Le « porcine respiratory disease complex » (PRDC) est un trouble multifactoriel dû à une infection simultanée ou séquentielle de divers micro-organismes pouvant intensifier ou prolonger les signes cliniques des porcs. On retrouve dans ce complexe Mycoplasma hyopneumoniae, un des agents initiateurs du PRDC et agent primaire de la pneumonie enzootique (EP) chez les porcs. Streptococcus suis est l’un des agents pathogènes secondaires du PRDC, c’est également un agent pathogène important induisant principalement des méningites, des septicémies et la mort subite des porcelets post-sevrés. Mycoplasma hyorhinis est également l’un des agents pathogènes secondaires du PRDC, et va induire des inflammations sérofibrineuses chez les porcelets. Comme ces trois pathogènes sont retrouvés au sein du PRDC et au niveau des voies respiratoires supérieures des porcs, il pourrait exister un effet positif des mycoplasmes sur la pathogénèse de S. suis. C’est pourquoi différentes expériences in vitro ont été réalisées avec les cellules épithéliales porcines (NPTr), les macrophages alvéolaires porcins (PAMs) et les cellules dendritiques porcines (BM-DCs) qui ont été pré-infectés par les mycoplasmes puis infectés avec S. suis. Il a été observé que la cytotoxicité et l’inflammation des cellules porcines ont été significativement augmentées lorsqu’elles ont été pré-infectées par les mycoplasmes puis infectées par S. suis. Cependant, la pré-infection des cellules n’a pas joué de rôle sur l’adhésion et l’invasion de S. suis, sur la phagocytose et la survie intracellulaire de la bactérie. Cette étude semble montrer que la pré-infection des mycoplasmes pourraient induire un contexte inflammatoire favorisant la pathogenèse de S. suis. / Porcine respiratory disease complex (PRDC) is a multifactorial disorder due to simultaneous or sequential infection with various microorganisms that can intensify or prolong clinical signs in pigs. Included in this complex is Mycoplasma hyopneumoniae, one of the initiating agents of PRDC and the primary agent of enzootic pneumonia (EP) in pigs. Streptococcus suis is one of the secondary pathogens of PRDC and is also an important pathogen, mainly causing meningitis, septicemia, and sudden death in post-weaned piglets. Mycoplasma hyorhinis is also one of the secondary pathogens of PRDC and will also induce serofibrinous inflammation in piglets. As all three pathogens are found in the PRDC and in the upper respiratory tract of pigs there may be a positive effect of mycoplasma on the pathogenesis of S. suis. Therefore, different in vitro experiments were performed with newborn pig tracheal cells (NPTr), primary porcine alveolar macrophages (PAMs) and porcine bone-marrow-derived dendritic cells (BM-DCs) that were pre-infected with mycoplasma and then infected with S. suis. It was observed that cytotoxicity and inflammation of pig cells were significantly increased when they were pre-infected with mycoplasma and then infected with S. suis. However, pre-infection of the cells did not play a role in the adhesion and invasion of S. suis and in the phagocytosis and intracellular survival of the bacteria. This study suggests that pre-infection with mycoplasma may induce an inflammatory context favoring the pathogenesis of S. suis.

Streptococcus suis (S. suis) is a common swine pathogen but also poses a threat to human health in causing meningitis and severe cases of streptococcal toxic shock-like syndrome (STSLS). Therefore, it is crucial to understand how S. suis interacts with the host immune system during bacteremia. As S. suis has the ability to introduce d-alanine into its lipoteichoic acids (LTAs), we investigated the working hypothesis that cell wall modification by LTA d-alanylation influences the interaction of S. suis with porcine blood immune cells. We created an isogenic mutant of S. suis strain 10 by in-frame deletion of the d-alanine d-alanyl carrier ligase (DltA). d-alanylation of LTAs was associated with reduced phagocytosis of S. suis by porcine granulocytes, reduced deposition of complement factor C3 on the bacterial surface, increased hydrophobicity of streptococci, and increased resistance to cationic antimicrobial peptides (CAMPs). At the same time, survival of S. suis was not significantly increased by LTA d-alanylation in whole blood of conventional piglets with specific IgG. However, we found a distinct cytokine pattern as IL-1β but not tumor necrosis factor (TNF)-α levels were significantly reduced in blood infected with the ΔdltA mutant. In contrast to TNF-α, activation and secretion of IL-1β are inflammasome-dependent, suggesting a possible influence of LTA d-alanylation on inflammasome regulation. Especially in the absence of specific antibodies, the association of S. suis with porcine monocytes was reduced by d-alanylation of its LTAs. This dltA-dependent phenotype was also observed with a non-encapsulated dltA double mutant indicating that it is independent of capsular polysaccharides. High antibody levels caused high levels of S. suis—monocyte—association followed by inflammatory cell death and strong production of both IL-1β and TNF-α, while the influence of LTA d-alanylation of the streptococci became less visible. In summary, the results of this study expand previous findings on d-alanylation of LTAs in S. suis and suggest that this pathogen specifically modulates association with blood leukocytes through this modification of its surface.

