Durant le développement préimplantatoire, les défauts de ségrégation des chromosomes conduisent à l'héritage d'un nombre incorrect de chromosomes, connu sous le nom d'aneuploïdie, qui provoque l'infertilité. L’imagerie à intervalle du développement préimplantatoire est introduite pour sélectionner le meilleur embryon et des efforts sont en cours pour utiliser l'imagerie non invasive pour identifier les ovocytes euploïdes en métaphase-II comme prédicteur de la viabilité future de l'embryon. Il est déjà bien établi que les ovocytes de mammifères en métaphase-II subissent des mouvements cytoplasmiques stéréotypés qui peuvent être visualisés par imagerie non invasive à fond clair à intervalle, appelée « flux cytoplasmique ». Ici, nous avons émis l'hypothèse que le flux cytoplasmique pourrait être affecté par le statut de ploïdie de l'ovule et donc être un outil de sélection utile pour sélectionner les ovules euploïdes de manière non invasive. Nous avons développé des conditions pour générer des ovules euploïdes et aneuploïdes à partir du même bassin d'ovocytes sains. Nous avons ensuite utilisé la microscopie d'imagerie en temps réel DIC, permettant de visualiser et de mesurer le flux cytoplasmique sans manipulation de l'ovule. Les mouvements cytoplasmiques ont été liés au statut de ploïdie pour chaque ovule individuel par immunofluorescence. Nos résultats montrent qu'il n'y a pas de différence de flux cytoplasmique entre les ovules euploïdes et aneuploïdes. Nos données démontrent que l'état de la ploïdie n'a pas d'impact sur les mouvements cytoplasmiques, suggérant que l'utilisation d'une imagerie non invasive pour essayer de distinguer l'état de la ploïdie entre des ovocytes autrement sains sera difficile. / Chromosome segregation errors during early development lead to inheritance of incorrect number of chromosomes, known as aneuploidy, which causes infertility and birth defects. Time-lapse microscopy of preimplantation development is being widely introduced with the aim of selecting the best embryo and efforts to use non-invasive brightfield imaging to identify euploid oocytes at metaphase-II as a predictor of future embryo viability are underway. It is already well established that mammalian metaphase-II oocytes undergo stereotyped cytoplasmic movements that can be visualised by non-invasive brightfield timelapse imaging, termed “cytoplasmic flow”. Here, we hypothesised that this cytoplasmic flow might be affected by ploidy status of the egg and therefore be a useful selection tool to select euploid eggs non-invasively. To address this, we developed conditions to generate euploid and aneuploid eggs from the same pool of otherwise healthy oocytes. We then used DIC live-imaging microscopy, which allowed us to visualise and measure flow without any manipulation to the egg. Importantly, individual eggs were scored for their ploidy status by immunofluorescence, so that cytoplasmic movements could be related to ploidy on an egg-by-egg basis. Our results show that there is no difference in cytoplasmic flow between euploid and aneuploid eggs. Therefore, our data demonstrates that ploidy status does not impact biologically relevant stereotyped cytoplasmic movements, suggesting that using non-invasive imaging to try to distinguish ploidy status between otherwise healthy oocytes will be challenging.

The avian immune system is a complex system of defence mechanisms that protect bird hosts against threats from ubiquitous pathogens. According to the co-evolutionary models, variability in immune traits of hosts is the key component providing ability to adapt and enhance their defence mechanisms in presence of constant selective pathogen pressure. Domestic chicken (Gallus gallus f. domestica) is used as a model organism in avian biology and also is one of the most important food-producing animals, not only for their meat but also for the egg production. Unfortunately, in research usually only inbred chicken lines are used and modern poultry husbandry is tight with unilateral breeding towards highly productive breeds. Those approaches decrease intra-population polymorphism in chickens. However, especially in case of farm animals, searching and extending the pool of immune variability and enhancing pathogen resistance is crucial for sustaining healthy and biologically secure populations and their products. Morphologically highly distinct traditional chicken breeds, which have evolved for hundreds years under different selective pressures, may represent this desirable immunological variability. In my thesis I described variability in chosen immunological traits, haematological parameters and proteomic...

