Seroprevalence of Foot and Mouth Disease virus antibodies in trade cattle (Bos indicus) in Kwara state of Nigeria

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1 Veterinary Wrld, EISSN: Available at RESEARCH ARTICLE Open Access Serprevalence f Ft and Muth Disease virus antibdies in trade cattle (Bs indicus) in Kwara state f Nigeria 1, O. H. Olabde, H. M. Kazeem, M. A. Raji and N. D. Ibrahim 1. Department f Veterinary Micrbilgy, Faculty f Veterinary Medicine, University f Abuja, Abuja, Nigeria 2. Department f Veterinary Micrbilgy, Faculty f Veterinary Medicine, Ahmadu Bell University, Zaria, Nigeria Crrespnding authr: O.H Olabde labdek@yah.cm, Telephne: Received: , Revised: , Accepted: , Published nline: di: /vetwrld Hw t cite this article: Olabde OH, Kazeem HM, Raji MA and Ibrahim ND (2013) Serprevalence f Ft and Muth Disease virus antibdies in trade cattle (Bs indicus) in Kwara state f Nigeria, Veterinary Wrld 6(10): Abstract Aim: A serlgical survey fr the detectin f Ft and Muth Disease virus antibdies in trade cattle was cnducted t determine the serprevalence f Ft and Muth Disease (FMD) in the Kwara state f Nigeria. Materials and Methds: A crss-sectinal study based n cnvenient sampling fr bvine bld was carried ut in 5 cattle markets (Offa, Bde Sadu, Ilesha baruba, Ilrin, Ajasse) acrss five lcal gvernment areas in Kwara State amngst cattle with unknwn histry f FMD vaccinatin ver a perid f August-September-Octber, The sera btained were screened R using NS-Blcking ELISA kit (PRIOCHECK ). Results: Out f the 450 cattle (268 cws and 182 bulls) screened fr FMD antibdies, ser-psitivity was highest amngst cws 62.13%. Percentage distributin f ser-psitive reactrs by lcatin shwed Ilesha baruba, Ilrin and Offa had 27.14%, 23.80% and 13.33% respectively, while serpsitivity amngst bulls was 37.86%, highest ccurrence in Offa 30.46% and least in Ilesha baruba 14.84%. FMD carriers accrding t breeds shwed 77.21% white Fulani (Bunaji) was psitive ver the ther breeds. The sex, lcatin and breed was significant by chi square p<0.05. FMD serpsitivity accrding t age indicated yung cattle within 6 mnths-2years grup had the highest ccurrence 75.14% ver the adult grup 24.85%. Cnclusin: The ccurrence thus cnfirms FMD endemicity and susceptibility f all sex, ages, and varius breed f unvaccinated trade cattle especially amngst white Fulani, yung cws within the lcal markets and thse frm neighbring West African cuntries. Thus, circulating FMDV strains respnsible fr this serpsitivity shuld be established in rder t design apprpriate cntrl strategy t limit FMD effect. Key wrds: Ft and Muth disease virus antibdies, Kwara state, serprevalence, trade cattle Intrductin Ft and muth disease (FMD), is a highly cntagius viral disease f bth dmestic and wild clven hfed animals characterized by high mrbidity and decreased livestck prductivity, with affected cuntries being excluded frm internatinal animal trade [1]. FMD virus (FMDV) which cnsists f a single-stranded, psitive-sense RNA genme, belnging t the family Picrnaviridae that encdes a large plyprtein, which is then cleaved int the structural prteins and nnstructural prteins [2]. The seven distinct sertypes include Type A, O, C (Eurpean types), Suth African types (SAT 1, SAT 2, SAT 3) and Asiatic Type. Cumulative incidence f FMD sertypes shwed that six f the seven sertypes (O, A, C, SAT-1, SAT-2, SAT-3) have ccurred in Africa [3]. In Nigeria, previus reprts cnfirm endemicity f FMD with serius ecnmic lsses due t sertypes A, and SAT 2 [4; 5], sertypes O, SAT1 and SAT2 [6]. SAT1 and SAT 2 sertypes antibdies were als demnstrated [7]. Reviews f FMD reprted sertypes Cpyright: The authrs. This article is an pen access article licensed under the terms f the Creative Cmmns Attributin License ( which permits unrestricted use, distributin and reprductin in any medium, prvided the wrk is prperly cited. A, O, SAT 1 and SAT 2 have been