Os objetivos gerais do presente estudo foram avaliar o efeito do polimorfismo genético da kappa-caseína, da raça e da estação do ano sobre as características físico-químicas (acidez, pH e crioscopia), composição (gordura, lactose, sólidos totais, contagem de células somáticas, uréia, proteína bruta, proteína verdadeira, nitrogênio não protéico e nitrogênio não caseinoso) e estabilidade do leite. Foram selecionados 11 rebanhos leiteiros comerciais, sendo cinco deles da raça Holandesa e seis da raça Girolanda, dos quais foram amostradas em média 122 vacas em lactação por rebanho, totalizando 1350 vacas amostradas em três períodos: 2 no período seco e 1 no período chuvoso. As vacas selecionadas foram analisadas quanto a composição e propriedades físico-químicas do leite, assim como para a determinação do polimorfismo de kappa-caseína. O presente trabalho foi dividido em dois estudos, no primeiro avaliou-se os efeitos da raça e estação do ano sobre características físico-químicas, composição e estabilidade do leite. Foi observado efeito de raça sobre acidez titulável, pH, lactose, uréia e estabilidade do leite. O efeito de sazonalidade mostrou-se significativo sobre pH, crioscopia, teores de lactose, uréia, proteína bruta, sólidos totais, contagem de células somáticas, proteína verdadeira, caseína, nitrogênio não protéico e estabilidade térmica do leite. Em animais da raça Girolanda, foram observados no período seco maiores teores de sólido totais. O ponto crioscópico do leite sofreu efeito do período de coleta somente em animais da raça Holandesa, onde valores mais altos foram observados durante o período seco. Os teores de lactose apresentaram maiores médias no período seco em animais da raça Holandesa. Os teores de uréia sofreram efeito de raça e período de coleta estudado. A proteína bruta do leite sofreu efeito dos fatores raça e período de coleta, sendo que vacas da raça Holandesa e Girolanda apresentam maiores teores no período seco. A concentração de caseína do leite sofreu efeito significativo do período de coleta. A estabilidade térmica do leite sofreu influência da raça, pois o leite de vacas da raça Holandesa mostraram-se mais estáveis do que os da raça Girolanda. A sazonalidade é um fator determinante para a estabilidade do leite, pois o leite mostrou-se mais estável no período chuvoso que em período seco. No segundo estudo foram avaliados o polimorfismo genético da kappa-caseína em vacas Holandesas e Girolandas e o efeito deste sobre características físico-químicas, de composição e estabilidade térmica do leite. A freqüência do alelo A foi maior do que a do alelo B no que diz respeito ao gene da kappa-caseína. Com relação às características físico-químicas do leite, não houve efeito do polimorfismo do gene da kappa-caseína sobre teores de gordura, sólidos totais, lactose, contagem de células somáticas e uréia do leite. Não foram observados efeitos do polimorfismo genético do gene para kappa-caseína sobre a composição protéica do leite, sendo que teores de proteína bruta, nitrogênio não protéico, nitrogênio não caseinoso e proteína verdadeira não sofrem influência. A estabilidade do leite frente à prova do álcool não sofreu influencia do polimorfismo genético de kappa-caseína / The objective of this study were to determine the effects of kappa-casein gene polymorphisms, breed and season on physical-chemical properties (acidity, pH and cryoscopy), composition (fat, lactose, total solids, somatic cells count, urea, crude protein, true protein, non protein nitrogen and non casein nitrogen) and stability of milk. For this aim 11 dairy herds were selected, six of them composed of Girolando cows and five from Holstein cows, in average milk samples were taken of 112 cows from each herd, collected three times: twice in dry season and once on rainy season. Each cow were analyzed for milk composition, physico-chemical properties, and to determine its kappa-casein polymorphism. This study was divided in two, the first one analyzed possible breed and season effects over milk physico-chemical characteristics, composition and stability. Breed effect was observed over acidity, pH, lactose, urea and milk stability. Season effects was significant for pH, cryoscopy, lactose, urea, CP, total solids, SCC, TP, casein, EqNPN and heat milk stability. In cows Girolando it was observed highest total solids in dry season. Milk cryoscopy had season effects only on Holstein cows, with highest results in dry season. Lactose concentration was greatest in Holstein cow in dry season. Urea concentration showed breed and season effect on this study. Crude protein was affected by breed and season, in dry season the concentration were highest for Holstein and Girolanda. Casein milk concentration demonstrated season effect. Heat milk stability showed breed stability, milk from Holstein cow were more stable that milk from Girolanda cows. Season is a determinant factor for milk stability, milk showed more stable in rainy season than during dry period. The second study analyzed: kappa-casein gene polymorphisms in Holstein and Girolando cows and its effects over milk physico-chemical characteristics, milk composition and milk heat stability. Kappa-casein allele A had a higher frequency, than allele B, in Girolando and Holstein cows in comparison with other polymorphisms. No effect of kappa-casein polymorphism was observed on milk phisico-chemical characteristics and on milk fat, total solids, lactose, SCC, and milk urea. There were no difference for milk protein composition (EqNPN, NCNC, TP, casein and EqNCN) for kappa-casein polymorphism. Milk alcohol stability did not showed effect of gene polymorphism

