Progress 09/01/20 to 04/21/25
Outputs
Target Audience:Note: This award is being transitioned from Kansas State University to Auburn University. The information in this final report represents the efforts completed at Kansas State University. In this project, we have delivered science-based, data-driven information on bovine anaplasmosis and strategies for managing this disease to a diversity of stakeholders. Our target audience includes our primary stakeholder (e.g. cattle producers, veterinarians), other professionals with interest in bovine anaplasmosis (e.g. researchers, policymakers, industry), and student trainees. Forums for these communication efforts have included formal programs (e.g. professional conferences/meetings, continuing education programs), producer-focused educationsl programs/outreach/extension activities (e.g. participation in producer meetings, serving as point of contact for anaplasmosisrelated questions); training opportunities for undergraduate students, professional students (e.g veteinary students seeking research opportunities), direct graduate student mentees and other graduate students looking for a diversity of training experiences, and publication of research results in peer-reviewed journals. Our project-related presentations at professional society meetings have reached the following target audiences: academic and governmental researchers and students interested in bovine anaplasmosis, tick-borne diseases of cattle, and cattle health. We have participated in continuing education events (e.g. formal and informal producer educational events)presenting on bovine anaplsmosis, tick-borne diseases of cattle, and controlling tick-borne diseases of cattle. Throughout this project we have worked with cattle producers, and this their helprecruitedcattle blood samples to assess actively circulating A. marginale genotypes. Recruitment of these samples has allowedus tocontinuously interface with two of our most important stakeholders (cattle producers and practicing veterinarians) on best practices to manage anaplasmosis within their own herds and has allowed us to anecdotally assess potential characteristics of A. marginale strains selected for assessment for inclusion into our experimental vaccine construct. Our producer-focused events and forums, outreach, and extension activities, describing our project-related data and outcomes have reached the following target audiences: cattle producers, practicing livestock veterinarians, livestock industry personnel, and people generally interested in cattle health, ticks, tick-borne diseases of livestock, and anaplasmosis control. This project has provided a platform for student training including graduate students, undergraduate students, and professional (DVM) students. These students have received hands-on training in developing research skills including labbased benchwork skills (e.g. sample handling/processing, diagnostic assays, microbiology techniques), animal handling techniques (e.g. safely working with cattle, bio-sample collection), communication skills (e.g. participation and/or presentation in lab meetings, local/regional meetings, national professional meetings), organizational skills and teamwork skills. Publication of research results in peer-reviewed publications also provided wider opportunities to communicate our projectrelated finds with various stakeholders in academia, government, industry, and other professional groups, or members of the general public interested in this work. Changes/Problems:PI is completing this final report early as instructed by NIFA so that this award may be moved from Kansas State University to Auburn University. This final report serves to summarize the activities and accomplishments of this project completed at Kansas State University. This award is intended tobe transferred to Auburn University (PI moved to Auburn University in Nov 2023) to complete project activities. Project PI changed institutions Nov 2023. Project funds have not be accessible since Nov 2023. An agreement to transfer this award from Kansas State University to Auburn University took several months. Additionally, prior to being able to initiate award transfer paperwork, an award extension needed to be approved so that the transfer process had sufficient time to occur. During this time period no spending could occur and project personnel (undergraduate and graduate students) left the project as spending ceased. These personnel have since graduated or are no longer available because PI moved institutions. We areworking with a collaborator to explore an alternative immunization strategy that would be both DIVA compliant and would still be informed by our A. marginale diversity evaluations and diagnostic assay development. This strategy will be evaluated upon transfer of the award to Auburn University. What opportunities for training and professional development has the project provided?While this award was at Kansas State University, this project has provided funding to train two Master's students (Flowers, Lovett), a doctoral student (Bickmeier). This project also provided opportunity for graduate students to attend and present their research at national meetings including the annual CRWAD meeting as well as other local and regional meetings. In addition, this project has provided training opportunies for 4 undergraduate students, 3 concurrent DVM students, and 2 Veterinary Summer Research Scholars. How have the results been disseminated to communities of interest?Project results have been shared via presentation at professional meetings (e.g. CRWAD). Overviews of bovine anaplasmosis disease, epidemiology, treatment and control, ticks and tick-borne diseases of cattle, and information on ongoing anaplasmosis studies (and opportunities to participate in those studies) have been shared with cattle producer stakeholers (e.g. individual consultations, cattlemens events, podcasts, interviews)and at livestock veterinarian continuing education events (e.g. local and state VMAs). These forums are wellreceived and have allowed us to engage and recruit producers to participate in on-going research efforts. Results have also been disseminated to communities of interest through peer-reviewed, open-access publications. What do you plan to do during the next reporting period to accomplish the goals?This award is being transferred from Kansas State University to Auburn University. This is the final report for activites performed at Kansas State University. Upon transfer and once funds are accessible, we anticipate performing the following: Objective 1: Continue to collect samples to examine the diversity of actively circulating A. marginale strains to inform stain selection for prototype anaplasmosis vaccine development design. Use the identified diversity to help refine selection of the A. marginale isolate to propogate for evaluation and inclusion in the prototype anaplasmosis vaccine. Work with our partners at SUA to identify and recruit a MS student to investigate diversity of A. marginale in Tanzania. Continue working on methods to streamline A. marginale msp1a genotype identification and analysis via massive parallel sequencing. Objective 2: Continue development of additional serologic-based diagnostic assays to characterize the immune response to specific A. marginale proteins. Select three of the recombinantly expressed conserved A. marginale proteins to generate polyclonal antibodies against. Perform Luminex assays to help characterize the cellular immune response to immunization against A. marginale in animals immunized with the existing available experimental vaccine and a prototype A. marginale vaccine. Continue revising protocols for generating sufficient whole killed A. marginale antigen material from stabiliates for preparation of the immunogen. Perform pilot immunization experiment with prepared whole killed A. marginale to evaluateimmunzation strategy prior to large multi-genotype cocktail immunziation experiment. Work with collaborators to test and compare a nanoparticle-based immunization strategy versus a whole-killed vaccine (if successful, this alternative strategy would still utilitze our A. marginale diversity data to inform antigens to incorporate and would also have the added benefit of being DIVA compliant). Objective 3: Work towards this objective will being once we confirm we have developed a protective immunogen in Objective 2. Consider alternative immunization strategies if challenges with generating sufficient whole-killed A. marginale material persists. Trainees: Recruit two additional graduate students (one Auburn University, one SUA) to help achieve project goals. This project will continue to support training opportunities with undergraduates and DVM students as additioanl efforts are needed. Dissemination of results: Results will be shared with interested stakeholders in the form of presentations by the PI or trainees at professional meetings (e.g. CRWAD) and stakeholder meetings (e.g. cattlemen/livestock association meetings). A new graduate student and additional technical help will be hired to assist in completing project objectives.
