Stable salmonella vaccine formulations

Abstract

The present disclosure provides formulations, kits, and vaccines directed to immunization of animals against Salmonella. Methods of usng the formulations, kits, and vaccines for protection of avians against one or more species of Salmonella are also provided.

Description

[0001] STABLE SALMONELLA VACCINE FORMULATIONS

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Application Serial No. 63 / 533,770, filed on August 21, 2023, the entire disclosure of which is incorporated herein by reference.

[0004] TECHNICAL FIELD

[0005] The disclosure relates to formulations, kits, and vaccines directed to immunization of animals against Salmonella. In one aspect, the present disclosure provides compositions and methods directed to protection of avians against one or more species of Salmonella.

[0006] BACKGROUND AND SUMMARY OF THE INVENTION

[0007] Salmonellosis is a common and usually fatal avian disease caused by the infection with Salmonella bacteria. Generally, the infection can be transmitted through means such as droppings and saliva when multiple birds gather together, such as at bird feeders. Salmonellosis is of large economic concern in the avian industry because it causes morbidity and mortality in many types of birds.

[0008] Vaccines to reduce the incidence of Salmonella infection are currently available. However, current vaccines could be improved if their availability to industry producers were increased. For instance, a Salmonella vaccine with improved stability would provide a longer duration of shelf life to utilize on avians. Moreover, vaccine formulations that include more doses per vial would provide an easier mechanism to vaccinate animals more quickly and efficiently. Finally, reformulation of vaccines to remove animal products would provide cost savings and an environmentally friendly process to aid in avian welfare. Thus, there exists a need for new vaccines directed to immunization of animals against Salmonella.

[0009] Accordingly, the present disclosure provides formulations and directed to immunization of animals against Salmonella. The exemplary embodiments of the present disclosure include several desirable features. First, the formualitons have improved stability in vials. For instance, embodiments of the present disclosure can provide an improved shelf life of 18-24 months at low temperatures. Second, the formulations are capable of increasing the number of vaccine doses per vial. For instance, as a result of the reduction in filling volume of the vials, vials can include up to 4000 doses. Furthermore, the reduced filing volume can provide a shorter freeze- drying program for the formulation to further improve stability. Moreover, the improved stability can also provide a beneficial release specification for viable bacterial counts. For instance, a reduction in the number of viable bacteria from currently 2.2 * 108CFU / dose per Salmonella strain to between 1.5-1.8 * 108CFU / dose can provide a shelf life of at least 18 months. Finally, the formualitons can advantageously be made without the inclusion of animal- derived products.

[0010] Various aspects of the disclosure are described more fully below. However, different aspects of the disclosure may be implemented in many ways and should not be construed as limited to the aspects set forth herein. The following detailed description is, therefore, not to be taken in a limiting sense.

[0011] BRIEF DESCRIPTION OF THE DRAWINGS

[0012] FIGURE 1 shows an exemplary process for producing the formulations of the present disclosure.

[0013] FIGURE 2 shows stability data for Salmonella Typhimurium in exemplary formulations (i.e., “L35-L54 Sal E 2000ds” and “L35-L57 Sal E 2000ds”) over a 21 month period.

[0014] FIGURE 3 shows stability data for Salmonella Enteritidis in exemplary formulations (i.e., “L35-L54 Sal E 2000ds” and “L35-L57 Sal E 2000ds”) over a 21 month period.

[0015] FIGURE 4 shows the course of viable bacterial count (VBC) for the Salmonella Enteritidis strain in each of three exemplified formulations. The Y-axis displays the CFU in x 108CFU per dose and the X-axis displays the time in months.

[0016] FIGURE 5 shows the course of viable bacterial count (VBC) for the Salmonella Typhimurium strain in each of three exemplified formulations. The Y-axis displays the CFU in x 10sCFU per dose and the X-axis displays the time in months.

[0017] DETAILED DESCRIPTION

[0018] Various embodiments are described herein as follows. In an illustrative aspect, a formulation comprising i) a bacteria composition and ii) a stabilizer composition is provided. In an embodiment, the bacteria composition comprises one or more bacteria of Salmonella serogroup C. Criteria for bacteria included in Salmonella serogroup C are known in the art, for instance serovars that are in subgroup Cl (e.g., the presence of 0:6,7 epitopes) and subgroup C2 (e.g., the presence of 0:6,8 or 0:8 epitopes).

[0019] In an embodiment, the bacteria composition comprises a Salmonella Enteritidis bacteria. In an embodiment, the Salmonella Enteritidis bacteria is a live attenuated Salmonella Enteritidis bacteria. In an embodiment, the live attenuated Salmonella Enteritidis bacteria is strain Sm24 / Rifl2 / Ssq.

[0020] In an embodiment, the bacteria composition comprises a Salmonella Typhimurium bacteria. In an embodiment, the Salmonella Typhimurium bacteria is a live attenuated Salmonella Typhimurium bacteria. In an embodiment, the live attenuated Salmonella Typhimurium bacteria is strain Nal2 / Rif9 / Rtt.

[0021] In an embodiment, the bacteria composition comprises a Salmonella Infantis bacteria. In an embodiment, the Salmonella Infantis bacteria is a live attenuated Salmonella Infantis bacteria. In an embodiment, the live attenuated Salmonella Infantis bacteria is strain Smid 4-22 / Rif2.