L'objectif de ce travail est de caractériser l'effet des levures vivantes Sc47 sur le métabolisme ruminal et l'utilisation digestive de la ration en lien avec le statut oxydo-réducteur du rumen chez des vaches laitières recevant des rations qui diffèrent dans leur composition. En effet, le milieu ruminal est très anaérobie et réducteur : les micro-organismes qui s'y développent sont à l'origine de ces conditions physico-chimiques particulières caractérisées par des valeurs de potentiel rédox (Eh) ou indice de Clark (rH) basses : respectivement dans cette étude entre –213 et –147 mV et entre 6.04 et 7.48 unités rH. L'impact de trois constituants du régime des vaches laitières d'une part sur le statut réducteur du milieu ruminal et d'autre part sur son métabolisme et/ou l'utilisation digestive de la ration a été testé en interaction avec l'addition de levures vivantes : un fourrage sec chez la vache tarie, deux concentrés azotés différant par le niveau de solubilité ruminal des protéines qu'il contient et enfin la nature de l'amidon, rapidement ou lentement dégradable dans le rumen chez la vache en lactation. Il apparaît nettement que le régime alimentaire ainsi que le niveau d'ingestion des animaux influencent directement le statut réducteur du rumen : un niveau réducteur bas étant favorable à l'activité d'une flore cellulolytique alors que des niveaux plus élevés s'avèrent être un indicateur de l'apparition de troubles métaboliques du rumen. En effet, le pouvoir réducteur ruminal est corrélé avec l'activité fermentaire qui s'y développe ainsi qu'à la structure des communautés bactériennes en présence. D'après nos résultats, l'effet de la levure vivante sur les conditions réductrices du rumen semble fortement conditionné par le niveau du statut réducteur induit par le régime alimentaire : les conditions réductrices ruminales peuvent être renforcées dans la mesure où le Eh initial se trouve, dans notre étude, entre –174 et –152 mV. Ce renforcement serait favorable à l'activité de la flore fibrolytique si celle-ci n'est pas dominante, ce groupe fonctionnel participant alors également au maintien des conditons réductrices. En outre, la levure vivante aurait un impact direct sur les bactéries protéolytiques favorisant la quantité de protéines "by pass" si bien que la valeur azotée de la ration peut être améliorée en particulier en termes de PDIA. / The objective of this work is to characterize the effect of live yeast Sc47 on ruminal metabolism and ration digestibility in relation to ruminal redox status of dairy cows fed diets that differed in their composition. Indeed, the rumen environment is very anaerobic and very reducing : the inhabiting micro-organisms are the main sources of particular physic-chemical conditions characterized by low values of redox potential (Eh) or Clark’s exponent (rH): in this study, values were between –213 and –147 mV and between 6.04 and 7.48 units for Eh and rH, respectively. The impact of three constituting ingredients of the diet of dairy cows was investigated. On the one hand, the reducing status of rumen was observed and on the other hand, the metabolism and/or digestibility of the diet with or without addition of live yeast was explored. The tested constituents were: hay for dry dairy cows, two concentrates differing in nitrogen levels of ruminal solubility, and two energetic concentrates differing in rate of ruminal degradation of starch – quickly or slowly degradable in the rumen of lactating dairy cows. It clearly appeared that the diet and the level of dry matter intakes of animals directly influenced the reducing ruminal status: low levels being favorable to the activity of cellulolytic microflora whereas higher levels appeared to be indicators of metabolic disorders occuring in the rumen. Indeed, the reducing power was correlated to rumen fermentative activity and the structure of bacterial communities involved. According to our results, the effect of live yeast on rumen reducing conditions appeared strongly influenced by the level of the intrinsic reducing status induced by diet: reducing conditions in the rumen could be strengthened to the extent that the original values ranged between –174 and –152 mV. This reinforced ruminal conditions would uphold the activity of fibrolytic microflora if it is not dominant, this functional group then participating to the maintenance of reducing conditions. In addition, live yeast has a direct impact on proteolytic bacteria promoting by-pass proteins so that the protein value of the ration may be improved especially in terms of PDIA.

