Progress 06/01/17 to 05/31/21
Outputs
Target Audience: Training of two full-time PhD Candidates and two Veterinary Research Scholars and one FFAR Veterinary Fellow. Preliminary results were presented to professional peers at national and international scientific meetings as abstracts, presentations and posters, and as work-in-progress seminars on the campus of the University of Missouri. Some of these reports were presented by students at multiple venues. Changes/Problems:Numerous pitfalls resulted in slower than expected progress of this project, most of these matters were described in previous annual reports. During the 2020-2021 time period, the most profound delay occured when both PhD candidates developed CoVID-19, one of whom required hospitalization. In addition to the student health scare, the PI alone was left to purchase and haul cattle feed, collect blood, feed ticks and to feed and clean the experimental cattle (twice per day) as they were kept in a concrete-floor barn. Afterward, time (i.e., Shahzad's visa) or funding had run out to support stipends for both PhD candidates, forcing them to compose and defend their dissertations prior to identification of tick antigens and publication of their results. The PI is preparing dissertation chapters for submission to peer-review journals, while working to characterize tick molecules uniquely reactive to protective immune sera. What opportunities for training and professional development has the project provided?Training activities included completion of research projects for two dissertations. Dr. Kyle Hoffman has completed the requirements for graduation with a PhD from the MU Molecular Pathogenesis and Therapeutics Graduate Program. Dr. Sammuel Shahzad has defended his dissertation and is preparing the final draft for submission to the graduate school, in order to complete the requirements for a PhD from the MU Pathobiology Area Program. For professional development, both PhD candidates participated in the 101st Conference of Research Workers in Animal Diseases (CRWAD), in addition to Bacterial Pathobiology Work in Progress Seminars at MU. Although they will be postdoctoral researchers, it is the PI's hope that Drs. Hoffman and Shahzad will attend the 103rd CRWAD in 2022. Several veterinary medical students were also trained in immunology and molecular biology methods during the summer of 2021. Kyle Cacciatorie (University of Illinois) and Bridgette Rogers worked in the lab as part of the MU Veterinary Research Scholars (VRSP, directed by Dr. Craig Franklin). In addition to benchwork, Cacciatore and Rogers also particpated in a field trip to collect blood and ticks from registered beef cows. Stefan Keller, another MU veterinary medical student, successfully competed for a Veterinary Student Research Fellowship from the Foundation for Food and Agricultural Research. All three students also had opportunities to participate in collections of wild ticks in Missouri, but they chose to continue benchwork instead. Cacciatore returned to UIUC in August, while Rogers and Keller continue part-time work in the lab. For professional development, all three of these students participated in the MU VRSP curriculum and in the online (pandemic derived) format of the 2021 National Veterinary Scholars Symposium. It is the PI's hope that these students will also be able to attend the 103rd (2022) CRWAD, if the SARS CoV-2 pandemic remains at a manageable state. How have the results been disseminated to communities of interest?The results were disseminated at meetings with other researchers, such as the Conference of Research Workers in Animal Diseases (CRWAD) and the National Veterinary Scholars Symposium (NVSS). At CRWAD, Dr. Hoffman was awarded the 2020 Graduate Student Presentation Award from the Society for Veterinary Tropical Medicine. Results were also disseminated in dissertations composed by Drs. Shahzad and Hoffman. Several manuscripts are in different phases of the publication process, the most relevant of these works are dissertation chapters that are in preparation for submission to peer-review journals. The PI provided several presentations to various groups, including cattle producers, bovine practitioners, electrical workers and dog breeders. Some of the past lectures were included in this report because they were inadvertently omitted from previous annual reports. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