[0022] In an embodiment, the bacteria composition is selected from the group consisting of a Salmonella Enteritidis bacteria, a Salmonella Typhimurium bacteria, a Salmonella Infantis bacteria, and any combination thereof. In an embodiment, the bacteria composition comprises a Salmonella Enteritidis bacteria and a Salmonella Typhimurium bacteria. In an embodiment, the bacteria composition comprises a Salmonella Enteritidis bacteria and a Salmonella Infantis bacteria. In an embodiment, the bacteria composition comprises a Salmonella Typhimurium bacteria and a Salmonella Infantis bacteria.

[0023] In an embodiment, the stabilizer composition comprises glycerol. Sources of glycerol are known in the art and glycerol could be used, for instance, for cryopreservation or as matrix former according to the formulations of the present disclosure.

[0024] In an embodiment, the stabilizer composition comprises sucrose. Sources of sucrose are known in the art and sucrose could be used, for instance, for cryopreservation or as matrix former according to the formulations of the present disclosure.

[0025] In an embodiment, the stabilizer composition comprises HEPES (i.e., 4-(2- hydroxyethyl)- 1 -piperazineethanesulfonic acid). HEPES is well known in the art as a zwitterionic sulfonic acid buffering agent. Sources of HEPES are known in the art and HEPES could be used, for instance, as a pH buffer according to the formulations of the present disclosure. In an embodiment, the stabilizer composition comprises glycerol, sucrose, and HEPES. In an embodiment, the stabilizer composition consists essentially of glycerol, sucrose, and HEPES. In an embodiment, the stabilizer composition consists of glycerol, sucrose, and HEPES.

[0026] In an embodiment, the stabilizer composition is substantially free of an animal- derived product. As used herein, the term “substantially free” can refer to a low number or a low concentration, such as less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.1%, and the like. In an embodiment, the stabilizer composition is substantially free of gelatin. In an embodiment, the stabilizer composition is substantially free of soy peptone. In an embodiment, the formulation further comprises sodium hydroxide.

[0027] In an embodiment, the stabilizer composition comprises glycerol at a concentration between 0.25-1.5% (w / w). In an embodiment, the stabilizer composition comprises glycerol at a concentration between 0.25-0.5% (w / w). In an embodiment, the stabilizer composition comprises glycerol at a concentration between 0.5-0.75% (w / w). In an embodiment, the stabilizer composition comprises glycerol at a concentration between 0.75- 1.0% (w / w). In an embodiment, the stabilizer composition comprises glycerol at a concentration between 1.0-1.25% (w / w). In an embodiment, the stabilizer composition comprises glycerol at a concentration between 1.25-1.5% (w / w). In an embodiment, the stabilizer composition comprises glycerol at a concentration between 0.75-1.25% (w / w). In an embodiment, the stabilizer composition comprises glycerol at a concentration between 0.75- 0.90% (w / w). In an embodiment, the stabilizer composition comprises glycerol at a concentration between 0.80-0.90% (w / w).

[0028] In an embodiment, the formulation is lyophilized. In an embodiment, the formulation is present in a vial. In an embodiment, the stabilizer composition comprises glycerol at about 70 mg per vial. In an embodiment, the bacteria composition and the stabilizer composition are present at a ratio of about 1:1.

[0029] In an illustrative aspect, a kit comprising i) a formulation comprising a bacteria composition and a stabilizer composition and ii) a vial is provided.

[0030] In an embodiment, the kit comprises up to 4000 doses of the formulation in the vial. In an embodiment, the formulation is lyophilized. In an embodiment, the kit is configured so that the formulation is stable for at least 18 months. In an embodiment, the stability is measured at a refrigerated temperature. In an embodiment, the stability is measured at a temperature between 2-8 °C. In an embodiment, the kit is configured so that the formulation is stable for at least 24 months. In an embodiment, the stability is measured at a refrigerated temperature. In an embodiment, the stability is measured at a temperature between 2-8 °C.

[0031] In an embodiment, the vial is configured for reconstitution of the formulation up to a volume of 8 mL. In an embodiment, the vial is configured for reconstitution of the formulation up to a volume of 10 mL. In an embodiment, the vial is configured for reconstitution of the formulation up to a volume of 12 mL.

[0032] In an embodiment, the bacteria composition comprises one or more bacteria of Salmonella serogroup C. In an embodiment, the bacteria composition comprises a Salmonella Enteritidis bacteria. In an embodiment, the Salmonella Enteritidis bacteria is a live attenuated Salmonella Enteritidis bacteria. In an embodiment, the live attenuated Salmonella Enteritidis bacteria is strain Sm24 / Rifl2 / Ssq.

[0033] In an embodiment, the bacteria composition comprises a Salmonella Typhimurium bacteria. In an embodiment, the Salmonella Typhimurium bacteria is a live attenuated Salmonella Typhimurium bacteria. In an embodiment, the live attenuated Salmonella Typhimurium bacteria is strain Nal2 / Rif9 / Rtt.