Die Mastitis beim Milchrind hat eine große ökonomische Bedeutung, daher liegt derzeit ein Forschungsschwerpunkt auf der Aktivierung und Stabilisierung der körpereigenen Abwehr zur Bekämpfung von Euterentzündungen. Besonders im peripartalen Zeitraum liegt eine Prädisposition für Infektionen vor, da eine physiologische Abwehrschwäche besteht. Daher erscheint die Förderung der Bildung von Antikörpern durch Impfungen sinnvoll. Getestet wurde der Einfluss von bestandsspezifischen E. coli-Vakzinen auf das Immunsystem, das Erregervorkommen in der Milch und die Eutergesundheit. Differenziert wurden Impfstoffe, die einerseits aus den Originalkulturen der antigenen Erreger (sogenannte large colony variants, LCV) oder aber aus den kleineren, intrazellulär persistierenden Erregern (small colony variants, SCV) hergestellt wurden. Letztlich zeigte sich bei Anwendung der Vakzinen an Milchrindern kein Unterschied zwischen LCV und SCV, bei beiden Impfstoffen war eine vakzinationsbedingte deutliche Steigerung der Antikörpertiter, welche einen Einfluss auf Erregervorkommen und Eutergesundheit hatte, erkennbar.

In der Literatur steht die Herzfrequenz (HF) in engem Zusammenhang mit dem Sauerstoffverbrauch und dem Energieumsatz sowohl von Menschen als auch von verschiedenen Tierarten. In der vorliegenden Arbeit sollte die Hypothese überprüft werden, ob Kühe mit einer hohen Milchleistung und einer hohen Energieaufnahme eine höhere HF infolge eines leistungsabhängig höheren Sauerstoffverbrauchs als niedriglaktierende Tiere haben. So könnte die HF als Indikator für quantitative Veränderungen des Energieumsatzes sowohl für die Einzelkuh als auch auf Bestandsebene genutzt werden. Material und Methoden In vorliegender Untersuchung wurde die HF von 32 Milchkühen der Rasse Deutsche Holstein gemessen. Die Herde erhielt während der Versuche betriebseigenes Grundfutter sowie Maisschrot, Sojaextraktionsschrot und Milchleistungsfutter II. Die tägliche Futter-, Energie- und Proteinaufnahme wurde bestimmt. Die HF-Messung der Kühe erfolgte in zwei Abschnitten. Abschnitt A: Bei 23 Kühen, unterteilt in unterschiedliche Reproduktions- und Leistungsstadien, wurde an fünf aufeinanderfolgenden Tagen die HF erfasst. Abschnitt B: Bei elf Einzeltieren wurde vom 6. bis 101. Laktationstag (LT) die HF in regelmäßigen Abständen alle 14 Tage und an drei aufeinanderfolgenden Tagen gemessen. Den Tieren wurde hierzu ein Polar Equine RSCX800 Science® Gurt mit integrierten Elektroden, ein dazugehöriger Sender und ein Empfänger (Polar Uhr RS800®) angelegt. Die HF wurde einmal pro Minute gespeichert und alle 24 h in das Computerprogramm Polar Pro Trainer Equine Edition 5® übertragen. Folgende weitere Daten wurden erhoben: Stalltemperatur, relative Luftfeuchtigkeit des Stalles, Body Condition Score, Körpermasse, Körperinnentemperatur, tägliche Milchmenge, Milchfett-, Milcheiweiß- und Milchharnstoffgehalt. Ergebnisse Abschnitt A: Die trockenstehenden Kühe hatten eine HF von 83±8 Schlägen/min, die laktierenden Kühe wiesen mit steigender Milchleistung (24,0; 37,1 und 47,7 kg fett- und eiweißkorrigierte Milch) HF-Werte von 84±3, 85±6 und 87±2 Schlägen/min auf. Abschnitt B: In den ersten 100 Tagen der Laktation verhielt sich die HF mit steigendem LT wie folgt: LT 8: 86±9 Schläge/min, LT 22: 86±11 Schläge/min, LT 36: 87±11 Schläge/min, LT 52: 83±10 Schläge/min, LT 66: 83±10 Schläge/min, LT 81: 83±10 Schläge/min, LT 95: 85±8 Schläge/min. Weder in Abhängigkeit von der Leistung (p=0,75) noch in Abhängigkeit vom LT (p=0,81) waren signifikante Unterschiede der HF zu verzeichnen. Die HF der trockenstehenden Kühe variierte im Vergleich zur HF der laktierenden Tiere nicht. Während ihres 24 h-Verlaufs unterlag die HF aller Gruppen einer großen Variation. Es war zu Ruhezeiten der Tiere eine leistungsabhängige Abstufung der HF zu erkennen. Die Umgebungstemperatur hatte in dieser Untersuchung keinen Einfluss auf die HF (r=0,01; p=0,95). Einen signifikanten negativen Zusammenhang gab es zwischen der HF und dem Alter der Tiere (r=-0,44; p<0,01). Dieser führte am 22. LT zu einem signifikanten Unterschied der HF der erstlaktierenden Kühe im Vergleich mit der HF der älteren Tiere (p=0,04). Ein hoher und signifikanter Zusammenhang