Impacts
What was accomplished under these goals?
Note:This award is being transferred from Kansas State University to Auburn University as PI moved to Auburn University. This is the final report represnts for accomplishments from activitiesperformedat Kansas State University. Access to funding ceased in Nov 2023 when PI moved Auburn University. Project objectives will be completed upon transfer of the award to Auburn University. Goal: The overall goal of this project is to develop a prototype, single-immunization anaplasmosis vaccine based regionallyrepresentative Anaplasma marginale antigen; and, to provide proof-of-concept data demonstrating protective efficacy against diverse A. marginale strain challenge in animals from different production system environments. Objectives: Objective 1 Accomplishments: We have recruitedand screened ~7,000 cattle bloodsamples from U.S. cattle herds. Additional herds from less represented regions of the U.S. have also been identified and we are awaiting access to project funds to collect and test these samples.These samples have been tested by both serologic and molecular assays for A. marginale. The A. marginale msp1a genotype has been identified from a representative subset of all positive samples. Multiple methods have been evaluated to improve throughput of identifying A. marginale genotypes.Six A. marginale strains have been characterized for potential inclusion in experimental vaccine construct. Of these strains, 4 are novel strains, isolated from naturally infected cattle in the U.S. that havenot been previously characterized. Objective 2 Accomplishments: We have identified and processed field-derived samples from participating producers that use the University Product anaplasmosis vaccine. Specifically, we evaluated whether a immunization-specific antibody response (anti-A. marginale Msp5, overall change in IgG titers) to cattle immunized with the University Products experimental anaplasmosis vaccine could be detected (the efficacy of the University Product anaplasmosis vaccine and the type of immunization response it generates will serve as a benchmark to directly compare the vaccine generated by this project. We have performed an initial study to evaluate the ability ofthe currently available experimental killed vaccine from University Products to generate a detectable immune response upon label-indicated prime boost administration and it'sefficacy to protect against (putative) homologous and heterologous A. marginale strain challenge.We have continued to work on developing additional diagnostic assays to evaluate cattle vaccinated with a wholekilled vaccine for other conserved A. marginale surface protein targets. Developing these other assays have involved cloning, expression, and purification of multiple conserved A. marginale proteins (some of these proteins have been challenging to produce). Three of these proteins will be selected to generate polyclonal antibody against. We continued working on refining our methodology to generate sufficient antigen material for immunization experiments and diagnostic assays. Objective 3 Accomplishments: Nothing to report on the prototype implant vaccine as this is dependent on generating a suffieniently immunogenenic and protective whole-killed immunogen. However, we are in active discussions with a collabortor to test a novel nanoparticle vaccine design that would be informed by our A. marginale genetic diversity analyses in a small pilot. This collaborator has extensive experience with this vaccine platform and protein expression.
Publications
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2025
Citation:
Reif KE. Bovine anaplasmosis: Transmission, Prevalence, & Management. Southeastern Food and Farm Veterinary Summit. March 2025. Opelika, AL.
