Antimicrobial Resistance in Campylobacter jejuni and Campylobacter coli

Antimicrobial Resistance in Campylobacter jejuni and Campylobacter coli Campylobacters are a common cause of bacterial gastroenteritis worldwide, with Campylobacter jejuni and C. coli being the most common species isolated in human infections. If antimicrobial treatment is required, the drugs of choice at the moment are the macrolides and fluoroquinolones. In this thesis, the in vitro resistance profiles of the C. jejuni and C. coli strains were evaluated with emphasis on multidrug resistance. The aim was also to evaluate the different resistance mechanisms against the macrolides. Further, the disk diffusion method was compared to agar dilution method and its repeatability was evaluated, since it has been widely used for the susceptibility testing of campylobacters. The results of the present study showed that resistance to the fluoroquinolones is common in strains isolated from Finnish patients, but resistance to the macrolides is still rare. Multidrug resistance was associated with resistance to both ciprofloxacin and erythromycin. Among the available per oral drugs, least resistance was observed to coamoxiclav There was no resistance to the carbapenems. Sitafloxacin and tigecycline were in vitro highly effective towards Campylobacter species. A point mutation A2059G of the 23S rRNA gene was the main mechanism behind the macrolide resistance, whereas the efflux pumps did not seem to play an important role when a strain had A2059G mutation. A five amino acids insertion, which has not been described previously, in the ribosomal protein L22 of one highly-resistant C. jejuni strain without mutation in the 23S rRNA gene was also detected. Concerning the disk diffusion method, there was variation in the repeatability In conclusion, macrolides still appear to be the first-choice alternative for suspected Campylobacter enteritis. The in vitro susceptibilities found suggest that co-amoxiclav might be a candidate for clinical trials on campylobacteriosis, but in life-threatening situations, a carbapenem may be the drug of choice. More studies are needed on whether the disk diffusion test method could be improved or whether all susceptibilities of campylobacters should be done using a MIC based method.

Use of pyrosequencing to identify streptococci and to detect mutations causing antimicrobial resistance

Rapid identification and resistance determination of pathogens in clinical specimens is vital for accurate treatment and monitoring of infectious diseases. Antimicrobial drug resistance is increasing globally and healthcare settings are facing this cost-intensive and even life-threatening problem. The incidence of resistant pathogens in Finland has remained relatively steady and manageable at least for the time being. DNA sequencing is the gold standard method for genotyping, mutation analysis, and identification of bacteria. Due to significant cost decrease in recent years, this technique is available to many research and clinical laboratories. Pyrosequencing technique, a rapid real-time DNA sequencing method especially suitable for analyzing fairly short stretches of DNA, was used in this study. Due to its robustness and versatility, pyrosequencing was applied in this study for identification of streptococci and detection of certain mutations causing antimicrobial resistance in different bacteria. Certain streptococcal species such as S. pneumoniae and S. pyogenes are significantly important clinical pathogens. S. pneumoniae causes e.g. pneumonia and otitis media and is one of the most important community-acquired pathogens. S. pyogenes, also known as group A streptococcus, causes e.g. angina and erysipelas. In contrast, the socalled alpha-haemolytic streptococci, such as S. mitis and S. oralis, belong to the normal microbiota, which are regarded to be non-pathogenic and are nearly impossible to identify by phenotypic methods. In this thesis, a pyrosequencing method was developed for identification of streptococcal species based on the 16S rRNA sequences. Almost all streptococcal species could be differentiated from one another by the developed method, including S. pneumoniae from its close relatives S. mitis and S. oralis . New resistance genes and their variants are constantly discovered and reported. In this study, new methods for detecting certain mutations causing macrolide resistance or extended spectrum beta-lactamase (ESBL) phenotype were developed. These resistance detection approaches are not only suitable for surveillance of mechanisms causing antimicrobial resistance but also for routine analysis of clinical samples particularly in epidemic settings. In conclusion, pyrosequencing was found to be an accurate, versatile, cost-effective, and rapid DNA sequencing method that is especially suitable for mutation analysis of short DNA fragments and identification of certain bacteria.

Resistance to potato virus A and potato virus Y in potato cultivars grown in Finland

Selostus: Suomessa viljeltävien perunalajikkeiden kestävyys A- ja Y-virusta vastaan

Pathogen derived resistance to Potato virus Y: mechanisms and risks

Selostus: Patogeenivälitteinen, siirtogeeninen kestävyys perunan Y-virusta vastaan: mekanismit ja riskit

Pressure to change and resistance against it: consumer perceptions and acceptance of nutritionally modified foods

Selostus: Muutospaineet ja muutosvastarinta: kuluttajien suhtautuminen ravitsemuksellisesti muunnettuihin elintarvikkeisiin

LM-vesiturbiinin tuotteistaminen

Tämän työn tarkoituksena on kehittää LM-vesiturbiini tuotantoon soveltuvaksi, käytöltään pitkäikäiseksi ja helposti huollettavaksi tuotesarjaksi. Työn ohjaavana runkona on käytetty VDI-Richtlinie 2221: 'Kehitystyön ja konstruoinnin yleinen kulku', mukaista järjestelmää. Lisäksi on käytetty valmistuksellista modulointia tuotteen osien suunnittelussa. Erityisesti on huomioitu tuotteen koko elinkaaren aikaiset kustannukset (LCC) ja sen vaatimukset mitoitukseen. LM-vesiturbiinistaon kehitetty kolmen erikokoisen laitteiston tuotesarja. Tuoteperheen yksittäisistä sylinteriryhmistä voidaan koota joustavasti erikokoisia ja -tehoisia energiantuotantoyksiköitä, jotka soveltuvat käyttöön hyvin pieniinkin virtauksiin ja putouskorkeuksiin. Piensarjatuotantoon hyvin soveltuvat materiaalit on valittu silmälläpitäen mahdollisimman edullista kokonaiskustannusta. Kiertomäntä laitteiston toiminnan kannalta tärkeimpänä ja siten vaativimpana osana on edellyttänyt laserhitsausta ja -leikkausta sekä koekappaleiden valmistusta, joiden perusteellalopulliseen rakenteeseen on päädytty. Mitoitus perustuu perinteisiin lujuuslaskentamenetelmiin, joiden avulla eri osien kestävyys käyttöolosuhteissa on pyrittysaamaan luotettavaksi vähintään 20 vuoden kestoiälle ja väsymiskestävyyteen käytön aikaisissa vaihtelevissa olosuhteissa. Kiertomäntä on ns. kevytrakenne koostuen ohutlevykennorakenteesta ja täytteenä olevasta kompound-materiaalista.