konnte zwischen der HF und dem Graviditätstag der trockenstehenden Kühe gezeigt werden (r=0,83; p=0,04). Die Tiere, die sich in einer negativen Energiebilanz befanden, neigten in dieser Untersuchung zu einer niedrigeren HF als die Kühe mit einer positiven Energiebilanz. Schlussfolgerung Telemetrische Systeme gewinnen beim Monitoring von Gesundheitsproblemen und Stoffwechselparametern eines Milchviehbestandes zunehmend an Bedeutung. Die HF kann hier jedoch nicht eingesetzt werden, da sie als Indikator für quantitative Veränderungen des Energieumsatzes weder für die Einzelkuh noch auf Bestandsebene geeignet ist. Nach vorliegenden Ergebnissen haben Kühe mit einer höheren Milchleistung und einer höheren Energieaufnahme keine höhere HF. / Previous studies have revealed a close relationship between heart rate (HR) and energy expenditure in humans and some other animal species. The present study examined whether dairy cows with a higher milk yield and greater energy intake have a higher HR due to greater oxygen consumption. HR might serve as an indicator of quantitative changes of energy expenditure in an individual or in a group of dairy cows at the herd level. Materials and methods The HRs of 32 dairy cows of the German Holstein breed were measured. The cows received forages and fresh beet pulp in combination with ground corn, soybean extracts, and a concentrate during the study. Daily feed, energy, and protein intake were determined. HR measurement was performed in two blocks. In Block A, cows were divided into four groups that differed in reproductive state and milk yield, and HR was measured for five consecutive days. In Block B, every 14 days, HR was recorded for three consecutive days in 11 cows from 6 to 101 days in milk (DIM). A Polar Equine RSCX800 Science® belt with integrated electrodes, a transmitter, and a receiver (Polar RS800®) was applied to the animals. HR was recorded once a minute and transferred every 24 h to the software Polar Pro Trainer Equine Edition 5®. The following data were also collected: stable temperature, stable relative humidity, body condition score, body weight, body temperature, daily milk yield, milk fat, milk protein, and milk urea. Results In Block A, dry cows had a HR of 83±8 beats/min, and lactating cows with increasing milk yield (24.0, 37.1, and 47.7 kg energy-corrected milk) showed HRs of 84±3, 85±6, and 87±2 beats/min, respectively. In Block B, the HRs were as follows: 8 DIM: 86±9 beats/min, 22 DIM: 86±11 beats/min, 36 DIM: 87±11 beats/min, 52 DIM: 83±10 beats/min, 66 DIM: 83±10 beats/min, 81 DIM: 83±10 beats/min, 95 DIM: 85±8 beats/min. There was not a significant difference in the HR according to milk yield (p=0.75) or days of lactation (p=0.81). The HR of dry cows did not differ from that of lactating cows. There was a large variation of HR in all groups during the day. During resting times, HR of the high-yielding cows tended to be higher and HR of the dry cows tended to be lower than the HR of the other groups. The ambient temperature had no influence on HR in this study (r=0.01, p=0.95). There was a significant negative correlation between HR and a cow’s age (r=–0.44, p<0.01). The HR of primiparous cows was higher than that of multiparous cows, but the difference was only significant when cows were 22 DIM (p=0.04). A high and significant correlation was found between HR and pregnancy status (r=0.83, p=0.04), with HR increasing during pregnancy. Dairy cows with a negative energy balance tended to have a lower HR than cows with a positive energy balance. Conclusion Telemetry systems have become increasingly important in monitoring health problems and the parameters of energy metabolism in dairy herds. The results of this study indicate that HR cannot be used as a monitoring tool, because HR is not a suitable indicator of quantitative changes of energy expenditure either at the individual or herd level. According to the results of this study, dairy cows with a higher milk yield and greater energy uptake do not have a higher HR.

