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Title: Effect of pH on desiccation survival of Salmonella
Creator: Chen, Fangyu
Date: 2019
Description: One intrinsic property of foods, pH, can affect Salmonella survival in high-moisture foods, but its effect in low-moisture foods is unknown....
Show moreOne intrinsic property of foods, pH, can affect Salmonella survival in high-moisture foods, but its effect in low-moisture foods is unknown. In this study, the effect of pH on desiccation and persistence of Salmonella was explored using two approaches. First, the pH range that affects survival in low-moisture environments was explored. The effect of acid adaptation in acidic low-moisture environments was also explored. Salmonella Anatum was grown on trypticase soy agar with 0.6% yeast extract (TSAYE). After harvest, cells were divided and one portion treated independently at pH 3, 4, 5, 7, and 8 for 30 min. Both portions were then desiccated on a cellulose filter in a biohazard cabinet (23±2°C) overnight (24±2 h). After desiccation, cells not previously pH treated were resuspended in buffers at pH 3, 4, 5, 7, and 8 for 30 min, and cells previously pH treated were resuspended in buffered peptone water (BPW). All suspensions were plated on TSAYE with ammonium iron citrate and sodium thiosulfate to determine surviving populations. In addition, S. Anatum was grown on TSAYE adjusted to pH 4, 5, 7, and 8. Cells were either harvested with buffer with the same pH of the growth media or with saline then treated at pH 4, 5, 7 and 8. All were desiccated as indicated before. Desiccated cells were stored at 20% RH at 25°C for up to 29 days. To determine the effect of prior acid adaptation on survival in acidic environments, Salmonella Anatum and Salmonella Agona were grown on agar with or without 1% glucose, harvested, then suspended in buffer at pH 4, 5, and 7. Each culture was desiccated on cellulose filters and stored at 30% RH at 25°C for up to 29 days. Harvested cells were also stored in buffers at the same pH held stored at 25oC for the same time periods. In addition, acid-adapted cells were harvested with saline, desiccated and stored as indicated for each type of cells on wheat flakes. Results from pH range finding experiments indicated pH did have an effect on the survival of Salmonella during desiccation. Desiccation prior to treatment will affect survival at different pH levels. However, prior pH adaptation did not result in increased survival under different pH conditions once cells were desiccated. Acid adaptation prior to desiccation at low pH adversely affected survival for S. Agona but not S. Anatum. Survival after desiccation at different pH levels was greater than survival in the same pH buffers. No advantage or differences in survival was observed with a commercial wheat flake product indicating results obtained in a model environment may be reduced or eliminated when food components are present.
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Title: CHARACTERIZATION OF NOVEL PULSED UV-LIGHT SYSTEMS FOR INACTIVATION OF LISTERIA MONOCYTOGENES IN APPLE JUICE AND ON APPLE SURFACE
Creator: Malik, Sargun
Date: 2018
Description: Pulsed light processing can effectively inactivate microorganisms from the surface of foods or in transparent liquid foods. Pulsed light...
Show morePulsed light processing can effectively inactivate microorganisms from the surface of foods or in transparent liquid foods. Pulsed light systems currently available in the market operate at a fixed pulse duration and frequency and might not be optimized for microbial inactivation. A novel pulsed light system (Model X 1100; Xenon Corporation, USA) enables the researchers to adjust various parameters including pulse duration (100-7000μsec), voltage(1000-3000V), frequency (0.1-20Hz),% of energy (0-100%), and energy (up to 9J /cm2 / pulse of optical energy or 2433J / pulse of electrical energy ) . This study evaluated the effect of various parameters (treatment time, voltage, frequency, energy / pulse) on inactivation of Listeria monocytogenes in buffered peptone water (BPW), apple juice, and apple surface. For liquids, a 4-mL of sample (4-mmdepth) artificially inoculated with Listeria monocytogenes was treated in a quartz Petri dish (5.5-cmdiameter). For solid food, the top surface (skin side) of a slice of apple (1×1×0.5cm）was inoculated and exposed to various pulsed light treatment conditions. The results indicated that the impact of these factors vary as many of these factors are inter-related. In general, increasing the frequency, input voltage, pulse duration, and percentage of energy, increased the microbial reduction at the tested conditions (p<0.05). For instance, reductions of 1.21and 5.47 log10 CFU/mL were obtained in BPW and reductions of 1.35 and 4.70 log10 CFU/mL was acquired in apple juice, at 0.1and 0.82Hz, respectively, for a 20-sec treatment at 2500V (50% energy,700 μsec pulse width). Increased energy per pulse resulted in increased microbial reduction. For example, reductions of 2.30, 5.59, 6.69, and 6.69 log10 CFU/mL were obtained at 645, 1241, 1837, and 2433J/ pulse of electrical energy, respectively, in apple juice. Similarly, reductions of 5.34, 6.45, 6.02, and 6.56 log10CFU/mL were obtained at 645, 1241, 1837, and 2433J/ pulse, respectively in BPW. Lower reduction was obtained from the skin surface of the apple, for instance, reductions of 0.70 and 1.19 log10 CFU/ slice were obtained at 0.10 and 0.82 Hz, respectively, after a 10 seconds treatment at 2500V (50%energy). Similarly, reductions of 2.44, 2.43, 3.39, and 3.48 log10 CFU/ slice were obtained at 645, 1241, 1837, and 2433J/ pulse of electrical energy, respectively, after a 15 seconds treatment at 3000V (0.2Hz). The results were similar to the pulsed light treatment with RC 800 system (Xenon Corporation, USA). Absorption of pulsed light energy resulted in temperature increase in the products. Temperature increase of up to 11°C was observed at the treated conditions. The results suggest that this novel pulsed light system can potentially be used for inactivation of Listeria monocytogenes.
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Title: Evaluation of Salmonella Proliferation on Alfalfa Sprouts during Storage at Different Temperatures
Creator: Lin, Chih Tso
Date: 2020
Description: Sprouts, a low-calorie vegetable rich in nutrition, have been a popular ingredient in many meals in the USA. They are grown either at...
