Microbiology Commons^™

Characterisation Of Bacterial Extracellular Polysaccharides (Eps) And Other Complex Carbohydrates With Prebiotic Potential, Padraigin Murphy

Theses

Prebiotics are selectively fermented ingredient that allows specific changes, both in the composition and/or activity in the gastrointestinal microbiota that confers benefits upon host well-being and health’ It is not the prebiotic by itself but rather the changes induced in microflora composition that is responsible for its effects. Several sources of prebiotic were used in this study, namely extracellular polysaccharides (EPS) originating from Lactic Acid Bacteria (LAB), barley, oat. Laminaria digitata. Laminaria hyperborea and Saccharomyces cerevisiae. EPS-producing LAB were isolated from a wide variety of environmental and gastrointestinal niches. The strains were typed and classified according to their EPS-producing ability …

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 …

The Evolution Of Bacteriophages E11/2 And E4/Lc For Control Of Verocytotxigenic E.Coli At Key Stages In The Beef Chain, Bríd Coffey

Theses

The objective of this thesis was lo evaluate the potential of bactcriophages e4/1 c and cl 1/2 ns biocontrol agents for E. coli O157:H7. Initially, e4/lc and el 1/2 were subjected to high-throughput 454 sequencing. The data generated revealed that e4/I c (47,112 bp) is very similar to the lytic E. coli phages RTP and JK06. Interestingly, differences within the tail module led to the identification of a putative site of attachment to E. coli O157:H?. Comparative genomic analysis indicated that el 1/2 (168,470 bp) is a novel member of the T-even group, with high degrees of similarity to T4 …

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.

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.

Identification And Molecular Characterisation Of Mycobacteriophages And Their Lysis Proteins, Marine Henry

Theses

Given their potential as specific and natural biocontrol agents, bacteriophages and their associated proteins have become the focus of renewed interest over the last decade. This study reviews the current state of mycobacterial diseases control worldwide and presents how phage therapy and the production of recombinant bacteriophage lysis proteins can be applied as novel biocontrol strategies for mycobacteria.

Chapter 11 presents an overview of the predicted LysA and LysB proteins in all the mycobacteriophages sequenced to date and proposes an in silico 3D predictive model of the lysin B of a recently isolated and characterised mycobacteriophage. Our data also confirms …

The Rapid Identification Of Novel Anti-Mycobacterial Drugs And The Transporters Involved In Their Resistance, James Carroll

Theses

Despite the success of anti-mycobacterial drugs over the past 70 years, mycobacterial disease, particularly tuberculosis is still responsible for millions of deaths worldwide. Additionally, the emergence of Multidrug Resistant (MDR-TB) and Extensively Drug Resistant (XDR-TB) Tuberculosis throughout the world has motivated calls by the World Health Organization for novel mycobacterial drugs, vaccines and diagnostic tests. Consequently, this study sought to identify and evaluate the efficacy of a range of anti-mycobacterial compounds against a representative cohort of pathogenic mycobacterial species. The development and employment of the rapid, robust and inexpensive microtitre alamarBlue assay (MABA) facilitated the assessment of a vast array …

Isolation And Molecular Characterization Of Mycobacterium Avium Subsp. Paratuberculosis In Ireland, Pierre Emmanuel Douarre

Theses

MAP is a proven animal pathogen causing Johne's disease, one of the most widespread and economically important diseases of ruminants. Since 1992 and the opening of the European market, the prevalence of MAP in Ireland has increased considerably. Improvements in diagnostic strategies for Ireland and elsewhere are urgently required. MAP infects a wide host range and has emerged as a potential threat to human health and has been associated with Crohn's disease. The control of paratuberculosis infection requires a better understanding of the genetic diversity of the pathogen. The objective of this study was to isolate and characterize this enteric …

Detection And Molecular Characterisation Of Rotavirus, Adenovirus And Bocavirus Circulating In Southern Ireland, Olivia Cashman

Theses

This study reports the detection and molecular characterisation of group A rotavirus circulating in the south of Ireland in both the bovine and human population. Characterisation of the group A rotavirus was carried out through the extraction of viral nucleic acid in faecal specimens collected from hospitals in the Cork and Waterford area and Cork regional veterinary laboratory from 2006-2009. Other gastroenteritis viruses characterised during this study include adenovirus type 41, and the newly identified gastrointestinal virus, human bocavirus, which was identified for the first time in Ireland during this study. The human bocaviruses identified in children in the south …