|
Progress 09/01/23 to 08/31/24
Outputs
Target Audience:In this reporting period, we have continued to deliver science-based, data-driven information on bovine anaplasmosis and strategies for managing this disease to a diversity of stakeholders. Our target audience includes our primary stakeholder (e.g. cattle producers, veterinarians), other professionals with interest in bovine anaplasmosis (e.g. researchers, policymakers, industry), and student trainees. Forums for these communication efforts have included formal programs (e.g. professional conferences/meetings, continuing education programs), producer-focused educationsl programs/outreach/extension activities (e.g. participation in producer meetings, serving as point of contact for anaplasmosisrelated questions); training opportunities for undergraduate students, professional students (e.g veteinary students seeking research opportunities), direct graduate student mentees and other graduate students looking for a diversity of training experiences, and publication of research results in peer-reviewed journals. Our project-related presentations at professional society meetings reach the following target audiences: academic and governmental researchers and students interested in bovine anaplasmosis, tick-borne diseases of cattle, and cattle health. We participated in a continuing education eventfor livestock veterinarians - the South Dakota VMA -presenting on bovine anaplsmosis, tick-borne diseases of cattle, and controlling tick-borne diseases of cattle. As in previous years, we continued to work with cattle producers to recruit samples to assess actively circulating A. marginale genotypes. Recruitment of these samples allows us to continuously interface with cattle producers and practicing veterinarians on best practices to manage anaplasmosis within their own herds. The events we have participated in,describing our project-related data and outcomes, have reached the following target audiences: cattle producers, practicing livestock veterinarians, livestock industry personnel, and people generally interested in cattle health, ticks, tick-borne diseases of livestock, and anaplasmosis control. This project has also provided a platform for student training including graduate students, undergraduate students, and professional (DVM) students. These students have received hands-on training in developing research skills including labbased benchwork skills (e.g. sample handling/processing, diagnostic assays, microbiology techniques), animal handling techniques (e.g. safely working with cattle, bio-sample collection), communication skills (e.g. participation and/or presentation in lab meetings, local/regional meetings, national professional meetings), organizational skills and teamwork skills. Publication of research results in peer-reviewed publications also provided wider opportunities to communicate our projectrelated finds with various stakeholders in academia, government, industry, and other professional groups, or members of the general public interested in this work. Changes/Problems:Project PI moved institutions during this project period (Sep 1, 2023 to Aug 31, 2024). Project funds have not be accessible since Nov 2023. During this project period (Sep 1, 2023 to Aug 31, 2024), there werelimited accomplishments because PI changed institutions. Access to project grant funds was restricted in Nov 2023 when PI began appointment with Auburn University. An agreementto transfer this award from Kansas State University to Auburn University tookseveral months. Additionally, prior to being able to initiate award transfer paperwork, an award extension needed to be approved so that the transfer process had sufficient time to occur. During this time period no spending could occur and project personnel (undergraduate and graduate students) left the project as spending ceased. These personnel have since graduated or are no longer available because PI moved institutions. Upon submission of this progress report, a final study report will be submitted to close the award at Kansas State University so that it may be transfered to Auburn University. We are still working with a collaborator to explore an alternative immunization strategy that would be both DIVA compliant and would still be informed by our A. marginale diversity evaluations and diagnostic assay development. This strategy will be evaluated upon transfer of the award to Auburn University. What opportunities for training and professional development has the project provided?During this reporting period, this project has provided funding to train a doctoral student (Bickmeier). This project also provided opportunity for the graduate student to attend and present her research at the annual CRWAD meeting. In addition, this project continued toprovided training opportunities for current KSU DVM student(Beltz). How have the results been disseminated to communities of interest?Preliminary results have been shared via presentation at professional meetings (e.g. CRWAD). Overviews of bovine anaplasmosis disease, epidemiology, treatment and control, ticks and tick-borne diseases of cattle, and information on on-going anaplasmosis studies (and opportunities to participate in those studies) have been shared with cattle producer stakeholers and at livestock veterinarian continuing education events (e.g. South Dakota VMA). These forums are well-received and have allowed us to engage and recruit producers to participate in on-going research efforts. What do you plan to do during the next reporting period to accomplish the goals?Of greatest priority is to complete transfer of award from Kansas State University to Auburn University and establish relevant project-related regulatory approvals at Auburn University. Upon transfer and once funds are accessible, we anticipate performing the following: Objective 1: Continue to collect samples to examine the diversity of actively circulating A. marginale strains to inform stain selection for prototype anaplasmosis vaccine development design. Use the identified diversity to help refine selection of the A. marginale isolate to propogate for evaluation and inclusion in the prototype anaplasmosis vaccine. Work with our partners at SUA to identify and recruit a MS student to investigate diversity of A. marginale in Tanzania. Continue working on methods to streamline A. marginale msp1a genotype identification and analysis via massive parallel sequencing. Objective 2: Continue development of additional serologic-based diagnostic assays to characterize the immune response to specific A. marginale proteins. Select three of the recombinantly expressed conserved A. marginale proteins to generate polyclonal antibodies against. Perform Luminex assays to help characterize the cellular immune response to immunization against A. marginale in animals immunized with the existing available experimental vaccine and a prototype A. marginale vaccine. Continue revising protocols for generating sufficient whole killed A. marginale antigen material from stabiliates for preparation of the immunogen. Perform pilot immunization experiment with prepared whole killed A. marginale to evaluate immunzation strategy prior to large multi-genotype cocktail immunziation experiment. Work with collaborators to test andcompare a nanoparticle-based immunization strategy versus a whole-killed vaccine (if successful, this alternative strategy would still utilitze our A. marginale diversity data to inform antigens to incorporate and would also have the added benefit of being DIVA compliant). Objective 3: Work towards this objective will being once we confirm we have developed a protective immunogen in Objective 2. Consider alternative immunization strategies if challenges with generating sufficient whole-killed A. marginale material persists. Trainees: Recruit two additional graduate students (one Auburn University, one SUA) to help achieve project goals. This project will continue to support training opportunities with undergraduates and DVM students as additioanl efforts are needed. Dissemination of results: Results will be shared with interested stakeholders in the form of presentations by the PI or trainees at professional meetings (e.g. CRWAD) and stakeholder meetings (e.g. cattlemen/livestock association meetings).
Impacts
What was accomplished under these goals?
Note: Project PI moved institutions during this project period (Sep 1, 2023 to Aug 31, 2024). Project funds have not be accessible since Nov 2023. PI institutionalmove, lack of access to project funds, and loss of personnel have impacted accomplishments during this period. Goal: The overall goal of this project is to develop a prototype, single-immunization anaplasmosis vaccine based regionallyrepresentative Anaplasma marginale antigen; and, to provide proof-of-concept data demonstrating protective efficacy against diverse A. marginale strain challenge in animals from different production system environments. Objectives: Objective 1 Accomplishments: We have recruited an additional ~50 cattle samples from U.S. cattle herds and have identified ~10 additional herds from across the country that are willing to submit samples. These samples await testing by both serologic and molecular assays for A. marginale and A. marginale msp1a genotype identification. We continued to work on methodology to streamline msp1a genotyping to improve workflow efficiency. Objective 2 Accomplishments: We have continued to work on developing additional diagnostic assays to evaluate cattle vaccinated with a wholekilled vaccine for other conserved A. marginale surface protein targets. Developing these other assays have involved cloning, expression, and purification of multiple conserved A. marginale proteins (some of these proteins have been challenging to produce). Three of these proteins will be selected to generate polyclonal antibody against. We continued working on refining our methodology to generate sufficient antigen material for immunization experiments and diagnostic assays. Objective 3 Accomplishments: Nothing to report on the prototype implant vaccine as this is dependent on generating a suffieniently immunogenenic and protective whole-killed immunogen. However, we are in active discussions with a collabortor to test a novel nanoparticle vaccine design that would be informed by our A. marginale genetic diversity analyses in a small pilot. This collaborator has extensive experience with this vaccine platform and protein expression.