<p>  </p> <p>Heat stress affects breeder ducks raised in North America and other parts of the world, but the effects of such stress on the progenies is not known. Therefore, the objectives of this study were to investigate: 1) The objectives of this thesis were to first investigate the effect of heat stress or exposure to exogenous glucocorticoid (GC) on fertility, production performance, egg biochemistry, egg quality, and welfare of breeder Pekin ducks. 2) the effects of maternal GC on phenotypic plasticity and behavior of the F1 generation. Three studies were carried out to investigate these objectives.</p> <p>The first experiment was conducted to test the hypothesis that chronic treatment with low levels of either corticosterone or cortisol would alter heterophil to lymphocyte ratio (HLR) and immune organ morphometrics. Further, we wanted to determine if chronic treatment with either GC would elicit an increase in cortisol levels in egg albumen. To test our hypotheses, we implanted silastic capsules subcutaneously under the skin of the neck of adult ducks (n = 5/sex/dose) using propofol anesthesia. Capsules contained corticosterone, cortisol, or empty capsules as controls. Over the course of 2 weeks, blood serum, blood smears, body weights, and egg quality data were collected. After 2 weeks, ducks were euthanized using pentobarbital (FatalPlus, 396 mg/ml/kg) and body weight, weights of spleens, livers, and the number of active follicles were recorded. Blood smears were analyzed for HLR by a lab unaware of the treatment groups. Albumen GC levels were assessed using mass spectrometry. Data were analyzed using a 2- or 3-way ANOVA as appropriate and <em>post hoc </em>with Fishers protected least squares difference (PLSD). There were no treatment effects on egg quality measures or body weight. Corticosterone treatment did elicit an increase in serum corticosterone (p < 0.05), but not cortisol levels, compared to controls in both sexes. Both cortisol and corticosterone treatments increased (p < 0.05) serum levels of cortisol compared to controls. Relative spleen weights were higher (p < 0.05) in hens following corticosterone but not cortisol treatment. No other organs showed any differences among the treatment groups. Both GCs elicited an increase (p < 0.001) in HLR in hens at all time-points over the 2-week treatment period compared to controls. Cortisol, not corticosterone, elicited an increase in HLR for drakes (p < 0.05) compared to controls at day 1 after implants. Chronic treatment with cortisol, but not corticosterone, elicited an increase (p < 0.01) in egg albumen cortisol levels compared to other groups. Corticosterone was not detected in any albumen samples.</p> <p>The goal of our second experiment was to test the hypothesis that heat stress (HS) would alter welfare, egg quality, and morphometrics of breeder ducks. Furthermore, we wanted to test if HS would increase cortisol levels in egg albumen due to recent exciting findings that cortisol, not corticosterone, is isolated in egg albumen. To test our hypothesis, adult Pekin ducks were randomly assigned to two different rooms at 85% lay with 60 hens and 20 drakes per room. Baseline data including body weight, body condition scores (BCS) (such as footpad quality, eyes, nostrils, feather cleanliness, and feather quality scores), and egg production/quality were collected the week preceding heat treatment. Ducks were subjected to cyclic HS of 350C for 10h/day and to 29.50C for the remaining 14h/day for 3 weeks while the control room was maintained at 220C. Eggs were collected daily, and body weights were taken on days 0 and 21 relative to the onset of heat treatment. BCS were collected weekly. Eggs were collected weekly for quality assessment and albumen glucocorticoid (GCs) levels assessment using mass spectrometry. One week before the exposure to HS, 10 hens and 5 drakes were euthanized and the same number again after 3 weeks of HS or control exposures using pentobarbital and birds necropsied. Body weight, weights of the liver, spleen, and the number of maturing follicles were recorded. Data analyses were done by 2- or 3-way ANOVA as appropriate with a Tukey-Kramer post hoc test. BCS were analyzed using a chi-squared test. A p value ≤ 0.05 was considered significant. Circulating levels of corticosterone were significantly (p < 0.01) elevated at week 1 only in the HS hens while there was no significant difference in the circulating levels of corticosterone in drakes compared to the controls. The circulating levels of cortisol increased significantly at week 1 (p < 0.05), week 2 (p < 0.05), and week 3 (p < 0.01) in the hens and at week 2 and 3 only (p < 0.05) in the drakes compared to the controls. Feather quality scores (p < 0.01), feather cleanliness scores (p < 0.001) and footpad quality scores (p < 0.05) increased