The species of Brucella were isolated and characterized almost 120 years ago and their genomes sequenced for almost 4 years. Compared to other bacterial pathogens relatively, little is known about the factors contributing to their persistence in hosts and multiplication within phagocytic cells. Also, many aspects of the interactions between Brucella and its host remain unclear. Molecular characterization of intracellular survival processes of Brucella will provide guidance for additional prevention and control measures. One of the features that distinguishes Brucella is that they do not express classic virulence factors. Thus identification of virulence factors has been elusive and some of the identified virulence genes are putative. Disruption of putative virulence genes and studying the consequent effect on attenuation in cell lines or mouse models is a widely used method. However, in most cases it is not apparent whether the mutated genes encode virulence factors or merely affect normal metabolic or biological functions. Some mutations in Brucella can be compensated by redundancy or backup mechanisms. One method for identifying putative virulence genes involved in pathogenesis is to express these genes in a nonpathogenic host and isolate recombinants with increased virulence or survival ability either in cell culture or animal model. We hypothesize that over-expression of Brucella putative virulence genes in the non-pathogenic and close phylogenic relative Ochrobactrum anthropi should enhance its survival in infection models in vivo. O. anthropi is one of the closest Brucella relatives based on DNA, rRNA, and protein analyses but it is unable to establish chronic infection and considered as opportunistic pathogen that, under certain circumstances, may produce disease in immunocompromised humans. Therefore, we established enhanced expression system in Brucella and Ochrobactrum to identify B. suis virulence genes. We created an enhanced expression system that can be used for cloning and expression of heterologous genes in Brucella and Ochrobactrum. We studied the transcriptional activity of several promoters and created some tools to enhance the expression, detection and purification of Brucella recombinant protein in Ochrobactrum. The presumable importance of alkyl hydroperoxide reductases encoded by ahpC and ahpD genes and their contribution to intracellular survival of Brucella were studied by over-expressing them. The recombinant O. anthropi expressing B. suis ahpC and ahpD genes were able to resist in vitro killing by H2O2 and or cumene hydroperoxide and survived longer in the macrophage J774 A.1 cell line. The control O. anthropi was cleared from BALB/c mice in five days while the recombinants were recovered from spleens, livers and lungs of infected mice up to eight days post-infection. We tested the contribution of B. suis cyclic glucan synthetase gene (cgs) to virulence by over-expressing it in O. anthropi. We studied the ability of the recombinant O. anthropi to resist killing in vitro and in vivo. We generated evidence that B. suis cgs when over-expressed in O. anthropi increased the amount of cyclic glucans synthesized and accumulated in the periplasmic space. This accumulation changed the virulence of the microorganism from a soil bacterium that cleared from mice in less than five days into a pathogenic organism that could survive up to 9 days and at higher doses killed the mice. In summary, several vectors have been constructed for gene expression and protein purification in Brucella and Ochrobactrum. Novel useful tools for enhancement of heterologous gene expression were created and demonstrated to work in Brucella and Ochrobactrum. Brucella putative virulence genes were studied in Ochrobactrum using the newly constructed vectors and tools. Ochrobactrum as a gain of function model for studying putative virulence genes of intracellular pathogens in general and for Brucella in particular proved to be a very useful model. / Ph. D.

