Food Science Commons^™

Investigating The Effect Of Agronomic Factors On Microbiome Of Horticulture Produce, Maryada Bohra

ORBioM (Open Research BioSciences Meeting)

The ranged microbiome in fruits and vegetables contributes substantially to the health of vegetation which, in effect, benefits human health. These microorganisms undertake an extensive variety of responsibilities including flavour creation, ripening and health preservation via the production of second-generation metabolites. There is limited information however on the influence of agronomic practices such as crop ripening stage at harvest, storage, packing material and how they impact the microbiome of the crop itself. The overall role of the crop microbiome in gut health is also not fully clear.

The Meta-Hort project will examine the effects of premature harvesting along with tray …

Genomic Diversity And Carbohydrate Utilisation In Human-Associated Bifidobacterial Isolates, Ortensia Catalano Gonzaga

ORBioM (Open Research BioSciences Meeting)

Bifidobacteria are beneficial commensals of the human gastrointestinal tract and their presence in the gut has been associated with positive health effects on the host. They account for a vast proportion of the infant gut microbiota, when the infant is fed on a milk-based diet, with their number progressively decreasing in adult and elderly. The gut microbiota and associated metabolic activities significantly impact on human health by promoting appropriate development of the infant immune system and contributing to the maintenance of the intestinal homeostasis. In recent years it has become clear that microbial colonization of the gut immediately following birth …

Growth Of Food-Borne Pathogens Listeria And Salmonella And Spore-Forming Paenibacillus And Bacillus In Commercial Plant-Based Milk Alternatives, Klaudia Bartula

ORBioM (Open Research BioSciences Meeting)

An increase in vegan diet preference, lactose intolerance, calorie concern and environmental awareness has led to a rise in the popularity of plant-based alternatives to bovine milk. However, there are still gaps in understanding how known bacterial food contaminants behave in plant-based beverages. The present study is the first to compare the growth of food-pathogens Listeria monocytogenes and Salmonella enterica, food spoilage Bacillus subtilis and an industrial milk product isolate, spore-forming Paenibacillus in commercially available ultrahigh temperature processed bovine milk and plant-based milk alternatives (coconut, almond, cashew). Beverage samples were inoculated with a strain cocktail or individual strains of either …

Valorisation Of Underutilised Dairy Waste Residues: Production Of Lactic Acid Through Microbial Fermentation, Chatan Rai Surana

ORBioM (Open Research BioSciences Meeting)

Annually, approximately 190 million tonnes of liquid waste or co-products is generated by the dairy sector across the globe. These waste streams are nutrient-rich and currently underutilised which gives scope to transform them through microbial fermentation to produce economically valuable products and reduce their negative environmental impact. Such an approach can contribute to the circular bioeconomy by making food production systems more sustainable.

In this study, whey from acid casein hydrolysis (acid whey) and salty whey from Cheddar cheese manufacture were evaluated as feedstocks for production of lactic acid. A total of 466 lactic acid bacteria strains were screened on …

Growth Of Foodborne Pathogens Listeria And Salmonella And Spore-Forming Paenibacillus In Commercial Plant-Based Milk Alternatives, Klaudia Bartula

ORBioM (Open Research BioSciences Meeting)

An increase in vegan diet preference, lactose intolerance, calorie concern and environmental awareness has led to a rise in the popularity of plant-based alternatives to bovine milk. Non-dairy products are a fast-growing food industry sector; however, there are still gaps in understanding how the known bacterial food contaminants behave in plant-based beverages. The present study is the first to compare the growth of food-pathogens Listeria monocytogenes and Salmonella enterica and an industrial milk product isolate, spore-forming Paenibacillus, in commercially available ultrahigh temperature processed (UHT) bovine milk and plant-based milk alternatives (coconut, almond, cashew). Beverage samples were inoculated with a strain …

Application Of Fructans-Degrading Lachancea Fermentati Fst5.1 As An Alternative To Baker’S Yeast In The Production Of A Low-Fodmap Whole Wheat Bread., Małgorzata Borowska

ORBioM (Open Research BioSciences Meeting)

FODMAPs are a group of poorly absorbed dietary carbohydrates comprised of easily fermentable oligo-, di-, monosaccharides and polyols. When consumed, they can exert gastrointestinal symptoms of pain, flatulence, or an irregular bowel movement in individuals with irritable bowel syndrome. Whole wheat bread is an excellent source of proteins, vitamins, polyphenols, and other nutrients, but its consumption is restricted in a low-FODMAP diet due to high fructans content. Although alternatives exist, these are primarily gluten-free, often inferior in terms of quality, nutritional profile, and consumer acceptance. Bioprocessing using microbial fermentation has a proven capacity to reduce FODMAPs in cereal products, but …

A Study On Plant-Derived Small Proteins And Peptides With Antiyeast Activity And Preliminary Investigations In Food Materials, Giulia Mignone