significantly in the HS group compared to controls, higher BCS indicate a decline in welfare. HS elicited a significant (p < 0.001) decrease in egg production at weeks 1 and 3 and a descriptive decrease in the number of fertile eggs upon candling at 10 days of incubation, numeric decrease hatchability and increase in the number of dead embryos in the HS group after the incubation period. Hens in the HS group showed a significantly decreased BW (p < 0.001), and number of ovarian follicles (p < 0.05) compared to controls. Shell weight decreased significantly at week 1 (p < 0.05) compared to controls. Yolk weight decreased significantly at week 3 (p < 0.01) compared to controls. HS elicited a significant increase in albumen cortisol levels at week 1 (p < 0.05) and week 3 (p < 0.05).</p> <p>The third experiment was conducted to determine if parental exposure to heat stress would impair performance, hypothalamic pituitary adrenal (HPA) axis response, welfare, or behavior of their offspring. To achieve these goals, we treated adult drakes and hens at peak lay to heat stress or control temperature for 3 weeks and incubated eggs collected from the last 3 days of the experiment. A total of 76 ducklings were placed into pens from each parental treatment group: control (CON-F1) and heat stress (HS-F1) and raised as grow-out ducks. Weekly data for body weights, body condition scores (BCS), and novel object test (NOT) were collected weekly. At 3 weeks of age, ducks (n = 6 per treatment group) were subjected to adrenocorticotropic hormone (ACTH) (ACTH/cosyntropin, 0.0625 mg/kg) challenge or vehicle as control. Blood samples were collected from the metatarsal vein into serum-separator tubes at 0, 1, 2, 3, and 4 hours relative to treatment for the determination of serum glucocorticoids. Blood smears were also produced from these same samples to determine heterophil to lymphocyte ratios (HLR). All injected birds were euthanized with pentobarbital on the second day relative to ACTH administration, spleen and bursa were removed and weighed immediately. Duck level analyses were completed using 1-, or 2 -way ANOVA as appropriate. BCS were analyzed using a chi-squared test. We observed that HS-F1 had a lower hatch weight (p < 0.05) compared to CON-F1. However, growth rates during the 5-week grow-out period were not significantly different between the two flocks. NOT (N = 4) analyses showed that the HS-F1 had a greater fear response (P< 0.001) compared to CON-F1. Similarly, an ACTH stimulation test showed that the HS-F1 ducks had significantly heightened corticosterone and HLR responses compared to CON-F1 ducks (p < 0.05). The HS-F1 showed altered baseline and ACTH-stimulated levels of cortisol compared to controls.</p> <p>In conclusion, GC elicit differential effects and although corticosterone has been stated to be the predominant GC in avian species, cortisol may provide critical information to further understand and improve welfare. HS decreased performance, fertility, and productivity of breeder ducks. In addition, HS and exogenous GC elicited a selective deposition of cortisol, not corticosterone, in the egg albumen. The maternal cortisol deposited in eggs alter the hypothalamic-pituitary adrenal (HPA) axis and behavioral responses of the F1 generation. This suggests that maternal hormones can alter the phenotypic plasticity of the offspring and can be used to produce offspring that have better adaptation to the rising temperatures as a result of climate change. Finally, the measure of cortisol in egg albumen can be used as a non-invasive marker of stress.</p>

The basement membrane is a specialized extracellular matrix (ECM) that is crucial for the development of epithelial tissues and organs. In Drosophila, the mechanical properties of the basement membrane play an important role in the proper elongation of the developing egg chamber; however, the molecular mechanisms contributing to basement membrane mechanical properties are not fully understood. Here, we systematically analyze the contributions of individual ECM components towards the molecular composition and mechanical properties of the basement membrane underlying the follicle epithelium of Drosophila egg chambers. We find that the Laminin and Collagen IV networks largely persist in the absence of the other components. Moreover, we show that Perlecan and Collagen IV, but not Laminin or Nidogen, contribute greatly towards egg chamber elongation. Similarly, Perlecan and Collagen, but not Laminin or Nidogen, contribute towards the resistance of egg chambers against osmotic stress. Finally, using atomic force microscopy we show that basement membrane stiffness mainly depends on Collagen IV. Our analysis reveals how single ECM components contribute to the mechanical properties of the basement membrane controlling tissue and organ shape.

