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Phase 4 (2009 Deadline)
Capacity Building, Epidemiology, and Risk Assessment of Endemic and Emerging Tick-Borne Disease in KPK and FATA, Pakistan
Mike Teglas, University of Nevada, Reno
Khalid Khan, Veterinary Research Institute Peshawar
Pakistani Funding (Department of State): $140,303
US Funding (Department of State): $172,186
Project Dates: November 15, 2010 - November 14, 2013 (Extended through September 30, 2014)
Project Overview
In Pakistan more than 75 percent of the population earns their livelihood from raising livestock. Tick-borne diseases often result in decreased meat, milk, and fiber production, spontaneous abortion, and death in livestock populations. Infection by tick-borne pathogens not only results in economic hardship for people relying on livestock for their survival but also can be transmitted to the farmers themselves. The Khyber Pakhtunkhwa (KPK) province of Pakistan and the adjacent Federally Administered Tribal Areas (FATA) share a poorly controlled border with Afghanistan, which increases the risk of transboundary transmission of diseases and parasites, yet there is little data regarding the distribution of ticks and tick-borne diseases in those regions. The long-term goal of this study is to establish a region-wide surveillance program for ticks and tick-borne diseases and to gain better understanding of key ecological factors required by ticks. Data collection efforts and analysis will serve as a source of hands-on training for Pakistani scientists and students and will serve as the platform for capacity building at the collaborating institutions. By the end of this project, the Pakistani partner institute will be able to offer molecular-based diagnostic services for tick-borne diseases and train and support local veterinary and human healthcare providers. An online database of ticks and tick-borne diseases in Pakistan will provide for better information dissemination and communication, and the undergraduate and graduate students participating in the project should also see their academic and career paths in the molecular biological sciences strengthened. The US partner university also plans to use the relationships developed through this cooperative program to recruit graduate students into a proposed International Vector Biology Training Program that would provide sustained training and support to vector-borne disease prevention efforts being developed in Pakistan and potentially other countries.
Quarterly Update
During the 3rd quarter of 2013, Sample collection from multiple areas of the province have been collected and processed for use in Drs. Khan and Ijaz’s upcoming training in US. The visa application process has been delayed considerably this year. A renewed DS 2019 Form was sent to Drs. Khan and Ijaz. They are currently waiting for approval of the visa application.
Tick borne disease ELIZA tests have been sent to Pakistan for us in project analyzing exposure in human agricultural workers from UNR. The two PIs will continue with a new project investigating the prevalence of zoonotic tick-borne disease (transmitted from animals to humans) in farmers and agriculturalists that live in close proximity to their livestock. The PIs expect this project to be a component of one or more graduate level student’s research project.
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2011 Show summary || Hide summary
The existing Parasitology Laboratory at VRI has been redesigned and renovated by the Khyber Pakhtunkhwa (KPK) Provincial Communication and Works Department via their Annual Development Program, thus leveraging funds available under this grant. A new BIO-Rad CFX 96 qPCR machine (paid for from this grant) has been purchased and installed, along with other items including a DNA slab gel system, a mini gel protein system, and a transblot transfer system, along with accessories and consumables. So far, the project has involved several other participants besides the PIs on both sides: a postdoc, three graduate students, and three undergraduates at the University of Nevada-Reno and two research associates and ten students at VRI. Both sides have also established linkages with other institutions to expand the scope of the project. For example, VRI is working with the KPK Agricultural University, Gomal College of Veterinary Sciences, and the University of Peshawar, as well as government agencies including the Livestock and Dairy Development Department, Central and Regional Veterinary Hospitals, and dairy farms managed by the military.
Collaborative work on the project took a big step forward when Dr. Khan made his first visit to Dr. Teglas’s lab for training on molecular genetic and spatial epidemiological techniques (September 30 - November 15, 2011). At the end of his visit, Dr. Khan received an electronic file containing multiple manuscripts detailing the molecular reagents and techniques needed to establish project-related assays at his laboratory in Pakistan. He also took with him some necessary laboratory equipment such as pipettes and consumables required for his planned research efforts. Meanwhile, the US partners have already published two papers based on work done under the project (one accessible here) and made one presentation. Two other joint papers were prepared during Dr. Khan’s visit and are being submitted for publication. One, entitled “The emergence and maintenance of vector-borne diseases in the Khyber Pakhtunkhwa Province and the Federally Administered Tribal Areas (FATA), Pakistan,” will serve as a review of the current status of vector-borne diseases in the provinces surrounding Peshawar. The second manuscript, entitled “Prevalence of tick-borne haemoprotozoan infections in ruminant livestock in the Peshawar district and periphery, Khyber Pakhtunkhwa (Pakistan),” describes the prevalence of veterinary pathogens in livestock in the region with a primary emphasis on those diseases that may be zoonotic for the Pakistani people that depend on them.