Publications
- Type:
Peer Reviewed Journal Articles
Status:
Published
Year Published:
2023
Citation:
Lovett AC, Reppert EJ, Huser S, Rob�rt BD, Jaeger JR, Kang Q, Liu R, Anantatat T, Armstrong CL, Reif KE. Kansas beef bulls with chronic anaplasmosis demonstrate satisfactory breeding soundess outcomes at breeding soundness examination. J Am Vet Med Assoc. 2023. Oct 13;262(1):53-60. doi: 10.2460/javma.23.05.0292.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2024
Citation:
Bickmeier NP, Toillion AR, Skinner BC, Anantatat T, Amachawadi RG, Liu C., Reppert EJ, Coetzee JF, Reif KE. Antimicrobial susceptibilities of enterococcus from calves treated with chlortetracycline for anaplasmosis control. Conference for Research Workers of Animal Diseases. Jan 20-23, 2024. Chicago, IL.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2024
Citation:
Reif KE. Biting into Vector-Borne Cattle Diseases: Anaplasmosis and Theileriosis
|
Progress 09/01/22 to 08/31/23
Outputs
Target Audience:In this reporting period, we have continued to deliver science-based, data-driven information on bovine anaplasmosis and strategies for managing this disease to a diversity of stakeholders. Our target audience includes our primary stakeholder (e.g. cattle producers, veterinarians), other professionals with interest in bovine anaplasmosis (e.g. researchers, policy-makers, industry), and student trainees. Forums for these communication efforts have included formal programs (e.g. professional conferences/meetings, continuing education programs), producer-focused educationsl programs/outreach/extension activities (e.g. participation in producer meetings, serving as point of contact for anaplasmosis-related questions); training opportunities for undergraduate students, professional students (e.g veteinary students seeking research opportunities), direct graduate student mentees and other graduate students looking for a diversity of training experiences, and publication of research results in peer-reviewed journals. Our project-related presentations at professional society meetings reach the following target audiences: academic and governmental researchers and students interested in bovine anaplasmosis, tick-borne diseases of cattle, and cattle health. We have participated in continuing education events (e.g. producer educational events - most notably, a series of three events for Ohio cattle producers that reached 150+ producers; plus a podcast for the SeconDairy podcast) presenting on bovine anaplsmosis, tick-borne diseases of cattle, and controlling tick-borne diseases of cattle. As in previous years, we continue to work with cattle producers to recruit samples to assess actively circulating A. marginale genotypes. Recruitment of these samples allows us to continuously interface with cattle producers and practicing veterinarians on best practices to manage anaplasmosis within their own herds. Our producer-focused events and forums, outreach, and extension activities, describing our project-related data and outcomes have reached the following target audiences: cattle producers, practicing livestock veterinarians, livestock industry personnel, and people generally interested in cattle health, ticks, tick-borne diseases of livestock, and anaplasmosis control. This project has also provided a platform for student training including graduate students, undergraduate students, and professional (DVM) students. These students have received hands-on training in developing research skills including lab-based benchwork skills (e.g. sample handling/processing, diagnostic assays, microbiology techniques), animal handling techniques (e.g. safely working with cattle, bio-sample collection), communication skills (e.g. participation and/or presentation in lab meetings, local/regional meetings, national professional meetings), organizational skills and teamwork skills. Publication of research results in peer-reviewed publications also provided wider opportunities to communicate our project-related finds with various stakeholders in academia, government, industry, and other professional groups, or members of the general public interested in this work. Changes/Problems:Some lingering effects from the COVID-19 pandemic (e.g. student recruitment, collaboration with international partners, supply chain issues, facility access/staffing) still affected the project during this reporting year. Our primary international collaborator also unexpectedly passed away. The primary graduate student working on this project required family leave time off during this reporting period. Outside of COVID-related and staffing issues, we are still struggling with production of sufficient antigen and generating a sufficient protective response. We are working with a collaborator to explore an alternative immunization strategy that would be both DIVA compliant and would still be informed by our A. marginale diversity evaluations and diagnostic assay development. What opportunities for training and professional development has the project provided?During this reporting period, this project has provided funding to train a doctoral student (Naemi Bickmeier). This project also provided opportunity for the graduate student to attend and present her research at the annual CRWAD meeting as well as other local and regional meetings. In addition, this project has provided training opportunies this reporting period for one master student and two undergraduate students interested in veterinary medicine (one of these students began their first year of DVM training during this period). How have the results been disseminated to communities of interest?Preliminary results have been shared via