Effect of administration of ultra-corn with bivalent Foot and Mouth disease oil vaccine in calves

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1 Effect of adminiration of ultra-corn wi bivalent Foot a Mou disease oil vaccine in calves Eman Mahmoud Mohamed, Ehab El-Sayed Ibrahim a Fatma Sayed Mohamed 1. Central Laboratory for evaluation of veterinary biologics, Abbasia, Cairo, Egypt; 2. Veterinary Serum a Vaccine Research Initute, P.O.Box:131, Abbasia, Cairo, Egypt Correspoing auor: Ehab El-Sayed Ibrahim, Fax: (202) , ehabelsayed@hotmail.com Received: , Revised: , Accepted: , Published online: How to cite is article: Mohamed EM, Ibrahim EE a Mohamed FS (2013) Effect of adminiration of ultracorn wi bivalent Foot a Mou disease oil vaccine in calves, Vet World 6(8): , doi: /vetworld Introduction Abract Aim: The present work was eablished in order to inveigate e effect of ultra-corn adminiration on e immune response of vaccinated calves wi FMD bivalent oil vaccine. Material a Meods: Forty calves; at a private farm in EL-Fayoum Governorate (Locality A); were divided into 4 groups rd where e fir group was vaccinated wi e locally produced FMD bivalent oil vaccine alone while e 2, 3 a 4 group were vaccinated wi e same vaccine simultaneously wi e inoculation of 1, 1.5 a 2mL/100kg body weight of ultra-corn respectively to eimate e antibody titer, e suitable dose a effect of ultra-corn as immunoimulant using SNT a ELISA. Also after at used e effective a lowe dose of ultra-corn simultaneously wi e vaccine in comparison wi e vaccine alone by using 26 calves (Locality B) to udy e efficacy of ultra-corn simultaneously wi vaccine a e vaccine alone via challenge te using e virulent FMDV serotype A,O. Results: Teed serum samples obtained on week intervals po vaccination of all calve groups were subjected for eimation of iuced FMD antibodies type A a O using serum neutralization te (SNT) a enzyme linked immune sorbent assay (ELISA). Bo tes iicate at 1.5mL, 2mL of ultra-corn enhanced e immune response of vaccinated calves exhibiting higher a longer immunity an ose received e vaccine alone. In addition 26 calves housed uer rerict hygienic measures at Veterinary Serum a Vaccine Research Initute, were divided into 4 groups where group-1 of 10 calves were vaccinated wi e bivalent FMD vaccine alone a group-2 was vaccinated wi e same vaccine simultaneously wi 1.5mL of ultra-corn while group 3 a 4 were kept as control for e challenge te. On e 4 week po vaccination group 1, 2 of ese animals was subdivided into 2 subgroups where e challenge te was carried out again type A in a subgroup a O in oer subgroup. SNT a ELISA showed similar results obtained from e vaccinated farm animals a challenge te via inoculation of virulent virus of FMD serotype A, O iicated at animals received ultra-corn showed 100% protection again bo types of FMD virus while ose did not receive ultra-corn showed 80% protection (vaccine alone). Conclusion: So it could be concluded at ultra-corn has a potentiate effect on e immunogenicity of bivalent oil FMD vaccine providing high immune levels of long duration. Keywords: bivalent, calves, FMD, immune response, serotype, vaccine This article is an open access article licensed uer e terms of e Creative Commons Attribution License ( org/licenses/by/2.0) which permits unrericted use, diribution a reproduction in any medium, provided e work is properly cited. Foot-a-mou disease (FMD) is considered one of e mo serious viral infections in liveock, because of its rapid transmission over a range of animal species. As FMD is ill eemic in large regions of e world, it is a highly infectious disease of ungulates primarily cattle, sheep, goats a pigs. It also affects wild animals such as buffaloes a deer [1]. Foot-a- mou disease virus (FMDV) is e etiologic agent of e diseases at can affect cloven-hoofed liveock. Infection wi FMDV characterized by fever, lameness a vesicular lesions on e feet, tongue, snout a teats, wi high morbidity but low