Syfte: Det övergripande syftet med uppsatsen är att förstå hur transaktionsplattformar ska lyckas i tidiga stadier. Detta genom att förstå hur en transkansaktionsplattform ska utformas för att kunna attrahera många användare.  Metod: Denna studie har använt en kvalitativ forskningsansats där intervjuerna genomfördes i tre faser. I studien genomfördes intervjuer med 24 svenska bolag och organisationer verksamma i energibranschen. Studien intervjuade två olika användargrupper vilket var beställare och utförare. Utöver det genomfördes även intervjuer med branschkunniga aktörer.  Resultat: Resultatavsnittet är indelat i två delar där första delen handlar om att skapa förståelse för sju faktorer som påverkar intentionen för potentiella användare att adoptera en transaktionsplattform. Här presenteras på vilket sätt dessa faktorer påverkar adoptionsbeslutet samt vilka handlingar som plattformsägaren kan vidta för att öka deras intention att adoptera. I andra delen av resultatavsnittet presenteras ett ramverk som plattformsägaren kan använda i utformningsstadiet. Ramverket handlar om att identifiera intressanta användargrupper samt adressera faktorer som påverkar adoptionsbeslutet.  Teoretiskt och praktiskt bidrag: Befintlig litteratur har sällan i samma studie inkluderat flera faser av plattformars livscykel, utan enbart fokuserat på en fas åt gången. Denna studie bidrar teoretiskt genom att undersöka hur utformningsfasen påverkar adoptionsfasen för transaktionsplattformar. Dessutom bidrar denna studie till litteraturen genom att undersöka tidiga stadier för transaktionsplattformar. Studien fokuserar på utformningsfasen, vilket sällan undersökts i befintlig litteratur. Studien bidrar även praktiskt genom att presentera ett ramverk som kan hjälpa chefer eller entreprenörer vid utformningen av en ny transaktionsplattform. Användning av ramverket kan underlätta hanteringen av höna-och-ägg-problemet som nya transaktionsplattformar alltid möts av.  Begränsningar och framtida forskning: Det finns framför allt två begränsningar med denna studie, vilket öppnar upp för framtida studier. Den första är att den undersökta intervjugruppen är relativt snäv vilket kan leda till branschspecifika resultat. Framtida studier kan därmed undersöka andra branscher och jämföra mot denna studies resultat för att öka generaliserbarheten. Den andra begränsningen är att denna studie enbart intervjuat potentiella användare av transaktionsplattformar och inte nuvarande användare – det vill säga adoptörer. Därmed var intervjuundersökning relativt hypotetisk och mycket fokus lades på vad intervjuobjekten kunde tänka sig. Framtida studier kan därmed även intervjua nuvarande användare av en transaktionsplattform för att vidareutveckla resultatet. / Purpose: The main purpose of the thesis is to understand how transaction platforms can become successful in early phases. This will be achieved by understanding how a transaction platform should be designed in order to attract many users to the platform. Method: This study had a qualitative research approach, and the interviews were performed in three phases. Interviews were made with 24 companies and organizations within the energy industry. Two different user groups were interviews, which was buyers and sellers. Besides that, people with high knowledge of the industry were interviewed. Result: The result section is divided in two parts, where the first part intends to create knowledge about seven factors that affects the intention for potential users to adopt a transaction platform. It is described in what way these factors affect the adoption decision and which action the platform owner can take to increase the intention to adopt. In the second part of the result section, a framework is presented that platform owners can use in the design phase. The framework is about identifying interesting user groups and addressing