Food Microbiology Commons^™

Controlling Brochothrix Thermosphacta As A Spoilage Risk Using In Package Atmospheric Cold Plasma, Apurva Patange, Daniela Boehm, Carmen Bueno-Ferrer, Patrick J. Cullen, Paula Bourke

Articles

Brochothrix thermosphacta is a predominant spoilage microorganism in meat and its control in processing environments is important to maintain meat product quality. Atmospheric cold plasma is of interest for control of pathogenic and spoilage microorganisms in foods. This study ascertained the potential of dielectric barrier discharge atmospheric cold plasma (DBD-ACP) for control of B. thermosphacta in response to key parameters such as treatment time, voltage level, interactions with media composition and post treatment storage conditions. Challenge populations were evaluated as suspensions in PBS, as biofilms in meat model medium and surface attached on raw lamb chops under MAP.

ACP treatment …

Comparative Genomic Analysis Of Two Serotype 1/2b Listeria Monocytogenes Isolates From Analogous Environmental Niches Demonstrates The Influence Of Hypervariable Hotspots In Defining Pathogenesis, Aidan Casey, Kieran Jordan, Aidan Coffey, Edward M. Fox, Olivia Mcauliffe

Department of Biological Sciences Publications

The vast majority of clinical human listeriosis cases are caused by serotype 1/2a, 1/2b, 1/2c, and 4b isolates of Listeria monocytogenes. The ability of L. monocytogenes to establish a systemic listeriosis infection within a host organism relies on a combination of genes that are involved in cell recognition, internalization, evasion of host defenses, and in vitro survival and growth. Recently, whole genome sequencing and comparative genomic analysis have proven to be powerful tools for the identification of these virulence-associated genes in L. monocytogenes. In this study, two serotype 1/2b strains of L. monocytogenes with analogous isolation sources, but …

A Portable And Automatic Biosensing Instrument For Detection Of Foodborne Pathogenic Bacteria In Food Samples, Zhuo Zhao

Graduate Theses and Dissertations

Foodborne diseases are a growing public health problem. In recent years, many rapid detection methods have been reported, but most of them are still in lab research and not practical for use in the field. In this study, a portable and automatic biosensing instrument was designed and constructed for separation and detection of target pathogens in food samples using nanobead-based magnetic separation and quantum dots (QDs)-labeled fluorescence measurement. The instrument consisted of a laptop with LabVIEW software, a data acquisition card (DAQ), a fluorescent detector, micro-pumps, stepper motors, and 3D printed tube holders. First, a sample in a syringe was …

Farm-To-Fork Fresh Produce Food Safety: An Evaluation Of Perceptions, Knowledge, And Implementation Of Good Agriculture Practices In Kentucky, Daniel J. Sinkel

Masters Theses & Specialist Projects

Farmers’ markets have increasingly become a popular venue for purchase of fresh, locally-grown produce, with the number of farmers’ markets in Kentucky reaching an all-time high of 159 in 2016. Good Agriculture Practices (GAPs) is a program created by the USDA’s Agriculture Marketing Service to function as a food safety audit for small-scale fresh produce growers, such as those who sell fresh produce at local farmers’ markets. However, under the provisions of the Food Safety Modernization Act of 2011, small-scale farmers who sell an average of $25,000 in annual fresh produce sales across the span of three years are exempt …

Fungi In The Healthy Human Gastrointestinal Tract, Heather E. Hallen-Adams, Mallory J. Suhr

Department of Food Science and Technology: Faculty Publications

Many species of fungi have been detected in the healthy human gut; however, nearly half of all taxa reported have only been found in one sample or one study. Fungi capable of growing in and colonizing the gut are limited to a small number of species, mostly Candida yeasts and yeasts in the family Dipodascaceae (Galactomyces, Geotrichum, Saprochaete). Malassezia and the filamentous fungus Cladosporium are potential colonizers; more work is needed to clarify their role. Other commonly-detected fungi come from the diet or environment but either cannot or do not colonize (Penicillium and Debaryomyces species, which are …

Mitigating Prevalence And Persistence Of L. Monocytogenes Using Novel Sanitation Strategies In Delis, Susan Rose Hammons