Phage Lysins - Structure, Genetics And Applications, Jennifer Garry

Theses

Antibiotics are of the utmost importance in the fight against pathogenic bacteria, however, in recent years there has been a continual increase in the development of antibiotic resistance among bacteria. As a result it has become essential to develop new technologies to combat these pathogens. One such technology is the application of phage lysins as antimicrobial agents against pathogenic infections. A promising phage lysin is LysK, a 495 amino-acid protein derived from Staphylococcal Phage K. LysK can lyse a wide range of staphylococcal species including MRSA. It is also capable of killing both live and dead cells increasing its potential …

Biosynthesis Of Exopolysaccharides From Lactic Acid Bacteria And Evaluation Of Their Prebiotic Potential, Colette Ann Forde

Theses

In this study, two strains of lactic acid bacteria (LAB) namely Weissella kimchii F28 and W. confusa MGl were used for the production of exopolysaccharides (EPS) from sucrose. Both strains produced glucan polysaccharides. In the case of W. kimchii F28, the EPS isolation procedure was performed six times, making modifications in order to optimize the yield of EPS. The maximum yield of EPS for this strain was 16 g per litre of culture. In the case of W. confusa MGl, the EPS isolation procedure was followed five times, resulting in a maximum yield of EPS for this strain of 34.4 …

Detection And Molecular Characterization Of Canine Enteric Viruses In Southern Ireland., Susan Mcelligott

Theses

Enteric viruses, such as canine parvovirus (CPV) and canine coronavirus (CCoV), are a major cause of gastroenteritis in dogs. More serious clinical symptoms occur in young pups, or when both of these viruses infect an animal simultaneously. Caliciviruses and rotaviruses can also cause gastroenteritis in dogs, but these viruses, while causing significant problems in other animal species and humans, are considered to be less significant pathogens of canines. However, on occasion, these viruses have been reported to produce severe clinical symptoms such as vomiting, haemorrhagic enteritis and severe dehydration.

Many viral vaccines for canines are formulated using old vaccine viral …

The Detection And Analysis Of Selected Pathogens In The Water Supply Of Dairy Cohort Herds., Niall Drummond

Theses

The issue of food safety is a growing concern to Irish agriculture and to the food industry. In the last twenty years, many novel pathogens, which can cause concern to the food industry, have been identified. Organisms such as Verocytotoxigenic Escherichia coli (VTEC) cause hemorrhagic colitis and Haemolytic Uremic Syndrome (HUS), which is the major cause of acute renal failure in children in many countries, and it’s presence in food is a cause of growing concern. Other pathogens causing concern are those implicated as causative agents of Acute Gastroenteritis (AGE) and these can be bacterial, viral and parasitic organisms.

Consumption …

The Molecular Identification Of Non Tuberculosis Mycobacteria (Ntm) From Clinical And Water Samples., Anne Curtin

Theses

Nontuberculosis Mycobacteria (NTM) are now considered a major source of opportunistic infection in immunocompromised patients. Reliable and rapid identification of NTM species is of the upmost importance for patient treatment regimes. Firstly, this study looked at the epidemiological trends of a small number of NTM isolates (n=50) from a large teaching hospital in Southern Ireland Cork University Hospital (C.U.H.). We found that 85% of clinical isolates were sampled from the respiratory tract, which is not unusual as the majority of samples analysed in the hospital lab originated from sputum samples. Also, the majority of patients were 59 years of age …

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 …

Molecular Characterization Of Gastroenteritis Viruses Among Equines And Porcines In Southern Ireland From 2005-2007., P. J, Collins

Theses

Rotaviruses are a major cause of gastroenteritis in humans and animals. To date, seven serogroups (A to G) of rotavirus have been determined. Group A Rotaviruses (GARVs) are enteric pathogens, causing acute watery dehydrating diarrhea in various host species, including birds and mammals. Rotaviruses account for ~611,000 child deaths each year, mainly in developing countries. Likewise, rotavirus-associated enteritis is a major problem in young calves, weaning and post-weaning piglets and foals. Vaccines against the most important serologic group of rotaviruses (GARVs) are available for the prevention of rotavirus diseases in cows and horses, and, more recently, they have been made …

Molecular Characterisation Of A Bovine-Like Rotavirus Detected From A Giraffe, Emily Mulherin, Jill Bryan, Marijke Beltman, Luke O'Grady, Eugene Pidgeon, Lucie Garon, Andrew Lloyd, John Bainbridge, Helen O'Shea, Paul Whyte, Séamus Fanning

Department of Biological Sciences Publications

Background

Rotavirus (RV), is a member of the Reoviridae family and an important etiological agent of acute viral gastroenteritis in the young. Rotaviruses have a wide host range infecting a broad range of animal species, however little is known about rotavirus infection in exotic animals. In this paper we report the first characterisation of a RV strain from a giraffe calf.

