Nutrition & Food Science Theses and Dissertations

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Three Clostridium species with Health Imparting Properties: In vitro Screening for Probiotic Potential
(2024) Mochama, Victor Moronge; Obanda, Diana; Nutrition; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
This research aimed to unlock the probiotic potential of the genus Clostridium, which is often overshadowed by the predominant focus on pathogenic species. The study specifically targeted three promising Clostridium species: C. disporicum, C. celatum, and C. vincentii, which have shown potential in mitigating diet-induced obesity. Despite the challenges presented by the anaerobic growth requirements of Clostridium bacteria, the study capitalized on their capacity to sporulate. This characteristic provides an avenue to use them as probiotics, with resilient and dormant spores capable of surviving food processing and harsh stomach conditions. The resilience of these spores was examined by exposing them to oxygen, heat, gastrointestinal juices, and bile salts. The spores survived oxygen exposure, exhibited resilience to both bile salts and gastric acids, and demonstrated a survival temperature of 70°C. When exposed to suitable germination conditions in vitro, the spores successfully germinated. The study assessed the colonization potential of the bacteria by evaluating their adhesion ability, and all bacteria were found to have the adhesion ability. Furthermore, a safety assessment was conducted by examining hemolytic activity and antibiotic susceptibility to selected antibiotics. The bacteria were found to be susceptible to the antibiotics and did not exhibit hemolytic activity. Bile salt hydrolase (BSH) activity and antibacterial activities were also assessed, and none of the bacteria exhibited BSH activity or antibacterial activity. Antioxidant tests revealed that C. vincentii had the highest antioxidant properties. Assessment of anti-inflammatory properties showed that C. celatum downregulated the gene expression of cytokine inflammation markers IL-6, IL-1, and iNOS while upregulating TGF-β expression. In summary all 3 bacterial species showed good probiotic potential from the in vitro tests. Particularly the formation of resistant spores that later germinated to vegetative cells that produced molecular patterns with antioxidant and anti-inflammatory properties. This necessitates further studies on their probiotic properties.
Investigation into the Impact of Food Matrix on Bacterial Survival during Gastric Digestion
(2024) Gao, Zhujun; Tikekar, Rohan V.; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Over the years, food safety research often focused on the bacterial survival during food processing and storage, whereas physiological studies extensively explored the host-pathogen interaction in gastrointestinal tract. There is a need to understand the intermediate step on pathogen survival during gastric digestion and the potential impact from its food carrier. This study utilized water-in-oil (W-O) and oil-in-water (O-W) emulsion as well as deionized water (DI) as the fundamental model food matrices to study the potential protection by food matrix during simulated gastric digestion. Using Salmonella enterica subsp. enterica serovar Typhimurium as a sample foodborne pathogen, this study investigated the survival kinetics of bacteria using various models of simulated gastric digestion. In a simplified static pH simulated gastric digestion model, inoculated W-O and O-W emulsion matrices were challenged with simulated gastric fluid (SGF) containing HCl and pepsin with mixing using a stomacher for two hours. W-O emulsion showed significant protection of Salmonella survival compared to O-W emulsion and DI water. This protective effect appeared to be matrix dependent regardless of the inoculation location of Salmonella (in dispersed phase vs. in continuous phase). Within the same emulsion type, inoculating Salmonella in water phase or oil phase did not show significant difference in its survivability during simulated gastric digestion. The study was then extended to an improved gastric digestion model where the chyme pH dropped from 4.0 to 1.5 over three hours, and the chyme mixing was achieved by an orbital shaker. In addition, the new SGF was modified to be HCl solution with pepsin, amano lipase A, mucin and NaCl. Under this digestion condition, there was no significant difference in Salmonella survival between W-O emulsion, O-W emulsion, and DI water. Moreover, the dispersed-continuous phase ratio of emulsion composition also showed no impact on Salmonella survival. The simulated gastric digestion model setup was also further analyzed including the role of individual digestive enzyme, the pH impact, and the mechanical mixing approach. In the dynamic pH simulated gastric digestion model, partial activity from lipase accelerated the disruption of emulsion structure for both W-O and O-W emulsion matrices. Mild mixing using an orbital shaker also showed difference in Salmonella survival compared to vigorous mixing using a stomacher. Lastly, this study expanded from using Salmonella as the single bacteria strain into a tailored natural microbiome community. Natural microbiome communities from Golden Delicious (GD) and Empire (EP) apples were manually enriched using bacteria culturing broth at pH 5 and pH 7, respectively. The enriched apple microbiome was then collected and analyzed using 16S rRNA sequencing to study the microbial composition. With a significant decrease in Alpha diversity, the culturable apple microbiome was successfully enriched from less than 3 log CFU/ml to more than 8 log CFU/ml. There was no known foodborne human pathogens detected in the enrichment, and the most abundant genera appeared to be potential plant growth promoting bacteria. The collected apple microbiome was then inoculated in various food matrices to study its survivability during dynamic pH simulated gastric digestion including DI water, apple sauce (AS), chicken puree (CK), sweet potato puree (SP), and W-O emulsion. The enriched apple microbiome showed remarkably high survivability in W-O emulsion throughout the full three-hour digestion treatment. CK also exhibited moderate protective effect compared to SP at the same condition. There was no significant difference between DI and AS on bacterial survivability. In addition, the apple microbiome enriched at two pH levels (5 & 7) showed similar bacteria inactivation kinetics. In conclusion, this study revealed the potential impact from food matrix on bacterial survival during simulated gastric digestion. W-O emulsion offered significant protection of certain bacteria strains or communities in specific simulated gastric digestion models. The parameters in gastric digestion models also affected bacterial survival. Future work should focus on exploring the potential impact from other types of food matrices, expanding the microbial survival study into other bacterial strains as well as a more complex microbiome community, and further comparing the various gastric digestion models.
Antiproliferative Activity of Soybean and Tempeh Extracts on Human Colorectal Cancer Cells
(2024) Fan, Rongjie; Lee, Seong-Ho; Wei, Cheng-I; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Tempeh, an indigenous Indonesian soybean product, is produced through a fermentation process of soybean that enhances the bioavailability of its beneficial nutrients such as proteins and phytoestrogens. Recent studies suggest that the fermentation process of tempeh may enhance the biofunctionality properties of soybeans including anticancer activity. The current study is designed to present a comparative analysis to see if defatted extracts of unfermented soybeans and tempeh (fermented soybeans) possess anti-proliferative activity in human colorectal cancer (CRC) cells. The experimental methods involve the production and extraction of soybeans extract (SE) and tempeh extract (TE) at a concentration of 35 g/100 mL (w/v) with 70% ethanol, followed by rotary evaporation and freeze-drying. MTT assays indicated that both SE and TE exhibited inhibitory activity in viability of human CRC cells, with TE demonstrating a more pronounced dose-dependent inhibition of cell growth compared to SE. Cell cycle analysis led to a significant increase of G1 arrest in both SE and TE-treated cells. The induction of apoptosis was observed from the cells treated with both SE and TE. Western blot analysis revealed an increase of PARP cleavage for both treatments, demonstrating activation of apoptotic pathways in SE and TE-treated cells.
