Source: LOUISIANA STATE UNIVERSITY submitted to NRP
BRUCELLA ABORTUS STRAIN 19 CAPRINE CHALLENGE MODEL FOR RUMINANT BRUCELLOSIS VACCINE RESEARCH
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
COMPLETE
Funding Source
ANIMAL HEALTH
Reporting Frequency
Annual
Accession No.
0229764
Grant No.
(N/A)
Cumulative Award Amt.
(N/A)
Proposal No.
(N/A)
Multistate No.
(N/A)
Project Start Date
Oct 1, 2012
Project End Date
Jun 30, 2017
Grant Year
(N/A)
Program Code
[(N/A)]- (N/A)
Recipient Organization
LOUISIANA STATE UNIVERSITY
202 HIMES HALL
BATON ROUGE,LA 70803-0100
Performing Department
School of Animal Science
Non Technical Summary
Brucellosis remains a worldwide human and animal health concern, especially in those countries where sheep and goats are a primary food source. The current caprine brucellosis virulent challenge model using select agent Brucella strains has proven to be an effective tool for cost-effective vaccine research. As a zoonotic disease, the best way to minimize the risk of human brucellosis is to reduce the incidence of the animal infection through the effective use of vaccination. However, the current federal select agent regulations which dictate who and how these virulent strains may be used have seriously impeded vaccine development. The ultimate goal of this project would establish a suitable challenge model using non-select agent brucellae, thus equipping researchers with an alternative pathway for vaccine candidate evaluation.
Animal Health Component
35%
Research Effort Categories
Basic
50%
Applied
35%
Developmental
15%
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3113820109060%
3113999110020%
3110899104020%
Knowledge Area
311 - Animal Diseases;

Subject Of Investigation
3999 - Animal research, general; 3820 - Goats, meat, and mohair; 0899 - Wildlife and natural fisheries, general/other;

Field Of Science
1040 - Molecular biology; 1090 - Immunology; 1100 - Bacteriology;
Goals / Objectives
The overall goal of the research is to establish a caprine brucellosis challenge model using Brucella abortus strain 19 as a non-select agent alternative for brucellosis vaccine development for domestic livestock and wildlife. Our hypothesis is that the once widely-used commercial vaccine B. abortus strain 19 (S19) may be used as a suitable virulent challenge to test potential vaccine candidates for protection against colonization and disease. Successful completion of the project would establish a caprine challenge model using non-select agent brucellae, thus equipping researchers with an alternative pathway for vaccine candidate evaluation.
Project Methods
1) Establish a colonization profile of B. abortus S19 in non-pregnant femalegoats from day three to day 42 post conjunctival inoculation 2) Establish the appropriate challenge dose of B. abortus S19 in the gravid goat to ensure colonization of the reproductive tract 3) Measure the protection afforded by the current bovine vaccine B. abortus strain RB51 or B. abortus strain 19 against challenge with B. abortus S19 in non-pregnant female goats (heterologous and homologous challenge) 4) Measure the protection afforded by the current bovine vaccine Brucella abortus strain RB51 or B. abortus strain 19 against challenge with B. abortus S19 in pregnant female goats (heterologous and homologous challenge)

Progress 10/01/12 to 06/30/17

Outputs
Target Audience:The target audience for our research includes federal and state regulatory agencies, livestock and wildlife managers, infectious disease scientists, vaccine manufacturers, and brucellosis researchers worldwide. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Through collaborative studies with other universities and the Department of Defense, the research team is continuously improving their technological and mentoring skills. Undergraduate and graduate students learn about brucellosis and infectious diseases through course work and invited lectures. A Master's student participating in the caprine studies successfully completed his thesis and received his degree. How have the results been disseminated to communities of interest?Research results are shared with collaborators and other brucellosis researchers at state, federal, and international meetings. A Master's thesis and a journal article using brucella media and culturing techniques were published. What do you plan to do during the next reporting period to accomplish the goals? Nothing Reported

Impacts
What was accomplished under these goals? Earlier studies with non-pregnant goats conjunctively challenged with either 7.5 x 10e9 cfu or 7.5 x 10e10 cfu of commercially available Brucella abortus strain 19 demonstrated that the goats were colonized and serological positive on the Rose Bengal Card test. Due to B. abortus Strain 19's ability to infect ruminants other than cattle, we hypothesized that a sufficient dose of vaccine strain could serve as a virulent challenge in pregnant goats. Goats were field bred by proven billies, and pregnancies confirmed by sonogram. Three groups of goats were infected with B. abortus strain 19 at approximately 100 days gestation: four goats received 1x10e9 cfu conjunctively; four goats received 1x10e10 cfu conjunctively; and the control group (5 goats) was infected with 1x10e7 cfu intravenously. None of the pregnant goats challenged with Strain 19 aborted; and no bacteria were recovered from milk, fetal lung, spleen, or abomasal fluid. B. abortus Strain 19 at the doses and routes used would not serve as an adequate virulent challenge in pregnant goats.