mortality [2]. There are seven types of FMD virus (FMDV) have been identified as; O, A, C, SAT 1, SAT 2, SAT 3 a Asia1 [3] a [4]. It was ated at vaccination again infectious diseases is e corner one to eradicate or even to control such diseases. Many trials were carried out in order to improve e potency of FMD inactivated vaccine using different adjuvant aiming to potentiate its immunogenicity a to increase e duration of iuced immunity in vaccinated animals [5]. Montanide ISA206 oil adjuvant quadric-valent FMD vaccine elicited a better immune response at any time an aluminum hydroxide gel vaccine, a is response was developed quicker. The animals maintained eir neutralizing antibody titers at >3 log 10 for e duration of e trial (90 days) [6]. The mean protective serum antibody titers again FMD in calves vaccinated wi double oil emulsion (Montanide ISA 206) evaluated by Enzyme Linked Immunosorbent Assay (ELISA) a Serum Nutralization Te (SNT) rd was arted at e 3 week po vaccination (WPV) reached e highe level at e 10 WPV a continued wi e protective level till e 32 WPV en arted to decline uer e protective level for bo FMD virus types O a A [7]

2 The immune response of vaccinated goats wi Cell culture: Baby Hamer kidney cell line (BHK21) alhydargel a double oil emulsion Montanide ISA Clone 13 maintained in FMD Department, Abbasia, 206 vaccines persied for 20 a 36 weeks po Cairo using Eagl's medium wi 8-10% erile bovine challenge, respectively [8]. Also [9,10] fou at such serum as described by [13], was used for application of vaccine iuced long laing immunity an at wi serum neutralization te. Alhydragel adjuvant. Vaccine: Local produced inactivated bivalent FMD The immune response eimated by using SNT vaccine including type (O 1/93/Aga) a type (A/ a ELISA revealed at e antibody titers of e FMD Egypt/2006) adjuvanted wi Montanide ISA 206 oil vaccines wi ultra-corn in calves appeared earlier a was supplied by Veterinary Serum a Vaccine Research higher at e fir six weeks po vaccination an at Initute to be used for vaccination of calves [14]. of FMD gel vaccine alone a e calves persied for 16 weeks po vaccination for bo [11]. Ultra-corn: The vaccinated calves wi bivalent FMD ISA - It is a product of Virbac Company, France. 206 inactivated oil vaccine should be revaccinated on - It is consied of Corynebacterium cutis lysate e 36 weeks po vaccination to avoid e decline of (deroyed a lysed by using ultrasonic device) e protective immune levels to e non-protective as 20mg/mL of e product. values [12]. - Ultra-corn is used for prevention of diarrhoea a The udy was aim to determine e effect of ultraadminiered to e dams at e late pregnancy, pneumonia infections of newborn animals when corn as an immune modulator to e locally produced bivalent FMD oil vaccine on e immune atus of prevention of placental infections in case of retained placenta when adminiered during late vaccinated calves. pregnancy a used as preventive measures or Materials a Meods supportive erapy in case of respiratory or digeive diseases due to contagious bacterial, Animals: The present work was subjected on 66 apparently healy calves of 6-8 mons of age from viral, a oer paogens. two localities a free from FMD type O /Aga/Egy/93 - Storage coitions : ore in e refrigerator (2-1 6 C). a A/Egy/1/2006 antibodies as screened by serum Vaccination: All vaccinated calves was vaccinated neutralization te a iirect ELISA. using inactivated bivalent (A,O) Foot a Mou *Locality (A): Disease Montanide ISA206 oil vaccine wi dose 2mL/ Includes 40 calves at private farm at El-Fayoum animal subcutaneously. Governorate divided into 4 groups (10 calves/ group) where group (1) was vaccinated wi e bivalent oil Challenge te: All vaccinated calves a positive FMD vaccine using a dose of 2mL/ animal inoculated control were inoculated wi e challenged FMDV subcutaneously while group 2, 3 a 4 were inoculated 4 eier serotype A or O wi titer 10 BID 50 at e base on wi 1, 1.5 a 2mL/100 kg body weight of ultra-corn