respnsible fr disease utbreaks in Nrthern Nigeria as these sertypes have been in circulatin in the last 54 years ( ) [8; 9]. The inaccurate reprted endemicity f the disease [4] is assciated t nne reprting f disease utbreaks [5] due t inefficient disease reprting system in the cuntry [10] and this has made planning fr preventin and cntrl f the disease very difficult [7]. Despite the endemicity f Ft and muth disease, there is paucity f reprted infrmatin as regards the dynamics and status f FMD in Kwara state where a high cncentratin f cattle herds is fund. This was assciated with insufficient cntrl effrts which had made the disease endemic with mild curse and mrtality [11]. Serlgical demnstratin f specific antibdies t nn-structural prteins in nnvaccinated animals, where a vesicular cnditin is present, is sufficient fr a psitive diagnsis. This is particularly useful in mild cases r in cases where epithelial tissue cannt be cllected [12]. The detectin f antibdies t the NSP 3ABC f FMDV has been shwn t be sensitive and specific methd t differentiate between infectin and vaccinatin [13-17]. NSPs, unlike structural prteins, are highly cnserved and therefre, are nt sertype specific and as a cnsequence, the detectin f these antibdies is Veterinary Wrld, EISSN:

2 Available at nt sertype restricted [18]. Enzyme-linked f the cattle marketers as earlier cnducted [22]. The immunsrbent assay (ELISA) test is a mre specific, ages were designated in Hausa as Marki (male)/ Marka sensitive, and fast and is independent f cell cultures (female)/wege (Fulani) as age frm 6 mnths-2years. and usually nt affected by anti-cmplement factrs Bijimi (male)/karsana (female) in Hausa while Nghari present in the cattle serum. The ELISA can be perfrmed (Fulani) as frm 3-5years abve. Age was gruped int with inactivated antigens, thus requiring less restrictive tw categries: 6 mnths - 2 years and 3 years and bicntainment facilities [19]. Hence serlgic abve. quantificatin f FMD antibdy level t 3-ABC nn- Sample cllectin: Five milliliters f bld was structural prtein can be used t establish the disease cllected frm each well restraint animal thrugh the burden [14]. This premised the use f PRIOCHECK jugular vein by using 10 ml syringe and 18G needle FMD-3ABC NS prtein ELISA t establish the int 5 ml sample vials and kept at rm temperature at prevalence f FMD in Kwara State which hsts the Ilesha baruba cattle internatinal market in Baruten an angle f 45 C fr 6-8 hrs t allw fr cltting f LGA that shares an internatinal brder with Benin bld and sera separatin. The sera were aspirated int Republic. new set f 5 ml plastic tubes, labeled, stred n ice pack and transprted t the labratry, Natinal Veterinary Materials and Methds Research Institute, Vm where it was kept at -20 C until Ethical apprval: The research prcedure was used. apprved by the staff and pstgraduates seminar Serlgical assay using FMDV-NS ELISA: FMDV NS cmmittee f the Faculty f Veterinary Medicine, fr in vitr detectin f antibdies against bvine Ahmadu Bell University Zaria, Kaduna State f FMDV in serum samples as described [16] was Nigeria. The samples were cllected, stred and R cnducted using PriCHECK FMDV-NS blcking analyzed using ELISA kit as apprved by the ELISA. The preparatin f wrking slutins was cmmittee in accrdance with the manufacturer s cnducted in advance, this include dilutin buffer (PRIOCHECKR) recmmendatin. wrking slutin, Additives, ELISA buffer, Cnjugate Study area: Kwara State is als referred t as the dilutin and Washing slutin as recmmended by the 'gateway' State because f its lcatin between the manufactures [Prinics Lelystad B.V.: The Netherlands]. Nrthern and Suthern parts f Nigeria. This State is a On the secnd day, the test plates were cated with first rder administrative divisin lcated n the 3ABC specific mnclnal antibdy (mab), using lngitude N 8 30' 0'' and latitude E 5 0' 0'' 8.5 / 5 (Ge 80µl ELISA buffer t