Show moreSprouts, a low-calorie vegetable rich in nutrition, have been a popular ingredient in many meals in the USA. They are grown either at commercial sprout farms or at home and served raw or lightly cooked. However, sprouts are also known as a source of foodborne illness outbreaks. FDA Food Code identifies raw sprouts as a time/temperature control for safety food. However, little information is known about the growth profile of foodborne pathogens in sprouts stored at different temperatures. This study aimed at evaluating the proliferation of Salmonella in alfalfa sprouts during storage at 4, 10, and 25℃ under two different contamination routes: 1) sprouts that were inoculated with Salmonella after harvest and 2) sprouts that were grown from contaminated seeds. Alfalfa sprouts grown from uninoculated seeds and harvested after 5 days of sprouting were divided into 25-g portions. Each portion was inoculated with a cocktail of five Salmonella serovars at levels of 10^1, 10^3 or 10^5 CFU/g prior to storage at 4, 10, or 25℃. Alternatively, sprouts grown for five days from seeds spiked with 1% of seeds previously inoculated with the Salmonella cocktail were divided into 25-g portions and stored at 4, 10, or 25℃. At defined time points (Days 0, 2, 4, 7, 14, and 21), levels of Salmonella and background microflora in stored sprouts were determined by plate count. Alfalfa sprouts appeared fresh during the 21 days of storage at 4 or 10℃ but started to show signs of spoilage after 4 days of storage at 25℃. The total plate counts maintained at a level above 9 log CFU/g throughout 21 d of storage at 4 and 10℃ or during the first 7 d of storage at 25℃. Storing sprouts at 4 or 10℃ could inhibit the proliferation of Salmonella. After 21 d of storage, the Salmonella counts in inoculated sprouts decreased slightly, by 0.88 or 0.93 log units, respectively. For sprouts stored at 25℃, the Salmonella growth profile differed depending on the route of contamination and the level of Salmonella at the start of storage. In sprouts inoculated at levels of 1.41, 2.83, and 4.75 log CFU/g, the Salmonella counts increased to 6.62, 6.86, and 6.68 log units, respectively, during the first 4-7 days of storage. For alfalfa sprouts grown from contaminated seeds, the Salmonella counts remained at a level similar to that in the harvested sprouts (8.16 log CFU/g) during the first 7 d. Results from this study further the understanding of pathogen growth in sprouts and will aid in the development of guidelines for proper storage of sprouts.
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Title: GROWTH KINETICS OF LISTERIA MONOCYTOGENES DURING REHYDRATION OF DEHYDRATED PLANT FOODS AND SUBSEQUENT STORAGE
Creator: Wu, Zihui
Date: 2020
Description: Dehydrated plant foods do not support the growth of pathogenic bacteria. However, once rehydrated, the high-water activity and neutral pH of...
Show moreDehydrated plant foods do not support the growth of pathogenic bacteria. However, once rehydrated, the high-water activity and neutral pH of these foods may support the growth of pathogens, such as L. monocytogenes, during storage. The goal of this study was to examine the growth kinetics of L. monocytogenes during 5, 10, and 25°C storage on potatoes, carrots, and onions after rehydration with 5 or 25°C water. Fresh plant foods were dehydrated at 140°F (60°C) for 24 h. A 4-strain rifampicin-resistant L. monocytogenes cocktail was inoculated onto dehydrated plant foods at 4 log CFU/g and dried for 24 h. Plant foods were rehydrated in 4-volumes of 5 or 25°C water for 24 h. At various timepoints during rehydration, 30 g of sample was removed and drained for 10 min. Samples were homogenized 1:10 with BLEB and the homogenate was plated onto BHIRif for enumeration. After rehydration, samples were drained and portioned into deli-style containers for storage at 5, 10, and 25°C and L. monocytogenes was enumerated at 1, 3, 5, and 7 d. Triplicate samples were assessed at each timepoint and three independent trials were conducted. Growth rates were determined using DMFit and data were statistically analyzed using Student t-test (α=0.05). Overall, the growth rates of L. monocytogenes during storage of potatoes and carrots were higher when rehydrated with 5°C water compared to 25°C water. The highest growth rate on potatoes was 3.51±0.43 log CUF/g per d with 5°C water rehydration and 25°C storage, resulting in a 1 log CFU/g increase in 0.29 d (7.0 h). When rehydrated with 25°C water and 25°C storage, the growth rate was significantly lower at 1.03±0.01 log CFU/g per d. The highest growth rate of L. monocytogenes on carrots was 0.68±0.07 log CUF/g per day when rehydrated with 5°C water and 10°C storage, resulting in a 1 log CFU/g increase in 1.47 d (35.3 h). For onion, L. monocytogenes was below the level of enumeration during storage at 5°C for both water rehydration temperatures and also for 10°C storage with 5°C water rehydration. The highest growth rate was 0.46±0.11 log CFU/g per d, resulting in a 1 log CFU/g increase in 2.17 d. The results of this study can aid in determining appropriate time and temperature control for safety for dehydrated potatoes, carrots and onions during rehydration and subsequent storage.
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Title: RECIPROCAL INTERACTIONS BETWEEN RED RASPBERRY POLYPHENOLS AND GUT MICROBIOME COMPOSITION AND METABOLIC HEALTH
Creator: Zhang, Xuhuiqun
Date: 2020
Description: Red raspberries (RRB) and fructo-oligosaccharides (FOS) have been associated with reduced risk of developing cardio-metabolic diseases. RRB...
Show moreRed raspberries (RRB) and fructo-oligosaccharides (FOS) have been associated with reduced risk of developing cardio-metabolic diseases. RRB are uniquely high in anthocyanin- and ellagitannin- type (poly)phenols, however, these (poly)phenols have low bioavailability. Gut microbiota can improve (poly)phenol bioavailability through fermentation processes generating absorbable metabolites and altering gut microbiota structure. However, clinical evidence on the effects of RRB intake on the gut microbiome, (poly)phenolic metabolites and metabolic health is lacking. Further, fermentable carbohydrates (FOS) that selectively stimulate gut microbiota growth may enhance metabolite generation. Therefore, the aim of this research is to investigate the interactions between the gut microbiome and RRB, and explore added effects of FOS as a possible nutritional strategy for improving metabolic health of at risk individuals with prediabetes and insulin resistance (PreDM-IR). Through a series of investigations drawn from a randomized clinical trial (RCT), the following hypotheses were tested: 1) Individuals with PreDM-IR will have a distinctive gut microbiome, and lower capacity to metabolize (poly)phenols compared to healthy individuals; 2) RRB intake for 4-week will increase microbial-derived (poly)phenolic metabolites and adding FOS will augment the effect; 3) RRB intake will improve metabolic risk factors in PreDM-IR and adding FOS will augment the RRB effect; 4) RRB and RRB+FOS supplementations will alter the structure of the gut microbiome explaining variances observed in metabolites and metabolic outcomes. In this single-blinded, crossover RCT, adults with PreDM-IR (n=26) and a healthy group (n=10) consumed 1 cup RRB (fresh weight equivalence) per day or RRB with 8g FOS per day for 4 weeks in random order separated by 4-week washout. Metabolic risk factors, (poly)phenolic metabolites and metagenomic profile were assessed before and after supplementation. Baseline characterization before supplementation revealed distinctive metabolites and metagenomics profiles related to metabolic status. After 4-week RRB, microbial (poly)phenolic metabolites, metabolic health indices and gut microbiome structure beneficially shifted in PreDM-IR group. Adding FOS increased specific microbial species and phenolic metabolites that correlated with β-cell function in PreDM-IR. Overall, nutritional strategies incorporating RRB and FOS may improve metabolic health of individuals with PreDM-IR through modulating gut microbiome composition and the capacity to metabolize RRB (poly)phenols.
