Food Safety, Great Plains Institute of

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Research from the Great Plains Institute of Food Safety. The institute website may be found at https://www.ag.ndsu.edu/foodsafety

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Analysis of Deoxynivalenol and Deoxynivalenol-3-glucoside in Wheat
(North Dakota State University, 2012) Burgess, Kimberly
Deoxynivalenol (DON), a mycotoxin produced in cereal grains infected by Fusarium Head Blight produced by Fusarium graminearium and Deoxynivalenol-3-β-D-glucopyranoside (DON-3G), were studied during processing using LC-MS-MS and GC. DON reduced significantly (P<0.05) 61.8% during milling into flour. Therefore, DON was concentrated mostly in the bran and germ. DON increased 40.8% during the fermentation stage of baking. DON increased in dough more than flour and mixed dough. Milling reduced by 23.7% but fermentation did not. But bread was significantly lower in DON-3G at 0.15 ppm than flour and dough at 0.31 ppm. The baking increased DON and decreased DON-3G showing a difference in stability of the mycotoxins during processing. Enzyme hydrolysis on DON using α-amylase, cellulase, protease, and xylanase, showed a significant increase with cellulase (20.8%), protease (11.4%), and xylanase (35.6%) compared to wheat composite. DON may be bound to the cell wall or protein component of the kernel.
Antimicrobial Resistance and Presence of Integrons in Salmonella Isolated from Animals and Humans in the United States of America and Uganda
(North Dakota State University, 2010) Mahero, Michael Wandanje
Salmonella has been cited as one of the leading causes of food borne illness world wide and in the United States (US), as well as an indicator organism for studying antimicrobial resistance (AMR) trends. The objective of this study was to characterise AMR patterns of Salmonella isolates from animals and humans in North Dakota, US, and Kampala, Uganda, and determine the association between the observed AMR and presence of class 1 integrons. Salmonella isolates were collected from the Veterinary Diagnostic Laboratory (VDL) at North Dakota State University and the North Dakota Department of Health respectively from 2003-2008. Samples were also retrieved from archives present at the Microbiology Department, Faculty of Veterinary Medicine at Makerere University in Kampala, Uganda. AMR profiles were determined using a panel of 15 antimicrobials as per the manufacturer's instructions (Sensitire, Trek Diagnostics System, Westlake, Ohio). Screening for the class 1 integrons was done using PCR with primers specific for the int 1. Out of 359 Salmonella isolates tested, 24. 79% were resistant to ~5 antimicrobials while 36.2% were resistant to at least 2. Pan susceptible isolates were mostly (65.05%) from human isolates. The most common multidrug resistant (MDR) phenotype among the tested isolates was the classic ACSSuT penta-resistance at 29.06% (50/172). The highest resistance frequency was seen against Tetracycline (39.6%) and Streptomycin (34.7 %), while 5.2% (17) of the isolates were resistant to Nalidixic acid and 56 (15.7%) to Ceftiofur. A total of20.7% (57/276) of the ND samples tested positive for presence of class 1 integrons. Class 1 integron was significantly associated (p< 0.05) with AMR to Ampicillin, Kanamycin, Tetracycline, Streptomycin and Sulfisoxazole. Of all Ugai.dan Salmonella isolates tested, 94.4% (68/72) were resistant to 2':2 antimicrobials. The highest resistance was observed against Sulfisoxazole and Trimethoprim-Sulphamethoxazole, and 45.8% of human and 46.2% of cattle isolates tested positive for presence of class 1 integrons. Presence of class 1 integron was significantly associated (p< 0.05) with AMR to Tetracycline and Amoxicillin. DNA sequencing of the class 1 integron variable regions identified several resistance genes including aadAJ, dfrA 7, and dfrA5 gene. The data indicated high AMR among antimicrobials widely used in veterinary and human medicine. Also, AMR was observed against drugs whose veterinary use is restricted, implying possible horizontal transmission. A good proportion (47.9% in Uganda and 29.85% in ND) of the Salmonella isolates from clinical cases of salmonellosis were MDR (resistant to 2':2) isolates bearing class 1 integron.
