Evaluation of Immunogenicity of Clostridium perfringens Type B Toxoid and Inactivated FMD (O) Virus with Adjuvant (ISA70-MF59).

Foot and mouth disease (FMD) and enterotoxemia are important diseases of hoofed animals. Vaccination against livestock pathogens, especially these two diseases, plays a key role in the prevention and control of these diseases. The use of combined vaccines with the aim of creating a better immune response and producing cheaper vaccines is a great contribution to Vaccine industry. This research aimed to compare the immunogenicity of FMD (O) and Clostridium perfringens type B toxoid along with adjuvant (MF59) and Montanide (ISA70) to create the best immunogenicity. To investigate the immune responses of vaccines, it was injected into an animal model, and the antibody titer was measured by enzyme-linked immunosorbent assay (ELISA) test and VN antibody titer. The results showed that the formulation with MF59 adjuvant brought more stable immunogenicity against FMD and Clostridium perfringens type B, and the length of the immunogenicity period also increased significantly. Therefore, the combined vaccine (Clostridium perfringens + FMD) could play a major role invaccine industry as an alternative vaccine against Clostridium perfringens and FMD in livestock.

Trials for preparation and evaluation of a combined inactivated reassorted H5N1 and Escherichia coli O157 vaccine in poultry.

BACKGROUND AND AIM: Avian influenza (AI), which is one of the major respiratory diseases of poultry, and Escherichia coli (E. coli) have caused major economic losses around the world, including in Egypt. Therefore, in this study, we aimed to produce a vaccine from E. coli O157 and AI H5N1 formulated with Montanide ISA70 for the protection of poultry against both diseases. MATERIALS AND METHODS: We divided one hundred 3-week-old chicks into four groups: Group 1 was vaccinated with prepared inactivated AI H5N1formulated with Montanide ISA70, Group 2 was vaccinated with inactivated E. coli formulated with Montanide ISA70, Group 3 was vaccinated with combined E. coli and AI H5N1 formulated with Montanide ISA70, and Group 4 was an unvaccinated control group. We measured the immune response using the HI (hemagglutination inhibition) test, enzyme-linked immunosorbent assay (ELISA), and the challenge test. RESULTS: We found the three vaccines to be safe and sterile during all periods of examination and observation. The HI test showed that Group 1 exhibited specific antibody titers of 2.3 log2, 4.3 log2, 7.5 log2, 7.8 log2, 8 log2, and 8.1 log2 from week 2 to week 7, respectively, post-vaccination. Group 3 exhibited antibody titers of 3.3 log2, 5.8 log2, 7.8 log2, 8 log2, 8.3 log2, and 8.3 log2 from week 2 to week 7, respectively, post-vaccination. The immune response in both groups reached a high titer at week 6. The combined inactivated E. coli and AI H5N1 vaccine generated a higher immune response than the inactivated AI H5N1 vaccine, and a significant difference exists between the two groups. For Groups 2 and 3, the ELISA antibody titer exhibited its lowest value, 1996.5 and 2036.7, respectively, at week 1 post-vaccination; whereas, both groups exhibited the highest titers, 2227.7 (for Group 2) and 2287.3 (for Group 3), in week 3 post-booster. The ELISA for the combined inactivated E. coli and AI H5N1 vaccine had a higher titer than did the inactivated E. coli vaccine, and a significant difference exists between the two groups. Moreover, the protection rate was higher in Group 3, with 100% for E. coli and 90% for the AI H5N1 vaccine. CONCLUSION: Our findings demonstrate that producing a combined vaccine using E. coli and AI H5N1 formulated with Montanide ISA70 is recommended for protection against both diseases.

Comparative Study on the Efficacy of MF 59, ISA70 VG, and Nano-Aluminum Hydroxide Adjuvants, Alone and with Nano-Selenium on Humoral Immunity Induced by a Bivalent Newcastle+Avian Influenza Vaccine in Chickens.

Newcastle disease (ND) and Avian influenza (AI) are the major problems and the most economically important viral diseases in the poultry industry; therefore, vaccination against these diseases is considered one of the most effective ways of prevention. Extensive studies have been conducted to improve the performance of vaccines, and one of the major achievements of these studies is the preparation of adjuvants as stimulants of the immune system and one of the most important compounds in killed vaccines. An immunogenicity comparison of three adjuvants including, ISA70VG, Nano-Aluminum Hydroxide (Nano-Alum), and MF59 alone or with Nano-Selenium (Nano-Se), was performed using bivalent Newcastle plus Avian Influenza (ND+AI) killed vaccine. In this study, 105 specific-pathogen-free chicks (Ross-308) were divided into 7 treatments, including T1 (control group), T2 (ISA70VG), T3 (ISA70VG plus Nano-Se), T4 (Nano-Alum Hydroxide), T5 (Nano-Alum+Nano-Se), T6 (MF59), and T7 (MF59+Nano-Se). The vaccine was injected subcutaneously on day 21 in the back of the neck area. The blood samples were taken on days 14, 21, 28, 35, 42, and 49 post-vaccination. Serums of the samples were titrated by the haemagglutination inhibition (HI) test against Newcastle and Avian influenza. Based on the results, the highest HI test titers were observed for the T2 and T3 treatments, while the T6 and T7 treatments had the lowest titers. Moreover, regardless of the type of the adjuvants, adding Nano-Se increased the antibody titer in the vaccinated groups. In conclusion, a combination of the ISA70VG adjuvant and Nano-Se induced excellent antibody titers using bivalent ND+AI killed vaccine.

