Pathogenic Microbiology Commons^™

Biowill - Characterising Willow Bark Bio-Actives For Skin Therapies, Arnold Marisa

ORBioM (Open Research BioSciences Meeting)

Willow bark is considered as a disposable by-product when processing willow for biomass. Willow (Salix) is known to contain high value bioactive compounds which include salicin and its derivatives, and other phytochemicals of interest such as polyphenols and flavonoids. The plant is historically known as the primary source of salicylates to which the well-known drug aspirin is derived from. The work forms part of the Interreg project BioWILL, which is focused on integrated “Zero Waste” biorefinery utilising all fractions of willow feedstock for the production of biochemicals and renewable energy. This project aims to investigate the crude and …

Investigation Of The Urobiome For The Production Of Novel Antimicrobials Against Uropathogenic E. Coli (Upec), Jennifer Jones

ORBioM (Open Research BioSciences Meeting)

BACKGROUND: Urinary tract infections (UTIs) are one of the most common bacterial infections globally with antibiotic treatment becoming increasingly less effective. The urobiome remains a relatively understudied niche as a source of potentially novel antimicrobials (e.g., bacteriocins). Improvements to bacterial culturing and sequencing techniques have highlighted the potentially rich source of alternative treatments and control strategies to target uropathogenic E. coli (UPEC).

MATERIALS AND METHODS: (a) BAGEL 4 was used for bioinformatic screening of the genomes of urobiome isolates to identify bacteriocin gene clusters (BGC).

(b) Expanded quantitative urine culture (EQUC) was used to culture mid-stream urine samples to isolate …

Elucidation Of Novel And Established Campylobacter Species With Clinical And Agricultural Significance Through Phenotypic, Genotypic, And Taxonomic Investigation, Caoimhe Lynch

Theses

Campylobacter spp. are well-established human, veterinary and economic pathogens, with a broad host range spanning from terrestrial and marine mammalian, avian and reptilian hosts. The scope of the study includes novel and notorious species within the genus, with reference to zoonotic agents Campylobacter jejuni and Campylobacter coli - the leading cause of human bacterial gastroenteritis in the EU and Ireland, Campylobacter fetus that represents the most common campylobacter causing bacteraemia and two novel species isolated from pigs not described in the literature previously. C. jejuni and C. coli isolates (n = 350) recovered from broiler caecal and neck skin samples, …

An Investigation Into The Prevalence Of Vancomycin-Resistant Enterococci Within An Irish Hospital, Dylan Casey

International Undergraduate Journal of Health Sciences

Vancomycin-resistant enterococci (VRE) are well-recognised nosocomial pathogens that pose a significant threat to public health. Associated with poorer clinical outcomes than their vancomycin-sensitive counterparts, the prevalence of VRE in Ireland has increased in recent times, with the European Antimicrobial Resistance Surveillance Network reporting that out of 29 countries, Ireland demonstrated the highest rates of vancomycin resistance among invasive Enterococcus faecium isolates between 2011 and 2014 (2011; 34.9%, 2012; 44.0%, 2013; 42.7%, 2014; 45.1%). Herein, we investigate VRE prevalence in the Mercy University Hospital (MUH) – an acute care hospital in southern Ireland. A total of 21 first-time VRE infections were …

A Rapid Viability And Drug‑Susceptibility Assay Utilizing Mycobacteriophage As An Indicator Of Drug Susceptibilities Of Anti‑Tb Drugs Against Mycobacterium Smegmatis Mc2 155, Gillian Catherine Crowley, Jim O'Mahony, Aidan Coffey, Riona G. Sayers, Paul D. Cotter

Department of Biological Sciences Publications

Background: A rapid in-house TM4 mycobacteriophage-based assay, to identify multidrug resistance against various anti-tuberculosis drugs, using the fast-growing Mycobacterium smegmatis mc² 155 in a microtiter plate format was evaluated, based on phage viability assays. Methods: A variety of parameters were optimized before the study including the minimum incubation time for the drugs, phage and M. smegmatis mc² 155 to be in contact. An increase in phage numbers over 2 h was indicative that M. smegmatis mc² 155 is resistant to the drugs under investigation, however when phage numbers remained static, M. smegmatis mc² 155 found to …

The Occurrence Of Listeria Monocytogenes In The Mushroom Production Chain And The Use Of Bacteriophage For Its Control, Vincenzo Pennone

