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tralizing antibody responses to foot-and-mouth disease quadrivalent (type O, A, C and Asia 1) vaccinesrowing calves with pre-existing maternal antibodies

Foot-and-mouth disease (FMD), caused by FMD virusDV) is one of the most contagious diseases of livestock

ubman and Baxt, 2004). The practice of regularphylactic vaccination is the method of choice fortrolling FMD in developing countries (Rodriguez and, 2011). In India, various reports have demonstrated

t oil adjuvanted vaccines elicit high immune responsesh in cattle and in small ruminants (Patil et al., 2002a,2b; Madhanmohan et al., 2012; Dar et al., 2013).ever, the presence of maternal antibodies pose a major

eat for eliciting protective immune responses to FMDcines in young, growing animals (Doel, 2003). Recently, et al. (2013) reported serological responses of growings that were vaccinated with oil adjuvanted, inactivatedD trivalent vaccines. The authors found that despite thesence of maternal antibodies in the vaccinated pigs, oiluvanted vaccine elicited high levels of serologicalponses. These results suggested that oil adjuvantedcines could overcome maternal antibodies for elicitingibody responses. As cows are the major natural hosts inian subcontinent, in view of above report, we aimed atparing the ability of inactivated quadrivalent (O, A, C

Asia 1) double-oil ISA 206 emulsified and aluminiumroxide adjuvanted FMD vaccines to elicit neutralizingibody responses in growing calves that had maternalibodies.Quadrivalent oil emulsified FMD vaccine was preparedng serotypes O (Ind R2/75), A22 (Ind 17/77), C (Ind 1/

and Asia 1 (Ind 63/72) as previously reported (Patill., 2002c). The viruses were concentrated by 8% PEG0 and inactivated by 0.001 M binary ethylenimine. All

four serotypes of inactivated FMDV were mixed to obtainthe antigen payload of 3.50 mg of 146S virus particles/dose/serotype and adjuvanted with Montanide ISA 206(Seppic, France) to form water-in-oil-water doubleemulsion. The aluminium hydroxide/saponin-adjuvantedquadrivalent vaccine containing the same antigen pay-load of above FMDV serotypes was used for thecomparison. Two to three weeks aged growing calves(n = 18, cross bred of Holstein-Friesian � Hallikar) born tovaccinated cows were randomly selected from dairyfarms. Calves were distributed into two groups, group I(n = 10) for oil-adjuvanted vaccines and group II (n = 8) foraluminium hydroxide-adjuvanted FMDV vaccines. GroupI animals were vaccinated with 2 ml of oil adjuvantedvaccine while group II was administered with 5 ml ofaluminium hydroxide-adjuvanted vaccines. Generalhealth, temperature, feeding and immune response weremonitored during the entire study period of 60 days.Animals from both groups were bled on 0 (pre-vaccina-tion), 15, 30 and 60 days post-vaccination and immuneresponse was studied by virus micro-neutralization test(Rweyemamu et al., 1978).

As shown in Fig. 1, both the groups of animals hadequivalent levels of neutralizing maternal antibodies toall four serotypes of FMDV (in the range of 2–2.86 Log10,irrespective of serotype). Thus, we ensured that the levelsof neutralizing antibody responses elicited in two groupsof calves following vaccination were not biased due topre-existing maternal antibodies. We found that despitethe presence of maternal antibodies, animals thatreceived oil adjuvanted vaccines (group I) showed a

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A B S T R A C T

The presence of maternal antibodies is a major obstacle for eliciting protective immune

responses to foot-and-mouth disease (FMD) vaccines in young, growing animals. In this

report, we analyzed the ability of inactivated quadrivalent oil emulsified and aluminium

hydroxide adjuvanted FMD vaccines to elicit neutralizing antibody responses in growing

calves that had maternal antibodies. Our results demonstrate that oil emulsified vaccines

but not aluminium hydroxide adjuvanted FMD vaccines could surmount maternal

antibodies to elicit strong and significant levels of neutralizing antibody responses in

growing claves.