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Title: HERBS AND SPICES ON ENDOTHELIAL FUNCTION OVER 24 HOURS: A RANDOMIZED, CONTROLLED TRIAL
Creator: Thorat, Rajrajeshwari Sunil
Date: 2021
Description: Modern-day eating patterns are characterized by readily available carbohydrates and/or fats and have consistently been shown to disturb...
Show moreModern-day eating patterns are characterized by readily available carbohydrates and/or fats and have consistently been shown to disturb endothelial function. Recent investigations suggest herb and spice blends have beneficial effects in reducing inflammation and increasing endothelial function in humans. This study was designed to characterize the effect of herbs and spices on endothelial function as measured by flow-mediated dilation (FMD) over 24 h using a challenge meal paradigm. In a randomized, single-blinded, 4-arm, crossover trial, sixteen overweight/obese adults (BMI = 28.4 ± 2.5 kg/m2; age = 39 ± 15 years) consumed a high carbohydrate high-fat meal (≈ 41% Fat and ≈46% Carbohydrate of total Kcal) with or without the spices combinations, including Italian herbs (rosemary, basil, thyme, oregano, and parsley), cinnamon, and pumpkin pie spice (cinnamon, ginger, nutmeg, and allspice) on four separate days at least three days apart. Meals provided to subjects were customized according to the individual's energy needs to maintain the body weight. The meal was composed of 35% of the daily estimated energy requirement. FMD was performed at 0, 1, 2, 4, 5.5, 7, and 24h. A pressure cuff was positioned below the elbow, artery diameter was measured before the pressure is applied and then inflated to 220 mmHg systolic pressure for 5 minutes. Immediately after cuff deflation, brachial artery vessel diameter was measured to obtain peak vessel relaxation. FMD was calculated as a percentage change in artery diameter before and after the release of the pressure. Baseline (t= 0 h) %FMD change was not significantly different between the treatments (p>0.05). There was no significant increase in the %FMD after consuming the control meal at all time points (p>0.05). Consumption of meals with Italian herb mix, cinnamon, and pumpkin pie spice significantly increased %FMD at 24 h compared to the control meal (mean ± standard error at 24 h, 7.83 ±0.89%, 7.98±0.95% and 8.44±0.70% vs 6.03±0.79%, respectively, p˂0.05). Furthermore, % FMD was significantly increased at t= 24h in Italian herb mix and cinnamon compared to their respective baselines 5.68 ± 0.37% to 7.83 ± 0.89% (p=0.01) and 6.43 ± 0.89% to 7.89 ± 0.95% (p=0.03), respectively. The results of this research indicate that spices may be beneficial for improving endothelial function after 24 h consumption, suggesting bioactivity of herbs and spices on endothelial function will be related to their time course of bioavailability of bioactive components. This was the first study that assessed the effect of herbs and spices over 24 hours using FMD. Understanding the mechanism of how these herbs and spices improve endothelial function will be the subject of future research.
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Title: POTENTIAL EXPOSURE TO SUBSTANCES IN POLYMER COMPOSITES USED AS FOOD PACKAGING MATERIALS
Creator: Shah, Saloni S.
Date: 2021
Description: In the food manufacturing, preservation, supply, and distribution chain, packaging plays a critical role. The fundamental goal of any...
Show moreIn the food manufacturing, preservation, supply, and distribution chain, packaging plays a critical role. The fundamental goal of any packaging method is to keep food contained and protected. There is an increasing demand for natural and "fresh-like" foods that are less processed and have a longer shelf life, necessitating a variety of packing strategies. With increasing demand, the biggest developments in the field of packaging technology have been innovative food packaging approaches, such as active packaging, intelligent packaging, and bioactive packaging, which include deliberate contact with the food or its surroundings and its effect on consumer health. Several research studies in the past few years have shown that nanocomposite materials have significant improvement in the strength, barrier characteristics, antimicrobial capabilities, and heat and cold stability of food packaging materials, but various studies have reported that these composites might be a source of engineered nanomaterials in the human diet or environment. It has also been reported in numerous studies that nanocomposites can migrate into the food during long-term storage. These studies use food simulants like acetic acid and water to mimic the food matrix. However, they raise issues regarding how ingredients in real foods could affect exposure. This research focuses on the migration of silver (Ag) ions into food matrix-like commercial beverages and demonstrating if the ingredients present in commercial food and beverages influence the migration process. For the study, polymer composites films and dogbones were made. Polymer composite films with 0.2%, 1%, and 5% of silver zeolite concentration in polylactic acid (PLA) were produced, and different media like water, Domino sugar, and Squirt were stored in packages manufactured from this material under accelerated room-temperature conditions. Polymer composite dogbones were made with low-density polyethylene (LDPE) and polypropylene (PP) with 1.25% and 2.51% of graphene and graphite. Further, these materials were characterized with the help of Thermogravimetric analysis (TGA), Fourier Transform Infrared Spectroscopy-Attenuated Total Reflection (FTIR-ATR), and inductively coupled plasma mass spectrometry (ICPMS). This hypothesis of this study was that, when polymer composites are employed in packaging applications, food and beverage components may impact dietary exposure to these particles, and the use of food simulants may underpredict the quantity of the migration in some cases
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Title: IDEOLOGICALLY MOTIVATED INTENTIONAL ADULTERATION: THEORY INTO INDUSTRIAL APPLICATION
Creator: DeVuyst, Adrian Jeffrey
Date: 2021
Description: Ideologically motivated intentional adulteration is an attempt to cause harm to consumers of food. Within the context of the United States of...