This project is focused on determining if host immunity to different tick tissue preparations can have different effects on transmission of a tick-borne disease agent between ticks and the vertebrate host. To the best of our knowledge, this work was the first to directly test the working hypotheses that (1) immunization of cattle with tick midgut would result in the highest reduction in acquisition of the tick-borne rickettsial pathogen, Anaplasma marginale, from immunized cattle; (2) immunization of cattle with tick salivary glands would have the highest reduction of tick transmission of A. marginale to susceptible immunized cattle; (3) denaturation of tick tissue preparations would increase efficacy with respect to interferance with transmission of A. marginale between ticks and cattle; and (4) differences in seroreactivity between protective and non-protective host sera with these tick-derived immunogens. All of these working hypotheses were tested, results from these experiments were presented at several conferences and described in more detail within two dissertations. Work is still underway in preparation to submit these results to peer-review journals and to identify tick antigens uniquely reactive with protective antisera. The following objectives were attained during this project. (a) Adaptation of anti-tick immunization procedures for experimental immunization of calves with tissue preparations from Dermacentor andersoni ticks. This work included dissection of ticks to isolate tick midgut and salivary gland organs, adapting previously reported methods for preparation of D. andersoni tissues preparatory to immunization of cattle, and experimental challenge infestations with adult stage D. andersoni ticks. To the best of our knowledge, this was the first adaptation of this anti-tick immunization system to a tick parasite of cattle indigenous to the USA. (b) Adaptation of the experimental bovine anaplasmosis model system to test efficacy of host immunization against D. andersoni tick molecules. These experiments expanded on the above-developed methods, to challenge immunized cattle with D. andersoni that had been experimentally infected with A. marginale and to feed previously unexposed D. andersoni on A. marginale-infected cattle that were immunized with different tick tissue preparations.To the best of our knowledge, this was the first adaptation of this anti-tick immunization system to the experimental bovine anaplasmosis model. (c) Adaptation of the experimental bovine anaplasmosis model system to test efficacy of host immunization against Dermacentor variabilis tick molecules. When our supplier decided to stop raearing D. andersoni, this unanticipated pitfall forced us to adapt this experimental system to D. variabilis, another tick vector that is indigenious to the USA. (d) Proteomics methodologies to identify molecules uniquely reactive to protective bovine immune sera. This work, which is still underway, involves using immune sera of bovine hosts in which ticks had reduced levels of performance with respect to feeding, acquisition of A. marginale from infected cattle or transmission of this pathogen to cattle. (e) Computer-based modeling of tertiary structures of serine protease inhibitors from tick saliva. This work was a collaboration with a coinvestigator of the current project, using a reverse approach for rational targeting of a native tick molecule to interfere with its biologic activity. (f) Ecology of A. marginale in Thailand and vector ticks in the USA. These were collaborative efforts to help establish preliminary data for future projects to better understand the natural history of ticks and adaptations of microbes to tick vectors. (g) Long read, high-quality genome sequence of a phenotypically novel Anaplasma marginale strain. Although this was not one the original objectives proposed for this project, we had a unique opportunity to generate, annotate and analyze the genome of an Illinois strain of A. marginale, which is novel in that this is the only strain described which is both i) reportedly not transmissible by Dermacentor ticks and ii) has prominent macromolecular structures (inclusion appendages) associated with the organisms inside of bovine erythrocytes.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2020
