Note:

Thesis (Ph. D.)--University of Wisconsin--Madison, 1974. / Typescript. Vita. Includes bibliographical references.

Strains of enteroaggregative E. scherichia colt (EAggEC), characterised by their pattern of adhesion to HEp-2 cells known as the `stacked brick' formation, are a significant cause of chronic diarrhoea in certain under-developed countries. Strains of EAggEC are detected either by a HEp-2 adhesion cell test or by an `aggregative adhesion' gene probe. The pathogenic mechanisms expressed by EAggEC are only poorly understood and the aim of the research described was to obtain a better understanding of how these bacteria cause disease. The adhesion of EAggEC to HEp-2 cells was shown in the majority of strains not to involve fimbriae and was thought to result from physical properties of strains such as charge, since EAggEC adhered to `fixed' HEp-2 cells and readily agglutinated a range of different erythrocytes. Certain strains of EAggEC, which also hybridised with a probe for diffuse adhesion, expressed membrane-associated proteins (MAPs) of 18 or 20 kDa responsible for HEp-2 adhesion. Divalent cations were essential for the expression of the MAPs, which did not contain disulphide bonds or have a quaternary structure. Strains of EAggEC did not express recognised subunit toxins such as Verocytotoxin or E. coil heat-labile toxin, and strains which hybridised with probes for enteroaggregativeh eat-stable toxin-1 did not produce E. coil heat-stable toxin detected by the infant mouse test. Some EAggEC strains (15%) had haemolytic properties. Certain strains expressed type II capsular polysaccharides and approximately 50% of strains expressed an aerobactin-mediated iron uptake system. It was concluded that strains of EAggEC belonged to a very diverse range of serotypes, and it was thought that this heterogeneity resulted from strains of E. coil readily acquiring the genes encoding the EAggEC phenotype. Strains of EAggEC were not associated with a single pathogenic phenotype and the ability of these bacteria to adhere to HEp-2 cells in a `stacked brick'-pattern remains the only common characteristic.

Thesis - Massachusetts Institute of Technology and Woods Hole Oceanographic Institution. / Bibliography: p. 126-134.

Thesis (M.Sc.) -- Memorial University of Newfoundland, 1987. / Restricted until November 1988. Typescript. Bibliography: leaves 102-110. Also available online.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 5 uppsatser.

Nanotechnology is a rapidly growing field focused on the manipulation and control of materials with dimensions under 100 nm. The novel electronic, optical and mechanical properties observed at the nanoscale have resulted in a number of applications in catalysis, light emitting devices, solar power, self-cleaning surfaces and medicine. Medical applications of nanotechnology (“nanomedicine”) are particularly promising for rapid clinical diagnosis and targeted treatments. Understanding the interactions of nanoparticles with living matter is of fundamental importance for all application areas: manufacture, use and disposal of the growing number of nanoproducts will result in increased environmental exposure in addition to direct exposure through nanomedical applications. However, there is a lack of standard methodologies for assessing these interactions. In this work the stability of silver-based nanoparticles was established by UV- Visible (UV-Vis) spectroscopy, atomic force microscopy (AFM) and transmission electron microscopy (TEM). The presence of a higher valence metal or polymer on the nanoparticle surface was demonstrated to improve stability. A standard methodology was developed to study nanoparticle-cell interactions: an “atlas” of the effects of known drugs on a cell is created, and compared with the effects of a nanoparticle. Escherichia coli was selected as a model organism and the effects of a range of antibiotics were characterised through a combination of microbiological assays and AFM. Susceptibility, population cell growth and individual heights, widths, lengths and volumes of bacteria were obtained on a 2% agarose substrate in air. The methodology was applied and adjusted for silver nanoparticles due to the interactions of silver with the bacterial growth medium. 10 and 30 nm silver nanoparticles and ions were found to kill E. coli through an internal mechanism of action, with a size-specific effect on the height of bacteria. Finally, a novel AFM characterisation method is described to examine the mechanical properties of live bacterial and human cells in liquid.