FUNCTIONAL FOODS THAT MANIPULATE THE GUT MICROBIOTA COMPOSITION AND REGULATE THE INTESTINAL IMMUNE SYSTEM-Urtica dioica a CASE STUDY
(2024) Fan, Si; Obanda, Diana; Nutrition; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
The prevalence of obesity and its comorbidities, such as insulin resistance and type 2 diabetes are major concerns. Diet-induced obesity and insulin resistance are associated with gut microbiota dysbiosis, specifically, a reduction in diversity and an increase in bacteria taxa linked with host tissue inflammation and extra energy harvest. Prevention strategies focusing on modification of diet and physical activity levels are beneficial, effective, and cost-efficient but difficult to maintain in the long term.Urtica dioica is widely used as a food in several cultures and its extract has been widely studied for intervention against several diseases, but the molecular mechanisms involved are unclear. In addition, most studies have focused on the extract but not the plant as a whole food. Because U. dioica exerts pleiotropic effects in several tissues, we hypothesized that its phytochemicals get into the systemic circulation to reach target tissues, such as abdominal adipose tissue and skeletal muscle but also impact obesity and insulin resistance through pathways involving the gut microbiota and ultimately the gut immune system. The data presented herein was acquired in a mouse model of obesity and insulin resistance. Supplementing the food with whole U. dioica vegetable, attenuated high fat (HF) diet-induced weight gain, fat accumulation, insulin resistance and changed the gut microbiota composition, by increasing diversity and promoting the proliferation of species from the genus Clostridium. This taxon has associated with lower body weight and activation of regulatory T cells (Tregs) in the intestinal immune system. In a second study, we confirmed that U. dioica induced T cells antigenic stimulation, promoted the activation of Tregs and overall protected against HF diet inflammation. Furthermore, besides attenuating HF diet induced fat accumulation, U. dioica promoted the browning or beiging of adipose tissue as evidenced by enhanced gene expression of key markers of this process. Brown or beige adipose tissue displays enhanced fat oxidation. Finally using enteroids derived from small intestinal tissue, we show that in presence of excess nutrients, supplementation with U. dioica reduces the amount fatty acids and glucose absorbed. In conclusion, supplementing a HF diet with U. dioica attenuates fat accumulation and insulin resistance via mechanisms involving the gut microbiota composition and function, the gut immune system and associated inflammation and moderation of amounts of macronutrients absorbed.
UNDERSTANDING THE INTERACTIONS OF COLD ATMOSPHERIC PLASMA AND THE PLANT STRESS RESPONSE TO IMPROVE FRESH PRODUCE SAFETY
(2023) Gilbert, Andrea Rochelle; Tikekar, Rohan V; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Cold atmospheric plasma (CAP) is a surface modification technology that produces oxidative species that inactivate microorganisms. This technology has been shown to be an effective sanitization technology for use on a variety of food products and leafy greens are a uniquely promising application. CAP products, such as reactive oxygen species (ROS), reactive nitrogen species (RNS), UV light, and ozone, are environmental stressors with antimicrobial effects that plants already encounter in their natural environment, and fresh produce will produce a stress response when treated by CAP and this stress response is not limited to directly treated tissue. The stress responses of baby spinach, red leaf lettuce, baby kale and live basil were evaluated post-CAP treatment. In all evaluated fresh produce CAP treatment resulted in a significant (p<0.05) increase in ROS. This increase in reactive oxygen species was not limited to the plant tissues directly treated by CAP. In live basil, ROS generation was detected on indirectly treated leaves sharing the same stem, and in spinach the increase in ROS levels continued for 12 hours post-treatment. To further understand how the elevated ROS levels affected plant tissues the flavonoid content was evaluated in baby spinach, red leaf lettuce and baby kale. Baby spinach CAP treatment resulted in a significant increase in flavonoids at 50w and 120w power level. Flavonoids in red leaf lettuce were only significantly affected in directly treated leaf tissue. In baby kale, CAP treatment significantly reduced flavonoid content in both direct and indirectly treated leaf tissue. To understand the differences between the two leafy greens species and the effect of ozone and UVB treatment, two products treated with CAP, were also evaluated on baby spinach and baby kale. The two non-CAP treatments resulted in the opposite effects in baby spinach and baby kale, and an examination into the response of the stress hormones salicylic acid and jasmonic acid was inconclusive. The novel interactions of this sanitization treatment with the plant stress response may provide useful opportunities to improve both the food safety and nutritional quality of fresh produce. In addition to the research project, I composed a needs assessment survey instrument to evaluate food safety compliance among small and very small food processors in the northeastern United States in cooperation with the Northeast Center for the Advancement of Food Safety. The survey instrument was sent to food safety professionals and regulators in the region. Results of the survey revealed that the food safety concepts that processors struggled to learn were often those cited by regulators during inspections.
EVALUATING THE EFFECTS OF MODIFIABLE LIFESTYLE AND CARDIOVASCULAR HEALTH FACTORS ON DIABETES LIFE EXPECTANCY IN NHANES AND BRAIN AGING IN UK BIOBANK
(2024) Feng, Li; Lei, David K.Y. DL; Ma, Tianzhou TM; Nutrition; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
This dissertation explored the impact of lifestyle and cardiovascular health factors on aging, particularly focusing on individuals with diabetes, the effects of blood pressure on brain aging, and the influence of cardiovascular health and genetic predispositions on brain white matter aging.The first study examined the trends in lifestyle quality among US adults with type 2 diabetes from 1999 to 2018 using NHANES data, involving 7,410 participants. A healthy lifestyle score encompassing smoking, drinking, physical activity, and diet showed a slight increased over the years. Notably, disparities remained significant by socioeconomic groups. The study found that adherence to low-risk lifestyle factors was associated with a 55%-57% lower risk of all-cause mortality, emphasizing the importance of lifestyle modification in diabetes management, and it was independent of cardiovascular risk control. The second study investigated the causal effect of elevated blood pressure on white matter brain aging in a cohort of 228,473 European ancestries aged 40-69 from the UK Biobank by using two-sample Mendelian randomization. Our result revealed that high blood pressure, particularly diastolic, accelerated the machine-learning-derived white matter brain age gap, based on white matter microstructure integrity measured by fractional anisotropy derived from diffusion tensor imaging data, with a causal effect evidence found in late middle-aged women. This underscores the importance of blood pressure control in preventing brain aging, especially in post-menopausal women. Lastly, the impact of Life's Essential 8 (LE8), a comprehensive measure of cardiovascular health (lifestyle part: diet, smoke, physical activity, sleep; health part: BMI, blood sugar, blood pressure, blood lipid), on white matter brain aging was assessed, with a particular focus on how the APOE4 genotype modifies the relationship. Analyzing data from 18,817 European ancestries aged 40-60 from the UK Biobank, the study revealed that higher LE8 scores correlated with a younger brain age. Interestingly, the effect varied significantly with APOE4 status, highlighting the need for personalized health strategies based on genetic profiles. In conclusion, these studies collectively highlight the crucial role of modifiable lifestyle and health factors in managing chronic diseases, controlling blood pressure, and maintaining brain health, with an emphasis on the integration of genetic profiles for personalized healthcare.