presentation at professional meetings (e.g. CRWAD). Overviews of bovine anaplasmosis disease, epidemiology, treatment and control, ticks and tick-borne diseases of cattle, and information regarding on-going anaplasmosis studies (and opportunities to participate in those studies) have been shared with cattle producer stakeholders and at cattle producer educational events. These forums are very well received and have allowed us to engage and recruite producers to participate in on-going research efforts. What do you plan to do during the next reporting period to accomplish the goals?Within the next reporting period, we anticipate performing the following: Objective 1: Continue to collect samples to examine the diversity of actively circulating A. marginale strains to inform stain selection for prototype anaplasmosis vaccine development design. Use the identified diversity to help refine selection of the A. marginale isolate to propogate for evaluation and inclusion in theprototype anaplasmosis vaccine. Work with our partners at SUA to identify and recruit a MS student to investigate diversity of A. marginale in Tanzania. Continue working on methods to streamline A. marginale msp1a genotype identification and analysis via massive parallel sequencing. Objective 2: Continue development of additional serologic-based diagnostic assays to characterize the immune response to specific A. marginale proteins. Select three of the recombinantly expressed conserved A. marginale proteins to generate polyclonal antibodies against. Perform Luminex assays to help characterize the cellular immune response to immunization against A. marginale in animals immunized with the existing available experimental vaccine and a prototype A. marginale vaccine. Continue revising protocols for generating sufficient whole killed A. marginale antigen material from stabiliates for preparation of the immunogen. Perform pilot immunization experiment with prepared whole killed A. marginale to evaluate immunzation strategy prior to large multi-genotype cocktail immunziation experiment. Work with collaborators to test and compare a nanoparticle-based immunization strategy versus a whole-killed vaccine (if successful, this alternative strategy would still utilitze our A. marginale diversity data to inform antigens to incorporate and would also have the added benefit of being DIVA compliant). Objective 3: Work towards this objective will being once we confirm we have developed a protective immunogen in Objective 2. Consider alternative immunization strategies if challenges with generating sufficient whole-killed A. marginale material persists. Trainees: Recruit twoadditional graduate students (one K-State, one SUA) to help achieve project goals. This project will continue to sponsor training opportunities with undergraduates and DVM students as additioanl efforts are needed. Dissemination of results: Results will be shared with interested stakeholders in the form of presentations by the PI or trainees at professional meetings (e.g. CRWAD) and stakeholder meetings (e.g. cattlemen/livestock association meetings).
Impacts
What was accomplished under these goals?
Goal:The overall goal of this project is to developa prototype, single-immunization anaplasmosis vaccine based regionally-representativeAnaplasma marginaleantigen; and, to provide proof-of-concept data demonstrating protective efficacy against diverseA. marginalestrain challenge in animals from different production system environments. Objectives: Objective 1 Accomplishments: We have recruited an additional ~700 cattle samples from U.S. cattle herds. These samples have been tested by both serologic and molecular assays for A. marginale. The A. marginale msp1a genotype has been identified from a representative subset of all positive samples. We also finished determining the msp1a genotype from arepresentative subset of samples collected during the previous reporting period. We continued to work on methodology to streamline msp1a genotyping to improve workflow efficiency. Objective 2 Accomplishments:We processed additional producer-derived samples from cattle vaccinated with the currently available experimental anaplasmosis vaccine to determine if this vaccine generates a specific anti-A. marginale Msp5 immune response. We have continued to work on developing additional diagnostic assays to evaluate cattle vaccinated with a whole-killed vaccine for other conserved A. marginale surface protein targets. Developing these other assays have involved cloning, expression, and purification of multiple conserved A. marginale proteins (some of these proteins have been challenging to produce). Three of these proteins will be selected to generate polyclonal antibody against. We continued working on refining our methodology to generate sufficient antigen material for immunization experiments and diagnostic assays. Objective 3 Accomplishments: Nothing to report on the prototype implant vaccine as this is dependent on generating a suffieniently immunogenenic and protective whole-killed immunogen. However, we are in active discussions with a collabortor to test a novel nanoparticle vaccine design that would be informed by our A. marginale genetic diversity analyses in a small pilot. This collaborator has extensive experience with this vaccine platform and protein expression.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2022
Citation:
Lovett AC, Reppert EJ, Jaeger JR, Kang Q, Flowers MR, Bickmeier NP, Anantatat T, ODay SC, Armstrong CL, Reif KE. Satisfactory breeding potential is transiently eliminated in beef bulls with clinical anaplasmosis. BMC Vet Res. 2022. Oct 29;18(1):381. doi: 10.1186/s12917-022-03470-7.