Theses

In Agaricus bisporus production, the occurrence of L. monocytogenes is still poorly understood. A number of studies have shown the presence of L. monocytogenes in the mushroom processing environment, suggesting its source from raw materials storage areas (Viswanath et al, 2013; Murugesan et al, 2015). In this study, 10 mushroom producers and three raw material (compost and casing) producers participated in two surveys on the occurrence of L. monocytogenes in the most high-risk areas of the production environment. In the first survey, floors, drains, platforms, crates, raw materials and mushrooms were tested for L. monocytogenes presence and the results were …

Crystallographic Structure Determination Of Bacteriophage-Encoded Enzymes That Specifically Target Pathogenic Bacteria, Marta Sanz Gaitero

Theses

Antibiotic resistance is becoming a serious public health concern. Infections that some decades ago could be treated with antibiotics now sometimes do not respond to traditional treatment, causing higher mortality and economic losses. An alternative to the use of antibiotics are bacteria's natural predators, bacteriophages (or phages), and specifically their lytic enzymes. These proteins are produced by phages to degrade bacterial peptidoglycan to inject their genetic material into the bacteria (virion-associated peptidoglycan hydrolases) or to release their progeny once the infection is finished (endolysins). They can be applied exogenously to lyse Gram-positive bacteria or be genetically engineered to lyse Gram-negative …

Mycobacterium Species Identification In An Acute Hospital: Towards Rapid Identification, And Improved Epidemiology, Using Novel Mass Spectrometry Analysis, James Anthony O'Connor

Theses

Mycobacteria are a heterogeneous group of bacteria that cause a wide spectrum of disease, including Tuberculosis (TB), the single biggest killer worldwide. The genus also incorporates over 170 nontuberculosis (NTM) species, many of which are opportunistic pathogens. The aim of this project was to ascertain the burden of mycobacterial disease in the region and to determine the efficacy of Matrix-Assisted Laser Desorption/Ionisation time-of-flight (MALDI-tof) mass spectrometry (MS) for novel uses as a rapid identification method for clinical mycobacteria and as a typing tool for Mycobacterium tuberculosis complex isolates. A five-year retrospective epidemiological study of the region was conducted, the first …

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 …

Clinical Coagulase-Negative Staphylococci; Identification, Characterisation And Treatment, Emma Gabriel

Theses

The work presented in this thesis focuses on clinically significant staphylococci (with an emphasis on coagulase-negative staphylococci) in terms of their isolation, identification, characterisation, and biocontrol.

CoNS are increasingly being recognised as true nosocomial pathogens yet remain under-characterised, particularly from an Irish clinical perspective. We examined a large cohort of clinically-derived CoNS, isolated from blood (n=l35), identifying them to species level. Extensive antibiotic resistance against nine clinically-relevant antibiotics was recorded. Additionally, widespread biofilm forming capabilities associated with increased virulence was detected both phenotypically and genotypically. We identified multiple species of CoNS in circulation in an Irish healthcare setting, and determined …

Phylogenetic Investigation Of Enteric Bovine Coronavirus In Ireland Reveals Partitioning Between European And Global Strains, Lynda Gunn, P. J. Collins, M. J. O'Connell, Helen O'Shea

Department of Biological Sciences Publications

Background

Bovine coronavirus is a primary cause of neonatal calf diarrhea worldwide, and is also associated with acute diarrhea in adult cattle during the winter season. There are no reports on molecular characterization of bovine coronavirus in Ireland, and little data exists apart from serological studies.

Findings

In this study, 11 neonatal (mean age 9 days) calf BCoV strains from the south of Ireland were collected over a one year period and characterized using molecular methods. The spike gene which encodes a protein involved in viral entry, infectivity and immune response shows the most variability amongst the isolates and was …

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 …

Identification Of The Molecular Mechanisms Governing The Osmotolerance Of The Emerging Gastrointestinal Pathogen Cronobacter Sakazakii, Audrey Feeney

Theses

Cronobacter sakazakii is an organism which has been associated with severe life- threatening diseases in infants and neonates. Mortality rates of up to 80% have been recorded in infants infected with Cronobacter sakazakii. Infants often suffer from septicaemia, necrotising enterocolitis and meningitis as a result of infection. A characteristic feature of the opportunistic foodborne pathogen Cronobacter sakazakii is its ability to survive in extremely arid environments such as powdered infant formula, making it a dangerous opportunistic pathogen of neonates. Herein, we provide a brief overview of the pathogen; clinical manifestations, environmental reservoirs and our current understanding of stress response mechanisms …