� 2014 Elsevier B.V. All rights reserved.

Contents lists available at ScienceDirect

Veterinary Microbiology

jo u rn al ho m epag e: ww w.els evier .c o m/lo cat e/vetmic

8-1135/$ – see front matter � 2014 Elsevier B.V. All rights reserved.

://dx.doi.org/10.1016/j.vetmic.2014.01.005

Letter to the Editor / Veterinary Microbiology 169 (2014) 233–235234

steady increase in neutralizing antibody responses up today 30 post-vaccination except for serotype O where thepeak response reached by day 15 (Fig. 1A). Also, theimmune response was quite uniform among the animals.However, irrespective of serotypes, the neutralizingantibody response declined after 30 days post vaccina-tion and reached the level corresponding to pre-immunization level by day 60.

In contrast to oil adjuvanted vaccines, aluminiumhydroxide-adjuvanted vaccines failed to induce neutraliz-ing antibody responses in growing calves that had pre-existing maternal antibodies. The neutralizing antibodytitres in group II continued to decline from day zero post-vaccination and were less than 1 log10 on day 60 post-vaccination (Fig. 1B). The differences in neutralizing

antibody responses between two groups of animalsfollowing vaccination were statistically significantthroughout the study period (P < 0.001).

Our results thus demonstrate that oil-adjuvanted FMDvaccine elicits strong and significant levels of neutralizingantibody responses in growing claves that had pre-existingmaternal antibodies. On the other hand, aluminiumhydroxide-adjuvanted vaccine failed to break the maternalantibody resistance to induce immune responses ingrowing calves. Thus our data along with that of Leeet al., confirm that oil adjuvanted vaccines could be used toelicit neutralizing antibody responses in growing clovenhooved animals irrespective of species. Considering theendemic nature of disease and high levels of FMDVneutralizing maternal antibodies, eight weeks appear to

Fig. 1. Neutralizing antibody responses in calves aged two to three weeks following vaccination with inactivated quadrivalent foot-and-mouth disease

vaccine adjuvanted with oil (A) or aluminium hydroxide (B). Neutralizing antibody response to individual FMDV serotypes in each animal was shown at day

zero (before vaccination) and 15, 30 and 60 days post-vaccination. The mean values are indicated by a horizontal bar. Statistical analysis was done by one-

way ANOVA and comparison between sets of results was assessed using Tukey post-test. Values of P < 0.05 were considered statistically significant

(*P < 0.05; ***P < 0.001 as compared to neutralizing antibody titres at day zero). Statistical analyses were performed using Prism 5 software.

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Letter to the Editor / Veterinary Microbiology 169 (2014) 233–235 235

ideal period in Indian subcontinent for FMDV vaccina- in growing calves that were born to immunized cows.others were not vaccinated, then vaccination could beertaken in calves as early as two weeks following birth,

protect the animals from disease and eventual FMDbreak. Further work is necessary to confirm theseults with newer vaccine candidates (Mohapatra et al.,8; Rudreshappa et al., 2012).

flicts of interest statement

None declared.

nowledgments

Authors are thankful to the Director, Indian Veterinaryearch Institute (IVRI) for providing the necessaryearch facilities. PKP is thankful to the Council ofntific and Industrial Research (CSIR), New Delhi for

financial support.

erences

P., Kalaivanan, R., Sied, N., Mamo, B., Kishore, S., Suryanarayana, V.V.,Kondabattula, G., 2013. Montanide ISATM 201 adjuvanted FMD vac-cine induces improved immune responses and protection in cattle.Vaccine 31, 3327–3332.l, T.R., 2003. FMD vaccines. Virus Res. 91, 81–99.bman, M.J., Baxt, B., 2004. Foot-and-mouth disease. Clin. Microbiol.Rev. 7, 465–493.