EVALUATION OF THE COPPER HOMEOSTASIS AND SILVER RESISTANCE ISLAND AND ITS ROLE IN PERSISTENCE OF SALMONELLA ENTERICA
(2023) Haendiges, Julie; Tikekar, Rohan; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Salmonella enterica is one of the leading bacterial cause of foodborne illness in the UnitedStates. Although there are many serovars, only a small subset causes human illness. Since Salmonella is ubiquitous in the environment, the Food and Drug Administration has established regulations for food processors to ensure that the products are free of contamination. Low-moisture foods are commonly ready-to-eat, and due to the low water activity do not promote growth of bacteria. However, Salmonella has been shown to persist in these foods. There havebeen two outbreaks and multiple recalls in the United States due to contaminated pistachios. Based on a retrospective study, results show that there is evidence of a contamination in the growing orchard and a significant number of Salmonella isolates from the environments contain the Copper Homeostasis and Silver Resistance Island (CHASRI) cassette. This raises several questions: what is the prevalence of CHASRI among different Salmonella isolates from food and environmental sources? Does presence of CHASRI enable Salmonella to survive better against copper stress? And whether presence of CHASRI provide cross-protection against other stresses such as desiccation and thermal treatment? This dissertation attempts to answer those questions. The prevalence of the CHASRI in Salmonella was determined by the use of publicly availablewhole genome sequencing data. The CHASRI was found in 61 different serovars and types of sources. The presence of the CHASRI in isolates from low-moisture foods that have caused previous outbreaks (peanut butter, nuts, spices) was interesting to note, and leads to future studies on correlations between this island and virulence. Based on results of phylogenetic analysis of CHASRI sequences from closed genomes, we determined there were four types of CHASRI found in Salmonella. Traditionally, the Salmonella Genomic Island-4 (SGI-4) is found but in addition the CHASRI can incorporate by itself, within a variant of SGI-4, or via a rare plasmid. Interestingly, the sequence of the CHASRI from SGI-4 and the variant SGI-4 were highly different. The high SNP differences in sequence along with the difference in the arsenic operon led to the conclusion that these variants arose independently. A Salmonella Senftenberg strain (CFSAN047523), isolated from pistachios, was used to createthree knockouts (∆cus, ∆pco, and ∆CHASRI). Previous studies have looked at the minimum inhibitory concentration (MIC) of strains with and without the CHASRI but have omitted the minimum bactericidal concentration (MBC). In this study, we used the knockouts to test for both MIC and MBC. While the MIC was similar for the strains, the MBC was greater in the wild type and partial CHASRI knockouts. Growth and inactivation kinetics were measured in different concentrations of copper sulfate. At higher levels of copper sulfate, the presence of the CHASRI made cells more resilient to inactivation by copper sulfate. Evidence shows that the stress response in Salmonella has the ability to crosstalk and provideprotection against multiple stresses. To investigate this phenomenon further, our isolates were tested against a multitude of stresses to evaluate for cross-protection that may be due to theCHASRI. Cells undergoing copper stress were better equipped to survive lethal copper concentrations and desiccation if the CHASRI was present. The presence of Salmonella in final pistachio products that have been fully processed identifies that some adaptation and stress response is occurring in the processing facility. Inoculated pistachios with the wild type and ∆CHASRI strain were thermally processed to test for survivors. This study showed that the presence of the CHASRI gave the isolate an advantage to survive thermal processing after desiccation. Overall, this study presents the prevalence of the CHASRI in Salmonella enterica as well as theimportant role it plays in copper tolerance. The evidence of cross-protection and tolerance to copper leads to future research regarding gene expression and virulence assessment.
FOOD SAFETY IN THE ERA OF NEXT-GENERATION SEQUENCING: GENOMIC CHARACTERIZATION OF SHIGA TOXIN-PRODUCING ESCHERICHIA COLI AND METAGENOMIC SURVEILLANCE OF IRRIGATION SURFACE WATER
(2023) Huang, Xinyang; Meng, Jianghong; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
In this study, we first utilized high-throughput next-generation sequencing (NGS) and bioinformatic analyses to characterize potential public health threats posed by non-top-7 Shiga toxin-producing Escherichia coli (STEC). NGS allowed us to detect virulence (n = 46) and antimicrobial resistance (AMR) (n = 27) factors within the genomes of the STEC strains, to make genome-wide comparisons with published human clinical isolates, and to characterize three novel O-antigen gene clusters. We found that the distribution of 33 virulence genes and 15 AMR determinants exhibited significant differences among serotypes (P < 0.05), and that 47 strains were genetically related to human clinical strains inferred from a pan-genome phylogenetic tree. We secondly developed a web tool, PhyloPlus, that allowed users to generate customized bacterial and archaeal phylogenies, which can be incorporated into their own microbial community studies. We also utilized two public datasets (human microbiome, n = 60; fermented food metagenomes, n = 62) to illustrate how application of phylogeny improved our analyses. We showed that the integration of phylogenies introduced alternative phylogeny-based diversity metrics and allowed more conservative null model constructions, thereby reducing potential inflation of type I errors. Finally, we employed deep metagenomic shotgun sequencing, and our developed web tool, to investigate on a collection of 404 surface water samples collected from four regions in Latin America. We reported the high detection rates of pathogenic and contaminant bacteria in these samples, including Salmonella (29.21%), Listeria (6.19%), and E. coli (35.64%), necessitating the monitoring and proper treatment on these surface waters. We also described the regional differences in terms of sample taxonomic composition and the resistome, and further presented key factors that drove the separation patterns for each sampling region. We utilized recent metagenomic assembly and binning algorithms to report the construction of 1,461 de-replicated metagenome-assembled genomes (MAGs) that were of at least medium quality. The incorporation of the MAGs into the taxonomic classifier Kraken2’s database led to a 12.85% increase in classifiable sequence reads. Additionally, we conducted network analysis on AMR genes and the genus-level taxonomy, based on assembled contigs, to provide information to better understand the dynamics of the transferring of AMR genes.