Inwieweit kann pflanzliches Futterfett als zusätzliche Energie oder anstelle anderer Energieträger zum Einsatz kommen? Die Energieversorgung der Wiederkäuer über Futterfett zu erhöhen, wird in jüngster Zeit in verschiedenen Ländern untersucht. Dabei zeigt sich, dass bei Fetteinsatz noch nicht alle Fragen der Stoffwechselbeeinflussung und Fütterungspraxis geklärt sind (MAHLKOW-NERGE, 2002). Aufgrund der z. T. noch offenen Fragen sollte in länderübergreifender Zusammenarbeit der Landesanstalten Sachsen (Köllitsch) und Sachsen-Anhalt (Iden) die Wirkung eines bereits vielerorts in der Milchkuhfütterung eingesetzten pansengeschützten Fettes charakterisiert werden. Es geht darum, die Kühe in der Hochleistungsphase (1. 150 Laktationstage) möglichst ausreichend mit Energie zu versorgen (Stoffwechselstabilisierung). Inwieweit dafür das pansengeschützte Pflanzenfett (Bergafat T 300) geeignet ist, wurde in Abstimmung mit der Sächsischen Landesanstalt für Landwirtschaft (LfL) in einem 140-tägigen Einzelfütterungsversuch der Landesanstalt für Landwirtschaft und Gartenbau Sachsen-Anhalt am Standort Iden untersucht.

La santé et la fertilité des vaches laitières sont au cœur des préoccupations de la filière professionnelle dans un objectif d’efficience, de quantité et de qualité de la production de lait. La mise en place d’une lactation performante se superpose aux profonds changements hormonaux et métaboliques de la période postpartum, se traduisant par une balance énergétique négative. Les conséquences en sont souvent une altération de la fertilité et une immunodépression qui accroit la susceptibilité aux pathologies. Dans les élevages, il existe encore une grande variabilité d’état général et de performances chez les vaches laitières malgré la sélection génomique. Il est proposé que des modifications de la méthylation de l’ADN puissent contribuer à cette variabilité phénotypique individuelle. En effet, la méthylation de l’ADN, en tant que processus épigénétique, est impliquée dans la régulation transcriptionnelle des gènes, et présente une certaine plasticité face aux contraintes environnementales. Nous avons fait l’hypothèse que des signatures épigénétiques portées par les cellules du sang, pourraient refléter l’état de santé des vaches et pourraient être modifiées en réponse à différents facteurs intrinsèques (parité, stades physiologiques…) et aux contraintes environnementales. Ces signatures ont été recherchées dans une population de cellules du sang particulière : les monocytes. Ces cellules, accessibles par prélèvements sanguins et purification en présence d’un anticorps spécifique, constituent la première ligne de réponse d’immunité innée face aux infections aigües participant à la dégradation de l’état de santé des vaches en postpartum. Pour tester l’hypothèse de signatures épigénétiques monocytaires, une analyse du méthylome par « Reduced Representation Bisulfite Sequencing », (RRBS) dans diverses situations d’élevage a été réalisée. En utilisant des ADN génomiques de vaches incluses dans plusieurs protocoles, 22 banques ont été construites et séquencées. Leur analyse a été réalisée en utilisant un pipeline d’analyses bioinformatique et biostatistique développé au laboratoire.En moyenne 1 250 000 CpG sont pris en considération et permettent l’identification et la localisation de cytosines différentiellement méthylées (DMC) : i) 27143 DMC en comparant les méthylomes de différents types cellulaires (monocytes versus fibroblastes et PBMC) ii) 4788 DMC en réponse à un challenge nutritionnel basé sur la distribution du complément alimentaire GENIAL®, fabriqué et distribué en élevage par nos partenaires PILARDIERE et XR-Repro. iii) 2615 et 4616 DMC en réponse au challenge infectieux pour le groupe de vaches témoins et le groupe en restriction alimentaire respectivement (protocole coordonné par Christine Leroux et José Pires (RUMINFLAME, INRA, Theix) combinant une restriction alimentaire et un challenge immunitaire, par injection de LipoPolySaccharide). iv) 4420 DMC issues de la comparaison entre méthylomes de vaches à génome constant (issues du transfert nucléaire, clones) et de vaches à génome variable mais de même âge et élevées dans les mêmes conditions que les clones. Pour certaines régions différentiellement méthylées (DMR) ciblant le promoteur de gènes, le statut de méthylation a été confirmé par conversion bisulfite