- Type:
Journal Articles
Status:
Accepted
Year Published:
2023
Citation:
Lovett AC, Reppert EJ, Huser S, Rob�rt BD, Jaeger JR, Kang Q, Liu R, Anantatat T, Armstrong CL, Reif KE. Kansas beef bulls with chronic anaplasmosis demonstrate satisfactory breeding soundess outcomes at breeding soundness examination. 2023. JAVMA.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Lovett AC, Reppert EJ, Huser S, Robert BD, Jaeger JR, Kang Q, Anantatat T, Armstrong CL, Reif KE. Cross-sectional study of Kansas beef bulls to examine associations between chronic bovine anaplasmosis and breeding soundness. American College of Veterinary Internal Medicine Annual Meeting. June 15-17, 2023. Philadelphia, PA. *KSU CVM Travel Award
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Bickmeier NP, Anantatat T, Beltz M, Eggers A, Weaver B, Almaraz J, Huser SM, Kang Q, Song X, Reif KE. Pregnancy success among eastern Kansas beef cows infected with Anaplasma marginale and/or Bovine Leukemia Virus. K-State Graduate Research, Arts, and Discovery Forum. March 9, 2023. Manhattan, KS.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Bickmeier NP, Anantatat T, Beltz M, Eggers A, Weaver B, Almaraz J, Huser SM, Kang Q, Song X, Reif KE. Pregnancy success among eastern Kansas beef cows infected with Anaplasma marginale and/or Bovine Leukemia Virus. K-State Phi Zeta Research Day. March 7, 2023. Manhattan, KS.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Bickmeier NP, Toillion AR, Skinner BC, Anantatat T, Amachawadi RG, Kang Q, Reppert EJ, Coetzee JF, Reif KE. Antimicrobial susceptibilities of E. coli from calves treated with chlortetracycline for anaplasmosis control. K-State Phi Zeta Research Day. March 7, 2023. Manhattan, KS.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Lovett AC, Reppert EJ, Huser S, Robert BD, Jaeger JR, Kang Q, Armstrong CL, Reif KE. Investigation of chronic Anaplasma marginale infection and breeding soundness in beef bulls. K-State Phi Zeta Research Day. March 7, 2023. Manhattan, KS.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Lovett AC, Reppert EJ, Jaeger JR, Kang Q, Bickmeier NP, Anantatat T, Flowers MR, ODay SC, Armstrong CL, Reif KE. Acute anaplasmosis reduces breeding soundness in experimentally infected beef bulls. March 7, 2023. Manhattan, KS. *2nd Place in Applied/Clinical Science, Large Animals Oral presentations
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Lovett AC, Reppert EJ, Huser S, Robert BD, Jaeger JR, Kang Q, Armstrong CL, Reif KE. Investigation of chronic Anaplasma marginale infection and breeding soundness in beef bulls. K-State Cattlemens Day. March 3, 2023. Manhattan, KS.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Bickmeier NP, Anantatat T, Beltz M, Eggers A, Weaver B, Almaraz J, Huser SM, Kang Q, Song X, Reif KE. Pregnancy success among eastern Kansas beef cows infected with Anaplasma marginale and/or Bovine Leukemia Virus. K-State Cattlemens Day. March 3, 2023. Manhattan, KS.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Bickmeier NP, Toillion AR, Skinner BC, Anantatat T, Amachawadi RG, Kang Q, Reppert EJ, Coetzee JF, Reif KE. Antimicrobial susceptibilities of E. coli from calves treated with chlortetracycline for anaplasmosis control. K-State Cattlemens Day. March 3, 2023. Manhattan, KS.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Lovett AC, Reppert EJ, Huser S, Robert BD, Jaeger JR, Kang Q, Anantatat T, Armstrong CL, Reif KE. Investigation of chronic Anaplasma marginale infection and breeding soundness in bulls. Conference of Research Workers in Animal Diseases Annual Meeting. January 21-24, 2023. Chicago, IL.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Lovett AC, Reppert EJ, Jaeger JR, Kang Q, Bickmeier NP, Anantatat T, Flowers MR, ODay SC, Armstrong CL, Reif KE. Acute anaplasmosis reduces breeding soundness in experimentally infected beef bulls. K-State Cattlemens Day. March 3, 2023. Manhattan, KS.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Bickmeier NP, Anantatat T, Beltz M, Eggers A, Weaver B, Almarez J, Huser SM, Kang Q, Song X, Reif KE. Pregnancy success among eastern Kansas beef cows infected with Anaplasma marginale and/or Bovine Leukemia Virus. Conference of Research Workers in Animal Diseases Annual Meeting. January 21-24, 2023. Chicago, IL. *Received K-State CVM Travel Award; Selected as the Society for Tropical Veterinary Medicine Best Student Presentation Award
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Bickmeier NP, Toillion AR, Skinner BC, Anantatat T, Amachawadi RG, Kang Q, Reppert EJ, Coetzee JF, Reif KE. Antimicrobial susceptibility changes in E. coli from calves treated with chlortetracycline for anaplasmosis control. Conference of Research Workers in Animal Diseases Annual Meeting. January 21-24, 2023. Chicago, IL.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2023
Citation:
Robbins G, Rogers LR, McDermott EG, Reif KE, Krumpleman B, Galloway D, Powell JG. Serial evaluation of A. marginale positivity rate within a single herd of mature beef cows. American Society of Animal Science Southern Section. January 21-24, 2023. Raleigh, NC.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
Huser SM, Almaraz JM, Taxis TM, Larson RL, Reif KE, Hanzlicek GA, Weaver BM, Amrine DE. Evaluation of herd and within-herd prevalence of Bovine Leukemia Virus in eastern Kansas beef cattle. American Association of Bovine Practitioners. September 22-24, 2022. Long Beach, California.
- Type:
Theses/Dissertations
Status:
Accepted
Year Published:
2023
Citation:
Lovett, Anne. Thesis title: Investigation of acute clinical and chronic anaplasmosis on breeding soundness in beef bulls. Published: August 2023.