2012 Show summary || Hide summary
Dr. Khan completed his first visit to the University of Nevada from September 30 through November 15, 2011. During his stay, he worked with Dr. Teglas and co-PI Dr. Nathan Nieto to learn various field sampling techniques, including field BSL-3 training in cooperation with the California Department of Public Health. He also received preliminary epidemiological statistical training and hands-on qPCR and traditional PCR training, including genomic extraction, assay development, and primer/probe creation, which will be very valuable in building molecular genetic diagnostic capacity at VRI. At the end of his visit, Dr. Khan received an electronic file containing multiple manuscripts detailing the molecular reagents and techniques needed to establish project-related assays at his laboratory in Pakistan. He also took with him some necessary laboratory supplies, such as pipettes and consumables required for his planned research efforts.
Back at VRI, the new CFX-96 qPCR (the same model being used at the U.S. partners’ lab) was purchased and installed, along with other equipment and supplies supported in the Year 1 budget for this project. These items included a DNA slab gel system, a mini gel protein system, and a transblot transfer system, along with accessories and consumables. Dr. Khan reports that the necessary infrastructure is now in place in his lab to begin providing state-of-the-art molecular diagnostic capabilities for tick-borne diseases of livestock. Two research associates have been hired (one male and one female), and field collection efforts are under way throughout Khyber Pakhtunkhwa to obtain ticks and livestock blood samples. Diagnostic assays for Anaplasma, Borrelia, Babesia, Mycoplasma, and Theilleria have been or are being developed. Plans for further exchanges of training visits in 2012 are currently being discussed.
By the end of July 2012, more than 5,000 tick and livestock blood samples had been collected from various parts of Khyber Pakhtunkhwa (KPK) and the Federally Administered Tribal Regions (FATA). However, the unreliability of the power supply at VRI has hampered preservation and processing of the samples. Although back-up generators ensure the refrigeration and long-term storage of samples, the inconsistency of the power supply complicates some types of testing such as PCR. Plans are being made for some samples to be brought by VRI researchers to UNR for testing, and Dr. Teglas is also shipping Dr. Khan some test kits that do not require the use of electricity or hard-to-obtain reagents.
Dr. Teglas and Dr. Nieto are still considering the possibility of visiting VRI to provide training to VRI personnel. In the meantime, they, Dr. Khan, and his colleague Dr. Ghufranullah of VRI participated in the 15th International Conference on Infectious Diseases, which was held in Bangkok June 13-16, 2012. They presented their paper on maintenance of endemic tick-borne relapsing fever (Borrelia hermsii) from the western United States and discussed plans for future sample collection and analysis. Another joint paper on vector-borne diseases in KPK and the FATA was published in July 2012 in the journal Frontiers in Systems Biology (abstract available online at http://www.frontiersin.org/Systems_Biology/10.3389/fphys.2012.00250/abstract). A follow-up training visit by Dr. Khan and another VRI researcher to UNR is being planned for the coming months.
2013 Show summary || Hide summary
Co-U.S. PI N. Nieto moved from UNR to Northern Arizona University (NAU), Flagstaff to begin a tenure track faculty position. The collaborators have made plans to continue their research work and incorporate the facilities at NAU into the training and research program as this institution has facilities and expertise that will nicely augment training already received at UNR. VRI Researchers, Drs Khan and Ijaz have submitted visa application and have had interviews with Homeland Security staff. They are currently waiting for the final approval in order to travel to UNR to continue training and strengthen research collaborations. Researchers are also expected to travel to NAU to obtain additional training in molecular genetics (and BSL-3 training) at their research facilities.
Sample collection from multiple areas of the province have been collected and processed for use in upcoming training in US. Importation permits to allow entry of extracted DNA from tick and livestock samples from Pakistan was obtained by US collaborators.
During spring 2013, 100 student interns have been trained by Pakistani PI at VRI in advanced pathogen detection systems, parasitology and tick borne diseases. 40 students in Veterinary Sciences from Sindh Agriculture University Tandojam and 60 interns from the Agriculture University Peshawar received orientation on molecular techniques in Dr. Khan’s lab.
Co-U.S. PI N. Nieto moved from UNR to Northern Arizona University (NAU), Flagstaff to begin a tenure track faculty position. The collaborators have made plans to continue their research work and incorporate the facilities at NAU into the training and research program as this institution has facilities and expertise that will nicely augment training already received at UNR. VRI Researchers, Drs Khan and Ijaz have submitted visa application and have had interviews with Homeland Security staff in the spring. They are currently waiting for the final approval in order to travel to UNR to continue training and strengthen research collaborations. Due to the delay of visa, multiple DS 2019 forms have had to be filed and submitted as the expected travel dates kept getting pushed back. Researchers are expected to travel to NAU to obtain additional training in molecular genetics (and BSL-3 training) at their research facilities.
Sample collection from multiple areas of the province have been collected and processed for use in upcoming training in US. Importation permits to allow entry of extracted DNA from tick and livestock samples from Pakistan was obtained by US collaborators
In the summer, 75 students (35 from Sindh Agricultural University Tandojam and 40 interns from the Agricultural University Peshawar) have been trained by Pakistani PI at VRI in advanced pathogen detection systems, parasitology and tick borne diseases.
The PIs are working together on developing a new project to investigate the prevalence of zoonotic tick-borne disease (transmitted from animals to humans) in farmers and agriculturalists that live in close proximity to their livestock. They expect this project to be a component of one or more graduate level students’ research project.
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