et pyroséquençage. L’expression des gènes associés a été étudiée. Une anti corrélation significative est observée entre méthylation et expression signant la fonctionnalité de ces régions.En comparant les 11 méthylomes monocytaires, il est montré que 21% des CpG sont extrêmement stables et ne présentent qu’une faible variation de méthylation entre échantillons ( 20%). L’ensemble de ces informations peut être pris en considération pour la conception d’un outil d’épigénotypage. A l’avenir, il serait aussi possible d’utiliser cet outil en routine afin d’appréhender les variations du méthylome monocytaire dans différentes conditions d’élevage. / In dairy breeding, the health and fertility of cows are the main concern with the aims to maintain milk quantity and quality and to reduce the interval between calving in a high competitive economical context. Postpartum period is marked by major hormonal and metabolic changes that affect productivity, immune responses and fertility. The consequences of immune response deterioration are an increasing susceptibility to diseases (mastitis, metritis, endometritis…). Genomic selection in livestock improves the performance of the population but does not exclude phenotypic variability at the level of livestock and the individual. It is proposed that DNA methylation could contribute to this individual phenotype variability. Indeed, DNA methylation is an epigenetic process involved in transcriptional regulation of genes displaying certain plasticity in front of environmental constraints.We assumed the epigenetic signatures carried by blood cells, could reflect overall health and could be modified in response to intrinsic factors (parity, stages …) and to environmental changes. These signatures were researched in a particular blood cells subpopulation: the monocytes. These cells, obtained by blood sampling and purification with a specific antibody, are the first line of defense against acute infections participating in health status deterioration of postpartum cows.To test the monocyte epigenetic signatures hypothesis, monocyte methylome were analyzed by « Reduced Representation Bisulfite Sequencing » (RRBS), in various breeding conditions. Using genomic DNA form cows included in several protocols, 22 libraries were constructed and sequenced. Their analyses were accomplished using a « homemade » pipeline which integrates bioinformatics and biostatistics analyses. On average, 1 250 000 CpGs were analyzed in order to identify differentially methylated cytosines (DMCs): i) 27143 DMC by comparison between different cells types (monocytes versus fibroblasts and PBMC) ii) 4788 DMCs in response to nutritional challenge based on the dietary supplement, GENIAL®, produced and distributed in breeding by our partner PILARDIERE and XR-Repro (in collaboration with Marion Boutinaud, INRA, Rennes). iii) 2615 and 4616 DMCs in response to infectious challenge with LipoPolySaccharide injection for control cows group fed normal diet and for dietary restriction cows group, respectively (collaboration with Christine Leroux and José Pires, RUMINFLAME, INRA, Theix; and Gilles Foucras (ENVT, Toulouse)). iv) 4420 DMCs from the comparison between constant genomic cow (Somatic cell nuclear transfer, clones) and variable genomic cows but with the same age and raised in the same conditions than clones.From DMCs, we identified differentially methylated regions (DMRs) defined as region with at least 3 DMCs inside 100 bp. For some DMRs targeting gene promoter, the methylation status was validated by bisulfite conversion and pyrosequencing. Gene associated expression were also investigated. A significant negative correlation has been observed between methylation and expression, highlighting the functional relevance of these DMRs in gene transcription control.By comparing the 11 monocyte methylomes, 21% of CpGs present a remarkable constant methylation level with weak variability between samples (20%).Taking together, these data can provide a list of relevant DMCs for an epigenetic tool conception. In the future, it would be possible to use this tool for a routine analysis in order to grasp monocyte methylome variations in different breeding management.

