Aim:A comparison study was conducted to explore the best internationally available adjuvant that could be used in production of a highly potent foot and mouth disease (FMD) vaccine, that could stimulate a strong immune response and possibly give greater protection against FMD.Materials and Methods:Four experimental batches of trivalent FMD vaccine were prepared with different available oil adjuvants which included Montanide ISA 201, 206, 61 and 50.Results:The results indicated that vaccines emulsified using Montanide ISA 201 and Montanide ISA 206 adjuvants elicited a protective humoral immune response from the 2nd week postvaccination (WPV) as for ISA 201 with serum neutralization test (SNT) and enzyme-linked immune sorbent assay (ELISA) antibody titers of 1.62±0.047a and 1.8±0.049a, 1.59±0.076a and 1.836±0.077a, and 1.71±0.06b and 1.96±0.074b for serotypes O, A, SAT2, respectively, and for ISA 206 at SNT and ELISA antibody titers of 1.5±0.082a and 1.84±0.084a, 1.56±0.037a and 1.818±0.052a, and 1.5±0.106a,b and 1.81±0.104a,b for FMD virus serotypes O, A and SAT2, respectively. For ISA 61 and ISA 50, the protective antibody titer appeared in the 3rd WPV. In the ISA 61 FMD vaccine, SNT and ELISA titer were 1.59±0.076a and 1.9±0.094a, 1.53±0.056a and 1.83±0.070a, and 1.5±0.082a and 1.84±0.094a for serotypes O, A and SAT2, respectively, and in the case of ISA 50 FMD vaccine, the SNT, and ELISA titer were recorded for serotypes O, A and SAT2 respectively, 1.59±0.037a and 1.8±0.030a, 1.68±0.056a,b and 1.916±0.065a,b, and 1.65±0.082a and 1.9±0.09a. On estimating the cellular immune response, the highest delta optical density levels for ISA 201 (0.395-0.460) and ISA 206 (0.375-0.428) were observed on 14 and 21 days post vaccination (DPV) respectively, while the highest levels of lymphoproliferation for ISA 61 (0.375-0.455) and ISA 50 (0.411-0.430) were on 21 and 28 DPV, respectively.Conclusion:The duration of immunity from Montanide ISA oils (201, 206, 61 and 50) FMD vaccines is a long-lived immunity which ranged between 32 and 38 weeks post vaccination but the Montanide ISA 201 FMD vaccine is superior to the others in the rapid cellular immune response of the vaccinated animals which showed its highest level within 14 days post vaccination.

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Aim: To determine the minimal effective dose of Foot and Mouth disease (FMD) serotypes (A, O, SAT2) according to antigenic content (146S) in order to produce a potent trivalent FMD vaccine.Materials and Methods: Monovalent ISA 206 vaccines were prepared with 3 final concentration of 146S (1.6, 2.2, 2.8 µg/dose). The vaccine potency was evaluated by the determination of guinea pig protective dose 50 (GPPD )for each 50 concentration of 146S for each type of FMD monovalent vaccine where a fourfold dilution of the vaccines was constructed and each dilution was inoculated as 0.5 ml S/C in each of 5 guinea pigs. Results:The obtained results revealed that by using 1.6 µg of 146S for type O Pan Asia-2, A Iran O5 and SAT/EGY/2012, the GPPD was 40.4, 19.75 and 31.6 respectively, while the use of 2.2 µg of 146S resulted in GPPD 78.6, 78.6 and 105.8 for the 50 50 three types respectively, and by using 2.8 µg of 146S resulted in GPPD 161.7, 105.8 and 161.7 for the three subtypes (A, O, 50 SAT2) respectively. So it is clear that the lowest 146S dilution inducing good protection (more than 72 GPPD ) was 2.2 µg for 50 each serotype of used FMD monovalent vaccines. Depending on this result, the trivalent vaccine was formulated as 2.2 µg of 146S payload from each virus type/dose with equal volume of montanide ISA 206 oil as adjuvant. For more confirmation the prepared trivalent vaccine potency was evaluated by Guinea pig protective dose 50 which was found to be 88 GPPD .Also 50 mean SNT antibody titer was detected in serum of the test Guinea pigs 1.56, 1.68 and 1.68 log /ml against FMDV serotype O 10Pan Asia-2, A Iran O5 and SAT/EGY/2012 respectively in a higher level than the recommended protective titer (PT=1.2). Also for further confirmation the formulated trivalent vaccine which contain 2.2 µg/serotype/dose were evaluated in cattle to measure the antibody titer against the three serotypes and the antibody against the three serotypes were found to be higher than the recommended titer (1.5) which extended for 32 WPV and these results came in parallel manner to the GPPD and antibody 50 titer of the Guinea pigs of the prepared trivalent vaccine with 146S (2.2 µg/serotype/dose). Conclusion:It could be concluded that the minimum content of antigenic 146S of FMDV serotype O pan Asia, A Iran O5 and SAT2/EGY/2012 should not be less than 2.2 µg/dose/ from each serotype in the trivalent vaccine aiming to induce the permissible protection in vaccinated livestock.

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