Location: Zoonotic and Emerging Disease Research
Project Number: 3022-32000-021-030-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Aug 1, 2026
End Date: Jul 31, 2027
Objective:
The overall objective of this project is to define tick-associated viral threats in livestock-linked ecosystems of Pakistan and use these data to improve understanding of CCHFV spillover drivers, transboundary movement risk, and potential implications for U.S. agriculture. Pakistan sits within the broad geographic range of CCHFV and contains large livestock populations, seasonal animal movement, and tick species known or suspected to support CCHFV ecology. However, the viral diversity, tick-host associations, and ecological conditions that shape CCHFV risk in key livestock corridors remain incompletely characterized. This information is directly relevant to U.S. agriculture because introduction of infected ticks, infected animals, or related viruses into new regions could threaten livestock systems, rural communities, animal movement networks, and agricultural continuity. Specific objectives are to: Characterize tick species and host associations in high-risk livestock and peri-domestic interfaces in Khyber Pakhtunkhwa, Pakistan. Environmental and host-attached ticks, with emphasis on Hyalomma anatolicum, Hyalomma kumari, and Amblyomma gervaisi, will be collected from livestock, wildlife, and relevant interface settings to define tick distribution, host use, and ecological overlap with agricultural systems.Determine whether collected ticks contain CCHFV or other emerging arboviruses of public or veterinary concern. Tick-derived nucleic acid will be tested using a broad multiplex bioscreening approach and CCHFV-specific real-time RT-PCR to identify known agents and prioritize samples for deeper characterization.Generate genomic data from positive or high-priority detections to assess viral diversity, relatedness, and movement across regional livestock corridors. Sequencing will be used to identify viral lineages, detect novel or divergent strains, and compare findings with available regional and global data.Integrate tick, host, pathogen, geospatial, and animal-movement data into models that identify ecological and agricultural drivers of spillover risk. Outputs will help define conditions that support CCHFV maintenance and spread abroad and estimate how similar pathways could affect U.S. livestock systems if CCHFV-associated ticks or viruses were introduced.
Approach:
This project will use a field-to-model framework that combines tick ecology, molecular testing, genomic characterization, and risk modeling. Sampling will focus on Khyber Pakhtunkhwa, Pakistan, a region with documented CCHFV activity, important livestock production, and frequent movement of animals and people across local and transboundary corridors. Approximately 500 ticks will be collected from livestock, wildlife, peri-domestic animals, and environmental sites where agricultural contact is likely. Priority tick groups will include Hyalomma species associated with CCHFV ecology and Amblyomma gervaisi from wildlife interfaces, while retaining flexibility to characterize additional tick species encountered during sampling. Ticks will be identified morphologically and, when needed, confirmed by molecular barcoding. For each specimen, metadata will be captured on host species, collection location, tick life stage, collection source, animal interface, and relevant ecological or management factors. These data will allow pathogen detections to be interpreted in the context of host use, livestock contact, and local movement patterns rather than as isolated laboratory findings. Tick samples will be processed using an inactivated split-sample workflow that supports broad molecular screening while maintaining appropriate biorisk controls for agents such as CCHFV. Extracted nucleic acid will be screened with a multiplex platform to detect selected high-consequence bacterial and viral agents and with CCHFV-specific real-time RT-PCR. Samples with positive, indeterminate, or otherwise high-priority results will be advanced for confirmatory testing and sequencing. Illumina and/or Oxford Nanopore sequencing will be used to generate viral sequence data when sufficient material is available. Resulting sequences will be assembled, quality checked, taxonomically classified, and compared with regional and global datasets to evaluate viral diversity, phylogeographic relationships, and potential evidence of transboundary movement. Analytical outputs will combine laboratory results with field metadata, tick species distribution, host associations, livestock density, animal movement patterns, land use, and geospatial features. Statistical and spatial models will identify factors associated with CCHFV or arboviral detection and will help distinguish risk linked to tick ecology, host availability, agricultural practices, and animal movement. These models will then be used to evaluate how CCHFV-associated ecological conditions abroad compare with U.S. livestock and wildlife interfaces. The final product will be a risk framework that supports prioritization of tick and livestock sampling, diagnostic preparedness, and agricultural biosecurity planning for high-consequence tick-borne viral threats and CCHF viral genomic data, closing critical gaps in our ability to assess diagnostic performance.