Fusobacterium necrophorum vaccine and method for making such vaccine

Abstract

The present invention relates to a method of producing a vaccine for the prevention of F. necrophorum bacterial infections, comprising isolating the F. necrophorum bacteria from a bovine species, growing the bacteria in a suitable growth medium for a period equal to between about 10 hours and about 18 hours so as to achieve a bacterial population equal to at least 1×10 5 CFU/mL, terminating the growth, and using a whole cell culture to form the vaccine. Additionally, the present invention relates to the vaccine comprised of a killed whole cell population of the F. necrophorum bacteria taken from a bovine. The present invention further relates to a method for preventing foot rot and liver abscesses caused by F. necrophorum bacteria.

What is claimed is:  
     
         1 . A method for producing a vaccine which establishes resistance to  F. necrophorum  bacteria in bovines immunized with said vaccine and substantially prevents pathogenic manifestations of the bacteria, such as foot rot and liver abscesses, comprising: 
 a) growing an isolate of  F. necrophorum  bacteria isolated from a bovine in a suitable growth medium for a period of time equal to between about 10 hours and about 18 hours to form an  F. necrophorum  bacteria culture, with said bacterial culture having a bacterial count population equal to at least 1×10 5  CFU/mL; and    b) terminating growth of said bacteria culture, thereby forming said vaccine.    
     
     
         2 . The method of  claim 1 , wherein said method comprises mixing said vaccine with an amount of diluent to form a vaccine inoculum.  
     
     
         3 . The method of  claim 2 , wherein said diluent is selected from the group consisting of filler, adjuvant and combinations thereof.  
     
     
         4 . The method of  claim 3 , wherein said adjuvant is mixed with said vaccine in amount equal to between 18% and 24% by volume of said vaccine.  
     
     
         5 . The method of  claim 4 , wherein said adjuvant is selected from the group consisting of oils, aluminum salts, polymers, dimethyldeodecylarnmonium bromide, poxvirus proteins, block co-polymers, triterpenoid glycosides, detergents and combinations thereof.  
     
     
         6 . The method of  claim 5 , wherein said oil adjuvant is selected from the group consisting of mineral oils, vegetable oils, animal oils and combinations thereof.  
     
     
         7 . The method of  claim 4 , wherein said adjuvant is a Suprlmm® oil.  
     
     
         8 . The method of  claim 1 , wherein said  F. necrophorum  bacteria are grown at a temperature ranging between about 33° C. and about 40° C.  
     
     
         9 . The method of  claim 1 , wherein Step (b) is accomplished by adding an amount of formaldehyde sufficient to terminate growth of said bacteria culture.  
     
     
         10 . The method of  claim 1 , wherein said  F. necrophorum  bacteria is a biotype A (FNN) strain.  
     
     
         11 . The method of  claim 9 , wherein said formaldehyde solution is added in an amount equal to about 0.4% by volume of said bacteria culture.  
     
     
         12 . The method of  claim 1 , wherein said bacteria is grown for a period of time equal to between 11 hours and 14 hours.  
     
     
         13 . The method of  claim 1 , wherein said bacteria culture has a bacterial count population equal to at least 1×10 8  CFU/mL.  
     
     
         14 . A method of producing a vaccine inoculum for immunizing bovines against a biotype A (FNN)  F. necrophorum  bacterial infection, comprising the steps of isolating a biotype A  F. necrophorum  bacteria from a bovine species member, growing successive generations of said  F. necrophorum  bacteria anaerobically in a broth comprised of brain-heart infusion broth growth medium at a temperature ranging between about 35° C. and about 38° C. for a period of time equal to between about 11 hours and about 14 hours until an optical bacterial cell density equivalent to 0.8 at 540 nm and a volume of at least 1×10 5  CFU/mL is reached, inactivating said bacterial culture by adding to said bacteria culture a formaldehyde solution to form a whole cell harvested bacteria culture, adding to said harvested whole-cell bacterial culture an amount of diluent comprised of an oil-based adjuvant, with said oil based adjuvant added in an amount equal to between 18% and about 24% by volume of said harvested bacterial culture to form said vaccine inoculum.  
     
     
         15 . A vaccine for establishing resistance to  F. necrophorum  bacteria in bovines, when members of the bovine species are inoculated with said vaccine, wherein said vaccine comprises, a whole cell culture of  F. necrophorum , with said culture derived from an isolate taken from a bovine species, with growth of said culture terminated and a last generation of said culture grown for a period of time equal to between about 10 hours and about 18 hours and a time sufficient for said culture to have a CFU count equal to at least 1×10 5  CFU/mL.  
     
     
         16 . The vaccine of  claim 15 , wherein said vaccine is further comprised of an amount of adjuvant added to said whole cell culture in an amount equal to between 18% and 24% by weight of said whole cell culture.  
     
     
         17 . The vaccine of  claim 16 , wherein said adjuvant is an oil based adjuvant.  
     
     
         18 . A method of preventing foot rot and liver abscesses in bovines caused by infection with  F. necrophorum  bacteria, wherein said method is comprised of: 
 a) growing an isolate of  F. necrophorum  bacteria, taken from a bovine species, for successive generations in a suitable growth medium for a period of time equal to between 10 hours and 18 hours to form an  F. necrophorum  bacteria whole cell culture, with said bacteria culture having a bacterial count population equal to at least 1×10 5  CFU/mL;    b) forming a vaccine by combining said bacteria culture with an amount of diluent; and    c) administering at least one dosage of said vaccine intramuscularly or subcutaneously to a bovine subject, with said dosage equal to between 1 and 5 mL.    
     
     
         19 . The method of  claim 18 , wherein two dosages of said vaccine are administered.  
     
     
         20 . A vaccine comprised of: 
 a) a killed biotype A (FNN)  F. necrophorum  isolate taken from a bovine species member of identified as ATCC Accession No. ______; and    b) a suitable carrier for said isolate.