SURVIVAL AND PERSISTENCE OF LISTERIA AND ESCHERICHIA COLI AND CHANGES IN PHYSICOCHEMICAL PARAMETERS IN AQUAPONICS SYSTEMS DURING LETTUCE PRODUCTION
(2023) ALHAMMAD, GHADAH ABDULRAHMAN; Lei, David K. Y.; Nutrition; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Aquaponic (AP) food production systems integrate aquaculture and hydroponics in soilless controlled environments while conserving space and natural resources (soil, water, air). Increasing global demand for high-quality quality nutritious, safe food continues to increase exponentially. Information specific to AP is needed to validate operational and handling practices for AP food safety. The first study is to assess the persistence of E. coli TVS 354 in bench-scale aquaponic systems (180 days after primary plant harvest). Methods: follow-up on the previous study and evaluate growth and persistence of E. coli TVS 354 at 180 days after primary plant harvest. Samples were collected from 10 independent aquaponic systems and bacterial growth was evaluated including aerobic plate count (APC), coliform bacteria (CB), heterotrophic plate count (HPC), and cultural enrichment for E. coli TVS 354 levels. Results: E. coli TVS 354 was not detected in samples from hydrotons (25 g), filters, fish feces, core and roots. The aerobic bacteria count of fish and plant tank samples ranged between 2.5 to 3 log CFU/mL for all treatments, while aerobic bacteria count on the core of the lettuce plant was significantly lower, ranging from 1.44 to 2.08 log CFU/mL. The second study is to evaluate the survival of Listeria innocua 2066 in effluent from fish tanks and plant tanks of aquaponic research units. Methods: four different treatments: fish tank water, fish tank water-replenished, plant tank water, and plant tank water-replenished. Each bottle was inoculated with Listeria innocua 2066, and bacterial growth and water chemistry conditions were evaluated over 7 days to identify physiochemical parameters associated with Listeria innocua 2066 survival and growth. Results: In 24 hours a significant reduction (p<0.05) in Listeria inccoua 2066 populations was observed for all treatments with a total reduction 3.9-3.7 log CFU/mL. Listeria innocua 2066 counts were lowest on day 7 for treatment groups, ranging from 1.1 log CFU/mL to 1.0 log CFU/mL. Treatments that were replenished daily from the aquaponic systems had higher counts of Listeria innocua 2066 compared to treatments that were non-replenished. The lowest APC on day 0, immediately after inoculation, ranged from 4.16 log CFU/mL to 4.32 log CFU/mL. A significant increase (P<0.05) in APC count was observed for all treatments in 24 hours while there is no significant difference in APC values between the treatments (p=0.35). The final and third study is to evaluate survival of inoculated Listeria innocua 2066 and nonpathogenic Escherichia coli and changes in AP physicochemical parameters. Methods: A four-week bench-scale AP experiment (n=12) with four goldfish (Carassius auratus)/aerated 37L tank, a 3-step biofilter, and four lettuce plants (Lactuca sativa var. Truchas)/37L hydroponic tank was conducted. Treatments were high and low-dose (6-log and 2-log CFU/mL, respectively) of Listeria innocua 2066-Er or E. coli-Rifr TVS 354, and uninoculated controls. Listeria innocua-2066-Er and E. coli-Rifr populations, mesophilic counts (APC), and physicochemical parameters (pH, temperature, dissolved oxygen, turbidity, ammonia, nitrite, and nitrate) were analyzed in plant tank water, and biofilters until plant harvest. Listeria innocua 2066-Er and E.coli-Rifr enumeration and APC (Petrifilm®) from lettuce shoots, roots and rockwool were determined at harvest. Results: Listeria innocua 2066 and E. coli TVS 354 populations declined significantly within 24 hours post-inoculation and were undetectable at day 14 and day 12 respectively (P <0.05). This decline was observed for both high and low-dose treatments. Listeria innocua 2066 and E. coli TVS 354 were detected in biofilters until week 4. At harvest, Listeria innocua 2066 and E. coli TVS 354 were recovered from lettuce roots, and rockwool, but not from plant leaves, and 100mL plant tank water grab samples. Lower leaves preharvest had a significantly higher APC (5.1- 6.4 log CFU/mL) relative to harvested lettuce upper leaves (2.8 - 4.2 log CFU/mL). Plant tank water pH had a significant effect (P<0.05) on Listeria innocua 2066 and E. coli TVS 354 survival. Significance: These results provide insights into the survival dynamics and sites of E. coli TVS 354 and Listeria innocua 2066 in AP and associated physicochemical conditions. These findings contribute to our understanding of potential food safety risks and associated risk factors such as inputs, physicochemical factors, and other environmental conditions in aquaponics systems.
DEVELOPMENT OF HORDEIN-PECTIN NANOPARTICLE COMPLEX FOR THE ENCAPSULATION OF BIOACTIVE COMPOUNDS FOR ENHANCED FUNCTIONALITIES
(2023) Tarwa, Kevin; Wang, Qin; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Nanoparticle delivery systems composed of food polymers are a sustainable and eco-friendly approach to protect functional ingredients and promote healthier food options. In this research, a hordein-pectin nanoparticle complex (HP-NPC) was fabricated using an anti-solvent precipitation and electrostatic deposition (pH 4) method for the encapsulation of hydrophobic bioactive compounds to enhance their functionalities. First, hordein was extracted from whole barley grains to obtain a dried powder to synthesize hordein nanoparticles (HNP). Then pectin with a degree of esterification (DE) around 71% was applied as a coating material. The average particle size of the freshly prepared nanoparticle complex was relatively small (~246 ± 11 nm), and Fourier transform infrared spectroscopy (FTIR) indicated that cationic hordein interacted with anionic pectin mainly though newly formed hydrogen bonds and electrostatic interaction as indicated by their opposite surface charges. Scanning electron microscopy (SEM) revealed that the morphology of the nanoparticle complex was spherical with a smooth surface. The pectin coating was shown to have a protective effect against pH (3.0-9.0), heat (80 °C for 0-120 mins), and salt (0-100 µM) which are all factors known to degrade proteins. Second, lutein, a hydrophobic bioactive xanthophyll was encapsulated into HP-NPC to develop a lutein-hordein/pectin nanoparticle complex (L-HP-NPC). Since lutein has low water solubility and low bioavailability in the gastrointestinal tract (GIT), the effect of the encapsulation system on the functional properties of lutein was investigated. The loading capacity (LC%) and encapsulation efficiency (EE%) was around 15.5 and 82%, respectively. In vitro digestion resulted in a higher bioaccessibility of lutein for encapsulated HP-NPC (~22.3%), which is defined as the percentage of lutein accessible for absorption in the simulated intestinal fluid (SIF) compared to lutein encapsulated into HNP (~9%). The ability of pectin to produce gels in acidic media was shown to have a significant effect against gastric enzymes that can degrade both hordein and lutein. Also, lyophilization (an important step in food processing) had no significant effect on the stability of L-HP-NPC. This encapsulation system could potentially be used as a functional ingredient in the food industry to develop healthy and nutritious foods for consumers. Third, carvacrol, a phenolic monoterpene known for its antimicrobial properties was encapsulated into HP-NPC to develop a carvacrol-hordein/pectin nanoparticle complex (CA-HP-NPC). Special focus was on the solubility of encapsulate carvacrol due to its known low solubility in aqueous solutions. The antimicrobial effectiveness of the encapsulated nanoparticle complex was tested against non-pathogenic gram-positive L. innocua and gram-negative E. coli K12. CA-HP-NPC was still able to maintain a relatively small particle size (~207 ± 8 nm) after being dispersed into water post-lyophilization. Carvacrol was shown to be effective against the two bacteria, however, CA-HP-NPC did not show antimicrobial effectiveness. Although carvacrol was successfully encapsulated into the nanoparticle complex, further studies on their release properties need to be investigated to further understand their functional properties for food applications.