Show moreIdeologically motivated intentional adulteration is an attempt to cause harm to consumers of food. Within the context of the United States of America (US), the current methods of addressing this risk are evolving in the modern post-Food Safety Modernization Act (FSMA) era. Currently, the US has the Food and Drug Administration (FDA), which requires companies to have a food defense plan with a risk assessment, mitigation strategies, and recordkeeping. Additional options from Global Food Safety Initiatives (GFSI) benchmarked standards offer additional options for a company. However, even with these standards companies are still being impacted by intentional adulteration. Historical examples from the poisoning of bread in Hong Kong during British occupation and spreading of bacteria on salad bars by the followers of Rajneesh, to more modern examples of putting needles in strawberries and urinating on production equipment show a food defense system that is not always able to address intentional adulteration. The question of why companies are still having intentional adulteration comes up. The lack of food defense events and primary research on the topic creates a system where individual companies must gather data. Evaluations and surveys at a manufacturing site, N=11, indicates that there is high confidence among front line workers about their level of knowledge, but workers are unable to articulate the basic principles of food defense. Each individual company is required to create a personalized food defense system in the status quo, but the results of the survey given suggests that the data they could gather may be insufficient to create an effective food defense system.
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Title: EFFECTIVENESS OF CLEANING STRATEGIES FOR REMOVING MILK CHOCOLATE FROM PILOT-SCALE PIPE/VALVE ASSEMBLY AND CHOCOLATE PROCESSING EQUIPMENT
Creator: Zhang, Liyun
Date: 2019
Description: Dark chocolate manufactured on shared processing lines with milk chocolate is a high-risk food for consumers with milk allergy. Inadequate...
Show moreDark chocolate manufactured on shared processing lines with milk chocolate is a high-risk food for consumers with milk allergy. Inadequate cleaning of shared chocolate manufacturing equipment can result in milk contamination of subsequent products, and product recalls. Limited information is available on the effectiveness of different cleaning procedures for preventing the transfer of milk to dark chocolate processed on shared equipment. Pilot-scale experiments investigated the effectiveness of three dry cleaning methods: 1) no cleaning, 2) pig purging, and 3) a cocoa butter flush (40°C, 1 hour) for removing milk chocolate residue from a heated (40ºC) standard (1.5” OD) sanitary stainless steel pipe (30.5 cm length) and attached butterfly or ball valve. After cleaning, milk-free dark chocolate (~27 kg, 40°C) was pumped through the pipe/valve combination. Dark chocolate push-through samples were collected and analyzed for milk concentrations with a Neogen Veratox total milk ELISA kit. Experiments with no cleaning resulted in initial milk concentrations up to 6,070 (9.6% CV) ppm milk and up to 14,900 (0.3% CV) ppm milk for the pipe/butterfly valve and the pipe/ball valve, respectively. Cocoa butter recirculation through the pipe/butterfly valve decreased initial milk concentrations to 680 (10.3% CV) – 2720 (2.6% CV) ppm milk. Use of a pig purging dramatically reduced milk levels to 45 (4.3% CV) – 180 (15.7% CV) for the pipe/butterfly valve and below limit of quantification of ELISA (LOQ, 2.5 ppm milk) for the pipe/ball valve. After most cleaning treatments, > 14 kg of dark chocolate push-through was required to obtain milk levels < LOQ.A second set of pilot-scale experiments determined the efficacy of cleaning procedures for removing milk chocolate from selected chocolate processing equipment. Three cleaning methods explored removal of milk chocolate from a ball mill and conche: 1) no cleaning, 2) a cocoa butter flush (40°C, 5 min), and 3) wet cleaning (detergent-rinse-air dry). After cleaning, three batches of milk-free dark chocolate (40°C) were processed in the ball mill (~0.35 kg) and conche (2.5 kg), and each batch was collected and analyzed for milk. Milk chocolate (1.5 kg) was processed on a 3-roll refiner, followed by push-through with dark chocolate (~8 kg), with 0.3 kg samples collected at 5-min intervals. Milk was not detected (Show less

Title: A NOVEL HYDROPONICS SYSTEM FOR PRODUCING SAFE AND HEALTHY SPROUTS
Creator: Azizinia, Mehdi
Date: 2019
Description: Sprouts can be considered as one of the most nutritious and cheap nutritional sources. Due to these advantages, sprouts consumption has...
Show moreSprouts can be considered as one of the most nutritious and cheap nutritional sources. Due to these advantages, sprouts consumption has increased significantly in recent decades. However, because of their susceptible nature to microbial growth, numerous outbreaks associated with this fresh produce have occurred and thus the safety of the sprout is of major concern. A novel kinetic hydroponics system (KHS) was developed to optimize an improve safe sprout production. In KHS, sprouting seeds are able to grow under water while air is continuously introduced. In this study, effect of various airflow rates and light on yield, germination percentage, and physical properties of sprout were examined. In addition, microbial growth during the shelf life of sprout grown, using conventional and KHS methods were monitored. Moreover, the efficacy of chlorine-based sanitizers for reducing microbial loads during KHS sprout production was tested. Results showed that air flow rate had a positive impact on yield. However, higher airflow (8 and 10 feet3/minute) significantly lowered yield. Also, KHS has a significant higher yield compare with conventional method (110.30±4.88 versus 66.19±2.66 g). KHS did not have positive impact on germination percentage. Germination percentage was almost the same in KHS and conventional method (80.67±1.15% versus 81.33±1.53%). Moreover, when various light wavelengths were used, germination percentage increased significantly in KHS (from 91±2.65 to 96±1% in various wavelengths). In terms of color, there were no significant differences in color of sprouts in both systems. In KHS, when dark conditions applied, stem length was significantly higher (31.32±3.55 mm) than those sprouts treated with light. For example, stem length in white light was 8.54±1.32 mm. In contrast, leaves length was significantly higher when light used (highest was 6.67±0.49 mm for combination of blue and red lights compare to 3.19±0.22 mm for dark KHS). Analyzing microbial background showed that sprouts produced in KHS had lower total aerobic counts compared with conventional system (7.24±0.49 versus 8.22±0.18 log CFU/g respectively). However, after 21 days of shelf study at 4°C sprouts in both systems almost had the same counts (10.02±0.70 versus 9.55±0.49 log CFU/g in KHS and conventional systems respectively).
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Title: EVALUATION OF LISTERIA MONOCYTOGENES ENRICHMENT AND COMPOSITING PROTOCOLS FROM ENVIRONMENTAL SAMPLES
Creator: Eckert, Christine
Date: 2019
Description: Environmental sampling in a food production plant is routinely conducted using devices, such as sponges or swabs, to verify cleaning...