Open Access Dissertations

Ready-to-eat (RTE) deli meats sliced at retail are predicted to cause 83% of deli meat-associated listeriosis cases annually. While L. monocytogenes is commonly found in delis (55-90%), environmental prevalence varies by store (0-40%). Study 1 evaluated efficacy of 10 standardized SSOPs by testing 28 food and non-food contact surfaces for L. monocytogenes over 6 months in 30 retail delis. Intervention strategies employed in this study significantly decreased L. monocytogenesprevalence on FCS and NFCS in stores with moderate prevalence (0.5%, 0.8% reduction). However, we found that L. monocytogenes could persist despite implementation of SSOP control strategies. Study 2 assessed an …

Listeria Monocytogenes Strain Variation: Starvation, Metabolism, And Macrophage Survival, Nathan A. Jarvis

Graduate Theses and Dissertations

Listeria spp. are often isolated from raw and processed food products. Listeria monocytogenes is responsible for death rates of 10 to 30% in immune compromised people; it can also infect a vast range of animals. There are numerous reports of poultry harboring or becoming infected with L. monocytogenes. First, this research assessed the ability of L. monocytogenes to replicate within chicken macrophages. Using the gentamicin-killing assay, a cell culture of chicken macrophage-like cells (HD11) were infected with two strains of L. monocytogenes individually (EGD-e and Scott A). Results show that unlike data on mouse macrophages, intracellular L. monocytogenes do not …

Assessing Stress Responses To Atmospheric Cold Plasma Exposure Using Escherichia Coli Knock Out Mutants, Lu Han, Daniela Boehm, Sonal Patil, Patrick J. Cullen, Paula Bourke

Articles

Aim: This study investigated the effect of Atmospheric cold plasma (ACP) exposure induced stress on microbial inactivation patterns and the regulation of genes involved in the microbial stress response in conjunction with key processing parameters of exposure time and post treatment storage time.

Methods and Results: Cell suspensions of Escherichia coli BW 25113 and its isogenic knock-out mutants in rpoS, soxR, soxS, oxyR and dnaK genes were treated with high voltage ACP in a sealed package for 1, 3 and 5 min followed by 0, 1 and 24 h post-treatment storage. ROS densities and colony formation were …

From Milpas To The Market: A Study On The Use Of Metal Silos For Safer And Better Storage Of Guatemalan Maize, José Rodrigo Mendoza

Department of Food Science and Technology: Dissertations, Theses, and Student Research

This project aimed to implement the use of metal silos to improve quality and safety of maize consumed by inhabitants of the highlands of Guatemala. This manuscript includes a literature review of the maize production chain in Guatemala, a survey about agricultural practices used in the region of study, as well as a characterization of the analyzed maize regarding its mycoflora, nutritional composition, and insect infestation. To better understand the current situation regarding agricultural practices and maize consumption, a survey was carried out. Sample consisted of 280 families representing 14 rural communities distributed in the townships of Todos Santos and …

A Novel And Rapid Staphylococcus Aureus Bacterial Identification Method Utilizing Immunomagnetic Beads And Single Cell Laser-Light Scattering, Kaylagh Hollen

All NMU Master's Theses

Staphylococcus aureus is the most commonly isolated human associated bacterial pathogen. It plays an important role in skin and soft-tissue infections, pneumonia, endocarditis, osteomyelitis, foreign-body infections, and sepsis. S. aureus diagnosis and treatment requires a minimum of 24-48. With this in mind, previous studies suggest that faster pathogen identification has been linked to improved patient outcomes. Improved patient outcomes including a reduction in hospitalization time, decreased risk of nosocomial infections, and decreased in medical costs. The impact of faster identification on patient outcome has led us to develop an alternative method of S. aureus identification via ImmunoMagnetic Separation (IMS) and …

Decay Of Macroalgae And Leaves And Their Relation To Detrital Food Webs, Megan E. Grandinetti

Masters Theses & Specialist Projects

This project addressed if decaying macroalgae and leaf detritus play a major role in the detrital pool of a 7th-order karst riverine system. Decay rates, macroinvertebrates colonization patterns, and change in δ13C values of Cladophora, Platanus occidentalis, and a mix of Acer negundo and A. saccharinum were tracked during summer and autumn months for portions of multiple years.