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 …

Gastroenteritis Viruses In Ireland: Epidemiology And Evolution, Lynda Gunn

Theses

Viral gastroenteritis is a common disease of both humans and animals. This thesis focuses on the use of classical virology, coupled with phylogenetics to study gastroenteritis viruses [Bocavirus (BoV), Coronavirus (CoV), and Rotavirus A (RVA)], isolated from selected samples from humans and animals. RVA was associated with outbreaks in elderly care facilities in Northern Ireland, isolates were genotyped and we reported the first detection of G2 lineage lie RVA in Ireland, with other common human RVA genotypes G1P[8], G2P[4] and G9P[8]. Evolutionary dynamics of human RVA VP7 genotypes was examined using codon models of evolution, results show heterogeneous selective pressure …

Detection, Identification And Characterization Of The Novel Campylobacter Species: Campylobacter Ureolyticus And Campylobacter Corcagiensis From Their Reservoirs., Monika Koziel

Theses

Campylobacter ureolyticus has been identified in a large proportion of diarrhoeal samples from patients in Ireland. However, the source of human infections has not been established and little is currently known about the potential reservoirs of C. ureolyticus in nature.

The aim of this project was to investigate a range of environmental and animal samples for the presence of this emerging pathogen using a molecular approach, to support the hypothesis of its zoonotic origin.

During the study, C. ureolyticus was detected in 30 out of 1,254 (2.4%) human diarrheic samples and isolated from 11 patients, describing, to the best of …

Characterisation Of The Enzybiotic Potential Of Selected Staphylococcus Aureus Bacteriophages, Ruth Keary

Theses

The work presented in this thesis focuses on harnessing the bactericidal potential of bacteriophage (phage) encoded peptidoglycan hydrolase enzymes to combat the significant pathogen Staphylococcus aureus. In order to identify novel peptidoglycan hydrolases lytic against S. aureus, the complete genomes of two staphylococcal temperate Siphoviridae bacteriophages, DW2 and CSl, were sequenced and annotated. Screening of these genomes identified genes for several putative peptidoglycan hydrolases including two endolysins and three virion-associated peptidoglycan hydrolases. The identification of putative genes relating to virulence and host fitness lends to the understanding of the role played by such temperate phages in the evolution of this …

Development Of A Recombinant Lactobacillus Salivarius For Prophylactic Applications Against Mycobacterium Avium Subspecies Paratuberculosis And Johne’S Disease, Christopher Johnston

Theses

Current whole-cell vaccines against Mycobacterium avium subsp. paratuberculosis, the etiological agent of Johne’s disease, are unable to provide complete protection against infection. Moreover, such whole-cell formulations interfere with diagnostic tests for bovine tuberculosis (Mycobacterium bovis). The development of subunit vaccines, those which are preferentially based on one, or a select few, immunodominant protective antigens is considered as the realistic next step in overcoming these issues. The (MAP) K-10 genome is a single circular chromosome of 4,829,781 base pairs and encodes 4,350 predicted ORFs. In order to identify MAP specific proteins for subsequent incorporation within a mucosal based vaccine development, a …

Identification And Characterisation Of Campylobacter Ureolyticus: A New And Emerging Gastrointestinal Pathogen, Susan Bullman

Theses

Campylobacter jejuni is universally recognized as the leading bacterial cause of human gastroenteritis. However, within recent years a growing number of non- jejuni/coli Campylobacter species have been acknowledged as emerging human and animal pathogens. Advances in molecular diagnostics and development of pioneering culture methodologies have led to the detection of a range of under-appreciated and nutritionally fastidious Campylobacter species, including C. ureolyticus. Herein, we provide the first report of C ureolyticus in the faeces of patients presenting with gastroenteritis and suggest a role for this organism as an emerging enteric pathogen.

Employing a two-tiered molecular study, we report C. ureolyticus …

The Isolation And Characterisation Of Acientobacter Baumannii Bacteriophage, Mohamed Farag M. Elbreki

Theses

Acinetobacter baumannii is a pathogen, which has an increasing prevalence of multidrug resistance. It exists widely in natural environments, and frequently in health-care settings where it has proven difficult to eradicate using antibiotic therapy. A possible alternative to conventional antibiotics is the use of bacteriophages (phages) as antibacterials. In this project, extensive screening of environmental samples such as soils, sewage and waters was undertaken to identify suitable phages. Three phages were successfully isolated from municipal waste in the Cork area. These were subsequently characterized in detail and examined by electron microscopy, which showed that they were all of the Myoviridae …

Antifungal Activity Of Lactobacillus Against Microsporum Canis, Microsporum Gypseum And Epidermophyton Floccosum, Jiahui Guo, Brid Brosnan, Ambrose Furey, Elke K. Arendt, Padraigin Murphy