Publications

  • Type: Theses/Dissertations Status: Published Year Published: 2017 Citation: McGee, Michael (2017)DESIGN AND HUMORAL ANALYSIS OF TWO EPITOPE-BASED BRUCELLA ABORTUS DNA VACCINES
  • Type: Journal Articles Status: Published Year Published: 2017 Citation: Bastian, F.O., J. Lynch, S. Hagius, X. Wu, G. McCormick, D. Luther, P.H. Elzer. (2017) Novel Spiroplasma spp. Cultured from Brains and Lymph Nodes from Ruminants Affected with Transmissible Spongiform Encephalopathy. J Neuropathol Exp Neurol doi: 10.1093/jnen/nlx 102: 1-10


Progress 10/01/15 to 09/30/16

Outputs
Target Audience:The target audience for this research project includes federal and state regulatory agencies, livestock and wildlife managers, brucellosis and other infectious disease researchers, and vaccine companies. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?A graduate student has been primarily responsible for the colonization study and has learned both laboratory and animal procedures involved in the project. Continued collaboration with other Universities and the Department of Defense has resulted in training of international scientists involved in brucellosis research. Interaction with US collaborators and the foreign researchers increased the expertise of the team. How have the results been disseminated to communities of interest?The project has been discussed with national and international collaborators. An article was published in an institutional magazine and three abstracts were presented at scientific conferences. The project director is involved in state and federal committees that focus on infectious diseases What do you plan to do during the next reporting period to accomplish the goals?The next step of the project is to perform a Brucella abortus strain 19 colonization study in pregnant goats in an attempt to develop a non-select agent vaccine challenge model.

Impacts
What was accomplished under these goals? Non-pregnant goats were conjunctively challenged with one of two doses of commercially available Brucella abortus strain 19 bovine brucellosis vaccine and necropsied at days 14 or day 28 post-infection. Blood and tissues were collected for serological and bacteriological evaluation. Goats receiving 7.5 x 109 cfu exhibited minimal colonization at both time points. The tissues from goats inoculated with 7.5 x 1010 cfu demonstrated higher levels of bacteria at 14 and 28 days post-infection. Sera were analyzed using the Rose Bengal Card test. At 14 days post-infection, 75% of the goats were positive while at 28 days post-infection 100% of the goats reacted positively on the card test.

Publications

  • Type: Other Status: Published Year Published: 2015 Citation: Louisiana Agriculture Magazine. AgCenter Scientists at the forefront of Brucellosis Eradication Vol.58, No. Fall 2015
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Hay, J., C.M. Farris, A. Andrushchenko, P. Elzer, S. Hagius, A. L. Richards, N. Maikanov, T. Ayazbayev. (2015) Biosurveillance for Tick-Borne Encephalitis Virus, Coxiella burnetii and Brucella spp. in Milk in Western Kazakhstan. International Society for Disease Surveillance, Denver, CO., December 9-10, 2015. http://ojphi.org * 8(1):e119, 2016.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Ajazbaev, T., Nina Mikhailyuk, N., Aizhan Yesmagambetova, A., Andryushchenko, A., Maykanov, N., Artemyeva, Y., Hagius, S. and Elzer, P. (2015) Serological evaluation of unpasteurized milk for bovine brucellosis detection in Western Kazakhstan Oblast. 68th Annual Brucellosis Conference, Chicago, IL, Dec 5-6, 2015. Abstract P4, p 43.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Tevdoradze, E., Antadze, I., Gunia, S., Skhirtladze, N., Balarjishvili, N. Kvachadze, L., Kotorashvili, A., Hagius, S., Elzer, P., Farlow, J., Kutateladze, M. (2015) Genetic determinants of homogeneity of Brucella bacteriophages. 68th Annual Brucellosis Conference, Chicago, IL, Dec 5-6, 2015. Abstract P24, p 58.


Progress 10/01/14 to 09/30/15

Outputs
Target Audience:Our target audience includes brucellosis researchers worldwide, federal and state regulatory agencies, livestock and wildlife managers, infectious disease scientists, and vaccine manufacturers. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?Through collaborative studies at other universities and in conjunction with the Department of Defense, training of international scientists in brucellosis serological and bacteriological techniques continues. Graduate and undergraduate students participate in infectious disease and immunology lectures involving brucellosis. A graduate student is being trained to participate in a caprine Brucella colonization and challenge dose study. How have the results been disseminated to communities of interest?Information is disseminated at national and international venues. The project director participates on state and federal committees that involve infectious diseases, especially brucellosis. A journal article was published, and three abstracts were prepared for presentations in December 2015. What do you plan to do during the next reporting period to accomplish the goals?A Brucella abortus Strain 19 colonization study in pregnant and non-pregnant animals will be performed to develop a non-select agent vaccine challenge model.