e tongue. All challenged animals were kept uer simultaneously wi e same dose of FMD vaccine. daily observation a clinically examined for 1 week *Locality (B): po infection (WPI), where body temperature a Includes 26 calves housed in Veterinary Serum a lesions on e gum a oral mucosa as well as foot pad Vaccine Research Initute, Abassia, Cairo. These were recorded. animals were divided into 4 groups where each of Serum neutralization te (SNT): It was performed group 1 a 2 includes 10 calves en divided equally using e micro titer technique as described by [15] to to be challenged wi FMDV serotype A,O, while each evaluate e levels of iuced FMD antibodies in of group 3 a 4 contains 3 calves. Group (1) was experimentally vaccinated calves. vaccinated wi FMD bivalent vaccine only a group (2) was inoculated wi 1.5mL of ultra-corn/100 Kg Iirect enzyme linked immune sorbent assay (i- ELISA): body weight animal simultaneously wi FMD vaccine Iirect ELISA was carried out according to while group 3 a 4 were kept wiout vaccination as [16] to follow up e immune response in animals. positive control challenged wi FMDV serotype A,O. Results a Discussion Eical approval: The experiment was as per e protocol Foot a Mou disease (FMD) is one of e mo of Initutional Animal Eics Committee, e auors wide spread diseases affecting cloven footed animals had taken permission of animal owners of private farm. of detrimental effects on meat a milk production, raer an convalescent animals became carriers a Viruses: Locally isolated FMDV type O 1/Aga/EGY/93 source of infection [17]. In countries where e disease 9 wi titter of 10 TCID 50 /ml a type A/EGY/1/2006 wi is eemic, draic measures of eradication are not 9 titer 10 TCID 50 /ml were supplied by Foot a Mou always economically feasible. Control is based on a Vaccine Research Department (FMDRD), Veterinary modified syem of vaccination a quarantine, using Serum a Vaccine Research Initute, Abbasia, Cairo. vaccines specific for e type a subtype of e viruses These viruses were used for challenge te. involved. Vaccines containing one or more immune 487

3 Table-1. Mean FMD serum neutralizing antibody titers in calves vaccinated wi bivalent FMD wi ultra-corn simultaneous (locality A) WPV calves receiving e Mean FMD serum neutralizing antibody titers (log 10/mL) in calves group vaccinated wi vaccine alone bivalent FMD vaccine simultaneously wi 1 ml ultra-corn/100kg 1.5 ml ultra-corn/100kg 2 ml ultra-corn/100kg body weight body weight body weight A* O** A O A O A O 0 0.2*** WPV: Weeks po vaccination, *A: FMDV serotype A, **O: FMDV serotype O, *** Antibody titers expressed in log 10. The recommeed protective levels of FMD antibody titers are 1.5 log 10 for SNT for bo of type A a O as eimated by SNT [4] Table-2. Mean FMD ELISA antibody titer of calves vaccinated wi bivalent FMD wi ultra-corn simultaneous (locality A) WPV calves receiving e Mean FMD ELISA titers (log 10/mL) in calves group vaccinated wi bivalent FMD vaccine alone vaccine simultaneously wi 1 ml ultra-corn/100kg 1.5 ml ultra-corn/100kg 2 ml ultra-corn/100kg body weight body weight body weight A* O** A O A O A O *** *A: FMDV serotype A, **O: FMDV serotype O, *** Antibody titers expressed in log 10 The recommeed protective levels of FMD antibody titers are 1.9 log 10 for ELISA for bo of type A a O as eimated by ELISA [4] types of e virus are produced in several countries. Gel WPV to record peak titers by e 16 WPV en began adjuvant vaccine is often less successful an to decline gradually by e 20 WPV till 36 WPV for vaccination of cattle a sheep by recently oil-adjuvant type O en became unprotected. On e oer side e vaccines which have been used wi excellent results. mean neutralizing antibody titers again type (A) in If e latter syem is used, annual revaccination is calves vaccinated wi bivalent oil FMD vaccine essential [18]. The progress in vaccine production is simultaneously wi 1 ml ultra-corn reached e directed towards e selection of e proper adjuvant