[0034] In an embodiment, the bacteria composition comprises a Salmonella Infantis bacteria. In an embodiment, the Salmonella Infantis bacteria is a live attenuated Salmonella Infantis bacteria. In an embodiment, the live attenuated Salmonella Infantis bacteria is strain Smid 4-22 / Rif2.

[0035] In an embodiment, the bacteria composition is selected from the group consisting of a Salmonella Enteritidis bacteria, a Salmonella Typhimurium bacteria, a Salmonella Infantis bacteria, and any combination thereof. In an embodiment, the bacteria composition comprises a Salmonella Enteritidis bacteria and a Salmonella Typhimurium bacteria. In an embodiment, the bacteria composition comprises a Salmonella Enteritidis bacteria and a Salmonella Infantis bacteria. In an embodiment, the bacteria composition comprises a Salmonella Typhimurium bacteria and a Salmonella Infantis bacteria.

[0036] In an embodiment, the Salmonella Enteritidis bacteria is present between 1 x 108colony forming units (CFU) and 6 x 10sCFU. In an embodiment, the Salmonella Enteritidis bacteria is present between 1 x 10sCFU and 2 x 10sCFU. In an embodiment, the Salmonella Enteritidis bacteria is present between 1.0 x 108CFU and 1.5 x 108CFU. In an embodiment, the Salmonella Enteritidis bacteria is present between 1.5 x 108CFU and 2 x 108CFU. In an embodiment, the Salmonella Enteritidis bacteria is present between 1.5 x 108CFU and 1.7 x 108CFU. In an embodiment, the Salmonella Typhimurium bacteria is present between 1 x 108CFU and 6 x 108CFU. In an embodiment, the Salmonella Typhimurium bacteria is present between 1 x 108CFU and 2 x 108CFU. In an embodiment, the Salmonella Typhimurium bacteria is present between 1.0 x 10sCFU and 1.5 x 108CFU. In an embodiment, the Salmonella Typhimurium bacteria is present between 1.5 x 108CFU and 2 x 108CFU. In an embodiment, the Salmonella Typhimurium bacteria is present between 1.5 x 108CFU and 1.7 x 10sCFU.

[0037] In an embodiment, the Salmonella Infantis bacteria is present between 1 x 108colony forming units (CFU) and 6 x 108CFU. In an embodiment, the Salmonella Infantis bacteria is present between 1 x 108CFU and 2 x 108CFU. In an embodiment, the Salmonella Infantis bacteria is present between 1.0 x 108CFU and 1.5 x 108CFU. In an embodiment, the Salmonella Infantis bacteria is present between 1.5 x 108CFU and 2 x 108CFU. In an embodiment, the Salmonella Infantis bacteria is present between 1.5 x 108CFU and 1.7 x 108CFU.

[0038] In an illustrative aspect, a vaccine comprising the composition or formulation of any of the embodiments of the present disclosure is provided.

[0039] In an illustrative aspect, a method of immunizing an animal comprising the step of providing the vaccine of any of the embodiments of the present disclosure to the animal is provided.

[0040] In an embodiment, the providing is via administration of drinking water of the animal. In an embodiment, the vaccine is reconstituted and placed in drinking water of the animal.

[0041] In an embodiment, the animal is an avian. In an embodiment, the avian is selected from the group consisting of a chicken, a duck, and a turkey. In an embodiment, the avian is a chicken. In an embodiment, the avian is a duck. In an embodiment, the avian is a turkey.

[0042] In an embodiment, the method provides a reduction in fecal excretion of Salmonella serogroup C in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Enteritidis in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Typhimurium in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Infantis in the animal.

[0043] In an embodiment, the method provides a reduction in fecal excretion of Salmonella Enteritidis and Salmonella Typhimurium in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Enteritidis and Salmonella Infantis in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Typhimurium and Salmonella Infantis in the animal.

[0044] In an embodiment, the method provides a reduction in organ colonization of Salmonella serogroup C in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Typhimurium in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Infantis in the animal.

[0045] In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Typhimurium in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Infantis in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Typhimurium and Salmonella Infantis in the animal.

[0046] In an embodiment, the method provides a reduction in organ colonization of Salmonella serogroup C in eggs from the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis in eggs from the animal. In an embodiment, the method provides a reduction in colonization of Salmonella Typhimurium in eggs from the animal. In an embodiment, the method provides a reduction in colonization of Salmonella Infantis in eggs from the animal.

[0047] In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Typhimurium in eggs from the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Infantis in eggs from the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Typhimurium and Salmonella Infantis in eggs from the animal.

[0048] In an illustrative aspect, a method of administering the vaccine to an animal is provided, wherein the vaccine is capable of inducing an immune response in the animal after at least 18 months from the manufacturing date of the vaccine.

[0049] In an embodiment, the administering is via administration of drinking water of the animal. In an embodiment, the vaccine is reconstituted and placed in drinking water of the animal.

[0050] In an embodiment, the animal is an avian. In an embodiment, the avian is selected from the group consisting of a chicken, a duck, and a turkey. In an embodiment, the avian is a chicken. In an embodiment, the avian is a duck. In an embodiment, the avian is a turkey.