Brucellosis is one of the neglected diseases in Nigeria. In Lagos, the commercial capital of Nigeria with about twenty one million people, a descriptive cross-sectional study was carried out in order to determine the sero-prevalence of brucellosis among people at risk in some selected abattoirs and secondary health care facilities (hospitals) in the state. Mixed sampling method was employed at the abattoir while convenient sampling method was used in sampling the respondents at the hospitals. Sera samples from three hundred and one (n=301) abattoir-based workers and traders; and one hundred and twenty one (n=121) hospital-based individuals which include people with febrile illnesses and blood donors were tested for brucellosis using Rose Bengal Plate test (RBPT), with indirect ELISA being used as a confirmatory test. Of the 301 abattoir-based workers and traders, 27 (8.97%) were sero-positive to the infection when Rose Bengal Plate test antigen was used. The twenty seven individuals consists of fifteen (15) butchers; four (4) veterinarians; two (2) meat transporters and bone/cow horn dealers each as well as one each of blood meal producer, abattoir engineer, water seller and meat supplier. When blood samples from the sero-positive individuals were subjected to ELISA, 3 (11.1%) were sero-positive to the brucellosis, while one is equivocal. These results confirm that agglutination observed on RBPT might be related to unknown cross-reactions and confirmation with a different test was necessary. None of the hospital-based respondents is sero-positive to the infection. The clinical signs significant for the infection in this study were fever, joint pain, lower backache, regular headache and miscarriage. Brucellosis awareness level among the respondents was very low. Data was analysed using (SPSS) version 20.0 at α0.05 significant level. The significant risk factors for human brucellosis according to this research are consumption of fura (unpasteurized milk) and wara (fresh cheese). The study revealed that brucellosis is not only an occupational disease but can also affect people who trade or live in proximity with infected animals. / Agriculture, Animal Health and Human Ecology / M. Sc. (Agriculture)

No description available.

No description available.

No description available.

L'état circulaire du chromosome bactérien pose un problème particulier lors de la réplication. Un nombre impair d'événements de recombinaison homologue donne des chromosomes dimères concaténés qui ne peuvent pas être divisés en cellules filles. Pour résoudre ce problème, les bactéries ont mis au point un mécanisme de résolution des dimères basé sur un système de recombinaison spécifique au site. Ceci est effectué par le système Xer/dif. Dans ce système, les protéines Xer effectuent une réaction de recombinaison dans le site dif au niveau du septum cellulaire immédiatement avant la division cellulaire. Dans la plupart des bactéries, cette réaction est effectuée par deux recombinases, XerC et XerD. Cependant, Streptococcus suis, un agent pathogène zoonotique important utilise un système de recombinaison différent, constitué d'une seule enzyme recombinase appelée XerS, qui catalyse la réaction de recombinaison dans un site dif non conventionnel. Pour caractériser le mode de clivage de XerS, des expériences EMSA ont été réalisées en utilisant des fragments de PCR marqués par HEX et des "suicide substrates". Nos données suggèrent que 1.) XerS est capable de lier la séquence entière de difSL; 2.) XerS lie plus efficacement le côté gauche des mutants difSL incomplets que le côté droit; 3.) XerS coupe les brins supérieur et inférieur du site difSL, avec une réaction plus efficace au bas. 4.) Modifications des nucléotides de la région la plus externe ou de la région centrale changent les préférences de clivage. 5.) XerS n'a montré aucune activité spécifique sur un autre site dif non conventionnel des Firmicutes, 6.) XerS interagit avec la sous-unité FtsK-y. L'ensemble des résultats présentés permet de mieux comprendre le fonctionnement de la recombinaison XerS dans le système de recombinase unique de Streptococcus et comment cette recombinaison est régulée par des facteurs de l'hôte. / The circular state of the bacterial chromosome presents a specific problem during replication. An odd number of homologous recombination events results in concatenated dimer chromosomes that cannot be partitioned into daughter cells. To solve this problem, bacteria have developed a mechanism of dimer resolution based on site-specific recombination system. This is performed by the Xer/dif system. In this system, the Xer proteins perform a recombination reaction in the dif site at the cell septum immediately prior to cell division. In most bacteria this reaction is performed by two recombinases, XerC and XerD. However, an important zoonotic pathogen; Streptococcus suis harbors a different recombination system, composed by a single recombinase enzyme called XerS, that catalyzes the recombination reaction in an unconventional dif site; difSL. A region characterized by two imperfect inverted repeat regions that flank a central region of 11 bp.To characterize the mode of cleavage of XerS, EMSA experiments were performed by using HEX-labelled PCR fragments and “nicked suicide substrates”. Our data suggests that; 1.) XerS is able to bind the entire difSL sequence; 2.) XerS binds more efficiently the left half side on incomplete difSL mutants than the right half side; 3.) XerS cleaves both the top and bottom strands of the difSL site, with a more efficient reaction at the bottom strand; 4.) Nucleotides at the outermost region of a T rich region seem to be determinant for binding selectivity and modifications of the extra spacing between the inverted repeat arms as well as length modifications of the central region change cleavage preference. 5.) XerS did not show any specific activity on another unconventional dif site in Firmicutes, as tested on difH. 6.) XerS interacts with FtsK-y subunit. This research aims to understand how XerS recombination works in the single recombinase system of Streptococcus and how this recombination is regulated by host factors. Exploration of these recombinases will provide a better understanding of the mechanisms of DNA exchange and genome stability in bacteria. It can also increase our knowledge of the evolution and speciation of recombinogenic bacteria.