protective level by e 2 WPV till 36 WPV en at can elaborate high a long laing immunity. The became unprotected level. Also mean neutralizing vaccine efficacy is measured by e level a duration antibody titers again type (O) in such calves showed of protective immune response a e proportion of e same manner but ill protective till e 40. The vaccinated animals which demonrate immunity mean neutralizing antibody titers again type (A) in (resi challenge) [19]. calves vaccinated wi bivalent oil FMD vaccine As obtained from e present work, Table-1 simultaneously wi 1.5 ml ultra-corn became showed at vaccinated calves wi bivalent FMD oil protective by e 2 WPV till 40 WPV where ey vaccine alone a smiltaniously inject wi eier 1, became unprotected by e 44 WPV. Similarly, mean 1.5 or 2 ml of ultra-corn exhibited specific neutralizing neutralizing antibody titers again type (O) in e same antibody again type A a type O arted from e 2 calves showed e same behavior iicating at type O WPV. On e use of FMD bivalent vaccine alone; it was antibodies were higher an ose of type A a las for noticed at e specific FMD neutralizing antibody 44 WPV. In calves vaccinated wi bivalent oil FMD titers reached a protective level arting from e 4 vaccine simultaneously wi 2 ml ultra-corn, FMD 488

4 Table-3. Mean FMD serum neutralizing antibody titers in calves vaccinated wi bivalent FMD a 1.5 ml ultra-corn/100kg body weight simultaneously (Locality B) Animals Treatment of animals Mean FMD serum neutralizing antibody titer (log 10/mL) challenge No. 0 WPV* 1 WPV 2WPV 3WPV 4WPV A** O*** A O A O A O A O 1 vaccinated wi A 2 FMD bivalent A 3 oil vaccine alone A A A O O O O O Mean vaccinated wi A 12 FMD bivalent oil A 13 vaccine a 1.5 cm A 14 of ultra corn A 15 simultaneously A O O O O O Mean control positive A A A A 24 control positive O O O O *WPV: Weeks po vaccination, **A: FMDV serotype A, ***O: FMDV serotype O The recommeed protective levels of FMD antibody titers are 1.5 log 10 for SNT for bo of type A a O as eimated by SNT [4] type A neutralizing antibodies became protective adminiration of ultra-corn iuced marked immune arting from e 2 WPV till e 40 WPV but e potentiating effect of e immune response developed means neutralizing antibody again type (O) in calves again Newcale disease vaccine in chickens vaccinated by e same manner became protective by especially when given simultaneously wi e e 2 WPV till e 44 WPV en became unprotected vaccine. level. Also e results of ELISA came parallel to ose As shown in Table-3 a 4, e challenge te was of SNT as shown in Table-2. These results agree wi carried out on e 26 calves on e 4 WPV, where e ose obtained by [7,9,10,20,21] ey reported at e mean antibody again type (A) in 5 calves vaccinated mean antibody titers again FMD vaccine rain wi bivalent oil FMD vaccine alone arted by e 1 O 1/3/93 were detected in sheep sera vaccinated wi WPV (0.75&0.98 log 10) to become protective at e 4 Alum-hydroxide gel vaccine following one week po WPV (1.53&1.81 log 10) by SNT a ELISA respectively vaccination (0.9 log 10) by SNT, whereas, e mean peak while ose again type (O) in oer 5 calves titers (1.9 log 10) by SNT were detected by e 16 week vaccinated wi bivalent oil FMD vaccine alone by 1 po vaccination. A also agree wi [11] who WPV were (0.96&1.196 log 10) to become protective at mentioned at e immune response eimated by e 4 WPV (1.63&2.01 log 10) by SNT a ELISA using SNT a ELISA revealed at e antibody titers respectively, in comparison wi e means antibody iuced by FMD vaccines wi ultra-corn in calves again type (A) in 5 calves vaccinated wi bivalent oil appeared earlier a higher at e fir six weeks po FMD vaccine simultaneously wi 1.5 ml ultra-corn vaccination an ose iuced by FMD gel vaccine arted by e 1 WPV (1.24&1.44 log 10) till become alone a persied for 16 weeks po vaccination in protective at e 4 WPV (1.88&2.074 log 10 calves for bo types. The significant increase in e ) by SNT immune response in calves vaccinated wi bivalent a