[0051] In an embodiment, the method provides a reduction in fecal excretion of Salmonella serogroup C in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Enteritidis in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Typhimurium in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Infantis in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Enteritidis and Salmonella Typhimurium in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Enteritidis and Salmonella Infantis in the animal. In an embodiment, the method provides a reduction in fecal excretion of Salmonella Typhimurium and Salmonella Infantis in the animal.

[0052] In an embodiment, the method provides a reduction in organ colonization of Salmonella serogroup C in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Typhimurium in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Infantis in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Typhimurium in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Infantis in the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Typhimurium and Salmonella Infantis in the animal.

[0053] In an embodiment, the method provides a reduction in organ colonization of Salmonella serogroup C in eggs from the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis in eggs from the animal. In an embodiment, the method provides a reduction in colonization of Salmonella Typhimurium in eggs from the animal. In an embodiment, the method provides a reduction in colonization of Salmonella Infantis in eggs from the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Typhimurium in eggs from the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Infantis in eggs from the animal. In an embodiment, the method provides a reduction in organ colonization of Salmonella Typhimurium and Salmonella Infantis in eggs from the animal. In an embodiment, the immune response is a protective immune response. As used herein, a protective immune response can refer to cell mediated and / or humoral (antibody) mediated immune response that will prevent or ameliorate a disease or infection in an animal. In an embodiment, the protective immune response is against Salmonella serogroup C. In an embodiment, the protective immune response is against Salmonella Enteritidis. In an embodiment, the protective immune response is against Salmonella Typhimurium. In an embodiment, the protective immune response is against Salmonella Infantis. In an embodiment, the protective immune response is against Salmonella Enteritidis and Salmonella Typhimurium. In an embodiment, the protective immune response is against Salmonella Enteritidis and Salmonella Infantis. In an embodiment, the protective immune response is against Salmonella Typhimurium and Salmonella Infantis.

[0054] In an embodiment, the vaccine is capable of inducing an immune response in the animal after at least 21 months from the manufacturing date of the vaccine. In an embodiment, the vaccine is capable of inducing an immune response in the animal after at least 24 months from the manufacturing date of the vaccine. In an embodiment, the vaccine is stored at room temperature. As used herein, room temperature generally refers to a temperature from about 20° C to about 25° C. In an embodiment, the vaccine is stored at refrigerated temperature.

[0055] The following numbered embodiments are contemplated and are non-limiting:

[0056] 1. A formulation comprising i) a bacteria composition and ii) a stabilizer composition.

[0057] 2. The formulation of clause 1 , any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises one or more bacteria of Salmonella serogroup C.

[0058] 3. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Enteritidis bacteria.

[0059] 4. The formulation of clause 4, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Enteritidis bacteria is a live attenuated Salmonella Enteritidis bacteria.

[0060] 5. The formulation of clause 5, any other suitable clause, or any combination of suitable clauses, wherein the live attenuated Salmonella Enteritidis bacteria is strain Sm24 / Rifl2 / Ssq. 6. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Typhimurium bacteria.

[0061] 7. The formulation of clause 6, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Typhimurium bacteria is a live attenuated Salmonella Typhimurium bacteria.

[0062] 8. The formulation of clause 7, any other suitable clause, or any combination of suitable clauses, wherein the live attenuated Salmonella Typhimurium bacteria is strain Nal2 / Rif9 / Rtt.

[0063] 9. The formulation of clause 1 , any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Inf antis bacteria.

[0064] 10. The formulation of clause 9, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Infantis bacteria is a live attenuated Salmonella Infantis bacteria.

[0065] 11. The formulation of clause 10, any other suitable clause, or any combination of suitable clauses, wherein the live attenuated Salmonella Infantis bacteria is strain Smid 4- 22 / Rif2.

[0066] 12. The formulation of clause 1 , any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition is selected from the group consisting of a Salmonella Enteritidis bacteria, a Salmonella Typhimurium bacteria, a Salmonella Infantis bacteria, and any combination thereof.

[0067] 13. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Enteritidis bacteria and a Salmonella Typhimurium bacteria.

[0068] 14. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Enteritidis bacteria and a Salmonella Infantis bacteria.

[0069] 15. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Typhimurium bacteria and a Salmonella Infantis bacteria.

[0070] 16. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol.

[0071] 17. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises sucrose. 18. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises HEPES.

[0072] 19. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol, sucrose, and HEPES.

[0073] 20. The formulation of clause 1 , any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition consists essentially of glycerol, sucrose, and HEPES.

[0074] 21. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition consists of glycerol, sucrose, and HEPES.

[0075] 22. The formulation of clause 1 , any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition is substantially free of an animal-derived product.

[0076] 23. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition is substantially free of gelatin.

[0077] 24. The formulation of clause 1 , any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition is substantially free of soy peptone.

[0078] 25. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the formulation further comprises sodium hydroxide.

[0079] 26. The formulation of clause 1 , any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol at a concentration between 0.25-1.5% (w / w).

[0080] 27. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol at a concentration between 0.25-0.5% (w / w).

[0081] 28. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol at a concentration between 0.5-0.75% (w / w).

[0082] 29. The formulation of clause 1 , any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol at a concentration between 0.75-1.0% (w / w).

[0083] 30. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol at a concentration between 1.0-1.25% (w / w). 31. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol at a concentration between 1.25-1.5% (w / w).