Impacts
What was accomplished under these goals? The evaluation of antibodies to Brucella in milk samples was continued utilizing ELISAs and Fluorescent Polarization to develop protocols for storage and testing. International scientists were trained in these protocols and assays. Goats have been procured for the Brucella abortus Strain 19 colonization study in pregnant and non-pregnant animals to develop a non-select agent vaccine challenge model.

Publications

  • Type: Journal Articles Status: Published Year Published: 2015 Citation: Sanodze, L, C.T. Bautista, N. Garuchava, S. Chubinidze, E. Tsertsvadze, M. Broladze, N. Chitadze, K. Sidamonidze, S. Tsanava, T. Akhvlediani, R.G. Rivard, R. Mody, M.J. Hepburn, P.H. Elzer, M.P. Nikolich, N. Trapaidze. (2015) Expansion of brucellosis detection in the country of Georgia by screening household members of cases and neighboring community members. BMC Public Health May 2;15(1):459.


Progress 10/01/13 to 09/30/14

Outputs
Target Audience: Our target audience includes brucellosis researchers worldwide as well as federal and state regulatory agencies, livestock and wildlife managers, and infectious disease scientists. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided? Graduate and undergraduate studies participate in infectious disease and immunology lectures involving brucellosis. Through collaborative studies at other universities and in conjunction with the Department of Defense, training of international scientists in brucellosis serological and bacteriological techniques is ongoing. How have the results been disseminated to communities of interest? The laboratory disseminated information at national and international venues. A book chapter and four abstracts were published. What do you plan to do during the next reporting period to accomplish the goals? If goats become available, acolonization study of both pregnant and nonpregnant goats using Brucella abortus Strain19 is planned.

Impacts
What was accomplished under these goals? Further evaluation of Brucella bacteriophages for strain identification and the incorporation of ELISAs and the Fluorescent Polarization Assay increased the laboratory's brucellosis diagnostic and identification capabilities. Problems associated with the detection of antibodies to Brucella in milk samples were studied to formulate the best procedure for storage and testing.

Publications

  • Type: Book Chapters Status: Published Year Published: 2014 Citation: Hagius, S.D., Q. P. Morgan, P.H. Elzer. (2014) Infectious Agents: Brucellosis. In Richard Hopper (Ed), Bovine Reproduction (pp533-540). Ames, IA: John Wiley &Sons, Inc.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Syzdykov ?.S., Kuznetsov ?.N., Yespembetov B.?., Duisenova ?.?., Zubova N.V., Blackburn J.K., Nikolich M.P., Elzer P.H. (2014) Spatial Distribution of Brucella melitensis Biovars in the South of Kazakhstan. ASM Biodefense and Emerging Diseases, Washington, D.C., Jan. 27-29, 2014. Abstract 200E.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Mamisashvili, E., S. Hagius, and P. Elzer. (2014) Milk sample testing by indirect ELISA and Fluorescent Polarization Assay. Brucellosis 2014 International Research Conference, Berlin, Germany, September, 2014. Abstract P60, p 124.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Tevdoradze, E., I. Antadze, S. Gunia, N. Skhirtladze, N. Balarjishvili, L. Kvachadze1, A. Kotorashvili, S. Hagius, P. Elzer, J. Farlow and M. Kutateladze. (2014) Phage-host interaction peculiarities in Brucella bacteriophages. Brucellosis 2014 International Research Conference, Berlin, Germany, September, 2014. Abstract P79, p 143.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2014 Citation: Karibayev, T., I. Sytnik, S. Tyulegenov, R. Seidakhmetova, A. Jailbekova, B. Kozhakova, A. Bizhanova, A. Tursunkulov, M.P. Nikolich, P.H. Elzer and J. Blackburn. (2014) Role of agricultural animals in brucellosis transmission in southern Kazakhstan. Brucellosis 2014 International Research Conference, Berlin, Germany, September, 2014. Abstract P117, p 197.


Progress 10/01/12 to 09/30/13

Outputs
Target Audience: Our target audience includes federal and state regulatory agencies, livestock and wildlife managers, and worldwide infectious disease researchers,especially brucellosis scientists. Changes/Problems: Sheep were used in a collaborative study so those samples were used this year instead of acquiring new goats. What opportunities for training and professional development has the project provided? A visiting scientist spent two weeks in the lab being trained on brucellosis diagnostics. A community college summer student learned various serological techniques used in the laboratory. How have the results been disseminated to communities of interest? The laboratory shared resultsat national and international venues through meetings with fellow scientists and researchers. Publications consisted of a journal publication and nine abstracts for posters or presentations. What do you plan to do during the next reporting period to accomplish the goals? Goats will be injected with strain 19 and will be evaluated bacteriologically and serologically.