Show moreEnvironmental sampling in a food production plant is routinely conducted using devices, such as sponges or swabs, to verify cleaning procedures and determine if any foodborne pathogens, such as Listeria monocytogenes (L. monocytogenes), are present. The devices used for environmental monitoring are enriched to improve pathogen detection. This study aims to 1) compare the limit of detection (LOD) of L. monocytogenes of two U.S. Food and Drug Administration (FDA) enrichment procedures (i.e., Bacteriological Analytical Manual (BAM) and Compliance Document) with and without food matrix, and to 2) assess the number of samples which can be wet and dry composited without loss of sensitivity from stainless steel. To compare the LOD of L. monocytogenes using UVM and BLEB, three inoculation levels (0.27±0.07, 0.59±0.05, and 1.00±0.15 CFU per 225 mL enrichment) with 30 enrichments each were used. Results showed that there was no significant difference between the number of samples where L. monocytogenes was detected for UVM and BLEB at any of the three inoculation levels. However, the limit of detection (LOD95%) for UVM/Fraser was higher than that of BLEB (2.13 and 1.44 CFU/mL, respectively). For wet compositing, 1.24±0.34 CFU of L. monocytogenes was inoculated into 45 enrichments of UVM or BLEB without food matrix and 7.2±0.18 CFU of L. monocytogenes was inoculated into 30 enrichments of UVM or BLEB with 4.13±0.12 log CFU of native microflora from Romaine lettuce wash (RLW). Secondary composite enrichments in Fraser broth were conducted at each of four different ratios: 1:1 (1 positive:1 negative), 1:2 (1 positive: 2 negative), 1:4 (1 positive: 4 negative), and 1:7 (1 positive:7 negative). There was no significant difference between the number of samples where L. monocytogenes was detected between BLEB and UVM with or without food matrix at any of the composite ratios. When comparing wet and dry compositing enrichments from stainless steel, 10.16 × 10.16 cm areas on stainless steel plates were inoculated with 464±22 CFU (2.67±0.24 log CFU) L. monocytogenes, dried for 24 h, and sponges were used to swab the surface of the plates. The sponges were then composited (into primary enrichments for dry compositing) or the secondary enrichments were composited (for wet compositing). Compositing was conducted with RLW containing 4.13±0.02 log CFU of background microflora. There was no significant difference between the number of samples where L. monocytogenes was detected for BLEB and UVM when comparing dry or wet compositing at any of the composite ratios tested. Results of this thesis will aid in determining if compositing of environmental samples is an option when L. monocytogenes is the target pathogen.
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Title: Fate of Listeria Monocytogenes on Hard-cooked Eggs Treated With Citric Acid
Creator: Zeng, Hui
Date: 2021
Description: Commercially-prepared hard-cooked eggs are available for foodservice and to the public in retail grocers. Potential contamination with...
Show moreCommercially-prepared hard-cooked eggs are available for foodservice and to the public in retail grocers. Potential contamination with Listeria monocytogenes during or after the cooking and peeling steps is of concern since this pathogen can proliferate at refrigeration temperatures. Citric acid is a common preservative used in the food industry to treat hard-boiled eggs (HBEs). The purpose of this project was to evaluate the efficacy of citric acid treatment of HBEs to reduce the population levels of L. monocytogenes during 24 h (treatment trials) and 28 d storage (storage trials) at 5 or 25°C. Fresh eggs were boiled for 12 min, cooled to 4°C, peeled, and stored at 5°C for 24 h prior to experiments. In treatment trials, HBEs were dip inoculated with a 4-strain cocktail of rifampicin-resistant L. monocytogenes resulting in either 4 (low) or 7 (high) log CFU/egg. Eggs were air-dried 10 min, followed by treatment with pH 2.5 citric acid (PHCA) or 0.2 M citric acid (calculated as the molarity resulting in pH 2.5: MCA) at 5 or 25°C for 24 h. In treatment-storage trials, citric acid treatment of HBEs occurred before or after inoculation, followed by 28-d storage at 5 or 25°C. L. monocytogenes populations were enumerated by homogenization of eggs with BLEB and cultivation on BHI/rifampicin agar. Enrichment in BLEB was conducted if the pathogen was below the level of enumeration. Significant differences in the populations of L. monocytogenes due to temperature of the acid treatment (5 or 25°C) or the two citric acids (MCA and PHCA) were determined using Student’s T-test and ANOVA with Tukey’s post-test, p ≤ 0.05. Overall, the largest L. monocytogenes reduction occurred after 6 h treatment of HBEs with PHCA at 25°C (1.59 ± 0.00 log CFU/egg) and after 24 h with MCA at 5°C (1.23 ± 0.54 log CFU/egg) when the pathogen was inoculated at the low and high levels, respectively. In treatment-storage trials, citric acid treatment after HBE contamination resulted in a fewer number of samples where the pathogen was detected compared to when treatment occurred before contamination. Citric acid treatment for 24 h also resulted in a greater number of samples where L. monocytogenes was not detected than the 1 h treatment. The results of this study determined that L. monocytogenes could survive on HBEs treated with citric acid, regardless of treatment or storage temperature and acid concentration (PHCA or MCA).
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Title: Growth Kinetics of Listeria monocytogenes and Salmonella enterica on Rehydrated Enoki and Wood Ear Mushrooms during Storage
Creator: George, Josephina
Date: 2023
Description: Plant foods, such as fruits and vegetables, that have been dehydrated do not support the growth of pathogenic microorganisms. Recent...
Show morePlant foods, such as fruits and vegetables, that have been dehydrated do not support the growth of pathogenic microorganisms. Recent listeriosis and salmonellosis outbreaks in the U.S. have been associated with imported specialty mushrooms. These mushrooms are commonly sold fresh or dehydrated. This study evaluated the survival and growth of two foodborne pathogens Listeria. monocytogenes and Salmonella. enterica on dehydrated mushrooms during both rehydration at 25 or 5℃ and storage at 5, 10, or 25℃. Fresh enoki and wood ear mushrooms were dehydrated for 24 h at 60°C. Dehydrated mushrooms were inoculated with a four-strain cocktail of S. enterica or L. monocytogenes at 4 log CFU/g. Mushrooms were dried for 1 h, followed by rehydration for 2 h with 5 or 25°C (water and air temperature). Rehydrated mushrooms were stored at 5, 10, or 25°C for up to 14 d. The pathogens were enumerated at 0, 1, 3, 6, 9 and 14 d. Three independent trials with triplicate samples at each time point were completed. Population differences were evaluated via Student’s t-test; p<0.05 was considered significant. The growth rates were determined by DMFit in Excel. Overall, the growth rates of L. monocytogenes and S. enterica on enoki mushrooms were significantly higher when the mushrooms were rehydrated at 25℃ and stored at 25℃ (P<0.05). The growth rates were 2.69 log CFU/g per day and 3.56 log CFU/g per day, for L. monocytogenes and S. enterica respectively. Since the growth of pathogens on wood ear mushrooms during rehydration and storage was considerably less and below the level of enumeration, enrichment of the pathogens was conducted. The pathogens could be suppressed during rehydration due to less nutrient contents and antimicrobial properties of wood ear. The result of this study outlines the importance of refrigerated storage temperature and time combination for safety during rehydration and subsequent storage of the mushrooms.
