US 12,403,187 B2
Bacterial vaccine
Henning Sørum, Oslo (NO)
Assigned to BELLEVACC AS, Ås (NO)
Appl. No. 17/419,855
Filed by BELLEVACC AS, Ås (NO)
PCT Filed Dec. 20, 2019, PCT No. PCT/EP2019/086639
§ 371(c)(1), (2) Date Jun. 30, 2021,
PCT Pub. No. WO2020/141103, PCT Pub. Date Jul. 9, 2020.
Claims priority of application No. 20181683 (NO), filed on Dec. 31, 2018.
Prior Publication US 2022/0072116 A1, Mar. 10, 2022
Int. Cl. A61K 39/085 (2006.01); A61K 39/02 (2006.01); A61K 39/09 (2006.01); A61P 31/04 (2006.01)
CPC A61K 39/085 (2013.01) [A61K 39/0208 (2013.01); A61K 39/092 (2013.01); A61K 39/107 (2013.01); A61P 31/04 (2018.01)] 16 Claims
OG exemplary drawing
 
1. A method for producing a bacterial vaccine composition against a bacterial disease in a tetrapod and/or a fish, wherein said bacterial vaccine composition comprises inactivated bacteria of the bacterium causing said bacterial disease, said method comprising:
i) preparing cultures A, B, C, D, F, G and H, or preparing all of cultures A to H, wherein:
culture A is prepared by first preparing a pre-culture “a” by inoculating said bacterium in 0.5 to 3.5% NaCl, and incubating before transferring said pre-culture “a” to a nutrient-rich bacterial growth medium with 0.5% to 3.5% NaCl, and incubating under microaerophilic conditions to prepare culture A;
culture B is prepared by first preparing a pre-culture “a” by inoculating said bacterium in 0.5 to 3.5% NaCl and incubating before transferring said pre-culture “a” to cell culture medium for fish or mammalian cells, and incubating under microaerophilic conditions to prepare culture B;
culture C is prepared by first preparing a pre-culture “b” by inoculating said bacterium in a cell culture medium for fish or mammalian cells, and incubating before transferring said pre-culture “b” to a solution typically comprising about 0.5 to 15% (w/v) gelatin with 0.05 to 1.5% glucose and incubating under microaerophilic conditions to prepare culture C;
culture D is prepared by first preparing a pre-culture “c” by inoculating said bacterium in blood, plasma or serum and incubating before transferring said pre-culture “c” to brain heart infusion medium with 0.05 to 1.5% glucose and incubating under microaerophilic conditions to prepare culture D;
culture E is prepared by inoculating said bacterium in enrichment broth and incubating under aerobic conditions to prepare culture E;
culture F is prepared by transferring bacteria from culture E or from enrichment medium containing for example blood, plasma and/or serum to 0.5 to 3.5% NaCl;
culture G is prepared by transferring bacteria from culture E or from enrichment medium containing for example blood, plasma and/or serum, to a cell culture medium for fish or mammalian cells;
culture H is prepared by transferring bacteria from culture E or from enrichment medium containing for example blood, plasma and/or serum to blood or plasma serum;
ii) inactivating the bacteria of the prepared cultures;
iii) optionally washing the inactivated cultures of step ii); and
iv) mixing the optionally washed and resuspended inactivated cultures to prepare the bacterial vaccine composition;
wherein step iv) of mixing optionally may be performed before step ii) or before step iii) instead of after step iii).