factors that affects the adoption decision. Practical and theoretical contribution: Existing literature has rarely focused on including several phases of the platform's life cycle. Previous studies has therefore only focused on one phase at a time. This study contributes to the literature by investigating how the design phase affects the adoption phase for transaction platforms. Additionally, this study contributes to the literature by investigating the early phase of a platform, which is rare to find in the existing literature. The study contributes practically by providing a framework that can help decision-makers and entrepreneurs to design a new transactional platform. Using the framework can help managers to cope the chicken-and-egg dilemma that new platforms often face.  Limitations and future studies: This study has two limitations that opens for future studies. Firstly, our interview group is relatively narrowed, which can lead to industry-specific findings. Future studies can therefore investigate other industries to compare those findings with this study to identify more general findings. The second limitation is that this study has exclusively done interviews with potential users. Therefore, the interview study was relatively hypothetical, and a lot of focus was on what the interviewees could imagine. Future studies can therefore do interviews with users of an existing transaction platform to develop our framework more.

The innate immune system in vertebrates is considered to lack specific memory. To investigate innate immune system based immunological protection mediated by cells that are not part of the acquired immune system the Tübingen recombination activation gene1 (rag1)t26683 mutant (MT) zebrafish was chosen. Molecular analysis demonstrated MT zebrafish kidney cells expressed Non-specific Cytotoxic cell receptor protein-1 (NCCRP-1) and Natural Killer cell (NK) lysin but lacked T cell receptor (TCR) and immunoglobulin (Ig) VH1, VH2, VH3 and VH4 expression. Differential counts of peripheral blood leukocytes indicated that MT fish had decreased lymphocyte populations (34.7%) compared to rag1+/+ wild-type (WT) fish (70.5%), and increased granulocyte populations (34.7%) compared to WT (17.6%). Further, endocytic functions of phagocytes from MT fish were compared to WT fish. No significant differences in the selective and non-selective mechanisms of uptake in phagocytes were observed between MT and WT zebrafish. For the first time it was shown that zebrafish phagocytes utilize macropinocytosis and Ca2+ dependant endocytosis mechanisms for antigen uptake. These characterization studies suggest that MT zebrafish provide a unique model for investigating innate immune responses because fully functional innate defenses are present without the influence of lymphocytes and lymphocyte associated acquired immune responses. To conduct such large scale investigations the first ongoing rag1t26683 mutant zebrafish breeding colony was established. To meet special husbandry needs of immunodeficient MT zebrafish, standard rearing protocols were advanced and the information was made available to the zebrafish community at: http://www.cvm.msstate.edu/zebrafish/index.html. Multiple trials were conducted to evaluate the potential for memory of the innate immune system. Significant reduction in mortality was observed in MT vaccinated zebrafish upon secondary exposure to Edwardsiella ictaluri when compared to unvaccinated, MT fish. This documents for the first time, that MT zebrafish, lacking an acquired immune system, are able to mount a protective immune response to Edwardsiella ictaluri and generate protection upon a repeated encounter to the same pathogen. The observed protection is long lasting and mediated by the innate immune system, but a specific mechanism is not yet defined.

No description available.

No description available.