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Title: Characterization and Migration of Silver Nanoparticles from Electron-Beam Irradiated Low-Density Polyethylene
Creator: Donovan, Dylan
Date: 2023
Description: Polymer nanocomposites (PNCs) and engineered nanomaterials (ENMs) may find use in a wide range of commercial applications, including food and...
Show morePolymer nanocomposites (PNCs) and engineered nanomaterials (ENMs) may find use in a wide range of commercial applications, including food and medical product packaging. Migration of nanofillers from polymer nanocomposites into food matrices could be a source of human dietary exposure to ENMs. Electron beam (e-beam) irradiation is a processing method used for microbial inactivation as well as for modifying properties of polymer films, such as stretch resistance and shrink tension. Process treatment of nanotechnology-based packaging materials either for sterilization or for strengthening of the polymer films may have a significant effect on the migration of ENMs into food matrices. The primary objective of this study is to investigate the effect of e-beam irradiation treatments of LDPE containing silver nanoparticles (AgNPs) and the subsequent migration of AgNPs into a food simulant under intended use conditions. The study observes a correlation between e-beam irradiation dose quantity and the release of AgNPs into a food simulant.
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Title: Efficacy of Power Ultrasound Technology on the Reduction of Listeria monocytogenes and Salmonella enterica on Produce Matrices
Creator: Biswas, Priya
Date: 2023
Description: Fresh produces are considered as ready-to-eat (RTE) and are minimally processed before the distribution to retailers and consumers. Fresh...
Show moreFresh produces are considered as ready-to-eat (RTE) and are minimally processed before the distribution to retailers and consumers. Fresh produce recalls are frequently linked with pathogenic bacteria like Salmonella enterica and Listeria monocytogenes because of minimal processing. This study evaluated the use of power ultrasound coupled with organic acids like citric, acetic, and lactic acid which are generally recognized as safe and often helps to maintain the quality and prolong the shelf life of fresh RTE fruits and vegetables.All the produce matrices which include cucumbers, romaine lettuce, tomatoes, and strawberry were inoculated with four-strain cocktails of rifampicin-resistant S. enterica or L. monocytogenes at approximately 8 log CFU/ matrix. The produce matrices were dried for 1 h and treated for 2 minutes using 2 % or 5 % citric, lactic, or malic acid. This treatment was conducted with or without power ultrasound treatment at 40 kHz. Samples were taken in sets of three and placed into a stomacher bags. The bag contained 225 ml of water or acid. Following a 2 min treatment period, the samples were placed in separate stomacher bags, each containing 225ml of BPB or BLEB, for S. enterica or L. monocytogenes respectively. Followed serial dilutions, samples were then plated on BHIARif plates. For each condition, triplicate samples were taken, and three separate trials were conducted. The use of Student's t-test allowed for the evaluation of population differences, with a significance level of p<0.05 being deemed significant. Cucumber, romaine lettuce, tomatoes, and strawberries treated with 5 % concentration of citric, lactic, and malic acids, with addition of ultrasound showed a greater result in reductions of S. enterica to populations of 5.54 ± 0.47, 4.54 ± 0.83, and 4.69 ± x 0.36, log CFU/cucumber, 6.66 ± 0.51, 4.12 ± 0.32, and 5.51 ± 0.68, log CFU/ lettuce, 4.38 ± 0. 47, 3.12, and 5.04 ± 0.37 log CFU/ tomato, 4.66 ± 0.49, 4.69 ± 0.06, and 6.22 ± 0.39, log CFU/ strawberries, respectively. For L. monocytogenes, 5 % concentration of acids with the addition of ultrasound resulted in populations of 7.69 ± 0.35, 6.04 ± 0.24, and 6.96 ± 0.41, log CFU/ cucumbers, 7.57 ± 0.12, 5.49 ± 0.55, and 5.78 ± 0.73 log CFU/ lettuce, 6.44 ± 0.13, 5.08 ± 0.12, and 6.04 ± 0.22 log CFU/ tomato, 6.16 ± 0.37, 5.18 ± 0.22, and 5.64 ± 0.50, log CFU/ strawberries, respectively. The most effective acid was lactic when compared with citric and malic acids. The objective of this study is to investigate the effectiveness of power ultrasound as a novel non-thermal processing technology, in order to contribute to the existing knowledge base on this topic.
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Title: Examination of Power Ultrasound and Organic Acid-based Hurdle Technology in the Reduction of Salmonella Enterica on Peaches and Apples
Creator: Mathias, Hina Valida
Date: 2023
Description: Fresh produce includes fruit matrices like whole peaches and apples that are minimally processed and are a popular choice among different...
Show moreFresh produce includes fruit matrices like whole peaches and apples that are minimally processed and are a popular choice among different types of demographics because of their nutrition content and health benefits. However, there have been increasing pathogen outbreaks in these matrices over the past few decades, which are majorly rooted in cross contamination either due to poor handling pre and post processing or the insufficient reduction of the pathogen at processing by the applied hurdle technology. While chemical sanitizers are a popular option in the food industry, the awareness and demand for green consumerism and sustainability have created a need for research to determine the efficacies of organic acids and non-thermal technologies like power ultrasound in the reduction of different pathogens on different food matrices. This study focusses on the S. enterica reduction capabilities of three organic acids – citric, malic, and lactic alone and in combination with 40 kHz power ultrasound at 1, 2 and 5% for treatment times of 2, 5 and 10 min on whole yellow peaches and gala apples. Peaches and apples were spot inoculated with a four-strain cocktail of S. enterica, resulting in 9 log CFU/fruit. Post air drying for 1 h, the fruits were treated with water, 1, 2, or 5% citric, lactic, or malic acid for 2, 5 or 10 min with and without power ultrasound treatment at 40 kHz. The population of S. enterica on the fruits was enumerated before and after treatment. Three independent trials with triplicate samples were performed for each condition. Population differences were evaluated via Student's t-test and ANOVA; p<0.05 was considered significant. The initial level of inoculum ranged from 8.67 ± 0.41 to 8.20 ± 0.26 log CFU/peach and 7.28 ± 0.60 to 8.17 ± 0.37 log CFU/apple in peaches and apples, respectively. Water treatments showed pathogen reduction as high as 1.22 log CFU/peach and 1.02 log CFU/apple. Citric acid treatments on peaches showed significant pathogen reduction at higher time increments at 5% with a reduction of S. enterica as high as 2.24 log CFU/peach after 10 min. Malic acid showed the highest recorded log reduction in peaches at 5% and 10 min being 4.20 log CFU/peach (n=1/9, samples above the enumeration limit) and apples at 5% and 5 min being 3.71 log CFU/apple (n=4/9, samples above the enumeration limit) both in combination with an ultrasound. Lactic acid, unlike the other two organic acids, showed a pathogen reduction of over 3 log CFU/fruit at 2% after 10 min, with the highest pathogen reductions of 3.76 log CFU/peach and >3.62 log CFU/apple at 5% and10 min. There was no particular trend with significant enhancement of pathogen reduction either with time increment or the addition of ultrasound and varied with the varying acids, treatment conditions and fruit matrices.