Results

This report describes the identification and detailed molecular characterisation of a rotavirus strain detected from a 14-day-old Giraffe (Giraffa camelopardalis), presenting with acute diarrhea. The RV strain detected from the giraffe was characterized molecularly as G10P[11]. …

Detection And Molecular Epidemiological Analysis Of Acute Gastroenteritis Viruses In The South Of Ireland, Grainne Lennon

Theses

Rotavirus infections are considered to be the most common cause of severe gastroenteritis in children under the age of 5 years, and are responsible for up to 611,000 deaths annually, mainly in developing countries. From 1997 to 1999, 16 hospital laboratories in the Republic of Ireland reported to detection of 4,643 cases of rotaviral disease, while in 2004 and 2005 and increase of 1,600 and 2,251 rotavirus cases were reported. These rotavirus cases have a significant impact on the healthcare system with the minimum cost per case being €728.40. A large percentage of the infections were reported in neonates, suggesting …

Mupirocin Resistance In Staphylococci: An Investigation Of Current Status And Potential Alternative Therapies, Susan O'Shea

Theses

Mupirocin is a topical antibiotic used in hospital infection control practices to control transmission of staphylococci, in particular MRSA, which has become a major problem in Irish hospitals and has a significant impact on patient morbidity and mortality. Overreliance on mupirocin to eradicate MRSA carriage and to prevent infection has led to mupirocin resistance among staphylococcal species. Two distinct types of resistance have developed within strains: low'-level mupirocin resistance (LMR) and high-level mupirocin resistance (HMR).

The aim of this study was to assess the prevalence and types of mupirocin resistance in a large population of clinical staphylococcal isolates including Methicillin …

Analysis Of Adenovirus And Bovine Coronavirus., Kevin Lane

Theses

This investigation involved the analysis of selected gastroenteritis viruses, adenovirus and bovine coronavirus, obtained from faecal samples, collected from hospital and diagnostic laboratories respectively in Cork city, Ireland. The viruses isolated from these samples were subjected to analysis using cell culture and nucleic acid extraction. In addition adenovirus was analysed using PCR and PCR followed by restriction endonuclease digest. Many samples contained more than one gastroenteritis virus, the virus of interest and a natural co-infection with rotavirus species. Rotavirus infections in Ireland have been extensively studied and we were interested in studying singly and doubly infect samples.

Bovine enteric coronavirus …

Epidemiology And Drug Resistant Profiles Of Candida Species, In Cork Hospitals., Emma Mccurtin

Theses

Candida causes superficial infections in healthy individuals; however in the immunocompromised host Candida can cause invasive, systemic infections and is a significant cause of morbidity and mortality. This is not due to a more virulent phenotype but to a weakened immune system unable to fight this opportunistic pathogen. The immunocompromised population is ever increasing, due to medical advancements and more aggressive treatments, and hence is the need for epidemiological data of these potentially life threatening infections. The aim of this study was to provide an epidemiological snapshot, of Candida infections from the immunocompromised population in Cork hospitals. Surveillance programmes from …

Design Of A Multiplex Pcr Assay For The Detection Of Neisseria Gonorrhoeae., Elizabeth J. O'Callaghan

Theses

The incidence of N. gonorrhoeae infections in Ireland has been steadily increasing since 2003. Culture is currently the preferred method of diagnosis, however the sensitivity of culture can be low. In recent years there has been increased interest in non-culture techniques such as polymerase chain reaction (PCR). Many of the PCR assays available have been shown to cross-react with commensal organisms, producing false positive results. This study aimed to improve the detection of N. gonorrhoeae by designing a multiplex PCR assay that used two N. gonorrhoeae genes as targets, thereby providing dual detection and confirmation of a positive result. PCR …

Clostridium Difficile: Comparative Evaluation Of Two Detection Techniques And Characterisation Of Clinical Isolates., Rebecca Dennehy

Theses

Clostridium difficile is one of the most common causes of antibiotic associated diarrhoea. The presence of C difficile in stool samples may be detected by a variety of means including enzyme immunoassays and culture. Enzyme immunoassays detect the presence of the toxins produced by the organism, toxin A and/or toxin B, but are known to lack sensitivity. The aim of this study was to compare the commercial ELISA assay currently used for the detection of C difficile toxins in Waterford Regional Hospital to culture for the organism to selective media. To compare the sensitivity of the two methods serial dilutions …