wells f the plates. Twenty Name Id : ) [20] with 16 Lcal Gvernment micrliter (20µl) negative cntrl was dispensed in Areas namely: Asa, Baruten, Edu, Ekiti, Ifeldun, wells A1 and B1, and 20ul f weak psitive cntrl was dispensed in wells C1 and D1 while 20µl f psitive Ilrin East, Ilrin West, Irepdun, Isin, Kaiama, Mr, cntrl was dispensed in wells E1 and F1. The test was Offa, Oke-Er, Oyun, Ilrin suth and Pategi. This perfrmed by dispensing the 20µl f test samples t the state shares cmmn bundaries with Niger and Kebbi remaining wells f a Test plates, sealed using enclsed States t the Nrth, Oy, Ond and Ed States t the plate sealer and shaked gently prir t vernight Suth, Benue, Plateau and Federal Capital Territry t incubatin at 22+3 C. After incubatin the plates were the East [21]. It maintains an internatinal bundary washed six times with µl f washing slutin with the Republic f Benin t the West. and 100ul cnjugate was added t all wells. The test Study design: A crss-sectinal study was designed plates were sealed using enclsed sealers and incubated using cnvenient sampling based n availability f fr 60 minutes at 22+3 C. FMDV NS specific antibdies, cattle and the willingness f the cattle wners at varius directed against the nn-structural prteins that may be lcatin cvered by this study. The sampling was present in the test samples bnd t the 3ABCprtein cnducted frtnightly ver a perid f three mnths and hence blcked the binding f the mab-hrpo. (August, September, and Octber, 2011). Five cattle After incubatin, the plates were washed six times with markets in different lcal gvernment areas was µl washing slutin (plates were tapped firmly cnsidered fr this study based n their lcatin and after the last washing) and 100µl chrmgen (TMB) prximity t ther States and internatinal brder substrate was dispensed t all the wells. After incubatin (Benin republic, Niger, Oy, Osun and Ekiti States). at rm temperature (22+3 C) fr 20 minutes the clr The markets include: Offa, Bde Sadu, Ilesha baruba, develpment was stpped, by adding 100µl f stp Ilrin and Ajasse in Offa, Mr, Baruten, Ilrin East, slutin and mixed well prir measuring. The clr and Irepdun LGAs respectively fr the cllectin f develpment was measured ptically at a wavelength bld frm trade cattle fr separatin int sera. Sex and f 450 nm t shw the presence f antibdies directed breed was being nted as bld samples were cllected against FMDV within 15 minutes after additin f stp fr risk factr analysis. Ageing f the cattle was slutin. The OD450 value f all samples including the cnducted as traditinally classified in Hausa/ Fulani cntrls were calculated and expressed as percentage languages during participatry epidemilgy in Inhibitin (PI) relative t the OD 450 max by first accrdance with lcal existing veterinary knwledge calculating the mean OD 450 value f wells A1 and B1 Veterinary Wrld, EISSN:

3 Available at Table-1. Serprevalence f FMD in cattle by sex in Kwara state Lcatin Female Male Psitive Psitive Negative Negative Ttal Ttal sampled sampled female male female male psitive negative Offa Bde Sadu Ilesha Ilrin Ajasse Ttal Table-2. Serprevalence f FMD in cattle by breed in Kwara state Type Psitive Negative Ttal White Fulani (Bunaji) Skt guduali (Bklji) Red Brr (Rahaji) Ttal 338 (75.11%)* * Prevalence f FMD carriers in cattle Table-3. Serprevalence f FMD in cattle by age in Kwara state Age Psitive Negative Ttal Yung (6 mths-2 years) Adult (3-5 years) Ttal 338 (75.11%)* * Prevalence f FMD carriers in cattle (Negative Cntrl= OD450 max). Then using the and least in Ilesha 14.84% (19/128). The cattle sex with frmula belw; lcatin als shwed statistical significance (P<0.05) 2 PI = 100 [OD test sample/od max] x 100 with sampling lcatins (LGAs) (χ =45.4, df=4) by chi A PI f < 50% was cnsidered negative and it was square analysis. interpreted that the animal tested had nt been expsed The distributin f ser-psitive FMD carriers