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Title: CHARACTERIZATION OF HERBS AND SPICES PHYTOCHEMICALS AND PHARMACOKINETIC PROFILE OVER 24-HOUR AFTER CONSUMPTION IN OVERWEIGHT/OBESE ADULTS
Creator: Huang, Yudai
Date: 2022
Description: The health benefits of herbs and spices (H/S) have been known since ancient times. They are a rich source of phytochemicals, such as phenolic...
Show moreThe health benefits of herbs and spices (H/S) have been known since ancient times. They are a rich source of phytochemicals, such as phenolic compounds and terpenoids. However, there is limited information on their absorption and metabolism in humans. Therefore, the primary objective of this study is to identify and characterize phytochemical compounds in H/S mixtures and their absorption and metabolism in the human body over 24 h. H/S and plasma samples used in this study were from a randomized, single-blinded, 4-arm, 24 h, multi-sampling, single-center crossover clinical trial (Clincaltrials.gov NCT03926442) conducted in obese or overweight adults (n=24, aged 37 ± 3 years, BMI=28.4 ± 0.6 kg/m2). Plasma samples were collected at baseline (t=0 h), 0.5, 1, 2, 4, 5.5, 7, and 24 h after consuming a high-fat high-carbohydrate (HFHC) meal with salt and pepper (control) or the control meal with 6 g of three different H/S mixtures (Italian herb: rosemary, basil, thyme, oregano, and parsley in the same ratio; cinnamon; and pumpkin pie spice containing cinnamon, ginger, nutmeg and allspice, the ratio unknown). The phytochemical compounds in the H/S mixtures and their metabolites in human plasma were tentatively identified and quantified by dynamic multiple reaction monitoring transitions on UHPLC-QQQ-MS/MS. Statistical analysis was conducted on SAS-PC 9.4 using non-parametric test via NPAR1WAY procedure. A total of 79 phytochemical compounds were quantified from samples of three H/S mixtures and pepper, of which 36 were flavonoids conpounds, 8 were terpenoids, 27 phenolic acids, and 9 were identified as other compounds. Acetone showed the highest extraction ability for both (poly)phenols and terpenoids in H/S compared to other organic solvents (50% and 80% methanol, ethyl acetate and chloroform). Italian herb contains 763.1 mg/100 g flavonoids, 879 mg/100 g phenolic acids, and 498.6 mg/100g terpenoids; cinnamon contains 981 mg/100 g flavonoids, 11.2 mg/100g phenolic acids, 292.3 mg/100g coumarin, and 1977.1 mg/100 g cinnamaldehyde; pumpkin pie spice contains 655.8 mg/100 g flavonoids, 17.1 mg/100 g phenolic acids, 226.5 mg/100 g coumarin, and 1633 mg/100 g cinnamaldehyde. A total of 47 metabolites were tentatively identified and quantified in plasma samples after H/S consumption over 24 h. Plasma concentrations of carnosic acid derivatives and the glucuronidation products increased after intake of Italian herb, and the Area under the curve (AUC0-24h) was significantly different from control (all P < 0.05) except carnosol glucuronide. Carnosic acid and carnosol had Tmax at 3.4±1.1 and 1.8±0.3 h, respectively, while both of their conjugated glucuronides kept increasing until 24 h. Coumarin glucuronide was increased by cinnamon and pumpkin pie spice consumption with peak concentrations reached at between 1.5-1.6 h. The AUC0-24h after both meals were significantly different from control meal, both P < 0.05. Our results suggest that H/S contain diverse categories of phytochemical compounds that are absorbed and metabolized in the human body into various metabolites in response to 3 different H/S test meals and their appearance in the blood starts as early as around 0.5 h and extends to as long as 24 h for select metabolites.
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Title: Factors Influencing the Level of Detection of Testing Listeria monocytogenes in Ice Cream
Creator: Chen, Bairu
Date: 2022
Description: The increasing evidence has shown that having a sensitive detection method for Listeria monocytogenes in food products is critical for public...
Show moreThe increasing evidence has shown that having a sensitive detection method for Listeria monocytogenes in food products is critical for public health as well as industrial economics. L. monocytogenes was associated with foodborne illness outbreaks linked to ice cream in the United States from 2010 to 2015, with another recent outbreak under investigation. The FDA Bacteriological Analytical Manual (BAM) method was commonly used for L. monocytogenes detection. However, the performance characteristics of the chromogenic methods (MOX, RLM, and R&F agars) remain to be elucidated. The factorial effect on Level of Detection (LOD) as an essential element of the International Organization for Standardization (ISO) approach for qualitative method validation was investigated in this study.For examining the LOD of L. monocytogenes in ice cream, fractional contaminated samples were prepared with the ice cream obtained from the 2015 outbreak and enumerated using the FDA BAM Most Probable Number (MPN) method for Listeria. The effect of test portion size was determined by comparing 10g and 25g using the BAM method with chromogenic agars (MOX, RLM, and R&F). The ISO single-lab validation requirement was followed for the factorial effect study, including four different factors: sample size (10g and 25g), ice cream types (commercially available regular vanilla ice cream and vanilla ice cream with low fat and no added sugar), re-freezing process (with re-freezing and without re-freezing process), and thawing process (slow thaw and fast thaw). LOD and relative LOD (RLOD) were computed using MiBiVal software to compare the sensitivity of the three chromogenic agars and the different factors. For all of the detection experiments, presumptive colonies were identified using the API listeria kit. The 2015 naturally contaminated ice cream was enumerated and resulted in an average contamination level of 2.15 MPN/g. At fractional levels of 0.25 MPN/10g and 0.75 MPN/10g, the positive rates of L. monocytogenes detected from 10g and 25g of sample portions were consistent with the statistically theoretical positive rates. The RLOD values for the reference method (MOX) and the alternative methods (RLM, R&F) were above 1 in both portion sizes, which suggested that MOX was slightly more sensitive than RLM and R&F. The factorial effect study indicated that the four factors have no significant influence on the LOD of L. monocytogenes detection at the fractional contamination levels. However, the test portion size of 25g provided more consistent results among the chromogenic media than the 10g portion size. Fat content was shown to have an effect on L. monocytogenes detection in a large test portion. The information from this study will be useful for the improvement of the reproducibility of a qualitative detection method and can also be used for data analysis standards such as ISO 16140 in method validation studies.