Packs of air-dried Cladophora, Acer, and P. occidentalis were placed in mesh bags and put in groups (n=4) in wire baskets. Seven baskets were submerged in riffle (0.5 m) and deeper run (2 m) habitats. Benthic organic matter was collected with each …

An Analysis Of Bacterial Contamination Of Chicken Eggs And Antimicrobial Resistance, Holly Spitzer

All College Thesis Program, 2016-2019

Chicken eggs are a major component of American diets, with an average yearly consumption of approximately 250 eggs per person (American Humane Society). While highly nutritious, eggs are also one of the leading causes of food poisoning and food borne illness in the United States. Eggs may become contaminated by a number of different types of bacteria during production, including Salmonella, a group of bacteria that, according to the CDC, causes more than 1.2 million cases of food borne illness in the United States every year. In an effort to decrease the frequency of bacterial contamination, many food producers …

Controlling Microbial Safety Challenges Of Meat Using High Voltage Atmospheric Cold Plasma, Lu Han, Dana Ziuzina, Caitlin Marie Heslin, Daniela Boehm, Apurva Patange, David Millan-Sango, Vasilis Valdramidis, Patrick J. Cullen, Paula Bourke

Articles

Atmospheric cold plasma (ACP) is a non-thermal technology, effective against a wide range of pathogenic microorganisms. Inactivation efficacy results from plasma generated reactive species. These may interact with any organic components in a test matrix including the target microorganism, thus food components may exert a protective effect against the antimicrobial mode of action. The effect of an in-package high voltage ACP process applied in conjunction with common meat processing MAP gas compositions as well as bacteria type and meat model media composition have been investigated to determine the applicability of this technology for decontamination of safety challenges associated with meat …

Influence Of Lactobacillus Rhamnosus Isolated From “Amabere Amaruranu” Cultured Milk On Adipogenesis, Justin E. Kotala

Electronic Theses and Dissertations

This study was performed to test the in vitro effects of a Lactobacillus rhamnosus isolate from “amabere amaruranu”, a traditional Kenyan cultured milk, on 3T3-L1 and Caco-2 cell lines. Cultures of fully mature 3T3-L1 adipocytes were treated with bacterial isolate cell extract (CE), filtered spent broth (FSB) from overnight bacterial culture, or with a PBS control. Expression levels of PPAR³1 and 2, C/EBP±, and ATGL proteins in 3T3-L1 cells were upregulated by FSB treatment. CE treatment did not affect protein expression levels. Expression of MTTP and SREBP-1c proteins in Caco-2 cells showed no change with either treatment. Optical density measurements …

A Tail Of Two Phages: Genomic And Functional Analysis Of Listeria Monocytogenes Phages Vb_Lmos_188 And Vb_Lmos_293 Reveal The Receptor-Binding Proteins Involved In Host Specificity, Aidan Casey, Kieran Jordan, Horst Neve, Aidan Coffey, Olivia Mcauliffe

Department of Biological Sciences Publications

The physical characteristics of bacteriophages establish them as viable candidates for downstream development of pathogen detection assays and biocontrol measures. To utilize phages for such purposes, a detailed knowledge of their host interaction mechanisms is a prerequisite. There is currently a wealth of knowledge available concerning Gram-negative phage-host interaction, but little by comparison for Gram-positive phages and Listeria phages in particular. In this research, the lytic spectrum of two recently isolated Listeria monocytogenes phages (vB_LmoS_188 and vB_LmoS_293) was determined, and the genomic basis for their observed serotype 4b/4e host-specificity was investigated using comparative genomics. The late tail genes of these …

Prokaryotic Diversity In The Rhizosphere Of Organic, Intensive, And Transitional Coffee Farms In Brazil, Adam Caldwell, Livia Silva, Cynthia Da Silva, Cleber Ouverney

Faculty Publications, Biological Sciences

Despite a continuous rise in consumption of coffee over the past 60 years and recent studies showing positive benefits linked to human health, intensive coffee farming practices have been associated with environmental damage, risks to human health, and reductions in biodiversity. In contrast, organic farming has become an increasingly popular alternative, with both environmental and health benefits. This study aimed to characterize and determine the differences in the prokaryotic soil microbiology of three Brazilian coffee farms: one practicing intensive farming, one practicing organic farming, and one undergoing a transition from intensive to organic practices. Soil samples were collected from 20 …