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<p dir="ltr">Inflammation is a vital aspect of the immune response and functions to maintain homeostasis and protect the host against tissue injury or infection. However, excessive or prolonged inflammation can lead to health disorders and production losses in dairy cows. inflammation is a normal part of the tissue response during parturition. However, exacerbated systemic inflammation in the early postpartum has been associated with decreased milk production, decreased reproductive efficiency, and increased disease severity. Several processes contribute to systemic inflammation in dairy cows, such as liver oxidative stress, bacterial or endotoxin translocation during increased gastrointestinal permeability (e.g., lipopolysaccharide; LPS), and postpartum adipose tissue and skeletal muscle mobilization. However, there is limited knowledge of the mechanisms underlying systemic inflammation in postpartum dairy cows. Several studies have identified potent lipid mediators known as oxylipins as direct mediators of inflammation. Some differences have been identified in dairy cows when comparing the oxylipin profile throughout lactation and between cows suffering from different disorders. In our first study, we evaluated the plasma oxylipin profile of dairy cows suffering from systemic inflammation in the first week after parturition. Cows were classified into one of four systemic inflammation categories based on plasma haptoglobin (Hp) concentrations and plasma oxylipin profiles were identified by mass spectrometry. The results from this study identified several oxylipins of interest that warrant further research to confirm their presence in dairy cows with systemic inflammation and to evaluate their biological function associated with the postpartum inflammatory process. Endocannabinoids are soluble mediators associated with inflammation in humans, mouse models, and, more recently, dairy cows. While studies have demonstrated endocannabinoids to contribute to inflammation in the adipose tissue of dairy cows, no studies have evaluated the circulating presence of these mediators in cows suffering from systemic inflammation. Therefore, our second study sought to further evaluate the plasma oxylipin and endocannabinoid profiles of dairy cows suffering from systemic inflammation. In this study, we compared the oxylipin and endocannabinoid profiles of dairy cows categorized into high- and low-inflammation groups based on plasma Hp concentrations. We identified oxylipins associated with inflammation and found differences in the plasma endocannabinoid profile of high-inflammation cows compared to low-inflammation cows. Platelets are well understood for their function in hemostasis, though more recently, they have been studied for their ability to modulate the immune and inflammatory response. Presently, no studies exist elucidating the role of platelets on systemic inflammation in dairy cows. In addition to evaluating plasma oxylipin and endocannabinoid profiles in our second study, we determined if changes in platelets are associated with systemic inflammation in postpartum dairy cows. Proteomic analysis was completed on isolated platelets and plasma collected from dairy cows with high and low inflammation based on plasma Hp concentrations. Results from this study provide the first evidence demonstrating differences in the platelet and plasma proteomes between cows suffering from high inflammation and cows with low inflammation. The lipidomic and proteomic profiles described in these studies identified differences between cows suffering from systemic inflammation and apparently healthy cows. The results from these studies suggest platelets could contribute to an inflammatory state and have identified specific oxylipins, endocannabinoids, and proteins that may contribute to systemic inflammation in dairy cows by impairing the immune response or directly exerting an inflammatory function. More research is needed to fully understand the <i>in vitro </i>and <i>in vivo </i>function of the many lipid mediators and proteins identified in these studies and their ability to contribute to an exacerbated systemic inflammatory response in dairy cows.</p>