Assessing Microbial Stability and Quality of Green Beans Using Various Home Canning Methods
(North Dakota State University, 2013) Kuchynski, Jenny
Today many consumers follow processing methods recommended either from family members or the internet, which they interpret as being safe. Processing temperature profiles, survival of B. stearothermophilus spores, texture, and color of green beans processed under four home canning methods were assessed. The products were processed using pressure, boiling water bath, steam, or oven canning methods. Pressure canning produced the greatest microbial reductions but this method resulted in the lowest bean quality. The boiling water bath, steam, and oven canning were found to be less safe because the product temperature never achieved 100°C and the resulting microbial counts, >1.7 log CFU/ml, were observed after processing. However, green bean quality was better than pressure canning, with beans from steam canning having the firmest texture and best green color. Although better green bean quality results were observed from internet or family based methods, their safety is questionable considering the high microbial survival.
Compensation for the Effectiveness of Risk Mitigation Strategies for Fusarium Head Blight(FHB) and Deoxynivalenol(DON)
(North Dakota State University, 2019) Lyonga, Nathalie Ndedi
Food safety related problems are one of the biggest challenges worldwide. DON is produced by Fusarium species which causes the well-known Fusarium Head Blight (FHB) of wheat and barley. FHB outbreaks have led to variability in yield and revenue losses over the years. The main objective of my thesis was to quantify risk premiums at the farm level and with industry impact, to determine the effectiveness of FHB/DON mitigation strategies over time from 1997 to 2014. Data on revenue losses ($million) were obtained from USDA-ERS and was simulated using a risk analysis software called @RISK 7.5. The sample data was simulated 10,000 times to obtain a population. Risk premiums were calculated for each year and for each crop over time and graphs were plotted. Trends in risk premiums showed an overall decrease from 1997 to 2014, indicating that variability of losses have reduced and that the management practices have been effective.
Detection and Molecular Typing of Methicillin-Susceptible Staphylococcus Aureus (MSSA) and Methicillin-Resistant Staphylococcus Aureus (MRSA)
(North Dakota State University, 2015) Velasco, Valeria
Methicillin-resistant (MRSA) and multidrug-resistant (MDR) Staphylococcus aureus, and the serotype (ST) 398 have been associated with human and livestock infections, being also detected in retail meat. The aim of this study was to determine the prevalence and molecular types of S. aureus strains from animals, retail raw meat, deli meat, and humans, determining the genetic similarity between the strains. A two-step selective enrichment followed by selective plating were used to isolate S. aureus from animals (n=167), retail raw meat (n=145), and deli meat (n=46). In addition, S. aureus from healthy people (n=550) was isolated by culture method. Positive isolates and MRSA isolates from clinical cases (n=108) were subjected to multiplex PCR (16S rRNA, mecA, and PVL genes), molecular typing and antimicrobial susceptibility testing. In addition, a real-time PCR assay was developed in order to decrease the time of detection of target genes of S. aureus in animal and meat samples, comparing the results with the standard culture/PCR method. The prevalence of S. aureus was 34.7% in animals, 47.6% in meat, and 13.0% in deli meat. The mecA gene was detected in S. aureus isolated from five pork meat samples and exhibited penicillin resistance. The ST398 was found in sheep, pigs, and pork meat. The S. aureus nasal carriage in healthy people was 7.6%. A total of 105 MRSA strains (97.2%) from clinical cases harbored the mecA gene and 11 (10.2%) the PVL gene. The rate of MDR was 70% in humans. A genetic similarity between strains from animals and meat, and from humans and meat was observed. Total agreement between the culture/PCR method and real-time PCR for detection of S. aureus was 68.9 to 97.8% (k=0.68-0.88), and the mecA gene, 86.7 to 98.7% (k=0-0.49). Therefore, the real-time PCR assay may be recommended as a rapid method for the detection of S. aureus, with confirmation of MRSA using the standard culture method. The presence of emerging S. aureus strains in the meat production chain and the genetic similarity between strains of different origin, suggests the contamination of meat, and a potential risk of transmission to humans.