A novel bivalent Pasteurellosis-RHD vaccine candidate adjuvanted with Montanide ISA70 protects rabbits from lethal challenge.

In the present study, a bivalent vaccine against Pasteurella multocida and rabbit hemorrhagic disease virus (RHDV) was formulated with Montanide™ ISA70 oil adjuvant (Seppic, Paris, France). Its efficacy was evaluated and compared to similar monovalent preparations and commercially available monovalent vaccines. White new Zeeland rabbit groups (n = 10) received 2 successive doses of the tested vaccines and were challenged 2 weeks after 2nd dose with Pasteurella multocida and RHDV or either pathogens according to their vaccination schedule. Challenged not-vaccinated group of rabbits (n = 10) was included as a control. The bivalent and monovalent ISA70 preparations were found stable, safe, sterile, pure and of low viscosity. Group 3 (GP3) which received bivalent vaccine showed the highest antibody geometric mean titers against Pasteurella multocida and RHDV evaluated by ELISA and hemagglutination inhibition (HI) respectively. Following virulent challenge; Gp3 rabbits were 90% protected from challenge over other groups that showed 80% protection. Detection of either pathogen in the livers of dead and euthanized rabbits had failed except for non-vaccinated controls. The bivalent vaccine candidate was fully protective. Immunization against both pathogens can be achieved by single vaccination.

Comparative safety and efficacy of two bivalent vaccines containing Newcastle disease LaSota and avian influenza H9N2 Sidrap isolate formulated with different oil adjuvants.

BACKGROUND AND AIM: Newcastle disease (ND) and avian influenza (AI) are two devastating diseases of poultry, which cause great economic losses to the poultry industry and disrupt food security in our country. The use of ND-AI inactive bivalent vaccine is very effective and economical to prevent and control ND and AI disease. Bivalent ND LaSota-AI H9N2 vaccine is not yet available in Indonesia. The inactivated vaccines used in poultry industry often require oil adjuvant to elicit a sufficient immune response. This study aimed to develop the bivalent inactive vaccines containing ND LaSota and AI H9N2 Sidrap isolate which are local isolates as poultry vaccine candidates, and formulated with two different commercial adjuvants, then compared. MATERIALS AND METHODS: Two vaccines bivalent were prepared by emulsifying inactivated Newcastle disease virus (LaSota strain) and AI H9N2 Sidrap isolate viruses with Marcol white mineral oil and Montanide ISA70 adjuvants. Both of bivalent vaccines were tested for safety (physical and histopathological at the injection site) and efficacy in specific-pathogen-free chickens. Parameters used for the evaluation of the efficacy were immunogenicity by hemagglutination inhibition and protection percentage. RESULTS: Both bivalent vaccines are safe to use. Post-vaccination (PV) immune response was observed using a hemagglutination inhibition test at 2, 3, 4, 5, 6, 7, and 8 weeks of PV. The bivalent vaccine B gives a better immune response to ND at 2, 3, and 4 weeks of PV (p<0.05) compared to the bivalent vaccine A, but in 5, 6, 7, and 8 weeks, the PV does not show differences in the immune response. The immune response to AI H9N2 showed differences at weeks 2 and 3 PV (p<0.05) with the bivalent vaccine B indicated higher immunity. A single immunization with both bivalent vaccines induces 100% protection in chickens that have been vaccinated against the deadly challenge with the virulent ND virus. CONCLUSION: Both of bivalent vaccines are safe to use and provide good efficacy against virulent ND viruses, but bivalent vaccine B (with Montanide ISA70 adjuvant) shows better immune response than bivalent vaccine A (Marcol white mineral oil adjuvant).

Comparative assessment of humoral immune responses of aluminum hydroxide and oil-emulsion adjuvants in Influenza (H9N2) and Newcastle inactive vaccines to chickens.

Context Adjuvants are compounds used in the preparation of inactive vaccines to enhance the immune response. Aluminum hydroxide (alum) is one of the first compounds approved by the Food and Drug Administration, which is used as adjuvants in vaccine products for humans. Montanide ISA 70 is an oil-emulsion adjuvant and is used in poultry inactive vaccines. Objective In this study, the effects of alum adjuvant on the efficiency and induction of immune response in inactive vaccines of Influenza and Newcastle are compared with those of ISA 70. Materials and methods Six groups of 7-d-old specific-pathogen-free chickens were inoculated with 0.3 ml of the prepared vaccines via the subcutaneous route in the neck. Immune response in each group after 7, 14, 21, 31, 41, and 45 d was evaluated using the technique of hemagglutination inhibition. Results The results were compared using SPSS software. Results showed that vaccines containing adjuvant ISA 70 depicted a higher increase in the immune response and adjuvant of 20% alum is similar to adjuvant of ISA 70 in boosting the immune system. There was no statistically significant difference between 10% and 20% alum, but these adjuvants are visibly different from ISA 70. Conclusion In conclusion, alum can be used as an easily accessible, harmless, and effective adjuvant; however, to increase the immune period using the inactive vaccines for poultry, more research would be necessary.