FERMENTING KALE VEGETABLE (Brassica oleracea Var Sabella) IMPROVES ITS PROPERTIES AS A FUNCTIONAL FOOD
(2023) Subedi, Ujjwol; Obanda, Diana; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
The properties of kale as a functional food are well known. Fermentation is a process that has been shown to improve the health impacts of foods. In this study, we sought to determine how fermentation further improves or augments the functional food properties of kale. We tested six different fermentation methods which included traditional practices and inoculation with different bacterial species and compared outcomes to the unfermented control. After 16 days of fermentation, we quantified (i) selected bioactive components and (ii) anti-nutritional factors. We then determined (i) the antioxidant capacity of the whole vegetable, (ii) the microbiota composition of the vegetable, and (iii) the anti-inflammation capacity of the ethanolic extract of the vegetable. Fermentation significantly increased (i) the quantities of total polyphenols from 8.54 to 10.71 mg GAE/g (ii) sulforaphane from 960.8 ± 41.76 to 1777 ± 45.95 μg/g, and (iii) antioxidant capacity from 61.99 to 67.37 % respectively, and antinutritional factors oxalate and tannin content significantly reduced by 49 % and 29.83 % respectively. Fermented kale extract exhibited potent anti-inflammatory effects in macrophages by reducing the iNOS expression by 84.3% and TNF-α, IL-1β, and IL-6 mRNA levels by 62, 68, and 85.5 %, respectively. Fermenting kale changed the surface microbiota by reducing the population of the inflammation-inducing Proteobacteria while increasing health-promoting Firmicutes; including Lactobacillus. All fermentation methods had a beneficial impact compared to the unfermented control, but the mixed culture of L. lactis and L. acidophilus was the most effective. In summary, fermenting enhanced the health benefits of kale by increasing the concentration of total polyphenol, sulforaphane content, antioxidant capacity, anti-inflammation capacity, and reducing the quantity of anti-nutritional factors. Furthermore, it promoted the prebiotic and/or probiotic vehicle properties of the vegetable by changing the proportion of beneficial bacteria and those associated with inflammation.
POPULATION AND GENETIC DIVERSITY ANALYSIS OF LISTERIA MONOCYTOGENES IN SELECT FOODS AND FOOD PROCESSING ENVIRONMENTS
(2023) Kwon, Hee Jin; Meng, Jianghong; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Listeria monocytogenes, a Gram-positive bacterium, is a foodborne pathogen that causes listeriosis in humans. L. monocytogenes can persist in various environmental conditions, including food-relevant conditions such as high salinity, refrigerated temperatures, and low moisture contents. Contaminated food products, including dairy products, deli meats, fresh produce, and soft cheeses, are the primary transmission vehicles for L. monocytogenes. The complex and dynamic population structure of L. monocytogenes complicates control efforts, particularly due to certain strains that may possess increased resistance to stress conditions and/or enhanced virulence. The advent of whole genome sequencing has facilitated comprehensive genomic analyses of L. monocytogenes, enabling a comprehensive understanding of its adaptation and survival characteristics over time and across various geographic locations. Understanding the population and genetic diversity of L. monocytogenes is crucial for the development of effective control measures, as it helps infer the spread and transmission pathways of L. monocytogenes through the integration of spatial-temporal factors. Furthermore, these analyses provide insights into the evolutionary relationships among L. monocytogenes strains. This dissertation aimed to investigate the population diversity of L. monocytogenes in various food sources and food processing facilities, utilizing the whole genome sequencing technology. The findings contribute valuable insights into the genetic diversity and population structure of L. monocytogenes, thereby aiding the understanding of the risk associated with L. monocytogenes contamination and the development of effective control measures to ensure food safety.