Citation:
K.S. Hoffman, S. Shahzad, A.A. AL-Hosary, S. Jittapalapong, G. Zhang, G. Johnson, P. Pithua, R. Stich. Protein antigens associated with seroreactivity of cattle producing reduced tick performance. 101st Conference of Research Workers in Animal Diseases, Chicago, IL, December 5-8, 2020. *Society for Tropical Veterinary Medicine (STVM)Student Presentation Award
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
Kyle J. Cacciatore, Sammuel Shahzad, Kyle Hoffman, Stefan H. Keller, Bridgette Rogers, Roger W. Stich. Seroreactivity of tick-immunized calves to five recombinant Dermacentor andersoni proteins. 2021 National Veterinary Scholars Symposium, August 4-6,2021
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
Bridgette M. Rogers, Sammuel Shahzad, Stefan H. Keller, Kyle J. Cacciatore and Roger W. Stich. Identification of tick salivary gland antigens recognized by protective immune sera using a cDNA library. 2021 National Veterinary Scholars Symposium, August 4-6,2021
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2021
Citation:
Stefan H. Keller, Sammuel Shahzad, Kyle J. Cacciatore, Bridgette M. Rogers, Roger W. Stich. Anti-tick vaccine discovery for control of Anaplasma marginale. 2021 National Veterinary Scholars Symposium, August 4-6,2021
- Type:
Theses/Dissertations
Status:
Published
Year Published:
2021
Citation:
Kyle Hoffman (Major Adviser: RW Stich), 2016-2021. University of Missouri Molecular Pathogenesis and Therapeutics Graduate Program. Area of study: Tick-borne Anaplasmataceae. Dissertation title: REFINEMENT OF AN ESTABLISHED LARGE ANIMAL MODEL TO UNDERSTAND THE TICK-PATHOGEN-HOST INTERFACE
- Type:
Theses/Dissertations
Status:
Under Review
Year Published:
2021
Citation:
Samuel Shahzad, DVM, MPP, Fulbright Fellow (Major Adviser: RW Stich), 2016-2021. Area of study: Tick-borne Anaplasmataceae. Dissertation title (Draft): INTERFERENCE WITH TRANSMISSION OF ANAPLASMA MARGINALE BY TARGETING TICK VECTORS WITH THE BOVINE IMMUNE RESPONSE
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Pongprayoon P, Niramitranon J, Kaewhom P, Kaewmongkol S , Suwan A, Stich RW, Jittapalapong S. 2020. Dynamic and structural insights into Ixodes ricinus serpin-2. Journal of Biomolecular Structure and Dynamics, 38:2296-2303. https://doi.org/10.1080/07391102.2019.1630003
- Type:
Journal Articles
Status:
Published
Year Published:
2020
Citation:
Saetiew N, Simking P, Saengow S, Herder S, Morand S, Sukprasert A, Stich RW, Jittapalapong S, Desquesnes M. 2020. Seasonal variation in Anaplasma marginale infections in beef cattle in previously flooded areas of Thailand. Agriculture and Natural Resources, 54(4):355-362. doi.org/10.34044/j.anres.2020.54.4.03
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Kamyingkird K, Jiyioong T, Amavisit P, Stich RW, Jittapalapong S. 2021. Molecular detection of Mycoplasma haemofelis, Candidatus Mycoplasma haemominutum and Candidatus Mycoplasma turicensis of stray cats residing in Bangkok monasteries, Thailand. Agriculture and Natural Resources, 55:423-430. https://doi.org/10.34044/j.anres.2021.55.3.12
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Spare M, Boorgula GD, Thomson D, Bai J, Goodin D, Anderson G, Stich RW, Hoobi A, Wilson S, Staggs A, Bowers A, Hamm A, Tatman J, Raghavan RK. 2021. Surveillance of host-seeking ticks in the Flint Hills region (USA) and associations with environmental determinants. Parasitologia 1: 137-147. https://doi.org/10.3390/parasitologia1030015
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Pongprayoon P, Kaewhom P, Kaewmongkol S , Eukote S, Stich RW, Wiriya B, Jittapalapong S. 2021. Structural dynamics of Rhipicephalus microplus Serpin-3. Molecular Simulation, 47: 1209-1216. https://doi.org/10.1080/08927022.2021.1962011
- Type:
Journal Articles
Status:
Published
Year Published:
2021
Citation:
Raghavan RK, Koestel Z, Boorgula GDY, Hroobi A, Ganta R, Harrington Jr J, Goodin D, Stich RW, Anderson G. 2021. Unexpected winter questing activity of ticks in the Central Midwestern United States. PLoS ONE 16(11): e0259769. https://doi.org/10.1371/journal.pone.0259769
