The scope and application of high throughput techniques has expanded from studying a single genome, transcriptome or proteome to understanding complex environments at a greater resolution with the help of novel computational frameworks. Comprehensive structural annotation i.e. description of all functional elements in the genome, is required for measuring genome response accurately, using high throughput methods. Annotation of genome sequences using high throughput data from RNA-seq and proteomics experiments complement computational methods for identifying functional elements and can help validate existing in silico annotation, correct annotation errors, and could potentially identify novel functional elements. Re-annotation studies in recent times have revealed shortcomings of automated methods and the necessity to validate existing annotations using experimental data. This dissertation elucidates re-annotation of Mannheimia haemolytica, Pasteurella multocida and Histophilus somni, bacterial pathogens associated with bovine respiratory disease in cattle. Experimental re-annotation of these bacterial genomes using RNA-seq and proteomics enabled the validation of existing annotation and discovery of novel functional elements that can be utilized in future functional genomics studies. We also addressed the need for developing an automated bioinformatics workflow that is broadly applicable for bacterial genome re-annotation, by developing open source Perl pipeline that can use RNA-seq and proteomics data as input. Simultaneous analysis of host and pathogen gene expression profiling using metatranscriptomics approaches is necessary to improve our understanding of infectious diseases. Traditional methods for analysis of RNA-seq data do not address the impact of cross-mapping of reads to multiple genomes for data originating from a metatranscriptomic study. Analysis of sequence conservation between species can help determine a metric for cross mapping to correct for signal vs. noise. We generated artificial RNA-seq data and evaluated the impact of read length and sequence conservation on cross-mapping. Comparative genomics was used to identify a core and pan-genome for quantifying gene expression. Our results show that cross mapping between genomes can directly be related to evolutionary distance between these genomes and that an increase in RNA-seq read length tends to negate cross mapping.

Avian cholera, caused by infection with Pasteurella multocida, is an important infectious disease of wild birds in North America Since it was first confirmed in 2005, annual outbreaks of avian cholera have had a dramatic effect on common eiders on East Bay Island, Nunavut, one of the largest breeding colonies of northern common eiders (Somateria mollissima borealis) in the eastern Arctic. I investigated potential avian and environmental reservoirs of P. multocida on East Bay Island and other locations in the eastern Canadian Arctic by collecting cloacal and oral swabs from live or harvested, apparently healthy, common eiders, lesser snow geese, Ross’s geese, king eiders, herring gulls, and snow buntings. Water and sediment from ponds on East Bay Island were sampled before and during outbreaks. Avian and environmental samples were tested using a real-time polymerase chain reaction (PCR) assay to detect P. multocida. PCR positive birds were found in every species except for snow buntings, and PCR positive common eiders were found in most locations, supporting the hypothesis that apparently healthy wild birds can act as a reservoir for avian cholera. In all years, P. multocida DNA was detected in ponds both before and after the avian cholera outbreak began each year, suggesting that the environment also plays a role in outbreak dynamics. Contrary to our expectations, model results revealed that ponds were generally more likely to be positive earlier in the season, before the outbreaks began. Whereas average air temperature at the beginning of the breeding season was not an important predictor for detecting P. multocida in ponds, eiders were more likely to be PCR positive under cooler conditions, pointing to an important link between disease and weather. Potential origins of P. multocida causing avian cholera in Arctic eider colonies were investigated by comparing eastern Arctic isolates of P. multocida to isolates from wild birds across Canada, and the central flyway in the United States. Using repetitive extragenic palindromic-PCR (REP-PCR) and multi-locus sequence typing (MLST), we detected a low degree of genetic diversity among isolates, and P. multocida genotypes were correlated with somatic serotype. Isolates from East Bay Island were distinct from P. multocida from eider colonies in the St. Lawrence Estuary, Quebec, however, East Bay Island isolates were indistinguishable from isolates collected from a 2007 pelagic avian cholera outbreak on the east coast of Canada. Isolates from East Bay Island and Nunavik shared sequence types, indicating possible transmission of isolates among eider colonies in the eastern Arctic. Previously, feather corticosterone in eiders was found to be significantly associated with environmental temperature during the moulting period. In my study, path analysis revealed that environmental conditions experienced during the moulting period had direct impacts on arrival date and pre-breeding body condition of common eiders during the subsequent breeding period on East Bay Island, with indirect impacts on both reproductive success and survival. Higher temperatures experienced during the fall moulting period appear to impose significant costs to eiders, with subsequent carry-over effects on both survival and reproduction many months later during avian cholera outbreaks. This thesis describes several important features of the host, agent and environmental dynamics of avian cholera in North America with a particular focus on the disease in the eastern Canadian Arctic. Continued exploration of infectious wildlife disease dynamics is needed to better predict, detect, manage, and mitigate disease emergence that can threaten human and animal health and species conservation.