Department of Biological Sciences Publications

A total of 220 lactic acid bacteria isolates were screened for antifungal activity using Aspergillus fumigatus and Aspergillus niger as the target strains. Four Lactobacillus strains exhibited strong inhibitory activity on agar surfaces. All four were also identified as having strong inhibitory activity against the human pathogenic fungi Microsporum canis, Microsporum gypseum and Epidermophyton floccosum. One of the four lactobacilli, namely Lb. reuteri ee1p exhibited the most inhibition against dermatophytes. Cell-free culture supernatants of Lb. reuteri ee1p and of the non-antifungal Lb. reuteri M13 were freeze-dried and used to access and compare antifungal activity in agar plate assays …

In Silico Prediction Of Protein-Protein Interactions Between Cronobacter Sakazakii And Human Host, Sean Held

Theses

The genus Cronobacter accommodates 16 biogroups of the emerging opportunistic pathogen known formerly as Enterobacter sakazakii. Cronobacter are occasional contaminants of milk powder and, consequently, powdered infant formula (PIT). Since it is a pathogen that is transmitted in infant formula it represents a significant health risk to neonates. These bacteria are opportunistic pathogens (can only cause disease when the host's resistance is low) and are linked with life-threatening infections in neonates. Some of the clinical symptoms of Cronohacter infection include necrotizing enterocolitis, bacteremia, and meningitis, with case fatality rates of 50-80% being reported (Healy et al. 2010). Infant formula manufacturers …

Examination Of The Prevalence And Persistence Of Pathogenic Staphylococci On Euro Currency, Emma Gabriel

Theses

Staphylococci are a diverse group of bacteria that cause diseases ranging from minor skin infections to life-threatening bacteremia. Methicillin-resistant S. aureus (MRSA) is the most common multidrug-resistant nosocomial pathogen in Europe estimated to cause 170,000 infections resulting in more than 5,000 deaths and additional healthcare costs of approximately €380 million annually (Kock et al., 201 1). Whilst rarely life-threatening, coagulase-negative staphylococci (CoNS), in particular S. epidermidis are responsible for the majority of indwelling medical device failures, causing intrinsically resistant biofilm infections, estimated to cost $2 billion per year in additional healthcare costs in the U.S. alone (Otto, 2009).

It is …

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 …

Enhanced Surveillance And Characterisation Of Three Significant Gram-Positive Pathogens, Annmarie Mollaghan

Theses

Clinical microbiology has been revolutionised in recent years by the introduction of molecular biology. Traditionally, the gold standard for diagnosis in the clinical microbiology laboratory has been culture, which in some cases, is gradually being replaced by molecular detection methods, particularly where the pathogen is difficult to grow or to identify, or when the significance of an organism is dependent on the detection of specific markers of pathogenicity, such as toxins. Many fastidious bacteria which are difficult to culture and present public health implications, i.e. Mycobacterium tuberculosis. Chlamydia trachomatis and Neisseria gonorrhoea, can now be detected using commercial molecular assays …

Purification, Characterisation And Applications Of A Cloned Staphylococcal Bacteriophage Lysin For The Control Of Multi-Drug Resistant Staphylococcus Aureus Including Mrsa, Mark Fenton

Theses

The work presented in this thesis focuses on the purification, characterisation and practical applications of an anti-staphylococcal lysin (LysK) and its single-domain truncated derivative (CHAP_k) against pathogenic staphylococci including meticillin resistant Staphylococcus aureus (MRSA). Both proteins were purified to greater than 90% homogeneity. Affinity chromatography was applied as the optimum method for LysK purification and ion-exchange chromatography was successfully applied to CHAP_k. The lytic spectrum of LysK was investigated by zymogram analysis where the enzyme exhibited strong activity against a range of staphylococci regardless of their origin, sequence type (ST) or antibiotic resistant profile. This analysis …

Campylobacter Ureolyticus: An Emerging Gastrointestinal Pathogen?, Susan Bullman, Daniel Corcoran, James O'Leary, Brigid Lucey, Deirdre Byrne, Roy D. Sleator

Department of Biological Sciences Publications

A total of 7194 faecal samples collected over a 1-year period from patients presenting with diarrhoea were screened for Campylobacter spp. using EntericBios, a multiplex-PCR system. Of 349 Campylobacter-positive samples, 23.8% were shown to be Campylobacter ureolyticus, using a combination of 16S rRNA gene analysis and highly specific primers targeting the HSP60 gene of this organism. This is, to the best of our knowledge, the first report of C. ureolyticus in the faeces of patients presenting with gastroenteritis and may suggest a role for this organism as an emerging enteric pathogen.