BACTERIAL GROWTH AND INACTIVATION IN HETEROGENEOUS ENVIRONMENT
(2023) Tsai, Shawn; Tikekar, Rohan V.; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
This dissertation explores the growth and inactivation of Salmonella enterica subsp. enterica Serovar Typhimurium in oil-in-water emulsion systems, with a particular focus on the effects of emulsifier type, oil fraction, temperature, molecular weight, and surface charge. First, investigation was conducted on the effects of emulsifier type (Tween 20, Tween 80, Triton X-100) and oil content (20%, 40%, 60%) on growth and thermal inactivation of S. Typhimurium in emulsions. The results indicated that while emulsifiers did not affect the bacterial growth rate or lag phase, the presence of high oil content (60%) prolonged the lag phase in emulsions. In addition, Tween 80 and Triton X-100 emulsifier solutions exhibited protective effects against thermal inactivation. Next chapter was focused on evaluating the impact of temperature on growth (7, 22, 37°C) and inactivation (55, 58, 60°C) kinetics of S. Typhimurium in emulsion systems with same emulsifier and oil fraction as previous chapter. The results indicated that temperature had a significant impact on bacterial kinetics, with increasing temperatures leading to faster growth and inactivation rates. Next, the effect of emulsifier molecular weight and surface charge on the growth and thermal inactivation of S. Typhimurium in emulsions was examined. To control the molecular weight with similar structure, whey protein was selected for experimentation. By adjusting the pH, it was possible to change the surface charge in whey protein. Results indicated that whey protein hydrolysate (WPH) with a lower molecular weight did not exhibit a lag phase in Salmonella growth. However, whey protein isolate (WPI) with higher molecular weight demonstrated no difference in the lag phase when compared to bacterial growth in TSB. Similar effects were observed with a positively charged emulsifier (WPI+). The findings suggest that the molecular weight of emulsifiers has a more significant impact on bacterial growth than their surface charge. Regarding the evaluation of inactivation, emulsifier solutions exhibited no significant difference compared to TSB, while emulsions stabilized by WPH and WPI+ showed some protective effects on S. Typhimurium. This observation can be attributed to the ability of positively charged emulsifiers to interact with the bacterial membrane, providing protective effects during thermal treatment. Lastly, to gain a comprehensive understanding of the mechanism concerned with the impact of emulsifier and oil inclusion on bacterial growth and inactivation behavior, S. Typhimurium was cultured in different emulsion-related environments and evaluated for nine stress-related genes (rpoE, rpoH, otsB, proV, fadA, fabA, dnaK, ibpA, ompC) after 20 hours of incubation at 37°C and after thermal treatment at 55°C for 45 min. It was found that ibpA was upregulated in all emulsifier environments, regardless of the presence of oil, indicating that IbpA was synthesized in emulsifier environments. Moreover, increased expression of fabA was also observed in Triton X-100 stabilized 60% emulsion, indicating poor heat resistance due to increased membrane fluidity. In the combination of gene expression data, our results showed that emulsifier solutions without oil exhibited a greater number of regulatory mechanisms compared to those containing oil, indicating that the presence of oil did not provide as much protection after thermal treatment. Based on these findings, the stress-related mechanism was constructed by the expression of those selected genes. Overall, this dissertation provides valuable insights into the factors influencing bacterial growth and inactivation in oil-in-water emulsion systems, as well as bacterial stress response in these systems. These findings provide important insights into the growth and inactivation behavior of S. Typhimurium in oil-in-water emulsion systems and the stress response mechanisms involved. Understanding these factors is crucial for developing effective control measures to ensure food safety and prevent foodborne illness outbreaks caused by this pathogen. This information can be used to optimize the formulation and processing of emulsion-based food products to minimize the risk of bacterial contamination and ensure their safety for consumption.
ANTICANCER ACTIVITY OF NATURAL PRODUCTS IN HUMULUS LUPULUS(HOPS) IN HUMAN COLORECTAL CANCER CELLS.
(2023) Tamia, Gillian; Lee, Seong-Ho; Nutrition; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Cancer is a major public health problem and the second-leading cause of death in the world. Colorectal cancer (CRC) is the third most diagnosed cancer in the U.S. CRC is highly associated with daily diet and eating patterns. A plant-based diet rich in phytochemicals has been known to be protective against the initiation and progression of CRC occurrence. The hop plant, a key ingredient in beer, contains a diverse form of bioactive compounds that possess biological benefits in tumorigenesis. Xanthohumol (XN), the most abundant prenylated flavonoid, has been used over the years to treat a broad range of chronic diseases such as diabetes, obesity, and cancer. Several derivatives of XN, including isoxanthohumol (IXN), 8-prenylnaringenin (8-PN), and tetrahydroxanthohumol (TXN), possess similar and greater biological benefits compared to XN. While XN's anti-cancer properties are well known, the effects of these derivatives have not been evaluated in human CRC models. Our study aimed to test the cancer-suppressive activities of these derivatives and elucidate anti-cancer mechanisms using human adenocarcinoma CRC cells. The results indicate that four hop compounds (XN, IXN, 8-PN, and TXN) significantly suppressed the proliferation of different types of human CRC cell lines. We selected TXN and XN for further studies due to their more significant and promising anti-proliferative activity compared with other forms. Flow cytometry analysis indicated that TXN and XN led to significant induction of S-phase and G2/M-phase arrest. An apoptotic assay showed a huge induction of early and late apoptosis in cells treated with TXN and XN at doses of 12 µM and 18 µM. Western blot data indicate that TXN and XN induce the cleavage of PARP and increase the expression of CHOP, IRE1α, and ATF4, indicating activation of caspase-dependent apoptosis and ER stress. In addition, a dose-dependent increase in intracellular ROS was observed in cells treated with 12 and 18 µM of TXN and XN, affecting mitochondrial dysfunction. Taken together, our current study proposes an anti-cancer mechanism by TXN and XN through their action on the induction of ROS release and mitochondrial dysfunction, ER stress, and apoptosis in human CRC cells.
GENOMIC FACTORS ASSOCIATED WITH PSEUDOMONAS VIRULENCE AND ANTAGONISM
(2023) Muruvanda, Tim; Meng, Jianghong; Nutrition; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
The longstanding problem with culture-dependent methods has been a reliance on pre-enrichment/enrichment steps which often comprise within them background microbial flora capable of outcompeting the target organism. Here, the genomic factors associated with Pseudomonas virulence and antagonism were studied in strains from selective media where it was shown to outcompete Salmonella, the target organism. 34 Pseudomonas strains of various species were sequenced and annotated. Results identified quorum sensing (QS) as the main factor of driving virulence and antagonism through the recruitment the type VI secretion system (T6SS). Additionally, two potential effectors corresponding to DUF 4225 and 4280 were found along with P.fluorescens strains containing the Pseudomonas virulence factor (PVF). These results may be valuable in the development of new strategies to better circumvent the effects of Pseudomonas in cultures.
Age, Nutrition, and Bone Metabolism: Analyses of Effects Using a Short-Term In Vivo Bone Model
(1987) Sinha, Rashmi; Soares, Jos; Nutritional Sciences Program; Digital Repository at the University of Maryland; University of Maryland (College Park, Md)
The preventative effects of dietary calcium, zinc, and vitamin D metabolites on the net loss of bone were assessed using rats of different ages. Biochemical changes were monitored in femurs, humeri, mandibles, scapulas, and tibias. In addition femora were sectioned into epiphyses-metaphyses and diaphyses to evaluate nutritional influences on the trabecular and cortical type bone. since measurable bone degradation due to aging and nutritional status requires long period of time, a short-term in vivo system was developed to simultaneously examine bone formation and resorption. The system consisted of subcutaneous implantation of demineralized (DB) and mineralized (MB) bone powders. There was evidence of bone formation and resorption in the DB and MB implants respectively, as assessed by marker enzyme (formation-alkaline phosphatase; resorption-acid phosphatase) activities, mineral concentrations, radioisotope incorporation, and histological studies. The results indicated that several different bone samples are required to adequately predict changes occurring in the skeletal system. The epiphysesmetaphyses of long bone is a useful sample site examining changes occurring in trabecular bone while the diaphysis can assess cortical bone status. There was decreased bone formation and resorption as assessed by alkaline and acid phosphatase activity in the MB and DB implants in the 24 month-old rats as compared to 2 month-old rats. Dietary calcium and zinc levels did not affect the overall status of the bones and implants in the aged rats. Conversely, in 2 month-old rats dietary calcium at 1.0% stimulated bone formation in the DB implant, whereas 0.2% calcium increased bone resorption in the MB implants. Furthermore, 75 ng dietary 1,25-dihydroxycholecalciferol (l,25(OH) 2 D) per day increased resorption in MB implant and inhibited mineralization of DB implants in the young rats. Dietary zinc at 300 ppm reduced bone calcification in 2 month-old. The results of these studies indicated that neither high levels of dietary calcium, nor zinc, act as prophylaxes to counteract bone loss due to aging. The dietary use of l,25(OH) 2 D in old animals needs to be investigated further, since results in young animals are contradictory with reports in older rats.