L’asthme équin, une maladie inflammatoire considérée non infectieuse, est néanmoins associé avec la présence de certains streptocoques et Pasteurellaceae. La similarité entre les espèces de Pasteurellaceae complexifie toutefois leur différenciation. L’objectif de cette étude est d’évaluer la présence de différents Pasteurellaceae dans les voies respiratoires de chevaux atteints d’asthme. Des amorces qPCR ont été optimisées pour [Pasteurella] caballi, Nicoletella semolina, Pasteurella multocida et Haemophilus influenzae, mais plusieurs espèces d'intérêt (notamment Actinobacillus spp.) n'ont pas pu être étudiées faute d'amorces spécifiques. Des lavages nasaux, oraux et bronchoalvéolaires de douze chevaux (six sains, six asthmatiques) gardés dans différents environnements ont été analysés. Pour N. semolina, les analyses ont été approfondies chez dix chevaux asthmatiques (phase II : lavages nasaux et bronchoalvéolaires), et chez dix chevaux sains (phase III : lavages nasaux). N. semolina a été identifié dans 0 à 66 % des échantillons et était retrouvé en plus forte quantité dans le nez de chevaux gardés à l’intérieur et nourris avec du foin. Dans les phases II et III, N. semolina a été identifié dans 90 à 100 % des échantillons nasaux, et dans six lavages bronchoalvéolaires. [P.] caballi a été identifié dans la cavité orale de 83 % des chevaux, indépendamment du statut de santé. H. influenzae et P. multocida n’ont pas été détectés. Aucun Pasteurellaceae analysé n’était associé à l’asthme équin sévère. N. semolina était fréquemment retrouvé dans les voies respiratoires et sa charge nasale était influencée par l’environnement. Les amorces synthétisées faciliteront de futures études sur les Pasteurellaceae. / Equine asthma, an inflammatory disease considered non-infectious, is associated with the presence of certain streptococci and Pasteurellaceae. Strong similarities between Pasteurellaceae species prevent specific differentiation. The objective of the study was to evaluate the presence of different Pasteurellaceae in the airways of horses with severe asthma. Quantitative PCR primers were optimized for [Pasteurella] caballi, Nicoletella semolina, Pasteurella multocida and Haemophilus influenzae. Several species of interest (in particular Actinobacillus spp.) could not be studied due to a lack in primer specificity. Nasal, oral and bronchoalveolar lavages from twelve horses (six healthy, six asthmatics) kept in different environments were analyzed. For N. semolina, further investigation was pursued through inclusion of ten asthmatic horses (phase II: nasal washes and bronchoalveolar lavages), and ten healthy horses (phase III: nasal washes). N. semolina was identified in 0 to 66 % of samples and was found in larger loads in the nose of horses kept inside and fed hay. In phases II and III, N. semolina was identified in 90 to 100 % of nasal samples and in six bronchoalveolar lavages. [P.] caballi was only identified in the oral cavity, with 83 % of positive horses regardless of health status. H. influenzae and P. multocida were not detected. No specific Pasteurellaceae studied here were associated with severe equine asthma. N. semolina was identified frequently in respiratory samples and its nasal load was influenced by the horse’s environment. The optimized primers will facilitate future studies on Pasteurellaceae.