Evaluation Of Four Novel Broad-Host-Range Antimicrobial Agents., Jiahui Guo

Theses

The overall objective of this thesis was to evaluate the antimicrobial effects of four novel broad-host-range antimicrobial agents. The first was a milk-derived lipoprotein developed by Westgate Biologicals with the support of Enterprise Ireland. This lipoprotein which was designated LactiSAL, was evaluated against a wide range of bacterial species associated with humans. Essentially, the LactiSAL was incorporated at a variety of concentrations into agar media, which were appropriate for cultivation of the different genera. Agars were examined for colony formation, after which it was possible to quantify the minimal inhibitory concentration for each genus. Subsequently, LactiSAL was incorporated at levels …

Accumulation Of Phycotoxins In The Mussel Mytilus Galloprovincialis From The Central Adriatic Sea, Maja Pavela-Vrančić, Ivana Ujević, Živana Ninčević Gladan, Ambrose Furey

Physical Sciences Publications

Surveys of DSP (Diarrheic Shellfish Poisoning) toxin profiles in the mussel Mytilus galloprovincialis from coastal waters of the central Adriatic Sea, over the years 1995 to 2001, demonstrate that incidents of shellfish toxicity in this area are dominated by the occurrence of okadaic acid (OA) and a PTX derivative, 7-epi-pectenotoxin-2-seco acid (7-epi-PTX-2SA). Toxin composition and the relative ratio of toxic components in shellfish is in correlation with the occurrence of DSP producing organisms from the Dinophysis species along with Prorocentrum micans and Lingulodinium polyedrum. The occurrence of D. sacculus shortly before the appearance of OA in shellfish implicates its possible …

Quantification Of Mrna Expression Of Her2/Neu, Estrogen Receptors Eralpha And Erbeta, And Progesterone Receptor Using Real-Time Reverse Transcription Pcr In Small Tumor Biopsies, In Human Breast Cancer, Evonne Marie. Mc Cabe

Theses

Early detection is an effective means of reducing cancer mortality. Early detection greatly improves treatment options and the chances for successful treatment. The prognosis for patients with breast cancer is determined by well-established biological features associated with biological aggressiveness, histological grade, tumour size, and nodal involvement. Breast cancer results from multiple genetic abnormalities, and cellular differentiation, tumour growth, and metastatic potential are the end result of these changes. Identification of molecular changes is of considerable value in enhancing the accuracy of prognostic indices, and more importantly, it may indicate various changes as putative targets for therapy. In recent years a …

Molecular Epidemiology Of Antibiotic Resistant Salmonella Enterica Strains From Different Animal And Human Sources In Ireland, David George Lee

Theses

In this study, 176 isolates of Salmonella were collected from different laboratories in Ireland. All isolates were confirmed to be of the genus Salmonella by performing bacteriological and biochemical tests. Briefly, all isolates were cultured onto XLD agar after which pure cultures were verified and stocked at this institute. Definitive identification was carried out using the API 20-E system and a separate urease test was also performed on all isolates. After confirmation of the genus, all isolates were prepared for antibiotic susceptibility testing and r-types were determined using sixteen antibiotics. Depending on the antibiotic resistance profiles, isolates were selected for …

The Analysis And Characterisation Of Rotavirus And Norovirus Circulating In The South Of Ireland, Norma Reidy

Theses

On average more than 3,000 cases of rotavirus diarrhea are detected annually amongst Irish children less than 3 years of age. Rotavirus disease occurs mainly during the winter months, with peak infections detected from February to April. The symptoms of disease are diarrhea and dehydration, with 80% of patients requiring hospitalisation, The mean duration of hospitalisation is 3-4 days, with an average cost per day of €409. Eighteen percent of infections were detected in infants less than 6 months of age, suggesting inadequate maternal derived protection from existing strains, or the existence of novel strains in Ireland with little or …

Validation Of An Immunomagnetic-Electrochemiluminescence Detection System For The Detection Of Escherichia Coli 0157 In Food And Comparison With An Iso Standard Method, Eimear Carney

Theses

Rapid and sensitive methods for detecting Escherichia coli 0157 in contaminated food, water and other biological samples are important in limiting and preventing the spread of this pathogen. Traditional culture based methods for bacterial identification and detection can be time consuming and labour intensive. Modem alternative methods should be faster whilst maintaining sensitivity. Immunomagnetic separation (IMS) methodology has been investigated and appears promising for rapid bacterial detection in unrelated food and environmental samples. In this study, a commercial sensor which combined IMS with electrochemiluminescence (ECL) was evaluated for the detection of E. coli 0157 in unrelated food matrices. Emitted signals …