coli O157:H7 and L. monocytogenes.

A Framework for Developing Research Protocols for Evaluation of Microbial Hazards and Controls during Production That Pertain to the Application of Untreated Soil Amendments of Animal Origin on Land Used To Grow Produce That May Be Consumed Raw

Application of manure or soil amendments of animal origin (untreated soil amendments; UTSAs) to agricultural land has been a long-standing practice to maintain or improve soil quality through addition of organic matter, nitrogen, and phosphorus. Much smaller quantities of these types of UTSAs are applied to land used for food crops than to land used for animal grain and forage. UTSAs can harbor zoonotic enteric pathogens that may survive for extended periods after application. Additional studies are needed to enhance our understanding of preharvest microbial food safety hazards and control measures pertaining to the application of UTSAs especially for land used to grow produce that may be consumed raw. This document is intended to provide an approach to study design and a framework for defining the scope and type of data required. This document also provides a tool for evaluating the strength of existing data and thus can aid the produce industry and regulatory authorities in identifying additional research needs. Ultimately, this framework provides a means by which researchers can increase consistency among and between studies and facilitates direct comparison of hazards and efficacy of controls applied to different regions, conditions, and practices.

Application of a novel antimicrobial coating on roast beef for inactivation and inhibition of Listeria monocytogenes during storage

The antilisterial efficacy of novel coating solutions made with organic acids, lauric arginate ester, and chitosan was evaluated in a three-stage study on inoculated roast beef for the first time. Ready-to-eat roast beef was specially ordered from the manufacturer. The meat surface was inoculated with five-strain Listeria monocytogenes cocktail inoculums at two different levels, ~3 and 6 Log CFU/cm(2) and treated with the stock solution (HAMS), the 1:5 diluted solution (MAMS), and the 1:10 diluted solution (LAMS) (stage 1). During the 20 min contact time, the antimicrobial coatings reduced the Listeria populations by approximately 0.9-0.3 Log CFU/cm(2). The higher the concentrations of the antimicrobial solution, the better the antilisterial effects were. The treated inoculated beef samples were then stored at 4 °C for 30 days. During storage, Listeria growth inhibition effects were seen. While no growth was seen from the HAMS-treated samples, a 1.6 Log CFU/cm(2) increase was seen for MAMS-treated samples, a 4.6 Log CFU/cm(2) increase was seen for LAMS-treated samples, and a 5.7 Log CFU/cm(2) increase was seen for NoAMS-treated samples on Day 30 (~3 Log CFU/cm(2) inoculation level). In the second stage, the impact of the roast beef storage time on solutions antilisterial effect was evaluated. Results showed that the effect of the antimicrobial solution was dependent on both the initial inoculation levels and storage times. In stage 3, the effect of the antimicrobial solution on roast beef quality was studied with both instrument measurement and sensory evaluation. Minor changes in color, pH, and water activity were found. However, only limited sensory differences were seen between the treated and untreated samples. When panels were able to accurately find color differences between samples, they preferred the treated samples. The findings of this research proved the antilisterial efficacy of the novel antimicrobial solution and showed its potential for being used as a roast beef cut surface coating to control Listeria contamination and for color protection.

Evaluating best practices for Campylobacter and Salmonella reduction in poultry processing plants

Poultry processing plants in the United States were surveyed on their current Campylobacter and Salmonella control practices. Following surveys, data were collected to develop a baseline for prevalence rates of Salmonella and Campylobacter; then changes in practices were implemented and evaluated for improvements in pathogen control. Surveys were sent to the plant Quality Assurance managers to determine production levels, antimicrobial interventions, and current pathogen testing practices. Initial sampling was performed at 6 plants with similar production volumes, at sites that included carcass samples before any pre-evisceration intervention, after exiting the inside-outside bird washer (IOBW), after exiting the pre-chiller, after exiting the primary chiller, and after exiting any post-chill intervention, as well as a water sample from each scalder, pre-chiller, primary chiller, and post-chill dip tank or finishing chiller. Enumerations and enrichments were performed for Campylobacter and Salmonella. Following the baseline sampling, changes in practices were suggested for each plant and a second sampling was conducted to determine their effectiveness. Results demonstrated that peracetic acid (PAA) was the most effective (P < 0.05) antimicrobial currently in use. The use of a post-chill antimicrobial immersion tank and/or use of a cetylpyridinium chloride (CPC) spray cabinet also displayed a further reduction in microbial levels (P < 0.05) when the primary chiller was not sufficient (P > 0.05). Microbial buildup in the immersion tanks demonstrates the need for effective cleaning, sanitation practices, and chiller maintenance to reduce contamination of poultry with Campylobacter and Salmonella.

Optimizing application parameters for lactic acid and sodium metasilicate against pathogens on fresh beef, pork and deli meats.

Lactic acid (LA) and sodium metasilicate (SM) have been approved for use as antimicrobials on meat. The objectives were to determine optimum concentrations, temperatures and hot-water dips of LA and SM for reduction of Escherichia coli O157:H7, non-O157 Shiga-toxin producing E. coli (STEC), Salmonella spp., and Listeria monocytogenes on beef, pork and deli meats. LA was applied at 1, 2, 3, and 4% and SM was applied at 2, 3, 4, and 5%. SM4 and LA4 were the lowest concentrations most effective against all pathogens. LA4 and SM4, the combination of the two (LASM), and distilled water control were applied at 4, 25, and 60°C. Temperature of application had no effect on pathogens. LA or SM alone were more effective in reduction of pathogens than LASM. Regardless of anti-microbial used in post-packaging lethality treatments, there were no differences in L. monocytogenes. Treating deli meats with LA or SM did not reduce L. monocytogenes. Both LA and SM can be applied to fresh beef and pork to decrease pathogens.

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

ABSTRACT 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 log10 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 log10 CFU/g for storage at 0, 4, and 10°C, respectively. Bacterial inactivation rates (IRs) estimated with the TMs (−0.245, −0.152, and −0.121 log10 CFU/day, respectively) were higher than those estimated with the MMs (−0.134, −0.0887, and −0.0732 log10 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 low-temperature 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.

Characterization and Survival of Environmental Escherichia coli O26 Isolates in Ground Beef and Environmental Samples

In addition to Escherichia coli O157:H7, shiga toxin-producing E. coli (STEC) O26 was added to the zero-tolerance adulterant list together with other 5 non-O157 STEC serogroups in 2012. Four farm O26 isolates were used in this study; they were obtained from a on-farm survey study conducted in Alabama. The presence of 3 major pathogenic genes (stx1, stx2, and eaeA) was determined through multiplex polymerase chain reaction (PCR). Two major pathogenic gene profiles were observed: 3 of the farm isolates contain only the eaeA gene whereas 1 farm isolate has both the eaeA and the stx1 genes. No significant difference was seen among the 4 farm isolates in the antibiotic resistance tests. To test their survival in ground beef and environmental samples, 2 inoculums were prepared and inoculated at various concentrations into samples of ground beef, bovine feces, bedding materials, and trough water. One inoculum was made of 3 farm isolates containing only the eaeA gene and another inoculum contained the isolate with both the eaeA and stx1 genes. Inoculated beef samples were stored at 4 °C for 10 d and the inoculated environmental samples were stored at ambient temperature for 30 d. Results showed that virulence gene profiles do not have an impact on O26s ability to survive in ground beef and in environment (P > 0.05). The inoculation levels, sample types as well as the storage times are the major factors that impact O26 survival (P < 0.05).