The Effects Of Electrostatic Spraying With Organic Acids In The Disintegration Of Biofilms Formed By E.Coli O157:H7 And Salmonella Typhimurium On Spinach And Cantaloupe, Ahmad Almasoud

Graduate Theses and Dissertations

Outbreaks from the consumption of fresh produce are a concern in the United States. The consumptions of fresh produce have increased recently which expose a large segment of society to such outbreaks. Spinach and cantaloupe are minimally heated or processed before consumption which makes them a possible source of foodborne illness. The objective of this research was to investigate the effect of organic acids alone and in combination to reduce attached Salmonella Typhimurium (S.T) and Escherichia coli O157:H7 (E.coli) on spinach and cantaloupe, and to disintegrate biofilm formed by these pathogens by electrostatically spraying with two organic acids. To quantify …

Characterization And Investigation Of Fungi Inhabiting The Gastrointestinal Tract Of Healthy And Diseased Humans, Mallory J. Suhr

Department of Food Science and Technology: Dissertations, Theses, and Student Research

Gastrointestinal microbiome studies have failed to include fungi in total community analyses. As a result, their diversity and function in the gut is poorly understood. Recent work has begun to uncover the role intestinal fungi play in diet, immune system development, interactions with other microorganisms in the gut, and pathogenesis of diseases. Advances in sequencing technologies allow for the ability to profile the fungal gut microbiome (“mycobiome”) in healthy and diseased states. This thesis explores the mycobiome in 1) healthy humans with a vegetarian diet and 2) pediatric small bowel transplant recipients that develop fungal bloodstream infections.

The gut mycobiome …

Functional Screening Of The Cronobacter Sakazakii Baa-894 Genome Reveals A Role For Prop (Esa_02131) In Carnitine Uptake, Audrey Feeney, Roy D. Sleator

Department of Biological Sciences Publications

Cronobacter sakazakii is a neonatal pathogen responsible for up to 80% of fatalities in infected infants. Low birth weight infants and neonates infected with C. sakazakii suffer necrotizing enterocolitis, bacteraemia and meningitis. The mode of transmission most often associated with infection is powdered infant formula (PIF) which, with an aw of ∼0.2, is too low to allow most microorganisms to persist. Survival of C. sakazakii in environments subject to extreme hyperosmotic stress has previously been attributed to the uptake of compatible solutes including proline and betaine. Herein, we report the construction and screening of a C. sakazakii genome bank and …

Concomitant Uptake Of Antimicrobials And Salmonella In Soil And Into Lettuce Following Wastewater Irrigation, J. Brett Sallach, Yuping Zhang, Laurie Hodges, Daniel D. Snow, Xu Li, Shannon L. Bartelt-Hunt

Department of Civil and Environmental Engineering: Faculty Publications

The use of wastewater for irrigation may introduce antimicrobials and human pathogens into the food supply through vegetative uptake. The objective of this study was to investigate the uptake of three antimicrobials and Salmonella in two lettuce cultivars. After repeated subirrigation with synthetic wastewater, lettuce leaves and soil were collected at three sequential harvests. The internalization frequency of Salmonella in lettuce was low. A soil horizon-influenced Salmonella concentration gradient was determined with concentrations in bottom soil 2 log CFU/g higher than in top soil. Lincomycin and sulfamethoxazole were recovered from lettuce leaves at concentrations as high as 822 ng/g and …

Antimicrobial Efficacy Of Natural Bioactive Compounds And High Pressure Processing Against Potential Pathogens In Infant Foods, Hayriye Cetin-Karaca

Theses and Dissertations--Animal and Food Sciences

This study investigated the antimicrobial efficacy of bioactive plant compounds along with high pressure processing (HPP) against pathogens Bacillus cereus and Cronobacter sakazakii in infant formula and infant rice cereal. The influence of these applications on antimicrobial activity, shelf-life and sensory attributes of infant foods were examined.