L'entre-deux-guerres commence à la fin de la Première Guerre mondiale et s'inscrit dans une volonté de changement, de paix durable et d'un nouvel ordre international. Cependant, la réalité du retour à la paix est compliquée par l'ampleur du conflit. L'espace public est transformé: monuments aux morts, destructions, veuves et orphelins. Il y a aussi une réflexion sur la durabilité de la civilisation occidentale : ses limites, ses modèles et ses dangers. Ce mémoire étudie la vision d’un artiste, Hermann-Paul, afin de connaitre les représentations d’un contemporain de l’entre-deux-guerres sur la France et les modèles de la civilisation occidentale. L’étude se concentre sur son travail dans la presse, plus particulièrement sur l’hebdomadaire Je suis partout qui offre le double avantage de pouvoir suivre l’artiste chaque semaine sur une décennie de décembre 1930 à février 1940. Plusieurs questions guident cette recherche. Quelle est la France d’Hermann-Paul? Pourquoi semble-t-elle en crise et quels en sont les modèles et les contre modèles? Les pacifistes sont des acteurs majeurs de la période qui défendent l’idée d’une paix durable. Par leur volonté de changement, ils participent aussi à cette angoisse civilisationnelle. Comment le caricaturiste les intègre-t-il dans sa vision française? Le mémoire se concentre également sur l’instrumentalisation du genre, et sousensemble qu’est la virilité. Il y a une carence d’études francophones dans ce domaine. Ce travail a donc pour objectif de participer à l’exploration du domaine de la virilité en histoire avec le cas d’Hermann-Paul. La caricature est une opportunité puisque ce format instrumentalise les codes du dessin et de la virilité afin de construire, justifier et en détourner les représentations. / The inter-war period began at the end of the First World War and was part of a desire for change, lasting peace and a new international order. However, the reality of a return to peace is complicated by the scale of the conflict. The public space is transformed: monuments to the dead, destruction, widows and orphans. There is also a reflection on the sustainability of Western civilization: its limits, its models and its dangers. This paper examines the vision of an artist, Hermann-Paul, in order to know contemporary representations of inter-war France and the models of Western civilization. The study focuses here on Hermann-Paul’s work in the press, particularly in the weekly magazine Je suis partout, which offers the advantage of being able to follow the artist every week over a decade from December 1930 to February 1940. Several questions guide this research. What is Hermann-Paul's France? Why does it seem to be in crisis and what are the models and counter-models? Peace activists, who defend the idea of a lasting peace, are major actors of the period. Through their desire for change, they also participate in this civilizational anguish. How does the caricaturist integrate them into his French vision? The paper also focuses on the instrumentalization of gender, and subset that is masculinity. There is still a lack of Francophone studies in this area. The objective of this study is to participate in the historical analysis of the field of virility, with the case of Hermann-Paul. The caricature is an opportunity since this format instrumentalizes the codes of drawing and virility in order to construct, justify and divert representations.