t FMD fr 40 days. A PI f > 50% was cnsidered accrding t breeds in this study (Table-2) shwed that psitive and recent expsure <40> days t FMD. Mre ut f the 306 white Fulani (Bunaji) sampled 77.21% specifically, a PI value f > 50% but < 70% was (261/338) was psitive which indicated the highest cnsidered a weak psitive result and a PI value f > ccurrence ver the ther breeds f cattle. Red brr 70% was cnsidered a strng psitive result [16]. (Rahaji) and Skt Gudali (Bklji) had 11.83% (40/338) and 10.94% (37/338) respectively. Hwever, Statistical analysis: The psitive reactrs as well as the the serprevalence f FMD in assciatin with breed assciated risk factrs were expressed as simple shwed statistical significance (P<0.05) with white descriptive statistics such as percentage/frequency and 2 fulani (χ =57.2, df=2) by chi square analysis. chats. The data was further subjected t chi square Table-3 shws the distributin f FMD analysis using SPSS 19 sftware. serpsitivity accrding t age classificatin based n Results existing traditinal knwledge f the marketers. The yung cattle within 6 mnths - 2years grup had The verall FMD serprevalence recrded in this highest ccurrence 75.14% (254/338), while the study was 75.11% (338/450) as shwn in Table-1. The ccurrence was least in the adult grup 24.85% distributin f ser-psitive reactrs by lcatin in (84/338). The serprevalence f FMD in assciatin different Lcal Gvernment Areas in the state with age shwed statistical significance (P<0.05) with indicated that Ilesha baruba LGA recrded the highest 2 the yung grup f cattle (6 mnths-2yrs) (χ =48.4, ccurrence f 22.48% (76/338). This was clsely df=1). fllwed by Ilrin East LGA 20.71% (70/338). Others Table-4 shws Percentage Inhibitin (PI) f sera include 19.82% (67/338), 19.23% (65/338) and samples frm FMD trade cattle carriers at Optical 17.75% (60/338) in Offa, Bde Sadu and Irepdun density (OD) f 450nm as 112 negative sera (PI% LGAs respectively. Chi square analysis shwed n <50%), 69 sera as weak psitive (PI% 50-70%) and statistical significance (P>0.05) f FMD in assciatin 269 strng psitive sera (PI% >70). with LGAs in the state especially in Offa, Bde Sadu, and Ajasse while Ilesha and Ilrin shwed assciatin Table-4. Percentage Inhibitin (PI) f sera samples frm with the ccurrence f FMD. Out f 450 cattle FMD carrier trade cattle at Optical density (OD) f 450nm screened, 268 were female (cws) and 182 was males Percentage inhibitin Frequency FMD carrier status (bulls). The highest number f females sampled was in <50% 112 Negative Ilesha (71) and Ilrin (63) respectively, while the 50-70% 69 Weak 70% > 269 Strng highest number f males sampled was in Offa (54) and Ttal 338 (75.11%)* Ajasse (52) respectively. The serpsitivity was highest * Prevalence f FMD carriers in cattle amngst female (cws) 62.13% (210/338) with highest Discussin ccurrence in Ilesha 27.14% (57/210), Ilrin 23.80% (50/210), and least 13.33% (28/210) in Offa. Hwever, The FMD psitive cattle in this market survey the serpsitivity amngst male (bulls) was 37.86% where either in the categry f Percentage Inhibitin (128/338) with the highest ccurrence in bulls in Offa (PI) PI f > 50%-70% and r PI value f > 70% that 30.46% (39/128) and 21.09% (27/128) respectively was cnsidered as weak - strng psitive respectively, Veterinary Wrld, EISSN:

4 Available at this cnfirmed that such cattle had been recently [26]. This variatin maybe attributed t high number f expsed t FMD within the minimum f <40> days as female sampled at varius lcatins as mre cws are previusly dcumented [16]. Thus cnsidered as FMD seemly fund in markets fllwing culling prbably carriers shwing n clinical signs but have evidence f due t failing reprductive perfrmance. detectable antibdies giving an verall prevalence f The distributin f ser-psitive FMD carriers 75.11% using ELISA kit (PRIOCHECK). This kit accrding t breeds in this study (Table II) shwed that