Trans-cinnamaldehyde (TC), (-)-Epigallocatechin gallate (EGCG) and [10]-Gingerol (GI) were incorporated (0.05%) in infant rice cereal reconstituted with infant formula. The cereal was inoculated with either B. cereus (ATCC 14579) or B. cereus spores (10⁷-10⁸ log CFU g^-1). All the samples were stored at 7, 23 or 37°C for …

Increasing The Nutritional Value Of Canola Meal Via Fungal Bioprocessing, Jason R. Croat

Electronic Theses and Dissertations

The main limitation of meals from canola and other Brassica spp. is the presence of glucosinolates (GLS), which are anti-nutritional and can even be toxic at high ingestion levels. Furthermore, large amounts of GLS can reduce palatability for livestock and thus reduce intake and growth rates. For this reason canola was bred to contain lower levels of GLS (<30 >μmol/g) and erucic acid (< 2%). However, feed inclusion rates are still limited to ~30%, and this reduces the value of canola meal. The goal of this thesis was to optimize a pretreatment and fungal conversion process to enhance the nutritional value of canola meal. Various combinations of physical/chemical pretreatments, fungal cultures, and incubation methods were investigated to metabolize GLS into cell mass, CO2, or other non-toxic components. These treatments also served to hydrolyze canola meal fiber into carbohydrates which were then metabolized by the fungi into single cell protein. Solid-state incubation conditions were initially tested, since filamentous fungi are well adapted to grow at lower moisture levels, and this would potentially reduce contamination issues with bacteria. Flask trials were performed using 50% moisture, hexane extracted (HE) or cold pressed (CP) canola meal, with incubation for 168 h at 30ºC. On HE canola meal Trichoderma reesei (NRRL-3653) achieved the greatest increase in protein content (23%), while having the lowest residual levels of sugar (8% w/w) and GLS (0.4 μM/g). On CP canola meal T. reesei (NRRL-3653), Aureobasidium pullulans (NRRL-58522), and …

Characterization Of The Ecological Role Of Genes Mediating Acid Resistance In Lactobacillus Reuteri During Colonization Of The Gastrointestinal Tract, Janina A. Krumbeck, Nathan L Marsteller, Steven Frese, Daniel A. Peterson, Amanda Ramer-Tait, Robert W. Hutkins, Jens Walter

Department of Food Science and Technology: Faculty Publications

Rodent-derived strains of Lactobacillus reuteri densely colonize the forestomach of mice and possess several genes whose predicted functions constitute adaptations towards an acidic environment. The objective of this study was to systematically determine which genes of L. reuteri 100-23 contribute to tolerance towards host gastric acid secretion. Genes predicted to be involved in acid resistance were inactivated, and their contribution to survival under acidic conditions was confirmed in model gastric juice. Fitness of five mutants that showed impaired in vitro acid resistance were then compared through competition experiments in ex-germ-free mice that were either treated with omeprazole, a proton-pump inhibitor …

Exploiting Bacteriophages And Associated Peptidoglycan Hydrolases With Potential For Biocontrol In Food Related Applications, Lorraine Endersen

Theses

The work presented in this thesis describes the isolation and characterisation of novel bacteriophages, and the subsequent exploitation of their bactericidal properties against two opportunistic food related pathogens, Mycobacterium avium subsp. paratuberculosis (MAP) and Cronobacter sakazakii.

MAP is a proven animal pathogen known to cause Johne’s disease in cattle but has been implicated as a causative agent of Crohn’s disease in humans. Despite the fact that this association has yet to be proven, significant focus has been directed towards evaluating the consequences of consuming milk contaminated with MAP. Accordingly, six mycobacteriophages were isolated and characterised in terms of temperature and …

Enhancement Of Antimicrobial Activity Of Naturally Occuring Phenolic Compounds By Nano-Particle Mediated Delivery Against Listeria Monocytogenes, Escherichia Coli O157:H7, And Salmonella Typhimurium, Madhuram Ravichandran

Graduate Theses and Dissertations

Food-borne illnesses are of major concern in the U.S. and worldwide. Salmonella,Listeria,E. coli O157:H7 continue to be some of the major foodborne pathogens. Recurring recalls due to these pathogens demand critical antimicrobial strategies to decontaminate the food through its procession from farm to fork. In our study, we combined the effectiveness of naturally occurring phenolic compounds individually, with/ without ethylenediamine tetraacetic acid (EDTA) and/or Nisin and nanoparticle delivery to improve the antimicrobial potential of the compounds against Listeria monocytogenes (L.m.), Salmonella Typhimurium (S.T.), and Escherichia coli O157:H7 (E.c.), in brain …

