• Vaccine Ontology ID: VO_0004004
• Type: Toxoid vaccine
• Adjuvant:
  • VO ID: VO_0000884
• Preparation: The vaccine used was a bacterin-toxoid vaccine prepared from a pure culture of C. botulinum type C, inactivated with formalin and combined with
  adjuvant (Martinez et al., 1999).
• Virulence: (Martinez et al., 1999)
• Description: Botulism, caused by Clostridium botulinum type C, is one of the most important diseases of wild waterfowl. During outbreaks, management usually consists of collection and disposal of sick and dead birds to reduce the amount of carcass material that can act as substrate for further toxin production. Immunization might be used to reduce the risk of re-intoxication in treated waterfowl. Immunization has been used to protect
  pheasants (Phasianus colchicus), broiler chickens, and other birds (Martinez et al., 1999).

• Vaccine Ontology ID: VO_0004084
• Type: Subunit vaccine
• Adjuvant:
  • VO ID: VO_0000127
  • Description: Type C and D toxins provoke botulism in many animal species, including birds. C. botulinum type C organisms have been isolated from the contents of the gastric tract of the carcass and environmental materials such as soil, maggots, food, and/or straw mats. At present, the most widely available vaccine for humans and animals is formalin-inactivated toxoids. Although these are very effective, they are expensive and time-consuming to prepare and are ssomewhat hazardous during detoxification. To solve these problems, a recombinant vaccine has been considered (Arimitsu et al., 2004).
• Preparation: The C. botulinum type C strain, C-Stockholm (C-St), and the type D strain, D-1873, were used for the production and purification of type C and D toxins, respectively (Arimitsu et al., 2004).
• Virulence: (Arimitsu et al., 2004)
• Description: Type C and D toxins provoke botulism in many animal species, including birds. C. botulinum type C organisms have been isolated from the contents of the gastric tract of the carcass and environmental materials such as soil, maggots, food, and/or straw mats. At present, the most widely available vaccine for humans and animals is formalin-inactivated toxoids. Although these are very effective, they are expensive and time-consuming to prepare and are ssomewhat hazardous during detoxification. To solve these problems, a recombinant vaccine has been considered (Arimitsu et al., 2004).

Mouse Response

• Host Strain: ddY
• Vaccination Protocol: Male 6- to 8-week-old ddY strain mice ,purchased from Shimizu Laboratory Supplies Co., Ltd. (Kyoto, Japan), were immunized according to protocol. As a negative control, PBS instead of the antigen was mixed with the adjuvant. Each antigen solution was injected s.c. into the dorsal side of the mice (0.1 ml). At 3 weeks post-immunization, a second immunization was performed. Partial bleeding was performed via the tail vein (mice) at 3 and 5 weeks after the primary immunization, and the specific antibody titers were checked by ELISA and Western blotting tests (Arimitsu et al., 2004).
• Persistence: (Arimitsu et al., 2004)
• Side Effects: None were noted (Arimitsu et al., 2004).
• Efficacy: The mice were challenged with lethal doses of the 16 S toxins. All 5 mice immunized with type C-H chain survived a 10^5 mouse i.p. MLD of C-16 S toxin with no symptoms. However, 4/6 mice challenged with a 10^6 mouse i.p. MLD died, and the 2 surviving mice showed severe botulism. On the other hand, all 5 mice immunized with type D-H chain were completely protected even though they were challenged with a 10^6 mouse i.p. MLD of D-16 S toxin. When the mice that survived the challenge with type C and D toxins were then cross-challenged with 10 mouse i.p. MLD of D and C toxins, respectively, no mice survived.
• Description: Since it appears to be difficult to prepare a large amount of recombinant whole neurotoxin, the study attempted to prepare recombinant HC containing the histidine (His) tag of types C and D, and the vaccine effects were analyzed in mice (Arimitsu et al., 2004).

Ducks Response

• Host Strain: mallards (Anas platyrhynchos) and Northern pintails (anas acuta)
• Vaccination Protocol: Three separate trials were conducted. In Trial I, 60 adult male mallards were divided randomly into two groups of 30. Each bird in the immunized group was injected on the dorsum of the lower neck with 1 ml of vaccine. The 30 control birds were injected with 1 ml of sterile 0.85% saline at the same site. On days 5, 10 and 15 post-immunization, a sub-group of 10 immunized birds and a subgroup of 10 control birds were moved to a separate room and each bird was given approximately 4.5 x 10^4 mouse lethal doses (MLD50) of type C botulinum toxin by gastric intubation. The birds were then observed several times each day. Clinical signs of botulism were recorded as Stage I (bird is able to walk but has paresis or ataxia), Stage II (bird has difficulty walking, often using the wings to assist, but is able to evade capture and can reach food and water), or Stage III (bird is prostrate and paralyzed). Birds in Stage III were euthanized by overexposure to anaesthetic, and a necropsy was performed (Martinez et al., 1999).
• Persistence: In Trials I and II, protection was not evident at 5 days but developed by 10 days pi and appeared to persist for 90 days (Martinez et al., 1999).
• Side Effects: None noted (Martinez et al., 1999)
• Efficacy: The results of Trials I and II confirm that a single dose of toxoid vaccine will provide significant protection to ducks against botulism. Approximately 85% of the immunized mallards and pintails did not have any clinical signs of botulism when challenged at 10 and 15 d, while all of the control birds had clinical botulism (Martinez et al., 1999).
• Description: The purpose of the study was to determine the protection afforded by a single dose of a commercially available vaccine given to ducks under experimental conditions, and to test the effectiveness of simultaneous administration of antitoxin and vaccine to intoxicated birds, as might be done under field conditions (Martinez et al., 1999).

Ducks Response

• Host Strain: The ducks were a cross of Japanese Mallard and Khaki Cambell, male and female, 3 weeks old, and were purchased from the Takahashi Hatching Farm (Osaka, Japan).
• Vaccination Protocol: Male and female 3-week-old ducks (a cross of Japanese Mallard and Khaki Cambell, purchased from the Takahashi Hatching Farm in Osaka, Japan) were immunized with 0.2 ml dorsal injections. As a negative control, PBS instead of the antigen was mixed with the adjuvant. At 3 weeks post-immunization, a second immunization was performed. Partial bleeding was performed via the basilic vein at 3 and 5 weeks after the primary immunization, and the specific antibody titers were checked by ELISA and Western blotting tests (Arimitsu et al., 2004).
• Persistence: (Arimitsu et al., 2004)
• Side Effects: None were noted (Arimitsu et al., 2004).
• Efficacy: All 7 immunized ducks resisted the challenge with 10 duck i.v. MLD, but the survival rate decreased to 5/7 (71.4%) and 4/7 (57.1%) when the birds were challenged with 10^2 and 10^3 duck i.v. MLD (Arimitsu et al., 2004).
• Description: Type C and D toxins provoke botulism in many animal species, including the avian form. In Japan, some farmers have used ducks, named "Aigamo" in Japanese, which are cross strain of Japanese Mallard and Khaki Campbell, for reducing the chemicals in the rice. Young ducks are released into a rice field to exterminate harmful insects or unwanted plants, grow up during the rice crop, and are used as meats after the completion of the harvest. However, a few hundred ducks died of botulism in a certain area of Ishikawa prefecture. These ducks showed symptoms of leg and wing paralysis and became weak and listless. C. botulinum type C organisms were isolated from the contents of the GI tract of the carcass and environmental materials such as soil, maggots, food, and/or straw mats. A study of vaccination in ducks ensued (Arimitsu et al., 2004).