detects antibdies directed against the nn-structural 77.21% white Fulani [Bunaji] was psitive which 3ABC prtein f FMDV, cnfirming FMDV infected indicated the highest ccurrence ver the ther breeds animals independent f animal vaccinatin status as f cattle such as Red brr [Rahaji] and Skt vaccines cnsist f (partly) purified structural prteins Gudali [Bklji]. This finding culd be assciated f FMD virus and therefre vaccinated animals nly with the predminance f white Fulani breed within elicits antibdies directed against the structural cattle markets and ultimately their availability at time prteins f the virus, which is the basis f this f sampling. Hence, this bservatin wuld require discriminatin between infected and vaccinated further elucidatin t establish the pssibility f any animals. breed predispsitin with FMD. Overall prevalence f 75.11% in this survey is Serpsitivity f FMD accrding t age (Table-3) higher than Natinal surveillance f FMD in which shwed that yung cattle within 6 mnths-2 years serprevalence f 55.5% was reprted [23], and als grup had the highest ccurrence 75.14% ver the 64.30% dcumented [24] in nmadic herds frm adult grup. These yung cattle were mst pised fr Plateau state as well as 56.3% in Js suth LGA in trade as their heavy availability in the markets culd be Plateau state [11]. The increased prevalence fund in assciated with affrdability and high turnver. This this study cmpared t previus studies may be may als have fast tracked migratin and cntact with assciated with extensive mvement f livestck,high ther FMD expsed cattle that suffered infectin frm rate f cntact between animals at livestck markets, multiple sertypes thus prducing antibdies against cmmn grazing places as well as at watering pint all sertypes f FMD. This serprevalence f FMD alng cattle rutes as earlier suggested [23] especially assciatin with age was similar t previus reprt [11] as this study area hst the cattle internal market at that cattle within the age f 1-2yrs are mre prne t Ilesha baruba, a Nigerian brder twn t Benin FMD. republic where these cattle are ff laded at Paraku and The ELISA emplyed in this study shwed that trek thrugh cattle rutes ver a three day jurney t the 112 sera was negative [PI% <50%], 69 sera as weak market fr n ward distributin t ther part f Nigeria. psitive [PI% 50-70%] and 269 strng psitive [PI% Hwever, the 72.62% evidence f FMD nn-structural >70] (Table-4). This High percentage ccurrence f prtein antibdies reprted fr cattle in sme states psitive sera indicated pr vaccinatin cntrl especially in the Nrth West and Nrth Eastern Nigeria attempts amngst Nigerian lcal and trade cattle alng [25] cnntes this finding prbably as thse areas were the West Africa sub regin as this technique screened Nigerian brder twns and first prtal f entry fr trade ff vaccinated cattle thus clarifying the dubtful cattle frm East and Central Africa. vaccinatin status f the cattle during this survey. This The distributin f ser-psitive reactrs by ppulatin f cattle with unknwn FMD vaccinatin lcatin in different Lcal Gvernment Areas in the histry had made the disease endemic with mild curse state (Fig.1) indicated that Ilesha baruba LGA recrded and mrtality as earlier reprted [11]. Previus studies the highest ccurrence f 22.48% (76/338). This was demnstrated antibdies t SAT1 and SAT 2 sertypes clsely fllwed by Ilrin East LGA 20.71% (70/338). [7] hwever, sertyping f psitive sera in this study is Ilesha and Ilrin shwed assciatin with the nging. ccurrence f FMD. This culd be attributed t influx Cnclusin and recmmendatin f large number f cattle t meet up high demand fr meat t satisfy the rbust human ppulatin in the state This study prvides preliminary infrmatin n capital (Ilrin) as well as heavy traffic and the prevalence and endemicity as well as risk factrs cngregatin f cattle frm different surces at the assciated with FMD as FMD carriers was fund in all Ilesha