Les processus de diversification adaptative, qui sont au cœur de la diversité du monde vivant, ont été étudiés grâce à une stratégie d'évolution expérimentale, initiée par le Pr Richard Lenski en 1988. Douze populations, fondées à partir d'un ancêtre commun d'Escherichia coli, sont propagées indépendamment depuis plus de 55 000 générations par transferts journaliers dans un milieu minimum limité en glucose. Un événement de diversification a émergé après 6500 générations d'évolution dans une seule des douze populations, appelée Ara-2, conduisant à deux lignées cellulaires différenciées, appelées S et L, qui continuent de co-exister depuis notamment grâce à des interactions négatives dépendant de leur fréquence. Deux propriétés confèrent à ce polymorphisme une grande originalité et donc un intérêt d'étude important : sa durée car il s'agit du plus long polymorphisme jamais identifié lors d'expériences d'évolution en laboratoire, et son unicité puisqu'il ne s'est produit qu'une seule fois au sein des douze populations initiées à partir d'un ancêtre commun. L'objectif de ce travail a été d'identifier les mécanismes du maintien au long terme des lignées S et L, ainsi que les bases génétiques de leur émergence. Le maintien du polymorphisme est lié à une forte dynamique des relations écologiques entre S et L, l'une des lignées envahissant systématiquement les niches écologiques de l'autre, qui réagit en conséquence pour éviter l'extinction. L'émergence de la lignée S est due à une succession précise de trois mutations, nécessaires et suffisantes pour établir les phénotypes de la lignée S. Les trois mutations affectent toutes des gènes codant des régulateurs globaux de la transcription, dont deux sont impliqués dans la régulation du métabolisme central. Pour l'un d'entre eux, l'allèle évolué altère les propriétés de liaison à l'ADN de la protéine évoluée. Bien que ce polymorphisme soit unique, ces trois gènes sont pourtant les cibles de la sélection naturelle dans la majorité des autres populations de l'expérience d'évolution. Pour deux d'entre eux, seul l'allèle substitué dans la population Ara-2 confère en fait les phénotypes de la lignée S. Ainsi, l'unicité de cet événement de diversification est liée à une succession d'événements mutationnels très précis, qui affectent par ailleurs les réseaux globaux de l'expression des gènes. Ces modifications graduelles ont ainsi conduit à l'émergence du plus long polymorphisme mis en évidence à ce jour dans des expériences d'évolution en laboratoire. / Adaptive diversification events that underly the diversity of the living world have been studied by an experimental evolution strategy initiated by Richard Lenski in 1988. Twelve populations founded from a common ancestor of Escherichia coli are propagated independently since more than 55,000 generations by daily transfer in a glucose-limited minimal medium. A diversification event emerged after 6500 generations of evolution in only one of the twelve populations, called Ara-2, resulting in two lineages of differentiated cells, called S and L, that coexist ever since owing to negative frequency-dependent interactions. Two properties make this polymorphism original and important: its length as the longest one ever observed in evolution experiments, and its uniqueness as it occurred only once in the twelve populations founded from the same ancestor. The aim of this work was to identify the mechanisms of the long-term coexistence of the S and L lineages, together with the genetic bases of their emergence. The maintenance of the polymorphism is characterized by a strong dynamic of the ecological relationships between S and L, with with L seeming to encroach over time on the niche of S, which reacts to avoid extinction. The emergence of the S lineage is due to the succession of three mutations, necessary and sufficient to establish its phenotypes. All three mutations affect genes encoding global transcriptional regulators, with two of them being involved in the regulation of central metabolism. For one of them, the evolved allele alters the DNA binding ability of the evolved protein. Although this polymorphism is unique, the same three genes are targets of natural selection in most other populations of the evolution experiment. For two of them, only the substituted allele of the Ara-2 population results in the phenotypes of the S lineage. Thus, the uniqueness of this diversification event is linked to a succession of precise mutational events that affect the global regulatory network in the cell. Those gradual modifications lead thus to the emergence of the longest polymorphism ever identified during evolution experiments in the laboratory.