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Progress 09/01/21 to 08/31/22
Outputs
Target Audience:We are continuing to work with cattle producers to recruit blood samples herds to assess actively circulating A. marignale genotypes. Through recruitment of these samples, we engage our primary stakeholder (U.S. cattle producers), informally engaging them broadly about the importance of bovine anaplasmosis, relaying our study objectives, and collecting feedback from them on their perceptions of the impact of this disease at the local level of their operation (including providing suggestions to reduce the impact of this disease in their own operation). We are still feeling the impacts of COVID-19 on collecting and processing these samples, but we have been able to continue collecting samples. Additionally, we have engaged cattle producers through presentations at cattle producer education events. Other target audience is the scientific community, with information shared through presentation of data and study objectives at scientific meetings. Changes/Problems:This project year was impacted by the continued affects of the COVID-19 pandemic (e.g. student recruitment, collaboration with international partners, supply chain issues, facility access/staffing). These impacts have delayed staffing and research productivity. However, despite this, we continue working towards project goals. Outside of COVID-related issues, in case antigen production becomes a limiting issue or our immunization doesn't elicit a sufficient protective response, we are actively exploring alternative immunization strategies that would also be DIVA compliant and would still be informed from our A. marginale diversity evaluations and diagnostic assaydevelopment. What opportunities for training and professional development has the project provided?This project has provided funding to recruite and train a doctoral student (Naemi Bickmeier). In addition, this project has provided training opportunities this year for two master's students, a second PhD student, two Veterinary Research Summer Program scholars, and four undergraduates interested in veterinary medicine. How have the results been disseminated to communities of interest?Preliminary results have been shared via presentation at professional meetings. Overviews of bovine anaplasmosis disease, epidemiology, treatment and control, and information about on-going anaplasmosis studies have been shared with cattle producer stakeholders and at cattle producer educational events. These forums are very well received and have also allowed us to identify and recruit producers to participate in on-going research efforts. What do you plan to do during the next reporting period to accomplish the goals?Within the next reporting period, we anticipate performing the following: Objective 1: Continue to collect samples to examine the diversity of actively circulating A. marginale strains to inform strain selection for prototype anaplasmosis vaccine development design. Use the identified diversity to help refine selection of A. marginale isolates to propogate for evaluation and inclusion in the prototype anaplasmosis vaccine. Work with our partners at SUA to identify and recruit a MS student to investigate diversity of A. marginale in Tanzania. Continue working on methods to streamline A. marginale msp1a genotype identification and analysis via massive parallel sequencing. Objective 2: Continue development of additional serologic-based diagnostic assays to characterize specific anti-A. marginale protein immune responses (this will be important as the currently available experimental vaccine does not appear to induce anti-Msp5 - common diagnostic target). Work on developing assays to evaluate cellular immune response to immunization. Refine protocol for generating sufficient whole killed A. marginale antigen material from stabilates for preparation of the immunogen. Perform pilot immunization with prepared whole killed A. marginale to evaluate immunzation strategy prior to large multi-group and cocktail immunization experiment. Objective 3: Work towards this objective will begin once we confirm we have developed a protective immunogen in Objective 2. Consider alternative immunization strategies if challenges with generating sufficient whole-killed A. marginale material persist. Trainees: Recruit an additional graduate student to help achieve project goals. This project will continue to sponsor training opportunities with undergraduates and DVM students as additional efforts are needed. Dissemination of results: Results will be shared with interested stakeholders in the form of presentations by the PI or trainees at professional meetings and stakeholder meetings (e.g. cattlemen/livestock association meetings).
Impacts
What was accomplished under these goals?
Objective 1Accomplishments:We have recruited an additional ~3,500 cattle samples from diverse locations in the U.S. These samples have been tested by both serologic and molecular assays for A. marginale. The A. marginalemsp1a genotype has been identified from a representative subset of all samples collected last year and about half of the samples collected this year. In addition to traditional genotype analysis procedures, we are working on methodology to streamlinethis process usingindividually barcoded samples and massive parallel sequencing technology. Characterized fivepotential A. marginale strains for inclusion in the prototype vaccine. Objective 2 Accomplishments: We have processed experimentally generated samples and producer-derived field samples for specific antibody response to immunization with this vaccine and protection against homologous and heterologous A. margiale strain challenge (challenge only-perfored in the controlled immunization study). We have also recruited blood samples from producers that elect to use the experimental anaplasmosis vaccine to determine if this vaccine generates a specific anti-Msp5 A. marginale immune response. We are also working to generate novel diagnostic assays against other conserved A. marginale surface proteins to support evaluation of specific anti-A. marginale immune responses (three recombinat A. marginale surface proteins are currently being expressed for this purpose). We are also working on refining methodology to generate a sufficient amount of antigen material for immunzation experiments and supporting diagnostic assays. Objective 3 Accomplishments: Nothing to report for this objective. This objective is dependent on Objective 1 and 2.
Publications
- Type:
Journal Articles
Status:
Submitted
Year Published:
2022
Citation:
Lovett AC, Reppert EJ, Jaeger JR, Kang Q, Flowers MR, Bickmeier NP, Anantatat T, ODay SC, Armstrong CL, Reif KE. Satisfactory breeding potential is transiently eliminated in beef bulls with clinical anaplasmosis. BMC Vet Res. 2022. Oct 29;18(1):381. doi: 10.1186/s12917-022-03470-7.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Curtis AK, Kleinhenz MD, Anantatat T, Martin MS, Magnin GC, Coetzee, Reif KE. Failure to eliminate persistent Anaplasma marginale infection from cattle using labeled doses of chlortetracycline and oxytetracycline antimicrobials. Conference of Research Workers in Animal Diseases. December 3-7, 2021. Chicago, Illinois.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Eggers A, Bickmeier N, Anantatat T, Beltz M, Almaraz J, Huser S, Reif K. Investigating the prevalence of Anaplasma marginale in breeding cows and the impact of chronic anaplasmosis on packed red blood cell volume and pregnancy rates. K-State Food Science Institute Undergraduate Research Symposium Fall 2021. December 14, 2021, Manhattan, KS.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Toillion AR, Reppert EJ, Apley MD, Amachawadi RG, Olson KC, Kleinhenz MD, Coetzee JF, Reif KE, Kang Q. Evaluation of chlortetracycline plasma concentrations relative to anaplasmosis status of naturally endemic beef cattle following free-choice administration of CTC-medicated mineral. K-State CVM Phi Zeta Research Day. Manhattan, KS. March 1, 2022.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
ODay SC, Bickmerier NP, Schmid IJ, Anantatat T?, McCann SM, Reif KE. Protective efficacy of the conditionally licensed anaplasmosis vaccine. K-State CVM Phi Zeta Research Day. Manhattan, KS. March 1, 2022.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
Lovett AC, Reppert EJ, Jaeger JR, Kang Q, Bickmeier NP, Anantatat T, Flowers MR, ODay SC, Armstrong CL, Reif KE. Acute anaplasmosis reduces breeding soundness in experimentally infected beef bulls. K-State CVM Phi Zeta Research Day. Manhattan, KS. March 1, 2022.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2022
Citation:
Lovett A, Reppert EJ, Flowers M, Bickmeier N, Jaeger J, Kang Q, Reif KE. Acute anaplasmosis reduces breeding soundness in experimentally-infected beef bulls. Capital Graduate Student Research Symposium. March 29, 2022. Topeka, Kansas.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Lovett A, Reppert EJ, Flowers M, Bickmeier N, Jaeger J, Kang Q, Reif KE. Acute anaplasmosis reduces breeding soundness in experimentally-infected beef bulls. American Association of Bovine Practitioners. October 7-9, 2021. Salt Lake City, Utah.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Toillion AR, Reppert EJ, Amachawadi RG, Olson KC, Coetzee JF, Kang Q, Reif KE. Effect of antimicrobial-based anaplasmosis control on E. coli chlortetracycline susceptibility from pastured cattle. Conference of Research Workers in Animal Diseases. December 3-7, 2021. Chicago, Illinois.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Flowers M, Anantatat T, Reppert EJ, Reif KE. Evaluation of enrofloxacin and oxytetracycline to eliminate persistent Anaplasma marginale infection in cattle. Conference of Research Workers in Animal Diseases. December 3-7, 2021. Chicago, Illinois.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Krueger SA, Anantatat T, Reif KE. Evaluation of Amblyomma americanum vector competence for Anaplasma marginale. Conference of Research Workers in Animal Diseases. December 3-7, 2021. Chicago, Illinois.