[0084] 32. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol at a concentration between 0.75-1.25% (w / w).

[0085] 33. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol at a concentration between 0.75-0.90% (w / w).

[0086] 34. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol at a concentration between 0.80-0.90% (w / w).

[0087] 35. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the formulation is lyophilized.

[0088] 36. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the formulation is present in a vial.

[0089] 37. The formulation of clause 36, any other suitable clause, or any combination of suitable clauses, wherein the stabilizer composition comprises glycerol at about 70 mg per vial.

[0090] 38. The formulation of clause 1, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition and the stabilizer composition are present at a ratio of about 1:1.

[0091] 39. A kit comprising i) a formulation comprising a bacteria composition and a stabilizer composition and ii) a vial.

[0092] 40. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the kit comprises up to 4000 doses of the formulation in the vial.

[0093] 41. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the formulation is lyophilized.

[0094] 42. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the kit is configured so that the formulation is stable for at least 18 months.

[0095] 43. The kit of clause 42, any other suitable clause, or any combination of suitable clauses, wherein the stability is measured at a refrigerated temperature.

[0096] 44. The kit of clause 42, any other suitable clause, or any combination of suitable clauses, wherein the stability is measured at a temperature between 2-8 °C. 45. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the kit is configured so that the formulation is stable for at least 24 months.

[0097] 46. The kit of clause 45, any other suitable clause, or any combination of suitable clauses, wherein the stability is measured at a refrigerated temperature.

[0098] 47. The kit of clause 45, any other suitable clause, or any combination of suitable clauses, wherein the stability is measured at a temperature between 2-8 °C.

[0099] 48. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the vial is configured for reconstitution of the formulation up to a volume of 8 mL.

[0100] 49. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the vial is configured for reconstitution of the formulation up to a volume of 10 mL.

[0101] 50. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the vial is configured for reconstitution of the formulation up to a volume of 12 mL.

[0102] 51. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises one or more bacteria of Salmonella serogroup C.

[0103] 52. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Enteritidis bacteria.

[0104] 53. The kit of clause 52, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Enteritidis bacteria is a live attenuated Salmonella Enteritidis bacteria.

[0105] 54. The kit of clause 53, any other suitable clause, or any combination of suitable clauses, wherein the live attenuated Salmonella Enteritidis bacteria is strain Sm24 / Rifl2 / Ssq.

[0106] 55. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Typhimurium bacteria.

[0107] 56. The kit of clause 55, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Typhimurium bacteria is a live attenuated Salmonella Typhimurium bacteria.

[0108] 57. The kit of clause 56, any other suitable clause, or any combination of suitable clauses, wherein the live attenuated Salmonella Typhimurium bacteria is strain Nal2 / Rif9 / Rtt.

[0109] 58. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Infantis bacteria. The kit of clause 58, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Infantis bacteria is a live attenuated Salmonella Infantis bacteria. The kit of clause 59, any other suitable clause, or any combination of suitable clauses, wherein the live attenuated Salmonella Infantis bacteria is strain Smid 4-22 / Rif2. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition is selected from the group consisting of a Salmonella Enter! tidis bacteria, a Salmonella Typhimurium bacteria, a Salmonella Infantis bacteria, and any combination thereof. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Enteritidis bacteria and a Salmonella Typhimurium bacteria. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Enteritidis bacteria and a Salmonella Infantis bacteria. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the bacteria composition comprises a Salmonella Typhimurium bacteria and a Salmonella Infantis bacteria. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Enteritidis bacteria is present between 1 x 108colony forming units (CFU) and 6 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Enteritidis bacteria is present between 1 x 108CFU and 2 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Enteritidis bacteria is present between 1.0 x 108CFU and 1.5 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Enteritidis bacteria is present between 1.5 x 10sCFU and 2 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Enteritidis bacteria is present between 1.5 x 108CFU and 1.7 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Typhimurium bacteria is present between 1 x 108CFU and 6 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Typhimurium bacteria is present between 1 x 108CFU and 2 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Typhimurium bacteria is present between 1.0 x 108CFU and 1.5 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Typhimurium bacteria is present between 1.5 x 108CFU and 2 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Typhimurium bacteria is present between 1.5 x 108CFU and 1.7 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Infantis bacteria is present between 1 x 108colony forming units (CFU) and 6 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Infantis bacteria is present between 1 x 108CFU and 2 x 10sCFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Infantis bacteria is present between 1.0 x 108CFU and 1.5 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Infantis bacteria is present between 1.5 x 108CFU and 2 x 108CFU. The kit of clause 39, any other suitable clause, or any combination of suitable clauses, wherein the Salmonella Infantis bacteria is present between 1.5 x 10sCFU and 1.7 x 108CFU. A vaccine comprising the composition or formulation of any one of clauses 1 to 38. A method of immunizing an animal comprising the step of providing the vaccine of clause 80 to the animal. 82. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the providing is via administration of drinking water of the animal.

[0110] 83. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the vaccine is reconstituted and placed in drinking water of the animal.

[0111] 84. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the animal is an avian.

[0112] 85. The method of clause 84, any other suitable clause, or any combination of suitable clauses, wherein the avian is selected from the group consisting of a chicken, a duck, and a turkey.