Streptococcus suis est une bactérie pathogène qui cause d'importantes pertes économiques dans l'industrie porcine à travers le monde. Comme il n’existe pas de vaccins commerciaux en Amérique du Nord, l'utilisation d'autovaccins administrés aux cochettes/truies pour induire des anticorps passifs chez les porcelets représente une alternative intéressante pour les producteurs. Cependant, il n’existe aucune production standardisée de ces vaccins et le produit final peut être très différent d'un laboratoire agréé à l'autre. Dans la présente étude, un vaccin autogène (« bacterin ») polyvalent contenant les sérotypes 1/2, 2, 5, 7 et 14 de S. suis a été préparé par un laboratoire agréé et utilisé dans un programme de trois doses administrées aux cochettes par voie intramusculaire. La réponse humorale (anticorps) chez les cochettes ainsi que le transfert passif d'anticorps aux porcelets ont été évalués. Contrairement à ce qui avait été publié précédemment avec un vaccin autogène produit par une autre compagnie, la réponse anticorps accrue observée chez les cochettes vaccinées était suffisante pour améliorer le transfert d'anticorps maternels aux porcelets âgés de 3 à 5 semaines. Cependant, les porcelets resteraient encore sensibles à la maladie à S. suis qui apparaît souvent pendant la deuxième partie de la période en pouponnière. Le niveau élevé d'anticorps n'a pas affecté l'excrétion de S. suis (ainsi que celle de sérotypes spécifiques de S. suis inclus dans le vaccin) chez les cochettes et les porcelets. Bien que tous les traitements antibiotiques aient été absents pendant l'essai, l'effet protecteur clinique du programme de vaccination avec le vaccin autogène n'a pas pu être évalué, car des cas limités d’infection à S. suis étaient présents pendant l'essai. D'autres essais pour évaluer l'utilité de la vaccination des cochettes/truies avec des vaccins autogènes pour protéger les porcelets de pouponnière devraient être réalisés. Il est nécessaire, pour les futurs essais sur le terrain, de toujours inclure un groupe témoin non vacciné, d'éliminer si possible tout traitement antimicrobien dans l'élevage et de confirmer l'étiologie des cas cliniques par un diagnostic en laboratoire lors de l'évaluation de l'effet protecteur de tels vaccins autogènes. / Streptococcus suis is a bacterial pathogen that causes important economic losses to the swine industry worldwide. Since there are no commercial vaccines available in North America, the use of autogenous vaccines applied to gilts/sows to induce maternal antibodies to protect piglets is an attractive alternative for producers. However, there is no universal standardization in the production of such vaccines and the final product may be highly different among licenced laboratories. In the present study, a polyvalent autogenous vaccine (“bacterin”) with S. suis serotypes 1/2, 2, 5, 7 and 14 was prepared by a licenced laboratory and used in a three-dose program given to gilts intramuscularily. The humoral (antibody) response in gilts as well as the passive transfer of antibodies to piglets were evaluated. Different from what was previously published with an autogenous vaccine produced by a different company, the increased response seen in vaccinated gilts when compared to non-vaccinated animals was sufficient to improve maternal antibody transfer to piglets of 3 to 5 weeks of age. However, piglets would still remain susceptible to S. suis disease that often appears during the second part of the nursery period. The high level of antibodies did not affect S. suis (as well as that of specific serotypes of S. suis included in the vaccine) shedding by both, gilts and piglets. Although all antibiotic treatments were absent during the trial, the clinical protective effect of the vaccination program with the autogenous vaccine could not be evaluated, since limited S. suis clinical cases were present during the trial. Further trials to evaluate the usefulness of gilt/sow vaccination with autogenous vaccines to protect nursery piglets should be done. There is a need, for future field trials, to always include a control non-vaccinated group, to eliminate if possible any antimicrobial treatment in the farm and to confirm the etiology of clinical cases by a diagnostic laboratory when evaluating the protective effect of such autogenous vaccines.