Antibacterial Activity Of Commercially Available Plant Extracts On Selected Campylobacter Jejuni Strains, Divya Oppath Sivasankaran

Graduate Theses and Dissertations

Campylobacter jejuni is recognized as a leading cause of foodborne illness worldwide with approximately 850,000 cases per year in the United Sates. A total of 18 species of Campylobacter have been identified worldwide to date. Foodborne transmission of C. jejuni is mainly through the consumption of unpasteurized milk, contaminated poultry meat and water. C. jejuni can survive in very diverse environments under stressful conditions in human and animals which is different from the specific conditions and media required in laboratory experiments. There has been increased research in recent years to identify naturally occurring antibacterial agents to control and eliminate C. …

Probiotic Potential Of Bacterial Isolates From ‘Amabere Amaruranu’ Cultured Milk, Blaise B. Boyiri

Electronic Theses and Dissertations

Probiotics are viable nonpathogenic microbes that positively affect host health. Probiotics inhibit infection, activate immunity, and promote mucosal-barrier development. Many microbes have probiotic activity. Nonetheless, the selection of stable strains and their specific mechanism(s) of action are not fully elucidated. Bacteria from ‘Amabere amaruranu’ cultured milk from Kenya were isolated and identified by PCR sequence analysis of the 16S rRNA gene. Isolates were examined for stability to acid and bile, antimicrobial activity, mucin production, and degradation and sensitivity to antibiotics, hence their potential for probiotics. Lactobacillus isolates were acid unstable, bile-stable, nonmucinolytic, and presented antibacterial activity. L. rhamnosus cell fractions …

Understanding The Factors Affecting Microbiological Quality Of Wheat Milled Products: From Wheat Fields To Milling Operations, Luis E. Sabillón Galeas

Department of Food Science and Technology: Dissertations, Theses, and Student Research

Although regarded as a low-risk commodity, wheat flour-based mixes have been implicated in several food safety incidents. The present thesis is a compilation of five scientific manuscripts on the effect of weather variation, milling steps and implementation of pre-milling interventions on the microbiological quality and safety of wheat and milled products. The first manuscript is a review of the microbiological quality and safety of wheat-based products. Despite the low water activity, wheat flour may harbor dormant but viable microorganisms, which could lead to safety concerns when flour is used to produce refrigerated dough products. The second manuscript illustrates the effect …

Investigating Antibiotic Resistance Levels Of Salmonella Internalized In Lettuce Leaves, Jason B. Thomas

McNair Scholars Research Journal

Contamination of food crops by the human pathogen Salmonella is a food safety threat worldwide. Though using treated wastewater for irrigation is a sustainable practice, it may introduce trace levels of Salmonella that may contaminate food crops. Salmonella could develop resistance to antibiotics present in wastewater. The overall goal of the project is to increase the understanding of the public health risk associated with the use of treated wastewater to irrigate food crops. The objective of this particular study is to determine the antibiotic resistance level of Salmonella internalized in lettuce leaves. In this experiment, thirty-six plants of the lettuce …

Transcriptome Analysis Of Listeria Monocytogenes Exposed To Biocide Stress Reveals A Multi-System Response Involving Cell Wall Synthesis, Sugar Uptake, And Motility, Aidan Casey, Edward M. Fox, Stephan Schmitz-Esser, Aidan Coffey, Olivia Mcauliffe, Kieran Jordan

Department of Biological Sciences Publications

Listeria monocytogenes is a virulent food-borne pathogen most often associated with the consumption of “ready-to-eat” foods. The organism is a common contaminant of food processing plants where it may persist for extended periods of time. A commonly used approach for the control of Listeria monocytogenes in the processing environment is the application of biocides such as quaternary ammonium compounds. In this study, the transcriptomic response of a persistent strain of L. monocytogenes (strain 6179) on exposure to a sub-lethal concentration of the quaternary ammonium compound benzethonium chloride (BZT) was assessed. Using RNA-Seq, gene expression levels were quantified by sequencing …