- Type:
Conference Papers and Presentations
Status:
Accepted
Year Published:
2021
Citation:
Bickmeier NP, Flowers MR, Anantatat T, Okafor C, Whitlock B, Strickland, L, Rhinehart J, Carter C, Reif KE. Genetic diversity of Anaplasma marginale in Tennessee beef cattle herds. Conference of Research Workers in Animal Diseases. December 3-7, 2021. Chicago, Illinois.
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Progress 09/01/20 to 08/31/21
Outputs
Target Audience:The initial objective of this studyinvolves recruitment of cattle blood samples from producers to assess actively circulating A. marginale genotypes. Through recruitment of these samples, we engage our primary stakeholder (U.S. cattle producers), informally engaging them broadly about the importance of bovine anaplasmosis, relaying our study objectives, and collecting feedback from them on their perceptions of the impact of this disease at the local level of their operation and more broadly for the U.S. cattle industry. Although our efforts to recruit samples were significantly limited due to COVID-19 restrictions we were able to initiate sample recruitment and are presently continuing these efforts. Changes/Problems:The first year of our project was severely impacted by COVID-19 pandemic regulations. These regulations limited and delayed our ability to recurit samples and fillstaffing roles. We are working towards our study objectives but at a delayed timeline due to pandemic restrictrions present during the majority of the first year of this project. What opportunities for training and professional development has the project provided?This project has provided funding to recruit and provide training for a dedicated doctoral graduate student (Naemi Bickmeier - recruited Jan 2021). Additionally, this study has provided opportunities for training of two undergraduate student research assistants during this evaluation period. Training has also been provided for one Veterinary Summer Research Scholar. Additional students and trainees will be hired during the next reporting period as COVID-19 related hiring restrictions are lifted. How have the results been disseminated to communities of interest?Preliminary results have been shared via presentation at professional meetings. What do you plan to do during the next reporting period to accomplish the goals?Within the next reporting period, we anticipate completing the following: Objective 1: Continue to collect samples to examine the diversity of actively circulatingA.marginalestrains to inform strain selection for prototype anaplasmosis vaccine development. Choose A. marginale isolates to propagate to prepare both vaccine and infection challenge material. Work with our partners at SUA to identify and recruit aMS student that will be perform a survey of A. marginale diversity in Tanzania. Objective 2: Prepare and evaluate prototype vaccine parenterally-administered formulation strategy to ensure that prototype vacccine induces appropriate immune reaction, does not contain viable A. marginale, and is not contaimated with residual red blood cell material. If the aforementioned is achievved, challenge immunized animals with homologus strain and evalute disease process. If successful, begin formulation of novel A. marginale vaccine cocktail informed by A. marginale diversity assessment in Objective 1, parenterally immunize animals with antigen forumlated with different antigens, monitor for appropriate immune response, and then challege with either homologous or heterologous A. marginale strains. If difficulties are encountered in any step, we will perform interative evaluations of the process to complete Objective 2 goals. Objective 3: Work towards this objective will begin once we confirm we have developed a protective immunogen in Objective 2. Trainees: In Year 02, an additional graduate student will be recruited to achieve project goals. Ths project will continue to sponsor training opportunities with undergraduates and DVM students as additional efforts are needed. Dissemination of results: Results will be shared with interested stakeholder in the form of presentations by the PI or trainees at professional meetings and stakeholder meetings (e.g. cattlemen/livestock association meetings).
Impacts
What was accomplished under these goals?
Objective 1 Accomplishments: We have recruited over 3.000 cattle samples and tested them for A. marginale. Positive samples are actively being genotyped to inform genotype selection for vaccine development. The graduate student assigned to this project is developing a high-throughput method of genotyping positive A. marginale samples. Objective 2 Accomplishement: We have performed an initialto evaluation ofthe presently available anaplasmosis vaccine (point of comparison once we develop our novel vaccine). Objective 3 Accomplishements: nothing to report(this objective requires completion of Objectives 1 and 2). (COVID-19 regualtions significantly reduced our ability to complete accomplishing Year 01 goals. We will continue working to complete Year 01 objectives in Year 02 as pandemic restrictions subside.)
Publications
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