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2020
Citation:
S. Shahzad, K.S. Hoffman, A.A. AL-Hosary, S. Jittapalapong, G. Zhang, G. Johnson, P. Pithua, R. Stich. Adaptation of a tick performance model system to cattle infested with Dermacentor variabilis. 101st Conference of Research Workers in Animal Diseases, Chicago, IL, December 5-8, 2020
|
Progress 06/01/19 to 05/31/20
Outputs
Target Audience:Training of one full-time PhD Candidate and one full-time PhD Student. Dr. Amira Al-Hosary, an Assistant Professor of Infectious Diseases at Assuit University, Egypt, was supported by the U.S.-Egypt Higher Education Initiative Graduate Scholarships for Professionals Program to participate in the project for four months as a Visiting Scholar. Preliminary results were presented to professional peers at national and international scientific meetings as abstracts, presentations and posters, and as work-in-progress seminars on the campus of the University of Missouri. Some of these reports were presented by students at multiple venues. Changes/Problems:We switched to a different tick species for the final three trials of this project. The university-wide CoVID-19 research ramp-down limited our ability to conduct bench work, including qPCR of ticks, host ELISA titers, identification oftick antigens andwork to confirm tick antigens identified with mass spectrometry. Therefore, we focused on completion of the immunization and challenge trials, while freezing and storing tick and serum samples for future analysis after the pandemic subsides. What opportunities for training and professional development has the project provided?The project provided training for both graduate students in humoral immunity of cattle, proteomics to identify tick antigens and bioinformatics. Dr. Al-Hosary, a Visiting Scholar from Assuit University, also received some training in bioinformatics and utilization of the bovine anaplasmosis model system. How have the results been disseminated to communities of interest?Results of research supported by this grant were represented in a total of four presentations by Hoffman (one),Shahzad (one) and Stich (two). Three abstracts were presented at the American Society for Rickettsiology Meeting (Santa Fe, NM) in June, 2019. One abstract was presented at the Conference of Research Workers in Animal Diseases in November, 2019. Hoffman and Shahzad also presented their work to the MU Molecular Pathogenesis and Therapeutics Graduate Program and the MU Bacterial Pathogenesis Works in Progress Seminars. Two manuscripts are in preparation for submission to peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?Objective 1. Finish immunization and challenge trials withD. variabilis. Objective 2. Finish identification oftick molecules uniquely reactive with sera from protected hosts. Objective 3. Test to confirm or refute tick antigens identified in Objective 2. Objective 4. Submit first two manuscripts for publication in peer-review journals. Objective 5. Prepare two more manuscripts for submission to peer-review journals.
Impacts
What was accomplished under these goals?
Two trials have been completed for Aim #1 with D. andersoni and 3 trials had been completed under Aim #2 with D. andersoni. However, because D. andersoni were no longer available, the final three trials had begun with D. variabilis. We also began work to use 2-Dimensional Western analysis to identify tick antigens uniquely reactive to protective bovine antisera. The CoVID-19 pandemic led to a university-wide research ramp-down in the middle of March, 2020. This research ramp-down slowed progress considerably and forced us to postpone the comprehensive exam of one of the PhD students. Every experimental procedure on campus and person in the lab had to be justified, approved and cleared, and as of May 2020, we were not allowed to conduct what were considered to be "elective experiments" at the bench. From mid-March through May 2020, our ramp-down work was restrictedto animal husbandry and tick maintenance. This was particularly problematic with respect to Dr. Al-Hosary's training as a Visiting Scholar (Dr. Al-Hosary arrived at MU in early March, 2020).