Theses

The discovery of novel antimicrobial molecules would benefit considerably different biotechnological fields, including medicine, agriculture and the food industry, on which this thesis work focuses. Specifically, here are presented fundamental studies on plant-derived antimicrobial proteins for the reduction of food spoilage caused by yeast microorganisms. Small proteins and peptides from the plant innate immune system possess several characteristics which could make them ideal candidates for a novel and natural antispoilage agent. The first Chapter reviews proteins families associated with the plant innate immune system that exhibit antiyeast activity and low molecular weight (<30 kDa), focusing on their structure and antiyeast mode of action. Chapter 2 investigates two peptides, Pn-AMP1 and Pn-AMP2 belonging to the hevein-type peptides family; peptides were synthetically generated using their native amino acid sequence. Synthetic Pn-AMP1 showed fast action activity against common food spoilage yeasts, it resulted potentially safe for consumption, and it was successfully incorporated in UHT milk and Fanta Orange, where it contained the growth of Kluyveromyces lactis and Zygosaccharomyces bailii, respectively. A different family of antimicrobial plant proteins (2S albumins) was studied in Chapters 3, 4, 5 and 6. Chapter 3 describes the selective extraction of an antiyeast protein (labelled WMS1 in this thesis work) of ~14 kDa from white mustard (Brassica hirta) seeds. The protein was purified in a relatively high yield, using a simple protocol that included only one chromatographic method step. In Chapter 4, bioinformatics tools and wet-lab techniques are used to study the structure and classify the antiyeast protein. The WMS1 antiyeast protein was identified as an isoform of the Napin protein Allergen Sin a 1 belonging to the 2S albumins family, characterised by high structural stability which donates to the protein resistance heat treatment and proteolytic degradation. Chapter 5 describes the antimicrobial spectrum of WMS1 , its antiyeast mechanism of action and its validity as preservative agent. Although WMS1 resulted potentially safe for consumption and inhibited the growth of Z. bailii in several beverage systems, its biotechnological potential is limited by its allergic properties. In Chapter 6, the protein WMS1 was cloned in a pET28a-MBP vector and expressed in an E. coli BL21 system. Interestedly, recombinant WMS1 and native WMS1 showed similar activity against the yeast Z. bailii. Thesis abstract 2 Chapter 7 includes a protocol for the rational design of ultra-short antimicrobial peptides. Since ultra-short antimicrobial sequences can be found within the sequence of any larger protein, the protocol was applied using WMS1 primary sequence, and seven ultra-short SinA-AMPs were chemically generated. Among the various SinA-AMPs, the peptide labelled SinA-pepIII had in vitro activity against several common yeast spoilage strains via membrane permeabilisation and it could be employed as a food preservative as it was pH- and heat- stable, potentially safe in terms of human consumption and it prevented the growth of Z. bailii in cranberry juice. The results obtained in this work offer new perspectives in the research for novel and clean-label food preservatives to fight the food waste phenomenon.

Is Tb Testing Associated With Increased Blood Interferon-Gamma Levels?, Aideen E. Kennedy, Jim O'Mahony, Noel Byrne, John Macsharry, Riona G. Sayers

Department of Biological Sciences Publications

The Republic of Ireland reports a relatively low prevalence of Johne’s disease (JD) compared to international counterparts. Postulated reasons for this include a lower average herd size and a grass-based production system. Ireland also engages in high levels of bovine tuberculosis (bTB) testing. As interferon-gamma (IFN-γ) is believed to play a key role in protecting against JD, it is our hypothesis that administration of purified protein derivative (PPD), as part of the bTB test, is associated with a systemic increase in IFN-γ production, which may potentially limit clinical progression of the disease. We studied 265 cows (202 Friesian and 63 …

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 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 …

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 …

Listeria Monocytogenes'. A Genomic Perspective On Its Pathogenesis, Persistence And Bacteriophages., Aidan Casey

Theses

The objective of this study was to investigate the genetic characteristics of Listeria monocytogenes that contribute to the threat it poses to the food processing industry. Transcriptomic analysis of a persistent L. monocytogenes isolate following exposure to an industrial detergent identified key upregulations in biological processes involved m peptidoglycan biosynthesis, chemotaxis and motility. Two bacteriophages against serotype 4b and 4e isolates of L. monocytogenes were isolated and sequenced. Comparative genomic and functional analysis identified the receptor binding proteins in each phage, whi]e transmission electron microscopy demonstrated a conserved baseplate architecture which also exists in other Gram­positive phages. Comparative genomics of …

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 …

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 …

Phages Of Non-Diary Lactococci: Isolation And Characterization Of Phi L47, A Phage Infecting The Grass Isolate Lactococcus Lactis Ssp Cremoris Dpc6860, Daniel Cavanagh, Caitríona M. Guinane, Horst Neve, Aidan Coffey, R. Paul Ross, Gerald F. Fitzgerald, Olivia Mcauliffe