H.S., Lee, N.H., Seo, M.G., Ko, Y.J., Kim, B., Lee, J.B., Kim, J.S., Park, S.,Shin, Y.K., 2013. Serological responses after vaccination of growingpigs with foot-and-mouth disease trivalent (type O, A and Asia 1)vaccine. Vet. Microbiol. 164, 239–245.hanmohan, M., Nagendrakumar, S.B., Kumar, R., Anilkumar, J., Man-ikumar, K., Yuvaraj, S., Srinivasan, V.A., 2012. Clinical protection, sub-clinical infection and persistence following vaccination with extinc-tion payloads of O1 Manisa Foot-and-Mouth Disease monovalentvaccine and challenge in goats and comparison with sheep. Res.Vet. Sci. 93, 1050–1059.apatra, J.K., Hemadri, D., Rao, T.V., Sreenivasa, B.P., Subramaniam, S.,

Sanyal, A., Periyasamy, T.R., Singh, N.K., Pattnaik, B., Venkataramanan,R., 2008. Assessment of suitability of two serotype A candidatevaccine strains for inclusion in FMD vaccine in India. Vet. Microbiol.131, 65–72.l, P.K., Bayry, J., Ramakrishna, C., Hugar, B., Misra, L.D., Natarajan, C.,2002a. Immune responses of goats against foot-and-mouth diseasequadrivalent vaccine: comparison of double oil emulsion and alumi-nium hydroxide gel vaccines in eliciting immunity. Vaccine 20, 2781–2789.l, P.K., Bayry, J., Ramakrishna, C., Hugar, B., Misra, L.D., Prabhudas, K.,Natarajan, C., 2002b. Immune responses of sheep to quadrivalent

double emulsion foot-and-mouth disease vaccines: rate ofdevelopment of immunity and variations among other ruminants.J. Clin. Microbiol. 40, 4367–4371.

Patil, P.K., Bayry, J., Nair, S.P., Gopalakrishna, S., Sajjanar, C.M., Misra, L.D.,Natarajan, C., 2002c. Early antibody responses of cattle for foot-and-mouth disease quadrivalent double oil emulsion vaccine. Vet. Micro-biol. 87, 103–109.

Rodriguez, L.L., Gay, C.G., 2011. Development of vaccines toward theglobal control and eradication of foot-and-mouth disease. ExpertRev. Vaccines 10, 377–387.

Rudreshappa, A.G., Sanyal, A., Mohapatra, J.K., Subramaniam, S., De, A.,Das, B., Singanallur, N.B., Jangam, A.K., Muthukrishnan, M., Villuppa-noor, S.A., Pattnaik, B., 2012. Emergence of antigenic variants with inserotype A foot and mouth disease virus in India and evaluation of anew vaccine candidate panel. Vet. Microbiol. 158, 405–409.

Rweyemamu, M.M., Booth, J.C., Head, M., Pay, T.W., 1978. Microneutra-lization tests for serological typing and subtyping of foot-and-mouthdisease virus strains. J. Hygiene (London) 81, 107–123.

Prasanna K. Patil*Indian Veterinary Research Institute, Hebbal,

Bangalore 560 024, India

Channabasavaraj M. SajjanarUniversity of Agricultural Sciences, Dharwad 580 005, India

Chitattor NatarajanIndian Veterinary Research Institute, Hebbal,

Bangalore 560 024, India

Jagadeesh Bayry**Institut National de la Sante et de la Recherche Medicale, Unite

1138, Paris F-75006, France

*Corresponding author. Present address: Division ofAquatic Animal Health and Environment, Central Institute

of Brackishwater Aquaculture (CIBA), Chennai 600 028,India.

Tel.: +91 44 24 61 69 48; fax: +91 44 24 61 75 23

**Corresponding author at: INSERM Unite 1138, Centre deRecherche des Cordeliers, 15 rue de l’Ecole de Medicine,

Paris F-75006, France.Tel.: +33 1 44 27 82 03; fax: +33 1 44 27 81 94

E-mail addresses: pkpatilvet@yahoo.com (P.K. Patil)jagadeesh.bayry@crc.jussieu.fr (J. Bayry)

7 October 2013