[0113] 86. The method of clause 84, any other suitable clause, or any combination of suitable clauses, wherein the avian is a chicken.

[0114] 87. The method of clause 84, any other suitable clause, or any combination of suitable clauses, wherein the avian is a duck.

[0115] 88. The method of clause 84, any other suitable clause, or any combination of suitable clauses, wherein the avian is a turkey.

[0116] 89. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella serogroup C in the animal.

[0117] 90. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Enteritidis in the animal.

[0118] 91. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Typhimurium in the animal.

[0119] 92. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Infantis in the animal.

[0120] 93. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Enteritidis and Salmonella Typhimurium in the animal.

[0121] 94. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Enteritidis and Salmonella Infantis in the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Typhimurium and Salmonella Infantis in the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella serogroup C in the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis in the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Typhimurium in the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Infantis in the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Typhimurium in the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Infantis in the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Typhimurium and Salmonella Infantis in the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella serogroup C in eggs from the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis in eggs from the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in colonization of Salmonella Typhimurium in eggs from the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in colonization of Salmonella Infantis in eggs from the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Typhimurium in eggs from the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Infantis in eggs from the animal. The method of clause 81, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Typhimurium and Salmonella Infantis in eggs from the animal. A method of administering the vaccine of clause 80 to an animal, wherein the vaccine is capable of inducing an immune response in the animal after at least 18 months from the manufacturing date of the vaccine. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the administering is via administration of drinking water of the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the vaccine is reconstituted and placed in drinking water of the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the animal is an avian. The method of clause 114, any other suitable clause, or any combination of suitable clauses, wherein the avian is selected from the group consisting of a chicken, a duck, and a turkey. The method of clause 114, any other suitable clause, or any combination of suitable clauses, wherein the avian is a chicken. The method of clause 114, any other suitable clause, or any combination of suitable clauses, wherein the avian is a duck. The method of clause 114, any other suitable clause, or any combination of suitable clauses, wherein the avian is a turkey. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella serogroup C in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Enteritidis in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Typhimurium in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Infantis in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Enteritidis and Salmonella Typhimurium in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Enteritidis and Salmonella Infantis in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in fecal excretion of Salmonella Typhimurium and Salmonella Infantis in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella serogroup C in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Typhimurium in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Infantis in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Typhimurium in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Infantis in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Typhimurium and Salmonella Infantis in the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella serogroup C in eggs from the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis in eggs from the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in colonization of Salmonella Typhimurium in eggs from the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in colonization of Salmonella Infantis in eggs from the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Typhimurium in eggs from the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis and Salmonella Infantis in eggs from the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the method provides a reduction in organ colonization of Salmonella Typhimurium and Salmonella Infantis in eggs from the animal. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the immune response is a protective immune response. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the protective immune response is against Salmonella serogroup C. 141. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the protective immune response is against Salmonella Enteritidis.

[0122] 142. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the protective immune response is against Salmonella Typhimurium.

[0123] 143. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the protective immune response is against Salmonella Infantis.

[0124] 144. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the protective immune response is against Salmonella Enteritidis and Salmonella Typhimurium.

[0125] 145. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the protective immune response is against Salmonella Enteritidis and Salmonella Infantis.

[0126] 146. The method of clause 139, any other suitable clause, or any combination of suitable clauses, wherein the protective immune response is against Salmonella Typhimurium and Salmonella Infantis.

[0127] 147. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the vaccine is capable of inducing an immune response in the animal after at least 21 months from the manufacturing date of the vaccine.

[0128] 148. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the vaccine is capable of inducing an immune response in the animal after at least 24 months from the manufacturing date of the vaccine.

[0129] 149. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the vaccine is stored at room temperature.

[0130] 150. The method of clause 110, any other suitable clause, or any combination of suitable clauses, wherein the vaccine is stored at refrigerated temperature.

[0131] EXAMPLE 1

[0132] Exemplary Process for Making Formulations

[0133] Exemplary formulations according to the present disclosure can be made according to the process shown in Figure 1. For instance, starting with a working Salmonella bacteria frozen culture, bacertia strains can be grown and processed according to the steps shown in Figure 1. For the instant example, a formulation using a stabilizer composition comprising glycerol, sucrose, and HEPES was prepared. First, the stabilizer composition was made by combining glycerol, sucrose, and HEPES. Then, concentrated fermenter broth comprising Salmonella strain(s) was added to the stabilizer composition. For the instant example, both Salmonella enteritidis strains were utilized.

[0134] The exemplary formulation was prepared as follows: i. HEPES and glycerol were added to sucrose solution and sterilized using a 0.2pm filter to form the stabilizer composition. The sterilized composition was directly transferred to the formulation tank. ii. Upon transfer, the formulation tank maintained stirring at lOOrpm. iii. After the stabilizer composition was transferred, stirring is continued for 5 minutes. iv. Approximately 200+0.5ml of a sample was drawn from the resultant mixture. From this sample, two further samples of 90+0.5ml were prepared and underwent sterility testing. v. The formulation tank was connected to the filtration skid, and the transfer line was steam sterilized for 30 minutes. After filtration was complete, the concentrate was transferred to the formulation tank (around 1 minute duration). vi. Stirring speed was increased to 350+25 rpm / minute. The formulation tank pH was brought to and maintained at 7.2+0.2 using NaOH as required. vii. Once the pH was achieved, stirring was maintained at > lOOrpm.