Department of Biological Sciences Publications

Lactococci isolated from non-dairy sources have been found to possess enhanced metabolic activity when compared to dairy strains. These capabilities may be harnessed through the use of these strains as starter or adjunct cultures to produce more diverse flavor profiles in cheese and other dairy products. To understand the interactions between these organisms and the phages that infect them, a number of phages were isolated against lactococcal strains of non-dairy origin. One such phage, ΦL47, was isolated from a sewage sample using the grass isolate L. lactis ssp. cremoris DPC6860 as a host. Visualization of phage virions by transmission electron …

A Single Point Mutation In The Listerial Betl Sigma(A)-Dependent Promoter Leads To Improved Osmo- And Chill-Tolerance And A Morphological Shift At Elevated Osmolarity, Ronald F. Hoffman, Susan Mclernon, Audrey Feeney, Colin Hill, Roy D. Sleator

Department of Biological Sciences Publications

Betaine uptake in Listeria monocytogenes is mediated by three independent transport systems, the simplest of which in genetic terms is the secondary transporter BetL. Using a random mutagenesis approach, based on the E. coli XL1 Red mutator strain, we identified a single point mutation in a putative promoter region upstream of the BetL coding region which leads to a significant increase in betL transcript levels under osmo- and chill-stress conditions and a concomitant increase in stress tolerance. Furthermore, the mutation appears to counter the heretofore unreported “twisted” cell morphology observed for L. monocytogenes grown at elevated osmolarities in tryptone soy …

Evaluation Of Novel Low Molecular Weight Antimicrobial Compounds Produced By Lactic Acid Bacteria., Jiahui Leona Guo

Theses

Fungal food spoilage plays a pivotal role in the deterioration of food and feed systems and some fungi are also able to produce compounds toxic for humans and animals. The mycotoxins produced by fungi can cause serious health hazards and include toxins which are cancerogenic, immunotoxic, teratogenic, neurotoxic, nephrotoxic or hepatotoxic effects and Kashin-Beck disease. In addition to this, fungal spoilage/pathogens cause losses of marketable quality and hygiene of foodstuffs, resulting in major economic problem throughout the world. Nowadays, food spoilage can be prevented using physical and chemical methods, but no efficient strategy has been proposed so far to reduce …

Emerging Dynamics Of Human Campylobacteriosis In Southern Ireland, Susan Bullman, Daniel Corcoran, James O'Leary, Derry O'Hare, Brigid Lucey, Roy D. Sleator

Department of Biological Sciences Publications

Infections with Campylobacter spp. pose a significant health burden worldwide. The significance of Campylobacter jejuni/Campylobacter coli infection is well appreciated but the contribution of non-C. jejuni/C. coli spp. to human gastroenteritis is largely unknown. In this study, we employed a two-tiered molecular study on 7194 patient faecal samples received by the Microbiology Department in Cork University Hospital during 2009. The first step, using EntericBio® (Serosep), a multiplex PCR system, detected Campylobacter to the genus level. The second step, utilizing Campylobacter species-specific PCR identified to the species level. A total of 340 samples were confirmed as Campylobacter genus positive, 329 of …

Probiotic Therapy - Recruiting Old Friends To Fight New Foes, Roy D. Sleator

Department of Biological Sciences Publications

Against a backdrop of increasing antibiotic resistance, and the emergence of new and evolving pathogens, clinicians are increasingly forced to consider alternative therapies - probiotics are one such alternative.

Development Of Pcr Assays To Detect And Characterise Salmonella Isolates From Cheese And Meat Products By Utilizing An Integrated Genomics Approach, Noelle Geraldine Mccarthy

Theses

Salmonella is an extremely important foodborne pathogen which is responsible for millions of cases of gastroenteritis annually. Salmonella outbreaks have been associated with numerous common food stuffs such as meat, dairy and seafood. Traditional culture based methods for the detection Salmonella are labourious and costly. Rapid detection and serovar identification would dramatically decrease the time required to identify Salmonella outbreaks and potential health risks.

In the US 99% of all Salmonella infections are caused by members of Salmonella enterica subspecies 1. Serovars within subspecies 1 can vary due to host specificity and pathogenicity. DNA sequencing projects have provided valuable genomic …

Integronlike Structures In Campylobacter Spp. Of Human And Animal Origin, Brigid Lucey, D. Crowley, P. Moloney, B. Cryan, M. Daly, F. O'Halloran, E. J. Threlfall, S. Fanning

Department of Biological Sciences Publications

Resistance to antimicrobial agents used to treat severe Campylobacter spp. gastroenteritis is increasing worldwide. We assessed the antimicrobial resistance patterns of Campylobacter spp. isolates of human and animal origin. More than half (n = 32) were resistant to sulphonamide, a feature known to be associated with the presence of integrons. Analysis of these integrons will further our understanding of Campylobacter spp. epidemiology.