L’amélioration du bien-être des vaches laitières est un réel enjeu face aux attentes sociétales. Cette thèse a pour objectifs d’identifier (i) les aspects les plus dégradés du bien-être des vaches, (ii)les indicateurs de risque et (iii) les facteurs associés à l’atteinte de la relation homme-animal. Ce projet repose sur une étude épidémiologique utilisant la méthode Welfare Quality R pour évaluer le bien-être des vaches. En général, les boiteries étaient peu fréquentes et les animaux avaient une bonne liberté de mouvement. L’absence de faim et de soif et l’expression des comportements étaient bien respectés mais variables entre élevages. Le confort de couchage, l’absence de blessures et de maladies, le comportement social, l’état émotionnel et la relation homme-animal étaient dégradés et variables. L’écornage était souvent réalisé sans antidouleurs. Les principaux indicateurs de risque pour le bien-être des vaches sont la localisation géographique (pour la soif), la race (faim,comportement social), le logement (confort de couchage, blessures), l’interaction logement-race (état émotionnel) et localisation géographique-système de traite (expression des comportements propres à l’espèce). La mise en place de plan d’actions doit donc tenir compte du type d’élevage. Aucune association entre le système d’élevage et la relation homme animal n’a été trouvée. Les conditions de vêlage, les représentations des éleveurs et le comportement social des vaches étaient par contre associés à ce critère. De plus, de fortes variations entre vaches ont été relevées, suggérant que le tempérament des animaux joue un rôle important. / Improving dairy cows’ welfare is a major challenge in response to citizen concerns. The present thesis aimed to identify (i) major welfare problems, (ii) which farm characteristics are associated with the impairment of welfare and (iii) which factors are associated with the impairment of human-animal relationships. We conducted an epidemiological survey and used the Welfare Quality R protocol to assess dairy cows’ welfare. In general, prevalence of lameness was low and cows had opportunities to walk. The absence of hunger and thirst and the expression of species-specific behaviors were achieved but variable between farms. Resting comfort, skin injuries, health, social behavior, cows’ emotional state and human-animal relationships were affected and variable. Disbudding was often performed without pain relief. The main risk indicators of poor cow welfare were farm location (for thirst), breed (hunger, social behavior), housing (resting comfort and skin injuries), the interaction housing-breed (emotional state) and location-milking system (expression of species-specific behaviors). Action plans should be designed according to farm types. The quality of the human-animal relationship was not associated with farm main characteristics, but to calving conditions, farmers’ attitudes and cows’ social behavior. In addition, we observed large variations between-cows in their responses to humans, suggesting that individual behavioral traits of cows play a major role.

Doctor of Philosophy / Department of Animal Sciences and Industry / Barry J. Bradford / The transition period in dairy cows is characterized by dramatic increases in nutrient requirements for lactation and substantial metabolic stress. The disturbed metabolic balance, coupled with suppressed immune function, contributes to markedly elevated incidence of health disorders. Several lines of evidence suggest that increased inflammation is common during the transition period. Unlike the classical inflammation associated with acute infection, the postpartum inflammatory state is low-grade and often of metabolic origin. This metabolic inflammation plays a key role in numerous disorders; an improved understanding of inflammatory pathways in transition cows may improve our ability to predict and prevent disorders. To mimic metabolic inflammation, in Experiment 1, we administered low amounts of recombinant bovine tumor necrosis factor-α (rbTNFα), a pro-inflammatory cytokine, to early lactation cows, and evaluated whether rbTNFα affects milk production, metabolism, and health. We found that rbTNFα administration increased systemic inflammation, decreased feed intake and milk yield, and increased incidence of disorders. Conversely, preventing excessive inflammation has the potential to improve productivity and health of dairy cows. To identify nutritional strategies that could enhance metabolism and immunity, we evaluated the efficacy of several feed additives. In Experiment 2, we evaluated effects of chromium propionate, rumen-protected lysine and methionine, or both on metabolism and immunity in lactating dairy cows, and found that supplementation of these nutrients may enhance neutrophil function. In Experiment 3, we determined whether supplementation of yeast product to transition cows could enhance production, metabolism, and immunity, and found that yeast product modulated feeding behavior, metabolism, immunity, and uterine inflammation. Overall, a greater understanding of the role of metabolic inflammation in the transition period and the nutritional strategies that could modulate these signals may improve the production and health of dairy cows.