Development of Real-Time PCR Method for Detection, Identification and Quantification of Five Different Fusarium Species
(North Dakota State University, 2010) Goswami, Kakolie
Fusarium head blight (FHB) is a fungal disease affecting cereal crops worldwide. FHB involves multiple Fusarium species that create food safety concerns by producing mycotoxins such as trichothecenes, zearalenone and moniliformin. Quantitative methods allowing rapid risk assessment of mycotoxigenic Fusarium species are needed to detect Fusarium spp. Real time quantitative PCR (qPCR) is a fast, sensitive and reliable alternative to conventional culture methods. An objective of the study was to develop TaqMan® based qPCR methods to detect, identify and quantify five different Fusarium species associated with FHB, namely F. graminearum, F culmorum, F avenceum, F poae and F sporotrichioides. The project was initiated with the selection of a protocol for Fusarium DNA extraction. Three different commercially available DNA extraction kits, including FastDNA ®, Qiagen® Blood & Tissue kit and Qiagen® DNeasy Plant kit were evaluated for speed, DNA yield and quality. The results showed that FastDNA ® kit gave the highest DNA yield in least time. TaqMan® Minor Groove Binder (MGB) probes for F culmorum, F poae and F sporotrichioides were developed using qPCR primers from a previous study. For F avenaceum and F. graminearum, TaqMan® based methods already available were used with modified conditions for improved detection. The qPCR methods were tested on a wheat system and the result was a qPCR system that rapidly identified and quantified Fusarium species associated with FHB.
Does Consumption of Beef from Cattle Administered Growth-Enhancing Technology Trigger Early Estrus in Pre-Pubertal Gilts?
(North Dakota State University, 2012) Anderson, Giovana Maranho
The objective was to determine if pre-pubertal gilts supplemented ground beef obtained from steers implanted with growth enhancing technology caused precocious puberty. Twenty-four gilts were selected for the same birth date from common parentage. Upon reaching 61 days of age, daily delivery of the low-estrogenicity base diet was supplemented with: 114 g beef natural patty (NAT), 114 g beef patty from steers that had received growth promoting implants (100 mg trenbolone acetate and 14 mg estradiol benzoate; IMP), 198 g tofu patty (TOFU), or the negative control (base diet only; CON). The estradiol equivalents (ng/kg) of the TOFU were approximately 500 fold times the NAT and 350 fold the IMP supplement. No differences (P = 0.55) were observed in number of days to reach estrus, feed efficiency (P > 0.19), live weight gain (P = 0.89), loin muscle development (P = 0.45), or subcutaneous fat deposition (P = 0.71).
Escherichia Coli in Bovine Calf Scours
(North Dakota State University, 2015) Starr, Crystal
Scours is caused by inflammation of the intestinal tract of ruminants leading to significant mortality and morbidity rates. It is predominately found in neonatal ruminants where the disease can occur 36 hours after birth. One of the most common infectious agents linked to scours is pathogenic Escherichia coli. Therefore, it is important to understand the virulence factors, diagnostic assays, age of the animals infected, and the co-factors associated with an E. coli scours outbreak. These factors are important in both scours disease pathogenesis and potential food safety-related postharvest pathogens. Using the most frequently identified virulence factors, a new scours diagnostic assay could be created to detect and prevent disease in cattle. The present study determined that virulence factors astA, fimC, fimH, int1, int2, irp2, papC were identified over 15% percent of the time and could be implemented into a more specific multiplex PCR test for pathogenic E. coli.
Fate of Deoxynivalenol and Deoxynivalenol-3-Glucoside during the Malting Process
(North Dakota State University, 2015) Jiang, Wei
Deoxynivalenol (DON) is commonly found on small grains and causes food safety issues. Deoxynivalenol-3-Glucoside (DON-3-G) is a conjugate, formed as a defense response by the host plant. Past studies have shown both to be present in Fusarium infected small grains, and processed products like beer, but there is limited information on DON-3-G in malt. Objectives were to determine the levels of DON-3-G in barley and wheat, and to study its fate during malting of inoculated and commercial samples. Commercial barley and wheat samples were used to determine levels in naturally infected grain. During malting, barley DON declined 48% on average, but DON-3-G increased by 115%. Both compounds increased in malted wheat. The genotype x crop year interactions were significant for both toxins, indicating that the genotypes did not respond similarly in the two years. The potential for large amounts of DON-3-G to be formed during malting has not been reported.