[0135] The process resulted in the following exemplary formulation shown in Table 1:

[0136] Table 1

[0137] The foundation process was considered to be robust if a processing time of two hours was not exceeded. It was determined that the proportion of concentrate and stabilizer composition could vary by up to 5% and after concentrating, the concentrate was harvested into a cooled tank for formulation. Alternativly, the stabilizer composition could be added to the concentrate while still circulating in the microfiltration mixing to reduce potential harvesting losses.

[0138] Table 2 shows exemplary formulation ranges for a typical 1+1 (50% :50%) ratio of formulation / mixing considering variability of the concentrate and stabilizer composition proportions.

[0139] Table 2 The volumetric formulation and mixing of concentrate and stabilizer composition takes place at temperatures between 4°C up to 25 °C.

[0140] In regards to freeze-drying, a higher biomass will result in a longer reconstitution time. For instance, initial slower freezing rates will lead to longer reconstitution times. Similarly, faster freezing ramps to faster reconstiution times.

[0141] If higher biomasses are targeted, the freezing ramp can be adapted to the change in biomass. In addition, higher biomass bulks result in lower residual moistures and, with the increase of biomass, the appearance of the cake increases positively. This can result in a better resistance against possible effects of glycerol, thus allowing the use of higher freeze-drying temperatures (e.g., an accleration of freeze-drying program).

[0142] EXAMPLE 2

[0143] Stability Evaluation

[0144] In the instant example, the stability of two exemplary formulations was evaluated. An exemplary formulation comprising Salmonella Typhimurium and Salmonella Enteritidis was evaluated. The exemplary formulatons comprised glycerol, sucrose, and HEPES and a vial containing 2000 doses was evaluated. The stability of the formulations was evaluated by measuring the CFU / dose of the Salmonella strains in the vials over a 21 month period. Without being bound by any theory, it is believed that the Salmonella strains should have a CFU / dose of at least 1 * 108for minimum release to provide vaccine efficacy.

[0145] Figure 2 shows stability data for Salmonella Typhimurium in the exemplary formulations (i.e., “L35-L54 Sal E 2000ds” and “L35-L57 Sal E 2000ds”). Figure 3 shows stability data for Salmonella Enteritidis in the exemplary formulations.

[0146] EXAMPLE 3

[0147] Analyses of Exemplary Formulations

[0148] Exemplary formulations according to the present disclosure can be prepared and analyzed. The instant example details the particulars for three exemplary formulations (F0261, F0554, and F0641) of the present disclosure. Over 7,000 vials were produced for each of the three exemplary formulations. Each vial was filled to a volume of 10 ml and included 4000 doses.

[0149] For each exemplary formulation, an ongoing stability examination was conducted over a period of up to 24 months. In particular, the stability of the formulations was evaluated by measuring the CFU / dose of the Salmonella Typhimurium and Salmonella Enteritidis strains in the vials. Residual moisture was also evaluated.

[0150] Table 3 shows the evaluation of the three exemplary formulations (F0261, F0554, and F0641).

[0151] Table 3 EXAMPLE 4

[0152] Stability Eylaution of Salmonella Strains in Exemplary Formulations

[0153] Exemplary formulations according to the present disclosure can be prepared and the stability of the various Salmonella can be analyzed. The instant example details the particulars for three exemplary formulations (F0261, F0554, and F0641) of the present disclosure. For each exemplary formulation, a model calculated the viable bacterial count (VBC) for each of Salmonella Enteritidis and Salmonella Typhimurium strains over a period for up to 21 months.

[0154] Table 4 shows the calculated slope (VBC) for Salmonella Enteritidis using an In model for a 21 month period. The caclualted slopes for each of the three exemplary formulations (F0261, F0554, and F0641) are shown to demonstrate the change in VBC per month.

[0155] Table 4.

[0156] Further, Figure 4 shows the course of VBC for the Salmonella Enteritidis strain in each of the three exemplified formulations.

[0157] Table 5 shows the calculated slope (VBC) for Salmonella Typhimurium using an In model for a 21 month period. The caclualted slopes for each of the three exemplary formulations (F0261, F0554, and F0641) are shown to demonstrate the change in VBC per month. Table 5.

[0158] Further, Figure 5 shows the course of VBC for the Salmonella Typhimurium strain in each of the three exemplified formulations.

Claims

WHAT IS CLAIMED IS:

1. A formulation comprising i) a bacteria composition and ii) a stabilizer composition, wherein the bacteria composition comprises one or more bacteria of Salmonella serogroup C.

2. The formulation of claim 1, wherein the bacteria composition comprises a Salmonella Enteritidis bacteria.

3. The formulation of claim 2, wherein the Salmonella Enteritidis bacteria is a live attenuated Salmonella Enteritidis bacteria.

4. The formulation of claim 3, wherein the live attenuated Salmonella Enteritidis bacteria is strain Sm24 / Rifl2 / Ssq.

5. The formulation of claim 1, wherein the bacteria composition comprises a Salmonella Typhimurium bacteria.

6. The formulation of claim 5, wherein the Salmonella Typhimurium bacteria is a live attenuated Salmonella Typhimurium bacteria.