Nous étudions la dynamique de nage de bactéries Escherichia coli dans différents environnements. Leur mouvement 3D est déterminé au moyen d'un système de suivi Lagrangien 3D automatisé pour suivre des objets fluorescents que nous avons développé. Les bactéries étudiées avec ce système présentent une dispersion du coefficient de diffusion rotationnel particulièrement large, ce qui contredit la vision communément admise de la dynamique "run-and-tumble" qui a été établie pour une bactérie qui nage. Ce résultat est interprété comme une conséquence de la distribution en loi de puissance des temps de "run" expérimentaux d'un flagelle individuel, qui jusqu'alors restait indépendant de la description cinématique. Dans des écoulements confinés, la migration vers l'amont d'E. coli sur les bords reste possible pour des taux de cisaillement bien plus grands que ceux de la surface plane. La vitesse des bactéries sur les bords n'est pas influencée par l'écoulement advectif. Le mouvement vers l'amont a lieu près des parois dans une "couche limite" dont la taille varie avec le taux de cisaillement imposé. La migration vers l'amont sous écoulement et persistance de direction se combinent lors du processus de contamination. Nous montrons que les bactéries peuvent contaminer des régions propres par nage vers l'amont dans des environnements confinés.Un modèle simple, qui prend en compte la statistique de rotation du moteur, décrit de manière satisfaisante les principales caractéristiques du processus de contamination, en faisant l'hypothèse d'une distribution en loi de puissance des temps de “run”. Le modèle échoue à reproduire la dynamique quanlitative lorsque l'on prend en compte la distribution classique de run-and-tumble. Nous en concluons que le transport macroscopique de bactéries est déterminé pour la statistique de rotation du moteur. / We present a study of the swimming dynamics of Escherichia coli bacteria in different physical conditions. Their 3D motion is assessed by means of a device for automated 3D Lagrangian tracking of fluorescent objects, that we developed for that purpose. Bacteria studied in that way display consistently large dispersion of the rotational diffusion coefficient, contradicting the standard vision of run-and-tumble dynamics established for an adapted bacterium. The result is interpreted as a consequence of the power law distribution of run times experimentally found for individual flagella, that up to now remained uncoupled with the motility description.We also study the bacterial swimming in polymeric suspensions, as well as in more concentrated active suspensions.In confined flows, upstream migration of E. coli at the edges remains possible at much larger flow rates compared to the motion at the flat surfaces. The bacteria speed at the edges is not influenced by the advective flow. Upstream motion takes place close to the edges in an “edge boundary layer” whose size varies with the applied flow rate. Upstream migration under flow and direction persistence combine during contamination processes. We show that bacteria can contaminate clean regions by upstream swimming in confined environments. A simple model considering the motor rotation statistics describes well the main features of the contamination process, assuming a power law distribution of run times. The model fails to reproduce the qualitative dynamics when the classical run-and-tumble distribution is determined by the motor rotation statistics.

The aim of this work was to go thoroughly throw the already existing HACCP system in the company and to evaluate if the hazard is treated in a good way and to find other more effective approach to prevent the hazard. An effort was focused on the primary processing milk and on the process of the treatment of acidified milk products. While investigating current condition the author has made several experiments focusing on the settled conditions of important process in the production (for ex. pasteurization). Chemical, physical, qualitative and microbial properties were observed on a concrete diary product. The main investigation {--} the simulation of disturbance of the cooling chain - was performed where products were exposed to various temperatures for various time periods. During this test were observed: the temperature, pH and microbial changes in exposed products. Due to results form all observations that where performed in this research its possible to proclaim that the HACCP system is settled in a good way and even if above mentioned situation happen the product is safe.