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The National Academies
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Phase 2 (2006 Deadline)
Building Molecular Biology Capacity for Preventing Tick-Transmitted Diseases in Pakistan
Thomas N. Mather, University of Rhode Island, Kingston
Abdullah G. Arijo, Sindh Agricultural University, Tando Jam
Pakistani Funding (HEC): $225,451
US Funding: $290,000
Project Dates on US Side: March 1, 2007 - October 31, 2011
In Pakistan more than 75 percent of the rural population practices livestock husbandry, and a majority of them depend on livestock for their subsistence. Crimean-Congo Hemorrhagic Fever virus and other significant tick-transmitted pathogens of humans and animals are endemic to certain regions of Pakistan and have the potential to cause significant human morbidity and mortality and impact Pakistan’s agricultural economy and the livelihood of its rural citizens. This project's goal was to establish a molecular entomology laboratory at Sindh Agricultural University (SAU) to build Pakistani capabilities to study and prevent tick-transmitted diseases in Pakistan. The project was also focused on developing high-throughput transcriptomic, functional genomic, and proteomic systems and strategies aimed at identifying tick salivary proteins that can produce strong delayed-type hypersensitivity responses, antibody responses, or a combination of both, that correlate with protection from tick-borne disease (TBD). This novel approach could accelerate anti-tick and TBD vaccine development by informing the vaccine candidate selection process. Moreover, functional genomic screens involving inhibitory RNA are expected to identify novel pharmaco-therapeutic targets for disrupting tick feeding and pathogen transmission.
Two views of the molecular entomology laboratory space at SAU—before the renovations (left) and after (right).
Dr. Mather and his colleagues at URI continued their studies on vaccines and small molecular targets to disrupt feeding of black-legged ticks (Ixodes scapularis) and transmission of the agents causing Lyme disease, anaplasmosis, and babesiosis. Dr. Arijo and his fellow researchers at SAU worked to (1) develop TBD surveillance and assessment capabilities to identify and prioritize vector tick species in Sindh Province; (2) establish a capacity at SAU to conduct TBD diagnostics using rapid polymerase chain reaction (PCR) and reverse transcription PCR assays without requiring specialized biocontainment facilities; and (3) focus vaccine and pharmaceutical discovery research on important Pakistani tick species identified by the activities described in aims 1 and 2.
In addition to these efforts on both sides, the project originally included plans for a series of hands-on workshops at SAU to train Pakistani faculty, researchers, and students in the cutting-edge techniques needed to build and advance an appropriate molecular biology capacity that can be applied to various programs for preventing tick-transmitted diseases in Pakistan. Unfortunately, due to the security situation in Pakistan and particularly in rural south Sindh, plans for these large-scale workshops were shelved, although Dr. Mather and his US colleagues have been able to deliver some of the training by videoconferencing and some by individual visits.
- Renovated SAU Tick Laboratory and installed new equipment
- Provided training to Pakistani and U.S. scientists involved in tick-bone disease prevention research on both sides
- Created normalized cDNA library from Pakistani ticks
- Piloted a joint undergraduate student exchange project between SAU and URI that provided students with opportunities to collaborate on zoonotic disease perception survey development and data analysis
- Provided training to more than 50 Pakistani students, researchers, and technicians in various seminars, workshops, and conferences.
Progress Report Summaries
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2010 Show summary || Hide summary || Download full report
2009 Show summary || Hide summary || Download full report
In 2009 considerable progress was made on establishing a pathogen-free tick (Hyalomma anatolicum) colony at SAU; this step was critical for developing well-characterized and bio-secure salivary gland material for transcriptomic sequencing and analysis. Through tele-video conferencing, the partners in this project continued to identify specific needs for customized molecular reagents and protocols with built-in biological safeguards. URI participants developed PCR primers specific for the most likely pathogens to be encountered in Pakistani Hyalomma ticks, as well as standardized protocols for bio-safe sample preparation, DNA extraction, and molecular pathogen detection. Additionally, URI developed appropriate animal research protocols for tick rearing at SAU and provided SAU with appropriate supplies to carry out tick colony development. These efforts were greatly facilitated by two exchange visits during the reporting period: SAU scientist Dr. Bachal Bhutto spent 7 weeks at URI in November-December 2009 focusing on tick surveillance, tick rearing, tick colony development, molecular pathogen detection, and genes to vaccines techniques. In November 2009 URI scientist Dr. Aftab Ahmed conducted proteomics training and hands-on workshops at SAU and UOK. Important inter-cultural and scientific interchange was ongoing through the period via live tele-video conference lab meetings, e-seminars, e-mail, and taped lectures. The group even piloted a joint undergraduate student exchange project between SAU and URI, as students at both institutions collaborated on zoonotic disease perception survey development and data analysis. Progress made during this reporting period will be critical for meeting the project milestone of obtaining transcriptomic information on Pakistani tick salivary glands from a characterized, pathogen-free source colony. Once available, these data will be used in a URI genes-to-vaccines strategy to identify anti-tick vaccine candidates for evaluation in Pakistan at SAU and UOK as well as at URI. A no-cost extension has been authorized to extend the project through April 2011.
2008 Show summary || Hide summary || Download full report
With outside support, third-year medical student Umar Rashid visited URI for 6 weeks in June-August 2008 for training on technical procedures, tick sampling and rearing, lab safety, and research ethics.
In additional to the laboratory research and field work under way in both institutions, in March 2008 Dr. Arijo held a mini-symposium to provide background and justification for developing this new Pakistani research capacity targeting molecular detection of veterinary parasites and anti-tick vaccination for detecting and preventing tick-borne diseases. Later in the year and he and colleagues conducted a pilot survey of stakeholders (farmers, veterinarians, etc.) to determine their perceptions of the tick problem and raise awareness of the importance of this issue for both animal and human health.
2007 Show summary || Hide summary || Download full report
In 2007 the Pakistani partners at SAU began rehabilitating a molecular entomology lab and animal space following suggested guidelines from URI scientists, who also provided input concerning proposed purchases of equipment and molecular kits and reagents. Renovations were completed by June 2008, and in the last quarter of that year the university received several critical pieces of equipment, including PCR thermal cyclers, a gel documentation system, a nanodrop spectrophotometer, and an incubator. SAU has hired and begun training a senior scientist, lab technician, computer operator, and four postgraduate students on the project, and efforts to collect ticks from livestock in various regions of Pakistan are ongoing. In addition, Dr. Arijo visited the URI molecular entomology lab in October 2007. He spent approximately one month receiving intensive training on experimental procedures and working with URI scientists to further develop training curricula and workshop plans.
Dr. Arijo working in the lab at URI during his fall 2007 visit.
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