Genetic and genomic investigations of hereditary, viral, and parasitic wildlife diseases in non-model organisms

本提案的目的是阐明野生犬科动物模型中与野生动物疾病相关的遗传/基因组因素。传染性和遗传性野生动物疾病造成经济和文化损失，并对人类健康构成潜在威胁。赤狐是一种非模式生物，非常适合对野生动物疾病的原因和后果进行分子遗传学和基因组研究。狐狸是“基因组启用”的可用家犬基因组组装，提供访问基因组资源，如微阵列和候选基因序列。纽芬兰红狐至少有三种重要的野生动物疾病，包括遗传性，病毒性和寄生性。纽芬兰和欧洲部分地区的养殖银经历了遗传性增生性牙龈炎（HHG），这是一种阻碍繁殖计划的牙龈疾病。使用犬微阵列，我们（HQP和我）已经确定了一组功能候选基因的发育遗传细胞信号转导途径，我们打算通过候选基因测序和定量PCR的特点，发现这种情况下的基因和致病突变的目标。将使用连锁分析鉴定和测试其他候选基因。全球体内寄生线虫法国心丝虫在北美的唯一地方性集中地是纽芬兰。这种传染病由于从狐狸传播到家犬，并越来越多地传播到土狼，因此令人担忧，预示着它可能会传播到加拿大其他地区。功能基因组方法将用于鉴定犬恶丝虫成功传播和宿主感染的相关性。狂犬病病毒的赤狐株在加拿大北方持续存在，并有可能入侵更广泛的地理区域，例如20世纪50年代中期在加拿大东部的传播，对公共卫生产生了明显的担忧。我建议回避狐狸在加拿大东北部的地理结构，使用全基因组单核苷酸多态性（SNP），以确定地理或其他障碍病毒传播和抗病性的遗传相关性。这项研究建立了纪念大学，毛皮饲养者协会和动物健康部门之间的合作，对加拿大和欧洲的毛皮农民，加拿大各地的野生动物管理官员和狗主人，人类疾病研究人员以及哺乳动物生长和发育的基础研究人员都很重要。例如，牙龈过度生长会导致牙脓肿，影响狐狸的进食能力，影响它们的生活质量和经济价值。因此，毛皮饲养者协会无法繁殖狐狸，而牙龈状况的遗传原因和遗传模式以及相关的毛皮质量尚不清楚。我们研究的一个预期成果是对这种疾病的诊断测试。HHG的基因和致病突变的发现也将为类似人类疾病的遗传学提供一个切入点，特别是以高脂血症为特征的形式;通过使用毛皮食肉动物疾病来模拟人类疾病，人类和动物的健康问题都得到了解决。与细胞周期相关的致病基因突变的鉴定也将为癌症研究提供信息。了解增生的性质有助于对生长和发育的基本知识。犬肺管圆线虫的工作将有助于建立一个寄生虫在野生犬科动物中传播和维持的时空模型，并有助于了解和预防这种严重感染在犬中的进一步传播。狂犬病也是一种严重的野生动物问题，对人类也有潜在的危害，了解其传播对于监测和预防这种病毒的传播至关重要。

The objectives of this proposal are to elucidate genetic/genomic factors associated with wildlife diseases in wild canid models. Infectious and inherited wildlife diseases cause economic and cultural losses and pose potential health threats to humans. The red fox is a non-model organism ideally suited for molecular genetic and genomic investigations into causes and consequences of wildlife diseases. Foxes are "genomically enabled" by the available domestic dog genome assembly, providing access to genomic resources such as microarrays and candidate gene sequences. Newfoundland red foxes harbour at least three significant wildlife diseases, including hereditary, viral, and parasitic. Farmed silver foxes in Newfoundland and parts of Europe experience hereditary hyperplastic gingivitis (HHG), a gum disease that hinders breeding programs. Using canine microarrays, we (HQP and I) have identified a set of functional candidate genes in developmental genetic cell signalling pathways which we intend to characterize via candidate gene sequencing and quantitative PCR, with the objective of discovering the gene and causative mutation of this condition. Additional candidate genes will be identified and tested using linkage analyses. The only endemic focus in North America of the global endoparasitic nematode French heartworm is in Newfoundland. This infectious disease is of concern due to transmission from foxes to domestic dogs, and increasingly to coyotes, foreshadowing its possible spread to other parts of Canada. Functional genomic approaches will be used to identify correlates of successful heartworm transmission and host infection. The red fox strain of rabies virus persists in northern Canada and threatens to invade a wider geographic area, as exemplified by a mid-1950s spread throughout eastern Canada, with obvious concerns for public health. I propose to eludicate fox phylogeographic structure in northeastern Canada, using genome-wide single nucleotide polymorphisms (SNPs) to identify geographic or other barriers to viral transmission and genetic correlates of disease resistance. This research forges collaborations among Memorial University, the Fur Breeder’s Association, and Animal Health Division, and will be important to fur farmers in Canada and Europe, wildlife management officials and dog owners throughout Canada, researchers of human disease, and basic researchers of mammalian growth and development. For example, the gum overgrowth causes dental abscess and compromises feeding ability of foxes, affecting their quality of life and economic value. The Fur Breeder's Association is therefore unable to propagate foxes while the genetic cause and inheritance pattern of the gum condition, and related fur quality, are unknown. An anticipated outcome of our research is a diagnostic test of this disease. The discovery of the gene and causative mutation of HHG will also provide an entry point into the genetics of a similar human disease, especially the form characterized by hypertrichosis; by using a furbearing carnivore disease to model a human one, both human and animal health issues are addressed. The identification of pathogenic mutations of genes associated with the cell cycle will also be informative to cancer research. Understanding the nature of the hyperplasia contributes to basic knowledge about growth and development. The canine pulmonary angiostrongylus work will help develop a spatio-temporal model of parasite spread and maintenance in wild canids and contributes to understanding and prevention of further spread of this severe infection in dogs. Rabies is also a serious wildlife concern and potentially harmful to humans as well, and understanding its transmission is vital to monitoring and preventing spread of this virus.