7. The formulation of claim 6, wherein the live attenuated Salmonella Typhimurium bacteria is strain Nal2 / Rif9 / Rtt.

8. The formulation of claim 1, wherein the bacteria composition comprises a Salmonella Infantis bacteria.

9. The formulation of claim 8, wherein the Salmonella Infantis bacteria is a live attenuated Salmonella Infantis bacteria.

10. The formulation of claim 9, wherein the live attenuated Salmonella Infantis bacteria is strain Smid 4-22 / Rif2.

11. The formulation of claim 1 , wherein the bacteria composition is selected from the group consisting of a Salmonella Enteritidis bacteria, a Salmonella Typhimurium bacteria, a Salmonella Infantis bacteria, and any combination thereof.

12. The formulation of claim 1, wherein the bacteria composition comprises a Salmonella Enteritidis bacteria and a Salmonella Typhimurium bacteria.

13. The formulation of claim 1, wherein the bacteria composition comprises a Salmonella Enteritidis bacteria and a Salmonella Infantis bacteria.

14. The formulation of claim 1, wherein the bacteria composition comprises a Salmonella Typhimurium bacteria and a Salmonella Infantis bacteria.

15. The formulation of claim 1, wherein the stabilizer composition comprises glycerol.

16. The formulation of claim 1, wherein the stabilizer composition comprises sucrose.

17. The formulation of claim 1, wherein the stabilizer composition comprisesHEPES.

18. The formulation of claim 1 , wherein the stabilizer composition comprises glycerol, sucrose, and HEPES.

19. The formulation of claim 1, wherein the stabilizer composition is substantially free of an animal-derived product.

20. The formulation of claim 1, wherein the stabilizer composition is substantially free of gelatin.

21. The formulation of claim 1, wherein the stabilizer composition is substantially free of soy peptone.

22. The formulation of claim 1, wherein the formulation is lyophilized.

23. The formulation of claim 1, wherein the bacteria composition and the stabilizer composition are present at a ratio of about 1:1.

24. A vaccine comprising the formulation of any one of claims 1 to 23.

25. A method of immunizing an animal comprising the step of providing the vaccine of claim 24 to the animal.

26. The method of claim 25, wherein the providing is via administration of drinking water of the animal.

27. The method of claim 25, wherein the vaccine is reconstituted and placed in drinking water of the animal.

28. The method of claim 25, wherein the animal is an avian.

29. The method of claim 28, wherein the avian is selected from the group consisting of a chicken, a duck, and a turkey.

30. The method of claim 28, wherein the avian is a chicken.

31. The method of claim 25, wherein the method provides a reduction in fecal excretion of Salmonella serogroup C in the animal.

32. The method of claim 25, wherein the method provides a reduction in fecal excretion of Salmonella Enteritidis in the animal.

33. The method of claim 25, wherein the method provides a reduction in fecal excretion of Salmonella Typhimurium in the animal.

34. The method of claim 25, wherein the method provides a reduction in fecal excretion of Salmonella Infantis in the animal.

35. The method of claim 25, wherein the method provides a reduction in organ colonization of Salmonella serogroup C in the animal.

36. The method of claim 25, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis in the animal.

37. The method of claim 25, wherein the method provides a reduction in organ colonization of Salmonella Typhimurium in the animal.

38. The method of claim 25, wherein the method provides a reduction in organ colonization of Salmonella Infantis in the animal.

39. The method of claim 25, wherein the method provides a reduction in organ colonization of Salmonella serogroup C in eggs from the animal.

40. The method of claim 25, wherein the method provides a reduction in organ colonization of Salmonella Enteritidis in eggs from the animal.

41. The method of claim 25, wherein the method provides a reduction in colonization of Salmonella Typhimurium in eggs from the animal.

42. The method of claim 25, wherein the method provides a reduction in colonization of Salmonella Infantis in eggs from the animal.

43. A method of administering the vaccine of claim 24 to an animal, wherein the vaccine is capable of inducing an immune response in the animal after at least 18 months from the manufacturing date of the vaccine.

44. The method of claim 43, wherein the animal is an avian.

45. The method of claim 44, wherein the avian is selected from the group consisting of a chicken, a duck, and a turkey.

46. The method of claim 44, wherein the avian is a chicken.

47. The method of claim 43, wherein the immune response is a protective immune response.

48. The method of claim 47, wherein the protective immune response is against Salmonella serogroup C.

49. The method of claim 47, wherein the protective immune response is against Salmonella Enteritidis.

50. The method of claim 47, wherein the protective immune response is against Salmonella Typhimurium.

51. The method of claim 47, wherein the protective immune response is against Salmonella Infantis.

52. The method of claim 43, wherein the vaccine is capable of inducing an immune response in the animal after at least 21 months from the manufacturing date of the vaccine.

53. The method of claim 43, wherein the vaccine is capable of inducing an immune response in the animal after at least 24 months from the manufacturing date of the vaccine.

54. The method of claim 43, wherein the vaccine is stored at room temperature.

55. The method of claim 43, wherein the vaccine is stored at refrigerated temperature.

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