STUDIES ON INTERACTIONS OF UV RADIATION WITH FOOD INGREDIENTS FOR IMPROVED QUALITY AND SAFETY
(2022) Montemayor, Abraham Moses; Tikekar, Rohan V; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Ultraviolet (UV) radiation has been used in the food industry during processing, and with increased demands for safer and higher quality foods, UV-A and UV-B are being explored as antimicrobial treatments. This project consisted of three studies: the first study investigated the production of reactive oxidative species (ROS) by the action of UV radiation on fructose. The second study focused on evaluating the impact of UV-A irradiated chitosan-gallic acid (CH-GA) antimicrobial film on the quality of strawberries. The third study evaluated the effects of using UV-C individually or in combination with UV-A and UV-B to improve the fruit color and safety, respectively, of Honeycrisp apples. It is known that fructose can generate ROS under thermal treatments and UV-C (254 nm) exposure. However, it is unknown whether UV-A or UV-B exposure can generate similar effects. For the first study, fluorescein, a fluorescent dye, was used as an indicator due to its known loss of fluorescence when exposed to ROS. Varying concentrations of fructose solutions combined with fluorescein were exposed to up to 1 J/cm2 of UV-A or UV-B radiation. Ascorbic acid (AA), a known ROS scavenger, was added to the fructose-fluorescein solutions prior to UV exposure to verify ROS generation. The fluorescence was measured at 485 nm (excitation) and 510 nm (emission), respectively. A storage study was done to determine whether ROS continued to generate following UV exposure. Fructose-fluorescein solutions were exposed to 0.1 J/cm2 of UV-B radiation and stored at 4°C or 37°C. The UV-B exposure of fructose-fluorescein showed a dose-dependent fluorescence decay, whereas UV-A did not elicit this response. Fluorescein degradation followed first-order kinetics, as indicated by the rate constants. The rate constants in the presence of 10-, 50-, and 100- mM fructose were 0.7±0.01 J/cm2, 4.3±0.6 J/cm2, and 0.3±0.03 J/cm2, respectively. However, in the presence of AA, fluorescein degradation deviated from first-order kinetics. The storage study indicated no significant difference between the UV-B exposed and control solutions, indicating ROS generation ceased after UV-B exposure. The results of the studies using control solutions were extrapolated to coconut water, a commonly consumed beverage. UV-B exposure did have a degradation effect on AA, but the ROS generated did not affect the AA. The ROS was produced only when fructose was exposed to UV-B. ROS can have adverse effects on the organoleptic properties of foods containing fructose, and the addition of AA can help quench ROS in a concentration-dependent manner. The second study evaluated quality parameters such as color, texture, pH, total soluble solids, and titratable acidity of strawberries coated with an edible chitosan-gallic acid (CH-GA) coating. The strawberries were dipped in the CH-GA solution and allowed to dry. The coated strawberries were exposed to UV-A with appropriate, unexposed controls also being used for comparison. Previous studies have indicated that the coating can exhibit moderate antimicrobial activity when irradiated with UV-A at 360nm. A 180-minute exposure reduced Escherichia coli (E. coli) O157:H7 on CH-GA coated strawberries by ~2-3-log CFU/mL. However, when the quality parameters were evaluated, it was found that the UV irradiated strawberries may have been initially affected with respect to color and texture, but the loss in quality slowed down over a 14-day refrigerated storage period. It was also seen that no significant differences were observed in color and firmness between the control and experimental groups on day 14. The third study (appendix 1) aims to evaluate UV-C radiation's efficacy on the inactivation of Listeria monocytogenes (L. monocytogenes) on apple surfaces. This study was performed within the broader aim of evaluating the effects of UV-A, UV-B, and UV-C and their combinations on the quality and safety of Honeycrisp apples. UV-C radiation can serve as an antimicrobial agent, while UV-A and UV-B radiations can affect the quality parameters such as color through the hormetic effect. Therefore, our goal was to identify optimum UV-A, UV-B, and UV-C radiation doses that can be applied to Honeycrisp apples to improve their coloration and microbial safety as the marketability of apples often depends on the redness of the fruit. The UV-C dose of 7.5 kJ/m2 resulted in a 1.2±0.06 log CFU/sample inactivation of L. monocytogenes on the apple surface. Interestingly, the additional UV-C dose exposure did not result in additional inactivation. This observed lack of dose-dependence could be the result of a) UV-C penetration interference from previously inactivated microbial cells resulting in a shadowing effect, b) the formation of a biofilm during ambient air drying and 4°C incubation that provided some protection during treatment, or c) higher resistance of L. monocytogenes sub-population against UV-C inactivation. This data will allow for future exploration of a synergistic treatment that can improve the color and appearance of Honeycrisp apples and improve their safety at the same time. UV radiation has shown promising antimicrobial activity and, through the studies carried out in this project, demonstrated potential beneficial or deleterious effects on food quality. The results from the first study showed the significance of understanding the interaction of food ingredients with UV radiation. The strawberry and apple studies show that UV radiation, when used at the correct dosage, can increase, or maintain the visual appearance of the fruits, making them more marketable. When used at the correct wavelength and for the appropriate duration, UV radiation can mitigate the prevalence of foodborne pathogens and contribute to food products' quality and shelf life.
VALIDATING THE EXPANDED FOOD SECURITY SCREENER AND HOME-DELIVERED MEAL PRIORITIZATION TOOL FOR ELIGIBLE INDIVIDUALS
(2022) Mendez, Izabelle; Sahyoun, Nadine; Nutrition; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
The Expanded Food Security Screener (FSS-Exp) is a screening tool for determining food insecurity risk based on individuals’ physical, economic, and social access to food. The FSS-Exp is the only tool that provides a simple and quick method for identifying which home-delivered meal (HDM) applicants are most vulnerable. However, further testing is required to determine the tool’s validity when compared with health and food-related measures associated with food security. Therefore, this study aimed to validate the FSS-Exp and HDM prioritization tool using concurrent and construct validity methods. To evaluate the concurrent validity, we examined the correlations between the FSS-Exp and other health and food-related tools such as SF-36, MOS-SSS, SWFL, SNAQ, and DST, which were associated with economic food insecurity and physical limitation To determine construct validity using the known-groups approach, we compared those health and food-related scores among participants grouped by different access barriers to food (priority levels). The results showed that the FSS-Exp correlated moderately (r=0.20 to 0.33) with other “gold standard” measures: SF-36, MOS-SSS, and SWFL. Furthermore, our results showed that individuals categorized as priority level A and B had significantly lower mean scores than priority level E for the SF-36 (HRQL) summary measures (p<0.05), SWFL (p<0.05), and the tangible social support ( p<0.01). Thus, the FSS-Exp was shown to be a valid measure in identifying and prioritizing older adults with physical and/or economic access to food, which is especially important for HDM programs with limited resources experiencing increased demand for services. For HDM programs which exist to serve vulnerable older adults, using the validated FSS-Exp tool can help identify those who are most vulnerable and in need of immediate service.