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Hoffman K, Shahzad S, Calcutt M, Brayton K, Foecking M and RW Stich. Sequence of a novel Anaplasma marginale genome determined with next generation PacBio sequencing technology. The 30th Meeting of the American Society for Rickettsiology, Santa Fe, NM, June 8-11, 2019
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2019
Citation:
Shahzad S, Hoffman K, Shen Z, Straka K, Daugherty E, Thompson D, Zhang M, Mitchell J, Zhang S and RW Stich. Rickettsial organisms associated with ticks collected from Missouri elk. The 30th Meeting of the American Society for Rickettsiology, Santa Fe, NM, June 8-11, 2019
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Stich RW, Hoffman K, Shahzad S, Zorn C, Scott S, Jittapalapong S, Johnson G, Pithua P, and G Zhang. Immune intervention targeting a tick vector of Anaplasma marginale. The 30th Meeting of the American Society for Rickettsiology, Santa Fe, NM, June 8-11, 2019
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Stich RW, Hoffman K, Shahzad S, Jittapalapong S, Johnson G, Pithua P, and G Zhang. Performance of Dermacentor andersoni ticks fed on calves immunized with denatured tick antigen preparations. 100th Conference of Research Workers in Animal Diseases, Chicago, IL, November 3-5, 2019
- Type:
Journal Articles
Status:
Other
Year Published:
2021
Citation:
Hoffman K, Shahzad S, Calcutt M, Brayton K, Foecking M, Jittapalapong S and RW Stich. 2020. Analysis of a novel Anaplasma marginale genome determined with next generation PacBio sequencing technology. Manuscript In Preparation.
- Type:
Journal Articles
Status:
Other
Year Published:
2021
Citation:
Hoffman K, Zorn C, Shahzad S, Jittapalapong S, Al-Hosary A, Zhang G, Johnson G, Pithua P, Stoffel R, Dhagat-Mehta B, Butcher M and RW Stich. 2020. A bovine model to identify tick molecules targeted by immune sera associated with reduced performance of Dermacentor andersoni ticks. Manuscript In Preparation.
|
Progress 06/01/18 to 05/31/19
Outputs
Target Audience:Continued training of one full-time PhD Candidate and one full-time PhD Student. Four Veterinary Medical (VM) Students were involved on a part-time basis; a VM-2 was a Veterinary Research Program Scholar during the summer intersession and the remaining VM-1, VM-2 and VM-4 students assisted with maintenance of experimental cattle. An undergraduate student from Harris-Stowe State University (St. Louis, MO) was also trained as a summer intern, under the auspices of the Missouri Louis Stokes Alliance for Minority Participation (MOLSAMP). Preliminary results were presented to professional peers at national and international scientific meetings as abstracts, presentations and posters, and as work-in-progress seminars on the campus of the University of Missouri. Some of these reports were presented by students at multiple venues. Changes/Problems:Adjustments were required to address unexpected pitfalls with respect to tick feeding and availability. For example, fewer ticks were available for challenge infestations, which required us to use smaller challenge group sizes. Also unexpected was the need to maintain a tick colony, because our supplier stopped selling the tick species chosen for this work. Because of these pitfalls, this study has required considerably more time and work than expected. What opportunities for training and professional development has the project provided?Two PhD students, Sammuel Shahzad and Kyle Hoffman, continue to work on this project. Both students are proficient with experimental calves, tick rearing, monitoring host blood for signs of anaplasmosis, PCR assays, ELISA and Western analysis. Both students presented research at the 2018 CRWAD, and Hoffman won the AAVP student presentation award for the second year in a row. In addition to this project, each student has an independent project to (1) sequence the genome of a novel A. marginale strain that is not transmissible by Dermacentor ticks (Hoffman) and (2) to identify ticks and associated tick-borne pathogens collected from elk in Missouri (Shahzad).Shahzad also visited the USDA ARS in Pullman, WA, to learn different approaches to artificial feeding of ixodid ticks. Hoffman is currently drafting his first manuscript and Shahzad is soon expected to take his comprehensive exam. Hillary Smith is an undergraduate student at Harris-Stokes State University (St. Louis, MO). Smith worked as an intern for the Missouri Louis Stokes Alliance for Minority Participation. Smith learned about ticks, bioinformatics, PCR and DNA sequence analysis while working in this lab. Sarah Scott is a participant in the Veterinary Research Scholar Program at the University of Missouri. Scott spent last summer learning how to perform ELISAs, which was used to analyze immune sera from calves immunized with different tick tissues. Kristin Manley, Ashley Otto and Nicole Bricker are veterinary medical students who assisted with the care of experimental calves. How have the results been disseminated to communities of interest?Results of research supported by this grant were represented in a total of 10 presentations by Smith (3), Hoffman (2), Scott (2), Shahzad (1), Manley (1) and Stich (1). Three abstracts were accepted for presentation at another conference in June, 2019. What do you plan to do during the next reporting period to accomplish the goals?Objective 1. Complete remaining immunization and tick challenge feeding trials. Objective 2. Identify tick molecules uniquely reactive with sera from protected hosts. Objective 3. Submit results and conclusions for publication in peer-review journals.