Three-dimensional printing technologies exhibit tremendous potential in the advancing fields of tissue engineering and regenerative medicine due to the precise spatial control over depositing the biomaterial. Despite their widespread utilization and numerous advantages, the development of suitable novel biomaterials for extrusion-based 3D printing of scaffolds that support cell attachment, proliferation, and vascularization remains a challenge. Multi-material composite hydrogels present incredible potential in this field. Thus, in this work, a multi-material composite hydrogel with a promising formulation of chitosan/gelatin functionalized with egg white was developed, which provides good printability and shape fidelity. In addition, a series of comparative analyses of different crosslinking agents and processes based on tripolyphosphate (TPP), genipin (GP), and glutaraldehyde (GTA) were investigated and compared to select the ideal crosslinking strategy to enhance the physicochemical and biological properties of the fabricated scaffolds. All of the results indicate that the composite hydrogel and the resulting scaffolds utilizing TPP crosslinking have great potential in tissue engineering, especially for supporting neo-vessel growth into the scaffold and promoting angiogenesis within engineered tissues.

Réalisé en cotutellle avec Gaétan Bourgeois et avec la participation de Guy Boivin, d'Agriculture et Agroalimentaire Canada, du Centre de Recherche et Développement en Horticulture à Saint-Jean-sur-Richelieu, QC J3B 3E6, Canada / On dénombre divers modèles de simulation de la mouche du chou (Delia radicum L.), mais plusieurs comportent d’importantes lacunes au niveau des différences génotypiques de l’insecte et des paramètres utilisés. L’objectif principal de ce projet est de rassembler les informations manquantes afin de créer dans le futur un modèle bioclimatique permettant de simuler efficacement la dynamique des populations de ce ravageur. L’effet de la température et de l’humidité du sol a été mesuré sur les œufs et les larves de la mouche du chou. L’humidité n’influence la survie des œufs qu’en dessous de 25% [m/m]. L’exposition graduelle des œufs à des températures élevées au dessus de 33°C affecte également la survie. La survie des larves augmente avec la hausse des températures et de l’humidité. Nous croyons que la mouche du chou est bien adaptée aux conditions des sols organiques au Québec, et nous recommandons l’intégration de la température du sol pour les stades au sol plutôt que de l’air dans l’élaboration d’un nouveau modèle. La ponte a également été étudiée à partir de différents critères préétablis pour chacun des génotypes hâtifs et tardifs, à différentes températures. Excepté pour la pré-oviposition qui est plus longue chez les hâtifs, aucune différence n’a été observée entre les génotypes. La majorité des critères, excepté la durée d’un épisode de ponte et la mortalité des œufs, a réagit à la température. Les nouvelles informations serviront à l’élaboration ou le perfectionnement d’un modèle de simulation de la dynamique de la ponte de la mouche du chou. / Few simulation models for the cabbage maggot (Delia radicum L.) had been seen, but several include significant weaknesses for the insect genotypic differences and parameters used. The main objective of this project is to collect the missing information to create, in the future, a bioclimatic model that will efficiently simulate the dynamics of the populations of this pest. The effect of soil temperature and moisture had been measured on eggs and larvae of the cabbage maggot. Humidity affects egg survival below 25% [w/w]. Gradual exposure of eggs to high temperature above 33°C also affects survival. Under tested conditions, larvae survival increases with rising temperatures and humidities. We believe that the maggot is well adapted to muck soil in Quebec, and we recommend to incorporated soil temperature rather than air temperature for above-ground stages into the development of a new model. Egg-laying activity has also been studied from various pre-established criterions for each of the early- and late-emerging genotypes, at different temperatures. Except for the pre-oviposition which is longer for the early genotype, no differences were observed between genotypes. The majority of criterions, except for the duration of oviposition bouts and egg mortality, responded to temperature. The new information will be used for the preparation and development of a simulation model of the egg-laying dynamic of the cabbage maggot.