ELISA respectively while e mean antibody titers FMD oil vaccine simultaneously wi ultra-corn could again type (O) in 5 calves vaccinated wi bivalent oil be attributed to e mode of action of ultra-corn as FMD vaccine alone arted by e 1 WPV (1.55 & immune imulant as explained by [22] who mentioned log 10) till become protective at 4 WPV (2.41 & at killed Corynebacterium cutis imulates e 2.22 log 10) by SNT a ELISA respectively. The activity of bo T a B lymphocytes which lead to antibody titers of control unvaccinated calves remain increase e antibody synesis, a [23] who fou unprotected till 4 weeks before arting challenge te. at corynaebactreium is one of e ronge iucers The clinical signs described in challenged vaccinated of lymphocytes trapping, [24] reported at e calves after 4 WPV (Table-5) included eier e 489

5 Table-4. Mean FMD ELISA antibody titer of calves vaccinated wi bivalent FMD a 1.5 ml/100kg body weight ultra-corn simultaneously (Locality B) Animals Treatment of animals Mean FMD ELISA antibody titer (log 10/mL) challenge No. 0 WPV* 1 WPV 2WPV 3WPV 4WPV A** O*** A O A O A O A O 1 vaccinated wi FMD A 2 bivalent oil A 3 vaccine alone A A A O O O O O Mean vaccinated wi FMD A 12 bivalent oil A 13 vaccine a A ml of ultra corn A 15 simultaneously A O O O O O Mean control positive A A A A 24 control positive O O O O *WPV: Weeks po vaccination, **A: FMDV serotype A, ***O: FMDV serotype O The recommeed protective levels of FMD antibody titers are 1.9 log 10 for ELISA for bo of type A a O as eimated by ELISA [4] Table-5. Determine FMD lesions a protection percentage in calves vaccinated wi bivalent FMD a 1.5 ml/ 100Kg body weight ultra-corn simultaneously after challenge again bo types of FMD virus Animals No. Treatment of animals Determined FMD lesions challenge Oral lesions Fore limbs Hi limbs Tongue Gum Lip Left Right Left Right 1 vaccinated wi FMD +* -** A 2 bivalent oil A 3 vaccine alone A A A O O O O O Protection 80% 11 vaccinated wi FMD A 12 bivalent oil vaccine A 13 a 1.5 ml of ultra corn A 14 simultaneously A A O O O O O Protection 100% 21 control positive A A A A Protection 0% 24 control positive O O O O Protection 0% *(+): means lesion **(-): no lesion 490

6 lesions on tongue epielial or on buccal mucosa a Charleon B. (2012) The importance of FMDV localisation feet. The lesions vary from erosion, vesicles a in lymphoid tissue. Veterinary Immunology a Immuno- paology, 148: ulceration. These signs appear to be characteriic for 3. Franki, R. I. B., Fauquet, C. M., Knudson, D. L. a Brown, FMD as ated by [2,4,12,13,25]. In e 10 calves F. (1991) Classification a nomenclature of viruses. 5 vaccinated wi FMD oil vaccine alone, one of e five Report of Int. Comm. on taxonomy of viruses suppl. 2. challenged calves wi serotype (A) showed 4. OIE (2010) Foot a mou disease, Chapter In generalized infection (means 80% protection again manual of aards for diagnoic tes a vaccine, 4 Ed. Paris, serotype A), also one of e five challenged calves wi 5. Barteling, J. (2002) Development a performance of serotype (O) showed generalized infection (means inactivated vaccines again foot a mou disease. Rev. 80% protection again serotype O) showing at e Sci. Tech., 21 (3): protection % for e FMD oil vaccine alone is 80%. On 6. Patil, P. K., Bayry, J., Ramakrishna, C., Hugar, B., Misra, L. D. a Natarajan, C. (2002) Immune response of goats e oer side, all of e 10 calves vaccinated wi FMD again FMD quadrivalent vaccine: comparison of double oil oil vaccine simultaneously wi 1.5 ml ultra-corn were emulsion a aluminum hydroxide gel vaccine in eliciting protected again bo types of FMD virus (A,O), immunity, Vaccine, 20: showing a protection % 100%. 