Impacts
What was accomplished under these goals?
Problems with the Dermacentor andersoni tick colony have delayed this work and limited the number of ticks available for challenge infestations, especially Anaplasma marginale-infected ticks for transmission challenge under Aim #1. Moreover, our supplier recently informed us that they will no longer provide this tick species for purchase. However, we now maintain our a D. andersoni colony, which is fed exclusively on cattle. Although our tick-feeding protocols are working, maintenance of this colony is time-consuming and requires additional calves. Thus, work has begun to investigate the feasibility of a continuous-flow artificial feeding apparatus for tick colony maintenance, analogous to those used to maintain mosquito and flea colonies. However, it is unlikely that an artificial feeding apparatus will be optimized in time to benefit the budget of this current project. Despite the above pitfalls, work has progressed since the previous report. Both of the graduate students associated with this project have become proficient in bovine and tick husbandry. The second trial was completed for Aim 2 and the the second trial for Aim 1 is nearly complete at the time of this report. Trial 3 for both Aims is underway. Calves were purchased for trial 4 of both Aims, which will begin this month. Although it is too soon to reach conclusions, the preliminary results are interesting and some of these results were unexpected.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Smith H, Hoffman K, Shahzad S and RW Stich. A molecular key for tick species. Undergraduate Research and Creative Achievements Forum, University of Missouri, Bond Life Sciences Center, July 26, 2018*
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Scott S, Hoffman K, Shahzad S and RW Stich. Bovine antibody response to immunization against Dermacentor andersoni midgut and salivary gland. National Veterinary Scholars Symposium: Veterinary Scientists in Global Health Research, Texas A&M University, August 2-4, 2018**
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Manley K, Shahzad S, Hoffman K and RW Stich. Quantification of rickettsial load during the acute phase of experimental bovine anaplasmosis. National Veterinary Scholars Symposium: Veterinary Scientists in Global Health Research, Texas A&M University, August 2-4, 2018
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Smith H, Hoffman K, Shahzad S and RW Stich. A molecular key for tick species. Annual Biomedical Research Conference for Minority Students, Indianapolis, IN, November 14-17, 2018*
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Hoffman K, Shahzad S, Calcutt M, Stich RW, Brayton K and M. Foecking. Sequence of a novel Anaplasma marginale genome determined with next generation PacBio sequencing technology. Vector-Borne and Parasitic Diseases Section, 99th Conference of Research Workers in Animal Diseases, Chicago, IL, December 2-4, 2018 (AAVP student presentation award)***
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Shahzad S, Hoffman K, Shen Z, Straka K, Daugherty E, Thompson D, Zhang M, Mitchell J, Zhang S and RW Stich. Detection of pathogens associated with different tick species collected from elk in Missouri. Vector-Borne and Parasitic Diseases Section, 99th Conference of Research Workers in Animal Diseases, Chicago, IL, December 2-4, 2018
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Hoffman K, Shahzad S, Smith H, Scott S, Thompson D, Daugherty E, Straka K, Zhang S and RW Stich. Preliminary development of molecular algorithm for identification of ixodid tick species. Vector-Borne and Parasitic Diseases Section, 99th Conference of Research Workers in Animal Diseases, Chicago, IL, December 2-4, 2018
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2018
Citation:
Stich RW, Hoffman K, Shahzad S, Scott S, Jittapalapong S, Johnson G, Pithua P, and G Zhang. Preliminary development of a bovine model to measure performance of Dermacentor andersoni ticks. Vector-Borne and Parasitic Diseases Section, 99th Conference of Research Workers in Animal Diseases, Chicago, IL, December 2-4, 2018
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2019
