الفهرس 
INTRODUCTIONOnly 14 pages are availabe for public view
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Abstract
Foot and mouth disease is a highly contagious viral disease of cloven-hoofed animals such as cattle, goats and sheep. Foot and mouth disease is one of the diseases that cause great economic losses in livestock. Vaccination is considered one of the most successful strategies for overcoming foot-and-mouth disease, especially in endemic areas. Choosing the suitable adjuvant is one of the most important steps affecting the efficiency of the vaccine. In Egypt, the Veterinary Serum and Vaccine Research Institute - the Ministry of Agriculture- produces the FMD vaccine against the strains detect in Egypt, and the Montanide oil ISA 206 is used as an adjuvant in the vaccine.
This study aims to search for other alternative sources of Montanide oil to try to reduce the cost of vaccine production by using clinoptilolite as an alternative to Montanide oil ISA 206 and as an adjuvant to enhance the efficiency of the trivalent of foot-and-mouth disease vaccine while reducing its production cost.
The experimental calves were used in our study split into four groups each group contains three animals and divided as follows:
• group 1 (Control): The animals were kept unvaccinated in an isolated area until the end of the experiment.
• group 2 (ISA 206): Animals were subcutaneously vaccinated with inactivated trivalent FMD vaccine with Montanide ISA 206 oil (w/w).
• group 3 (ISA 206+CLINP): the calves vaccinated with inactivated trivalent FMD vaccine mixed with 50% of Montanide ISA 206 oil and 1μg/dose of clinoptilolite(w/ ½ w oil / 1μg clinoptilolite)/dose.
group 4 (CLINP): Calves were vaccinated with inactivated trivalent FMD vaccine containing 1μg/dose of clinoptilolite.
All calves vaccinated by injecting with 3 ml per dose of the FMD vaccine subcutaneously in the neck.
The present study was performed for the evaluation of inactivated FMD vaccines with different adjuvants. This study included:
Evaluation of the cell-mediated immune response of the prepared vaccines in vivo using lymphocyte transformation assay, interleukin-2, and interleukin-6 in experimental calves.
Evaluation of the humoral immune response of all vaccinated animals using SNT and ELISA.
Determination of hematological parameters includes hemoglobin concentration (Hb), white blood cells (WBCs), red blood cells (RBCs), and platelets.
Estimation of biochemical parameters including urea, creatinine, alanine aminotransferase (ALT) aspartate aminotransferase (AST) activities, total proteins, albumin, globulins, and albumin/globulins ratio.
1) Evaluation of the cell-mediated immune response
- The results showed that the FMD vaccine with (ISA 206+CLINP) elicited a dramatically higher level of cellular immune response (lymphocyte transformation assay) than the (CLINP) or (ISA 206) vaccine.
- The results indicated a significant increase (p≤0.05) in the production of Interleukin-2 (pg/mL) in calves vaccinated with FMD with (ISA 206+CLINP) or (CLINP) vaccines starting from 3rd DPV,
- The results showed a significant increase in the production of Interleukin-2 (pg/mL) in the calves’ vaccinated group with FMD with (ISA 206) vaccine starting from 7th DPV.
- The results showed the significantly increase in concentration of IL-6 in lymphocytes stimulated by the recombinant FMD (ISA 206), (ISA 206+CLINP), and (CLINP) vaccine starting from 3rd DPV.
- The group vaccinated with the FMD vaccine with (ISA206+CLINP) recorded a high concentration of IL-6 compared with other groups.
2) Evaluation of the humoral immune response using SNT and ELISA.
- Our results showed that all vaccinated groups elicited significantly (p ≤ 0.05) higher levels of antibodies compared to control
- The group vaccinated with inactivated FMD with (ISA 206+CLINP) vaccine reports a maximum period of protective to humoral immune response started from 1st WPV and extended to 38thWPV which is the longer time for protective.
- Also, the calves vaccinated with (ISA 206) inactivated FMD vaccine showed that the protective humoral immune response started from 2nd WPV and extended to 34thWPV.
- However, the calves vaccinated with (CLINP) inactivated FMD vaccine showed that the protective humoral immune response started from 1st WPV and extended to 26th WPV.
3) Determination of hematological parameters
- The results clarified that there were no significant differences (p≤0.05) in Hb concentrations, red cell count, and platelets in both the FMD vaccinated calves groups (ISA 206), (ISA 206+CLINP), and (CLINP) and unvaccinated groups.
- The resulted showed that a significant elevations (p≤0.05) in WBCs in all FMD vaccinated groups compared to the control group starting from 3rd DPV until 7th DPV.
4) Estimation of biochemical parameters
(A) Urea and Creatinine concentration:
- Kidney function tests were qualified by urea (mg/dl) and creatinine (mg/dl) concentration.
- The results indicate that the urea concentrations showed a significantly higher (P≤ 0.05) after vaccination from the 2nd week and decrease until the 10th WPV in the group vaccinated with (ISA 206) FMD vaccine compared to control. This increase was at normal levels.
- While the FMD vaccinated groups with (CLINP) or (ISA 206+CLINP) vaccine showed that no significant difference in urea concentrations compared to the control.
- Also, our results showed a high increase (P≤ 0.05) in creatinine levels in group vaccinated with FMD vaccine with (ISA 206) at the 2nd WPV which is considered above normal level and started to decrease gradually till 6th WPV.
- While the calves vaccinated with inactivated FMD vaccine with (ISA 206+CLINP) or (CLINP) showed no statistically significant creatinine concentration compared to control.
(B) Aspartate aminotransferase AST (U/L) and Alanine aminotransferase (ALT) activity:
- The obtained data showed that there were significant increases in AST activities in serum of both (ISA 206) and (ISA 206+CLINP) vaccinated animals at 1st WPV until 3rd WPV.
- In addition, the vaccinated group with FMD with (CLINP) showed no significant difference compared to the unvaccinated group.
- Also, there were no significant differences in ALT activity in all vaccinated and unvaccinated groups.
(C) Total protein and albumin concentration:
- The results clarified that the three vaccinated groups (ISA 206), (ISA 206 +CLINP), and (CLINP), were significantly higher in total protein and after vaccination from the 1st WPV until the 10th WPV than in control.
- Specially, the group vaccinated with inactivated FMD vaccine with (ISA 206+CLINP) was reported a highest value of total proteins after vaccination.
- Conversely, albumin was no significant differences among the three vaccinated groups compared with control during the same period.
(D) Globulins concentration and A/G ratio
- The results indicate that serum values of globulins were significantly increased (p ≤0.05) and the A/G ratio was decreased in all vaccinated groups compared to control.
- Specifically, (ISA 206+CLINP) vaccinated calves demonstrated significantly highest values of globulins and lowest ratio of A/G compared to other vaccinated groups.
Conclusion
In conclusion, results clearly declared that the use of clinoptilolite as an adjuvant with ISA 206 oil (ISA 206 + CLINP) in inactivated FMD trivalent vaccine induces long-lasting immunity than that induced with oil adjuvant alone or with clinoptilolite alone and improve humoral and cellular immunity. It also did not have any effect on the biochemical parameters in calves.
Furthermore, it is recommended to use the FMD inactivated vaccine adjuvant with oil and clinoptilolite (1:1) in the companying of vaccination to control FMD. We also recommended increasing the studies on the effect of clinoptilolite as an antioxidant in veterinary.