7. Gamil, M. A (2010) Studies on e immune response of calves vaccinated again inactivated bivalent FMD virus The present obtained results iicate at 1.5 vaccine type O1 a A Egypt M.V.Sc in Veterinary ml/100 kg body weight ultra-corn enhances e Science (virology). Benha University. immune response of vaccinated calves to FMD oil 8. Faia, A. M. (2003) Vaccination of goats wi FMD bivalent vaccine a could be used as an immune vaccines. MVSc Thesis, University of Alexaria. potentiation to improve e immunogenicity of FMD 9. Sonia, A. M. (2007) Studies on e preparation of an improved Foot a Mou Disease oil vaccine. Ph. D. Thesis, vaccine a elongate e immune duration. University of Cairo. Conclusion 10. Selim, A. M. A., Abouzeid, N. Z., Aggour, A. M. a Sobhy, N. M. (2010) Comparative Study for Immune Efficacy of As per our results, it is concluded at ultra-corn Two Different Adjuvants Bivalent FMD Vaccines in Sheep. has a potential effect on e immunogenicity of Journal of American Science 6(10): Fadia, S. Hussein, Roushdy, O. H. a Halima, El-Watany bivalent oil FMD vaccine providing high immune (2000) Effect of simultaneous adminiration of ultra-corn levels of long duration. wi FMD gel vaccine in calves. J. Egypt Vet. Med. Ass. 60: Auors contribution (4): El-Sayed, E., Mossad, W., Ali, S. M. a Shawky, M. (2012): EMM helped in import of ultracorn from company, Studies on e duration of immunity iuced in cattle after done serological tes, prepared challenged virus of natural FMD infection a po vaccination wi bivalent oil vaccine, Vet World, 5(10): FMD, drafted e manuscript. EEI implemented e 13. Huang, Xuan, Yong, Li, Hui, Fang a Congyi, Zheng udy design, communicate wi e company a (2011) Eablishment of persient infection wi foot a mailing em to import e material of ultracorn, mou disease virus in BHK-21 cells. Virology Journal, 8: 169. preparation of bivalent inactivated FMD oil vaccine, 14. El-Sayed,E., Wael Mossad Gamal El-Din, Sonia Ahmed done serological tes, prepared challenged virus of Rizk a Magdi Abd El-Aty (2012) Effect of Different Storage Temperatures on e Efficacy of e Bivalent Foot FMD, drafted e manuscript, follow up wi journal. a Mou Disease Oil Vaccine. Journal of Advanced FSM done serological tes, drafted a revised e Veterinary Research, 2: manuscript. All auors read a approved e final 15. Ferreira, M. E. V. (1976) Prubade microneutralization manuscript. poraeudies de anticueropos de la fibre aftosa. 13 Centropanamericano Fiebre Aftosa, (21/22): Acknowledgements 16. Ferris, N. P., Kitching, R. P., Oxtoby, J. M., Philpot, R.M. a Rele, R. (1990) Use of inactivated foot-a-mou disease Auors are ankful to Prof. Dr. Seham Abd EL- virus antigen in liquid-phase blocking ELISA. Journal of Rashid Director of VSVRI, Prof. Dr. Sayed Zedan Virological Meods, 29: Deputy of VSVRI a all members of FMD 17. Depa, P. M., Dimri, U., Sharma, M. C. a Tiwari, R. (2012) department specially Prof. Dr. Magdy Abd El-Aty head Update on epidemiology a control of Foot a Mou Disease - A menace to international trade a global animal of FMD department, VSVRI. Also anks to Prof. Dr. enterprise, Vet World, 5(11): Mohamed Hassan Khoudier, Prof. Dr. Khayrat Abdel 18. Jamal, S. M., Ahmed, S., Hussains, M. a Ali, Q., (2008) Mageed Elian for reviewing is work. This work was Status of Foot a Mou disease in Pakian. The Global fued by Veterinary Serum a Vaccine Research control of FMD - Tools, ideas a ideals Erice, Italy Initute, Abbasia, Cario, Egypt. October Iyer, A.V., Ghosh, S., Singh, S. N. a Deshmukh, R. A. Competing interes (2001) Evaluation of ree ready to formulate oil adjuvants Auors declare at ey have no competing intere. for foot a mou disease vaccine production. Vaccine, 19(9-10): References 20. Abu Bakr, A. M. A. (2010) Trial for production of bivalent oil adjuvant FMD vaccine. Ph.D. (Virology), Fac. Vet. Med., 1. Paton, D. J., Sumption, K. J. a Charleon, B., (2009) Alex. University. Options for control of foot a-mou disease: knowledge, 21. El-Sayed, E (2011) Advanced udies on Foot a Mou capability a policy. Philos. Trans. R. Soc. B, 364: Disease Vaccines of sheep in Egypt. Ph. D. (Infectious , Diseases), Fac. Vet. Med., Cairo University. 2. Juleff N. D., Maree, F. F., Waters, R., Bengis, R. G. a 22. 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