Citation:
Scott S, Hoffman K, Shahzad S, Jittapalapong S and RW Stich. Bovine antibody response to immunization against Dermacentor andersoni midgut and salivary gland. 42nd Annual CVM Research Day, Columbia, MO, May 10, 2019**
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2019
Citation:
Hoffman K, Shahzad S, Calcutt M, Brayton K, Foecking M and RW Stich. Sequence of a novel Anaplasma marginale genome determined with next generation PacBio sequencing technology. The 30th Meeting of the American Society for Rickettsiology, Santa Fe, NM, June 8-11, 2019
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2019
Citation:
Shahzad S, Hoffman K, Shen Z, Straka K, Daugherty E, Thompson D, Zhang M, Mitchell J, Zhang S and RW Stich. Rickettsial organisms associated with ticks collected from Missouri elk. The 30th Meeting of the American Society for Rickettsiology, Santa Fe, NM, June 8-11, 2019
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2019
Citation:
Stich RW, Hoffman K, Shahzad S, Zorn C, Scott S, Jittapalapong S, Johnson G, Pithua P, and G Zhang. Immune intervention targeting a tick vector of Anaplasma marginale. The 30th Meeting of the American Society for Rickettsiology, Santa Fe, NM, June 8-11, 2019
|
Progress 06/01/17 to 05/31/18
Outputs
Target Audience:Laboratory instruction and training of two full-time PhD students and one part-time undergraduate student. A freshman-level Initiative for Maximizing Student Diversity (IMSD) Exposure to Research for Science Students (EXPRESS) participant observed bench methods while working with graduate students. Preliminary results presented to professional peers in seminar settings. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?As the project began, Professor Sathaporn Jittapalapong (Kastsart University, Bangkok, Thailand) visited the performance site (University of Missouri-Columbia) for one month. During this visit, Dr. Jittapalapong (1) helped train two PhD students assigned to the project, (2) assisted with tick dissection to separate different tissues, (3) advised the project director regarding materials and methods (e.g., adjuvant and tick tissue processing for immunization), (4) spear-headed a new Memorandum of Understanding between Kasetsart University and the University of Missouri and (5) met with other scientists in the area regarding related projects such as surveillence of tick-borne pathogens in Missouri (Missouri Department of Health and Senior Services and the US Centers for Disease Control and Prevention) and a novel treatment for trypanosomiasis. As previously mentioned, this project made it possible to mentor and to train two PhD students. Both students, Sammuel Shahzad and Kyle Hoffman, received extensive training in students in bovine (calf) husbandry and tick biology. Kyle Hoffman, a third-year student in the University of MIssouri Molecular Pathogenesis and Therapeutics (MPT) Graduate Program, has presented his work internally at three Infectious Disease Work in Progress Seminars (WIPS) and one MPT Student Seminar. Kyle also passed his comprehensive examination, so he is now a PhD Candidate. Kyle also won a student competition for his presentation at the Conference of Research Workers in Animal Diseases in 2017. Kyle has become adept at 2-D electrophoresis, is currently learning electron microscopy, ELISA and PCR methodologies, and he recently outlined his first manuscript for submission to a peer-review journal. Sammuel Shahzad, a second-year Fulbright Student in the University of MIssouri Pathobiology Area Graduate Program, had his first committee meeting. Sam has grown considerably as a student, at the bench as well as in his didactic and manuscript-driven coursework. Sam is now proficient with the conventional PCR technique, and he constructed a plasmid to use as a positive control for quantitative PCR. Although he possesses a strong proclivity for molecular diagnostics, Sam is also being trained in protein chemistry and electron microscopy. In addition to his presentation at the 2017 Conference of Research Workers in Animal Diseases, Sam has also given three Infectious Disease WIPS presentations. Katherine Weir was