Location: Animal Disease Research
Project Number: 2090-32000-031-00-D
Project Type: In-House Appropriated
Start Date: Oct 18, 2011
End Date: Oct 17, 2016
Objective 1: Identification of host genetic markers associated with reduced transmission and replication of ovine progressive pneumonia virus. Subobjective 1.1A: Identification of genetic markers for reduced odds of OPPV infection in domestic sheep. Subobjective 1.1B: Validation of genetic markers for reduced odds of OPPV infection in domestic sheep. Subobjective 1.2A: Identification of genetic markers for reduced OPPV proviral replication in domestic sheep. Subobjective 1.2B: Validation of genetic markers for reduced OPPV proviral replication in domestic sheep. Objective 2: Identification of host genetic factors in domestic and bighorn sheep associated with reduced transmission and disease due to Mannheimia haemolytica. Subobjective 2.1A: Identification of one or more genetic markers in a major gene region for reduced nasal shedding of Mh in domestic sheep. Subobjective 2.1B: Produce a high-throughput diagnostic test for quantifying nasal shedding of Mh in domestic sheep that has high concordance with nasal culture. Subobjective 2.1C: Validation of genetic markers from a major gene locus for reduced nasal shedding of Mh in domestic sheep. Subobjective 2.1D: Genomewide identification of genetic markers for reduced nasal shedding of Mh in domestic sheep. Subobjective 2.2A: Develop comparative transcriptome sequences for domestic versus bighorn sheep neutrophils (PMNs) both with and without exposure to Mh leukotoxin. Subobjective 2.2B: Develop a draft genome assembly for bighorn sheep. Objective 3: Develop intervention strategies to mitigate transmission of Mannheimia haemolytica from domestic sheep to bighorn sheep which will reduce respiratory disease in domestic and bighorn sheep. Subobjective 3.1: Quantify nasal, pharyngeal, pulmonary and serological antibody levels to leukotoxin-bearing Mh serotype A2 [MhA2(+LktA)] in adult domestic sheep with natural nasopharyngeal colonization. Subobjective 3.2: Intranasally immunize domestic and bighorn lambs with genetically modified MhA2 expressing an inactive form of Lkt and compare antibody responses to those measured in subobjective 3.1. Subobjective 3.3: Challenge MhA2(Lkt-mod) immunized domestic and bighorn lambs with wildtype MhA2.
Ovine respiratory disease presents many challenges to U.S. agriculture. Respiratory pathogens of domestic sheep like ovine progressive pneumonia virus (OPPV) and Mannheimia haemolytica (Mh) lead to millions of dollars in production losses annually. In addition to production losses in domestic sheep, Mh is an important bacterial pathogen of bighorn sheep. Data indicate that domestic sheep transmit pathogenic subtypes of Mh to bighorn sheep, and a currently debated intervention is separation of domestic from bighorn sheep areas. However, removal of domestic sheep grazing rights threatens a large fraction of the U.S. sheep industry, and additional intervention strategies are urgently needed. Existing data suggest that transmission of both OPPV and Mh has a genetic component in domestic sheep. The development of genetic markers will allow marker-assisted selection for reduced transmission and disease in domestic sheep, as well as reduced transmission to bighorn sheep. Furthermore, there are essentially no genomic and transcriptomic resources for bighorn sheep. However, the development of such resources will allow the formulation of specific biological hypotheses related to differences between bighorn and closely related domestic sheep. Protective immune responses to Mh in domestic and bighorn sheep have not been well-characterized. Documenting the characteristics of protective antibody mediated immunity in domestic sheep is an important first step toward development of a vaccine capable of providing preimmunity in bighorn sheep populations and improving the control of Mh mediated respiratory disease in domestic lambs. A goal of this research project plan is development of an intranasal vaccine utilizing a modified-live Mh expressing an inactive form of leukotoxin.