Food Safety Culture: An Underlying Cause for Success and Failures of Food Safety Management Systems
(North Dakota State University, 2018) Vashisht, Ashwini Kumar
Food Safety Culture (FSC) is an emerging behavior-based food-safety management concept. FSC has been linked as an underlying cause for food-safety management-system failures during foodborne outbreaks and recall investigations. This paper reviews the available literature on FSC for the origin, definitions, factors, barriers, and dimensions that influence the FSC’s performance and measurements. Data were obtained from peer-reviewed journals as well as publicly available information on the World Health Organization (WHO), Food and Drug Administration (FDA), and Center for Disease Control and Prevention (CDC) websites. The roles of organizational leadership and communication, food-handler behaviors, risk perception, regulatory authorities, and technological advancements are evaluated for FSC development and enhancement. It can be concluded that there is a need for a strong FSC within food manufacturing and service organizations because it enhances the food-safety management systems’ performance and may also reduce the global burden of foodborne illnesses and diseases.
Foreign Material Identification and Removal in the Food Safety Industry
(North Dakota State University, 2018) Dumas, Daniel Richard
Changes in the food industry including the mass production in response to the globalization of the food chain have led to additional food safety and quality challenges for the food industry. One aspect of these challenges is the effective identification and removal of foreign materials from products. The development and implementation of new manufacturing processes, materials, and techniques can produce many types of foreign materials or contribute to the challenges to their removal. These challenges facing the food industry are requiring more levels of protection along with advances in technologies to help reduce the potential safety and quality concerns of foreign materials. Foreign material identification is important for consumer safety as well as product image and sales. The technologies available for these purposes will be reviewed as well as some traditional methods used as well as some of short comings or problems facing these technologies in the food industry.
Growth of Fusarium graminearum and Production of Trichothecenes During the Malting of Winter Rye and Triticale
(North Dakota State University, 2019) Tang, Ruoling
There is growing interest in malting and brewing with rye. However, previous research has shown a propensity for the development of deoxynivalenol (DON) in rye malts, even when levels on the grain is low. The main objective of this study was to assess the growth of F. graminearum and development of trichothecenes during malting of rye. Infected samples were obtained from 2016 variety trails in Minnesota. While DON levels were generally below 0.2 mg/kg, an average increase of 41 % was seen after malting. The most significant increases in DON were at three days of germination. Fusarium Tri5 DNA levels were observed to increase at two days. When single kernels were tested, most were free from DON. Levels in the bulk grain sample were due to a small number of highly contaminated kernels. In the malted samples, a greater portion of kernels contained DON, and overall levels were much higher.
High Pressure Processing and Nisin as Possible Nonthermal Treatments for Control of Listeria and Impact on Quality Attributes in Cold Smoked Pacific Sockeye Salmon
(North Dakota State University, 2018) Desrocher, Lisa Dawn
Cold smoked salmon products are considered high risk for Listeria monocytogenes contamination by the U.S. Food and Drug Administration due to lack of validated kill step. Currently, there are no commercialized post-packaging control measures to mitigate Listeria spp. in cold smoked salmon processing. Nisin applied during cold smoked salmon processing has been reported to reduce L. monocytogenes with no change in final product organoleptic properties. High pressure processing of cold smoked salmon post-packaging has also been reported to mitigate L. monocytogenes. However, the pressure and time required have adverse effects on cold smoked salmon such as lightening of flesh color. The effectiveness of nisin and high-pressure processing on the survival of Listeria innocua in cold smoked salmon was recently reported. The combination of Nisin and HPP was found to be more effective for controlling L. innocua than either treatment alone while maintaining desirable consumer attributes.
The Impact of Bacterial Spoilage and Foodborne Pathogens on Beef Industry and Application of Antimicrobial Interventions
(North Dakota State University, 2019) Khadem, Enas Abdal Hadi
The beef industry continues to face concerns regarding the hygiene and the safety of its products. A wide range of microorganisms from various sources can grow on meat surfaces that are rich in fluid and nutrients. This paper was conducted to better understand the common spoilage microflora and the most threatening foodborne pathogens (E. coli O157:H7 and Salmonella spp.) in ground beef and the role of the virulence factors that allow pathogens to persist in the host. In addition to the above, this paper addresses the effects of using antimicrobial interventions on the ground beef products. Despite using innovative antimicrobial interventions to eliminate or reduce spoilage bacteria and common foodborne pathogens, there is still a need for new antimicrobial technologies to control the industry’s sanitary hurdles and to understand their affects on meat quality and sensory characteristics.