a junior-level part-time undergraduate student employee, who received training in animal care. Aaron McGhee was a freshman-levelInitiative for Maximizing Student Diversity (IMSD) Exposure to Research for Science Students (EXPRESS) participant, who observed bench methods while working with graduate students. How have the results been disseminated to communities of interest?The following press release was picked up by several media outlets: https://munews.missouri.edu/news-releases/2017/0828-usda-grants-mu-460000-to-develop-immunizations-for-tick-borne-disease/ What do you plan to do during the next reporting period to accomplish the goals?Objective 1: to finish the first trial of Aim 1. Objective 2: raise a tick colony with selection pressure for faster attachment and feeding on cattle. Objective 3: to complete the remaining tick feeding trials for Aims 1 and 2. The initial delays in acquiring facilities and animals are not expected to be a problem this year, the new approach to tick rearing will hopefully result in faster feeding rates for the ticks, and fully trained graduate students are expected to make this work progress faster.
Impacts
What was accomplished under these goals?
The overarching objective of this project is to determine if a novel approach to exposing cattle to tick tissues which directly interface with the host immune response (tick gut and salivary glands) will interfere with disease-causing organisms inside of the ticks. This work is expected to contribute to development of sustainable approaches to prevent existing, emerging or foreign tick-borne diseases among US livestock. This project is largely based on previous studies with foreign ticks and tick-borne diseases. To date, a bovine model has been adapted for this work, utilizing a tick species and a tick-borne disease that are found in the USA. Benchwork methodologies such as two-dimensional protein electrophoresis and PCR-based assays were also adapted to this project, which will allow in-depth analysis of the host immune response and its effect on the infection in the ticks, respectively. Tactically, facilities were arranged to conduct the project, experimental cattle were purchased and the first trial began for both specific aims. One unexpected challenge came in feeding the colony-reared ticks on these cattle, which proved more difficult than in years past, possibly due to high relative humidity at the time of the initial tick feeding. Another possibility is that these colony-reared ticks had become more adapted to rabbits and sheep. Another unexpected development was a 200% price increase for the Dermacentor andersoni tick species selected for this project. Based on the latter two issues, the decision has since been made to begin work to establish our own D. andersoni tick colony, which will be reared on cattle. Once ticks began to feed on experimental cattle, the next task was to prepare immunogen from tissues dissected from ticks at feeding days one through six. This was a laborious process, which was especially time-consuming with new graduate students to train. Sufficient immunogen was prepared for the first trial of each specific aim. Calves were immunized, and the first trial was completed for Aim 1. Trial 1 for Aim 2 is underway at the time of this report.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Shahzad S, Shen Z, Straka K, Daugherty E, Thompson D, Zhang M, Mitchell J, Zhang S and RW Stich. Preliminary development of a PCR-based assay panel to screen ticks collected from elk in Missouri. Vector-Borne and Parasitic Diseases Section, 98th Conference of Research Workers in Animal Diseases, Chicago, IL, December 3-5, 2017.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Hoffman, K, Zorn C, Shahzad S, Jittapalapong S, Zhang G, Johnson G, Pithua P, Stoffel R, Dhagat-Mehta B, Butcher M and RW Stich. Development of a bovine model to identify tick molecules targeted by immune sera associated with reduced performance of Dermacentor andersoni ticks. Vector-Borne and Parasitic Diseases Section, 98th Conference of Research Workers in Animal Diseases, Chicago, IL, December 3-5, 2017.
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