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Title: Growth kinetics of Salmonella enterica and Listeria monocytogenes during rehydration of dehydrated corn and subsequent storage
Creator: Mate, Madhuri
Date: 2022
Description: Dehydrated vegetables, including corn, are often used in restaurants and retail grocers. They do not support the growth of pathogens as their...
Show moreDehydrated vegetables, including corn, are often used in restaurants and retail grocers. They do not support the growth of pathogens as their moisture content is very low. After rehydration, these food products attain high water activity values suitable with neutral pH for the survival and proliferation of foodborne pathogens, including Salmonella enterica and Listeria monocytogenes. The purpose of the study was to examine the extent to which dehydrated corn supports the growth of S. enterica and L. monocytogenes during rehydration at 5 or 25°C water and following storage at 5, 10, and 25°C temperatures at 1, 3, 5 and 7 d intervals. Fresh corn was dehydrated at 60°C for 24 h. Dehydrated corn was inoculated with a 4-strain cocktail of either S. enterica or rifampicin-resistant L. monocytogenes, resulting in 4 log CFU/g, and held at ambient temperature for 24 h. This corn was then rehydrated using either 5 or 25°C water for 24 h. Throughout rehydration, corn samples were removed at intervals and enumerated. To enumerate S. enterica and L. monocytogenes, the samples were homogenized with BPB and BLEB respectively and cultivated on TSAYE with overlaid XLD or BHIARif200, respectively. Rehydrated corn was then stored at 5, 10, or 25°C and enumerated at intervals 1,3,5 and 7 d. Triplicate samples were assessed at each timepoint and three independent experiments were conducted for each rehydration water temperature. Growth rates were determined by DMFit and statistically analyzed using Student t-test. A p-value ≤0.05 was considered significant. Overall the growth rate of S. enterica was higher when rehydrated in 5°C water temperature and then stored at 25°C and was determined to be 0.61 ± 0.23 log CFU/g per d. This timepoint was also the shortest time required to increase by 1 log which was: 1.64 d, i.e. 39 h. For L. monocytogenes, the 25°C water rehydration showed the fastest growth rate when stored at 25°C. It took only 1.58 d or 37.8 h for 1 log increase in the population. After 5°C water rehydration of corn the highest populations of mesophilic bacteria and yeasts and molds were observed for 25°C storage ranging from 8.43 to 9.39 log CFU/g and 4.75 to 7.87 log CFU/g, respectively. After 25°C water rehydration, the highest population of mesophilic bacteria, 8.88 log CFU/g, was observed at 5°C storage at 5 d; yeasts and molds were 8.70 log CFU/g for 25°C storage on the same day. The results of this study determined that S. enterica and L.monocytogenes could survive and grow in dehydrated plant foods during rehydration and storage, highlighting the need for product assessments for these types of foods.
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Title: AN IMPROVED VALIDATED METHOD FOR THE DETERMINATION OF SHORT-CHAIN FATTY ACIDS IN HUMAN FECAL SAMPLES BY GC-FID
Creator: Freeman, Morganne M
Date: 2022
Description: Short-chain fatty acids (SCFAs) are metabolites produced by the gut microbiota through the fermentation of non-digestible carbohydrates....
Show moreShort-chain fatty acids (SCFAs) are metabolites produced by the gut microbiota through the fermentation of non-digestible carbohydrates. Recent studies suggest that gut microbiota composition, diet and metabolic status play an important role in the production of SCFAs. Current methods for the analysis of SCFAs are complex and inconsistent between research studies. The primary objective of this study was to develop a simplified method for standardized SCFA analysis in human fecal samples by gas chromatography with flame ionization detection (GC-FID). A secondary objective was to apply the method to fecal samples from a previous randomized, crossover clinical trial comparing participants with pre-diabetes mellitus and insulin resistance (IR-group, n=20) to a metabolically healthy reference group (R-group, n=9) after daily consumption of a red raspberry smoothie (RRB, 1 cup fresh-weight equivalent) with or without fructo-oligosaccharide (RRB + FOS, 1 cup RRB + 8g FOS) over a 4-week intervention period. Extraction parameters, including solvent selection and water content of the sample, were investigated before finalizing the method. Freeze-dried fecal samples (0.5 g) were suspended in 5 mL of milli-Q water, vortexed and centrifuged at 3,214 x g for 10 minutes. The supernatant was transferred to a clean tube, acidified with 5.0 M HCl and centrifuged again at 12,857 x g for 5 minutes. The resulting supernatant was transferred to a GC vial for analysis by GC-FID. Linear regression data for standards at concentrations 5-2000 ppm ranged from 0.99994-0.99998. Limit of detection (LOD) ranged from 0.02-0.23 µg/mL. Limit of quantification (LOQ) ranged from 0.08-0.78 µg/mL. The validated method was then applied to fecal samples collected from a previously conducted study. Nine SCFAs were identified and quantified (acetic, propionic, iso-butyric, butyric, iso-valeric, valeric, 4-methyl valeric, hexanoic and heptanoic acids). Statistical analysis (Student’s t-test, ANCOVA) was performed on PC-SAS 9.4 (SAS Institute). Acetic acid was significantly lower in the IR-group compared to the R-group before starting intervention (baseline, Week 0, IR v R-group, p=0.014). Intervention analysis comparing RRB to RRB + FOS at 4 weeks (WK4) showed a significant difference in 4-methyl valeric acid (p = 0.040) in the R-group. Trends of decreased SCFA content after 4-weeks of RRB and RRB + FOS compared to baseline were observed in both groups, though changes were not significantly different between dietary interventions at 4 weeks (p>0.05). Metabolic status and dietary intervention are discussed in relation to their impact on SCFA content in fecal samples and mechanisms of biological use as a metabolite. Limitations of the study include sample size and using only feces and not other biological samples for SCFAs analysis, which may be considered for future research.
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