An In-vitro Co-Culture Model to Study the Disruption of the Blood Brain Barrier by Cronobacter sakazakii. (formerly Enterobacter sakazakii)
(North Dakota State University, 2010) Koval, Erin Louise
Cronobacter spp. (formerly Enterobacter sakazakii) is an opportunistic pathogen associated with contaminated powdered infant formula. It causes necrotizing enterocolitis (NEC) and sepsis, which can develop into severe meningitis and brain abscess formation in infants. Very little is known about the specific pathogenic mechanisms of this organism. In this study, we propose a model to investigate the ability of Cronobacter spp. isolates to disrupt the blood-brain barrier. Biofilm formation of the Cronobacter spp. isolates used in the model was also assayed. Secondary mouse endothelial and astrocyte cells were grown to con.fluency on polyethylene terephtalate (PET) membranes (1.0 μm pore size) in 24-well plate hanging culture inserts (Millipore® Billerica, MA) to achieve a culture system similar to the physiological structure of the blood brain barrier in vivo. Initially, a monoculture system was tested containing only endothelial cells and then a co-culture system was developed with both cell types. Selected Cronobacter spp. isolates were added to the cell culture systems. Escherichia coli KI and Kl2 strains were used as positive and negative controls, respectively. Some of the treatments did not have bacteria added to them to serve as cell controls, and membranes without cells were included as media control blanks. The transendothelial electrical resistance (TEER) was measured across the cells to determine if the cell barrier was disrupted. Initially, measurements were taken at 0, 6, 24, and 48 hours after adding bacteria. Due to overall loss of cell integrity at 48 hours, a second experiment was performed where measurements were taken at 0, 6, and 24 hours after adding the bacteria. Biofilm formation was analyzed using a method described by O'Toole (72) and Skyberg (86). The monoculture and co-culture systems worked based on TEER measurements. The positive control (E.coli Kl) and Cronobacter sakazakii isolates disrupted the tight junctions between cells evidenced by significant decrease in TEER over time. The negative control (E.coli K12) did not have any significant effect on the cells. The cell control and negative control (E.coli Kl2) maintained the highest resistance values in both the monoculture and co-culture experiments. The positive control (E. coli Kl) and the Cronobacter sakazakii isolates caused the resistance across the cells to significantly decrease over time in both experiments. According to the results of our biofilm assay, none of Cronobacter spp. isolates used in the culture models formed biofilms. Further testing would need to be done using other biofilm detection procedures to confirm this conclusion Overall, we successfully constructed a co-culture model to depict the BBB. The selected Cronobacter sakazakii decreased resistance in this model of the BBB similar to the positive control Kl E.coli isolate. This assay can be used in future studies to test the potential pathogenicity of Cronobacter spp. bacteria as well as other bacteria involved with central nervous system diseases.
Listeria Monocytogenes in the Retail Food Service Environment
(North Dakota State University, 2018) Walpuck, David Andrew
Listeria monocytogenes is one of the biggest microbial concerns affecting today’s food industry. It is a ubiquitous liability with a high mortality rate, unique characteristics of growth and survival in many environmental conditions, making the pathogen a true risk to consumer health. Recent outbreaks of listeriosis have caused fatalities, massive well-publicized recalls costing the food industry heavy financial losses and damaged reputation. L. monocytogenes is the forefront of impediment and educational efforts from private industry and government agencies. The purpose of this study is to assess the features and concern of L. monocytogenes in the retail food service environment and its impact on operations. Regulatory surveillance of testing environmental samples and food products for L. monocytogenes highlight prevention. Organizations in the retail food service industry need a separate plan for training, food handling, sanitation and financial allocation to combat the potential threat of L. monocytogenes contamination.
Occurrence of Salmonella and Listeria Monocytogenes in Ready to Eat Meats in the United States: 2000-2010
(North Dakota State University, 2012) Ntaate, Anthony Wilfred
The purpose of this review is to characterize the occurrence of Salmonella and Listeria monocytogenes in ready to eat (RTE) meats in the United States between the years 2000-2010. Data were obtained from the CDC foodborne outbreak online database, morbidity and mortality weekly reports, summary of notifiable diseases and the foodborne outbreaks page. Additional information was obtained from peer reviewed journals. RTE roast pork, turkey deli meat, and Italian type salami meats were the vehicles in the Salmonella outbreaks reported. Half of the eight outbreaks reviewed were multistate in nature affecting many states and the rest were sporadic. The Salmonella serotypes isolated were Salmonella Uganda, Salmonella Hadar, Salmonella Montevideo, whereas the L. monocytogenes serotypes were 1/2a and 4b. The major risk factors for listeriosis and salmonellosis outbreaks were being elderly and having an underlying immunocompromising medical condition. Pregnant women were particularly at risk for listeriosis.