Enhancement of thermal processing with food-grade antimicrobial compounds in low-moisture food matrix to improve food safety
(2022) Ding, Qiao; Tikekar, Rohan V; Food Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Heat resistant foodborne pathogens have been a concern in low-moisture foods and ingredients (LMFs). Increased thermal resistance of pathogens such as Escherichia coli O157:H7 and Salmonella Typhimurium at low water activity (aw) reduces the efficiency of thermal treatment in LMFs. Alternative methods are therefore needed to augment thermal processing and reduce food safety risk. This study investigated the enhancement of thermal treatment efficiency against pathogenic bacteria in LMF matrices at different aw by inclusion of food-grade antimicrobial compounds. Based on their similar target sites in cells, it is hypothesized that antimicrobial compounds may work synergistically with heat treatment in LMF matrices. The treatment effect may be dependent on both aw and matrix compositions. Physiological and transcriptional changes may take place within cells adapted to different environments and contribute to the varied bacterial resistance. A combination of Butylparaben (BP), a known antimicrobial and thermal treatment was tested to enhance inactivation rates of S. Typhimurium and E. coli O157:H7 in meat and bone meal (MBM) equilibrated to water activity (aw) of 0.4 and 0.7. Presence of BP significantly accelerated thermal inactivation of E. coli O157:H7 in MBM at both 0.4 and 0.7 aw. However, inactivation rate of S. Typhimurium was not affected by the combined treatment at either 0.4 or 0.7 aw. BP accelerated the inactivation of the thermal-resistant subpopulation in E. coli O157:H7 but not S. Typhimurium. Therefore, further studies were conducted to identify alternative antimicrobial compounds to enhance thermal treatment against S. Typhimurium in LMF matrices and investigate the resistance mechanisms. Trans-cinnamaldehyde (CA) and eugenol (EG) were selected from a screening study to assist thermal treatment against S. Typhimurium adapted to different aw in whey protein (WP), corn starch (CS) or peanut oil (PO). Although addition of CA or EG significantly accelerated thermal inactivation of S. Typhimurium in water and LMF components at 0.9 aw, similar effect was not observed in bacteria adapted to lower (0.4) aw in any of those matrices. The matrix effect on bacterial thermal resistance was observed at 0.9 aw and were ranked as WP > PO > CS. The combined heat treatments reduced bacterial metabolic activity, which was partially dependent on the food components as well. S. Typhimurium had lower membrane fluidity because of membrane fatty acid composition change when adapted to a lower aw, which contributed to resistance against the combined heat treatments. Based on the aw- and matrix- dependent bacterial resistance and physiological changes observed in the two previous studies, transcriptional analysis was conducted to further understand the molecular mechanism behind the bacterial resistance. Expression of nine stress-related genes in S. Typhimurium adapted to different aw in LMF components were analyzed with or without the CA-assisted heat treatment. The upregulation of rpoH and dnaK in S. Typhimurium was induced by the stress during bacterial adaptation to the low aw environments as well as the combined heat treatment, which contributed to the bacterial resistance to both desiccation and the combined treatment. Although its link to the desiccation response in bacteria is not fully understood, the downregulation of ompC during the combined treatment also partially contributed to the treatment resistance. The upregulation of rpoE, otsB, proV and fadA in S. Typhimurium was induced by the desiccation stress during incubation at the low aw environment but was not a major contributor to the resistance during the combined treatment. The observed upregulation of fabA and downregulation of ibpA could not be directly linked to either bacterial resistance to desiccation or the combined heat treatment. Differential expressions were also observed among different aw levels or in different matrices at the same aw. The inconsistencies between the expression profiles of dnaK and ompC and bacterial resistance during the combined treatment suggested the presence of additional stress-response pathways.In conclusion, the results have demonstrated the potential of using food-grade antimicrobial compounds to complement thermal treatment in LMF during processes that start with a relatively high aw (such as dehydration). However, their effectiveness may subside at low aw environments. Future works should focus on optimizing the parameters of the CA/ EG-assisted heat treatment for higher efficiency, studying the molecular mechanism behind the bacterial resistance at global scale, and exploring alternative non-thermal processing technologies to enhance thermal treatment efficiency in LMFs without raising thermal treatment intensity.
Determinants of Food Insecurity in the League of Arab States
(2022) Sheikomar, Olfat Bakur; Sahyoun, Nadine R; Nutrition; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)
Background Food insecurity (FI) is widespread in the League of Arab States (LAS) due to factors including gender inequality, conflict, and political turmoil. However, limited data are available on its prevalence and determinants in that region. This dissertation aims to 1) validate the Food Insecurity Experience Scale (FIES), developed by the UN Food and Agriculture Organization, and assess the prevalence of FI in that region; 2) examine the association between FI, physical health, and mental well-being by sex and whether social support modifies this association in women; 3) examine the relationship between Palestinian live-in grandparents (GP) and the health and well-being of their grandchildren.Methods Rasch modeling was applied to the Gallup World Poll data to validate FIES in 19 countries of the region. Descriptive and logistic regression analyses were applied to data from 62,261 respondents aged 19 and over to examine determinants of FI. Logistic regression was also used to analyze the data of 2707 households and 8,034 children ages 0-17 from two surveys of Palestinian refugees. Results FIES met the Rasch assumptions indicating good internal validity. The prevalence of severe FI was 15.7% and women were at highest risk compared to men (17.6% vs. 14.1%, respectively, p<0.0001). Older age, living in rural areas, and high dependency ratio were associated with severe FI in women. High negative experience and low income were associated with severe FI, but good physical health and high positive experience with lower prevalence of severe FI. Having a live-in GP in the household was associated with grandchildren’s lower odds of experiencing an acute disease and higher odds of attending school. Conclusion FIES is a valid tool; however, cognitive testing of some items and omission of correlated ones may improve the scale. FI is highly prevalent and should be monitored to develop multi-sectoral intervention strategies. Mixed method studies are needed to better understand the relationship between grandparents and their grandchildren.

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