Impacts
What was accomplished under these goals? Goats injected with brucella proteins mounted an immune response to the specific proteins but did not react on the classical diagnostic tests; this bodes well for future vaccine studies. Sheep injected with Brucella abortus strain 19 were monitored weekly for their humoral response which will helpmodel the caprine study immunological assays.

Publications

  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Syzdykov, M.S., A. N. Kuznetsov, X. Huang, P.H. Elzer, B. A. Espembetov, S. F. Daulbayeva, J. K. Blackburn, M.P. Nikolich. (2013) Evaluation of spatial patterns of brucellosis in Southern Kazakhstan using GIS technologies. Proc. 94th Annual Meet. Conf. Res. Work. Anim. Dis., Abst. P39.
  • Type: Journal Articles Status: Published Year Published: 2013 Citation: Mamisashvili, E, I. Kracalik, T. Onashvili , L. Kerdzevadze, K. Goginashvili, T. Tigilauri, M.Donduashvili, M. Nikolaishvili, I. Beradze, M. Zakareishvili, M. Kokhreidze, M. Gelashvili, N. Vepkhvadze, S. E. R�cz, P. H. Elzer, M. P. Nikolich, J. K. Blackburn. (2013) Seroprevalence of brucellosis in livestock within three endemic regions of the country of Georgia. Prev Vet Med 110(3-4):554-7.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Zhgenti, E., K. Sidamonidze, M. Zakalashvili,, T. Akhvlediani, , X. Huang, T. Onashvili, , R. Rivard, P.Elzer, M. Nikolich, N. Trapaidze. (2013) Genetic Diversity of Brucella Isolates in Georgia. IMED 2013, Vienna, Austria, Feb. 15-18, 2013.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Trapaidze, N., T. Akhvlediani, L. Sanodze, N. Garuchava, I. Kokaia, M. Nikolich, P. Elzer, R. Mody, CT. Bautista, R. Rivard. (2013) Human brucellosis in Georgia: prevalence and distribution of the infection. IMED 2013, Vienna, Austria, Feb. 15-18, 2013.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Malania, L., M. Grdzelidze, M. Ramishvili, T. Akhvlediani, I. Kokaia, S. Tsanava, P. Imnadze, R. Rivard, P.Elzer, M. Nikolich, N. Trapaidze. (2013) Antibiotic Susceptibility of Human Brucella Isolates in Georgia. ASM Biodefense and Emerging Diseases, Washington, D.C., Feb. 25-27, 2013.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sidamonidze, K., E. Zhgenti1, M. Zakalashvili, L. Malania, M. Ramishvili, M. Grdzelidze, T. Akhvlediani, N. Kokaia, S. Tsanava, X. Huang, T. Onashvili, E. Mamisahsvili, R. Rivard, P. Elzer, M. Nikolich, N. Trapaidze. (2013) Molecular Typing of Human and Animal Brucella Isolates from Georgia. ASM, Denver, CO., May 18-21, 2013
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Omasheva, G., A. Aikimbayev, Sh. Zhandossov, A. Tuleuov, S. Hagius, P. Elzer, M. Nikolich, J. Blackburn. ( 2013). Brucellosis in Kazakhstan. 66th Annual Brucellosis Conference, Chicago, IL, Dec 7-8, 2013. Abstract P10.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Syzdykov, M.S., A. N. Kuznetsov, X. Huang, P.H. Elzer, B. A. Espembetov, S. F. Daulbayeva, J. K. Blackburn, M.P. Nikolich. (2013) Evaluation of spatial patterns of brucellosis in Southern Kazakhstan using GIS technologies. 66th Annual Brucellosis Conference, Chicago, IL, Dec 7-8, 2013. Abstract P11.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sytnik, I., S. Tyulegenov, T. Karibayev,A. Dzhailbekova, A. Shcherbakov, R. Seidakhmetova, A. Abenova, M.Nikolich, P. Elzer, J.K. Blackburn, X.Huang. (2013) Ecology of Brucella Biotypes in Southern Kazakhstan. 66th Annual Brucellosis Conference, Chicago, IL, Dec 7-8, 2013. Abstract P12.
  • Type: Conference Papers and Presentations Status: Published Year Published: 2013 Citation: Sidamonidze, K., E. Zhgeti,W. Su, M. Kokhreidze, N. Vepkhvadze, T. Akhvlediani, R. G. Rivard, P. Elzer, N. Trapaidze1, M. Nikolich. (2013) Implementation of the Bruce-ladder PCR assay for differentiation of Brucella species in the Country of Georgia. 66th Annual Brucellosis Conference, Chicago, IL, Dec 7-8, 2013. Abstract P14.