Aaron's
Introduction:
            The search for a safe Brucellosis vaccine with sufficient immune activation continues today with no candidates changing the game. Alternative vaccines have focused on presenting antigens such as outer membrane proteins, superoxide dismutases, cell envelopes, and lipopolysaccharides (LPS), but these have failed to gain traction and commercial use in Brucellosis prevention (Saez 2012). As intracellular pathogens, Brucella species rely heavily on their specialized (LPS) to evade host detection, making it a targeted portion of the bacteria. Mutated strains with “rough” LPS have been the source of wide scale vaccination. These live attenuated species of Brucella are safe for cattle 4 months and older, but unsafe for less than 3-month-old calves and pregnant cattle.  (Lalsiamthara 2017). The RB51 strain is infectious to humans, and with modified LPS can elude diagnostic detection. The current system of using an attenuated strain lacks efficiency and dependability.
Our preliminary data [RW1] is based on the possible immune activating capability of a Type V protein named Hia in Brucella Strain RB51, an adhesin homolog of Heamophilus. Our group is using this gene inside a nicin induced expression system (NICE) plasmid present in the Strain Lactococcus lactis. Using a live food grade bacterium with recombinant DNA as a vaccination method is not a new idea, in fact this has been proposed since the late 90s, but has never been implemented as a standard vaccine (Schurig 2002). Our group aims to use Lactococcus lactis as a vector for Brucella antigen, and offer mucosal immunity via a live oral vaccine. L. lactis is a common bacterium that is used to make yogurt, and other lactose products. This food safe probiotic rarely colonizes the host, and would not upset the rumen of the vaccinated animals. The NICE plasmid inside L. lactis will encode the Brucella antigen, along with chloramphenicol resistance, allowing us to test the L. lactis for proper plasmid uptake.  For this to be a vaccine candidate, it must show a willingness to grow in the rumen conditions of the cattle. The chloramphenicol resistance gene, if spread, could be a source for pathogenic antibiotic resistance. This aspect of the vaccine is being evaluated and characterized in depth, ensuring safe passage through the gastrointestinal tract of the vaccinated animal. To be effaceable, the probiotic must retain its genetic information.
The live vaccine in the form of Lactococcos lactis is a possible route to immunity without the risk of fetal abortion or human infection. Current methods require intravenous procedures, and unnecessary risk to those administering. The ability to use an oral vaccine would extend the range of those immunized to possibly wild elk and bison interacting with the cattle. Understanding the role a rumen environment will have on the Strain is required to ensure a stable vector capable of supplying immune response. Growth curves will show the effects a rumen environment and its activity can cause.
 
 
Empirical Model: 
Object, Hypothesis, and Specific Aims:
Methods:
Timeline: 
Frozen Stalk list 
  1. E. coli, Sept 21, 2017
  2. B. subtillus, Sept 21, 2017
  3.    
Scope: Vaccination in ungulates both domesticated and wild (cattle and elk).