cattle market, especially thse frm West sex, ages and breeds f unvaccinated trade cattle in African trade rute thus cntaminating envirnment kwara State f Nigeria as well as thse imprted by with FMDV. The variatin f ccurrence by lcatin cattle trading partners alng the West African cast. was similar t previus reprt [11]. This cnfirms FMD as a transbundary cattle disease. Sex distributin indicated serpsitivity was Thus, circulating FMDV strains respnsible fr this highest amngst female (cws) 62.13% (210/338) with serpsitivity shuld be established in rder t design highest ccurrence in Ilesha 27.14% (57/210), Ilrin apprpriate cntrl strategy t limit FMD effect % (50/210) which shwed assciatin with It is therefre, recmmended that cntinuus FMDV antibdy reactrs. Hwever, cntrast previus surveillance and quarantine f cattle within herds and study that stated n difference in risk assciatin with inter-state and natinal brders shuld be encuraged. FMD transmissin between male and female animals Eradicatin strategy thrugh cmpulsry vaccinatins Veterinary Wrld, EISSN:

5 Available at using cst-effective vaccines made f circulating disease in cattle in Js suth lcal gvernment area f Plateau strains are als advcated. state. Vm J. Vet. Sci. 8: Hamblin, C., Barnett, I. T. R. and Hedger, R.S. (1986) A new Authrs cntributins enzyme linked immunsrbent assay (ELISA) fr the OHO: This research wrk was part f dctrate detectin f antibdies against ft-and-muth disease virus, I. Develpment and methd f ELISA, J. Immunl. Meth. 93: investigatin. HMK, MAR and NDI: designed, planned and rganized the study as well as interpretatin f 13. Bergmann I. E., de Mell P.A., Neitzert E., Beck E. and Gmes I. (1993) Diagnsis f persistent aphthvirus result, discussin and review f manuscript. HMK and infectin and its differentiatin frm vaccinatin respnse in MAR als cntributed in sample cllectin. All authrs cattle by use f enzyme-linked immunelectrtransfer blt read and apprved the final manuscript. analysis with biengineered nnstructural viral antigens. Am. J. Vet. Res. 54: Acknwledgements 14. De Dieg, M., Brcchi, E., Mackay, D. and De Simne, F. (1997) The nnstructural plyprtein 3ABC f ft-and- Authrs are grateful t the Executive Directr, muth disease virus as a diagnstic antigen in ELISA t Natinal Veterinary Research Institute, Vm, Dr. differentiate infected frm vaccinated cattle. Arch. Virl. Ahmed, M.S fr granting us the permissin t stre and 142: analyze the sera samples as well as Drs. Ularamu, H.G, 15. Mackay, D. K., Frsyth, M.A., Davies, P.R., Berlinzani, A., Belsham, G.J., Flint, M. and Ryan, M.D. (1998) Wungak Y.S and Mr. Agm, D fr their assistance Differentiating infectin frm vaccinatin in ft-andduring the labratry analysis. Persnal fund f the muth disease using a panel f recmbinant, nn-structural authr (OHO) was used fr the executin f this prteins in ELISA. Vaccine. 16: survey. 16. Srensen, K. J., Madsen, K.G., Madsen, E.S., Salt, J.S., Cmpeting interests Nqindi, J. and Mackay, D.K. 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(1987) Studies n the epidemilgy f ft and internatinal trade Trp. Anim. Health. Prd. 40: and muth disease in Nigeria. PhD Thesis. Ahmadu Bell 23. Abegunde A.A., Ezekli C.D., Umh J.U., and Add P.B. University, Zaria: Pp. (1988) Epidemilgy f Ft and muth disease in Nigerian 7. Chukwued A. A., Nimzing L., Olabde, A.O. and cattle. Viral Diseases f animals in Africa. OAU/STRC Abegunde, A. (2008) Prevalence f Ft and Muth disease Scientific Publicatin Lags Nigeria, pp Virus, SAT 1 and SAT 2 sertype antibdies in Nigerian 24. Ehizibl, D.O., Ajgi, I., Umh, J.U., Kazeem, H.M., cattle. Anim. Prd. Res. Adv. 4 (2): Ehizibl, P.O, Perez, A.M. and Metwally, S.A. (2010) 8. Nwanta, J.A. and Ojemiren, F.O. (2000) Epidemilgy f Serlgical survey f Ft and Muth Disease (FMD) 3d Ft and Muth Disease (FMD) in nrthern states f Nigeria. 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