The Occurrence of Shiga-Toxin Producing Escherichia Coli (Stec) and Salmonella Species in Cattle Feedlot Runoff
(North Dakota State University, 2013) Tabe, Nessie Nanyongo
Zoonotic foodborne pathogens such as shiga-toxin producing Escherichia coli (STEC) and Salmonella on farm environments can potentially contaminate organic manure or agricultural watersheds and subsequently fresh produce during fertilization or irrigation. This study investigated the occurrence of STEC and Salmonella serotypes in cattle feedlot runoff samples in two feedlots in North Dakota. Using standard laboratory culture methods this study reported a 39% prevalence of STEC O45, 33 % (O103), 31 % (O157), 27 % (O121), 16 % (O26), 10% (O111), 10% (O113), 10 % (O145) and 39.7 % Salmonella. Additionally, occurrence of some STEC serotypes was influenced by feedlot (O111 and O121), sampling location in relation to vegetative filter strips (O157), and sampling time (O45 and O121). Although this study was the first to report occurrence of STEC serotypes including non-O157 serotypes in cattle feedlot runoff, further studies are needed to quantify the pathogen load in runoff prior to disposal.
Ochratoxin A and Ochratoxigenic Fungi in Freshly Harvested and Stored Barley and Wheat
(North Dakota State University, 2014) Kuruc, Julie
Ochratoxin A (OTA) is a toxin produced both prior to harvest and during storage by Penicillium and Aspergillus species in a variety of commodities. Although several studies have been conducted in Europe and Canada examining the occurrence and concentration of OTA in cereal grains, data is lacking for the United States, where guidance levels and regulations do not exist. This study aims to fill in the knowledge gaps surrounding OTA and ochratoxigenic fungi in barley and durum and hard red spring wheat grown in the northwestern and Upper Great Plains regions of the United States. In total 2.7% (n = 37) of the 1370 samples taken over 2 consecutive years had detectable levels of OTA (0.15-9.11 ng/g) directly after harvest. The number of positive samples was significantly greater in 2012 compared to 2011. This difference may be due to weather conditions during the planting and growing seasons or simply natural variation between years. Stored barley and wheat (N = 262) had a higher prevalence (12.2%) and greater range (0.16-185.24 ng/g) of OTA compard to freshly harvested samples. Although 81.3% of the OTA-positive samples had been stored for ≥6 months, samples that had been stored for as short as 1 month also tested positive. These results underline the importance of proper storage conditions in minimizing OTA contamination. P. verrucosum was found to be the primary ochratoxigenic species in these samples. Of the 110 isolates tested, 64.7% were confirmed OTA producers. Samples containing >1 ng/g OTA had significantly more OTA-producing P. verrucosum strains than samples with undetectable OTA. Infestation rate did not correlate with OTA level. Additionally, OTA concentration did not correlate with otanpsPN, an OTA biosynthesis gene. This indicates that the concentration of P. verrucosum in a sample may increase the likelihood of contamination but is not a reliable indicator of OTA level.
Presence of Escherichia Coli and Salmonella Enterica on Wheat and Possible Control Measures
(North Dakota State University, 2018) Anderson, Cassie Alexandra Palm
Wheat (Triticum spp.) is one of the most important cereal crops grown in the U.S., with an average of 50 million acres planted on an annual basis. Wheat is milled into flour or semolina, which is used to make bread, cookies, noodles, and pasta. Because some consumers eat raw flour, it is necessary for it to be free of pathogens including Escherichia coli and Salmonella enterica. The fecal matter of cattle and poultry often contains these bacteria and can contaminate wheat. Currently, there are no requirements for controlling E. coli and S. enterica in wheat, which has resulted in outbreaks of both pathogens. Thus, future research must focus on the development of processes that control these pathogens in wheat. Possibilities include feeding livestock probiotics, pasteurization, irradiation, and non-thermal plasma processing. The further development and implementation of these processes would decrease the safety risks associated with consuming raw wheat.

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