research article occurrence of newcastle disease and...

Research ArticleOccurrence of Newcastle Disease and Infectious Bursal DiseaseVirus Antibodies in Double-Spurred Francolins in Nigeria

Daniel Oladimeji Oluwayelu1 Adebowale Idris Adebiyi1 Ibukunoluwa Olaniyan1

Phyllis Ezewele1 and Oluwasanmi Aina2

1 Department of Veterinary Microbiology and Parasitology University of Ibadan Ibadan 20005 Nigeria2 Department of Veterinary Anatomy University of Ibadan Ibadan 20005 Nigeria

Correspondence should be addressed to Daniel Oladimeji Oluwayelu ogloryusyahoocom

Received 19 August 2014 Revised 23 October 2014 Accepted 6 November 2014 Published 18 November 2014

Academic Editor Carlos Gonzalez-Rey

Copyright copy 2014 Daniel Oladimeji Oluwayelu et al This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

The double-spurred francolin Francolinus bicalcaratus has been identified as a good candidate for future domestication due tothe universal acceptability of its meat and its adaptability to anthropogenically altered environments Therefore in investigatingthe diseases to which they are susceptible serum samples from 56 francolins in a major live-bird market (LBM) in Ibadansouthwestern Nigeria were screened for antibodies against Newcastle disease (ND) and infectious bursal disease (IBD) virusesHaemagglutination inhibition (HI) test and enzyme-linked immunosorbent assay (ELISA) revealed 250 and 357 prevalenceof ND virus (NDV) antibodies respectively while 54 and 571 prevalence of IBD virus (IBDV) antibodies was detected byagar gel precipitation test (AGPT) and ELISA respectively This first report on the occurrence of NDV and IBDV antibodies inapparently healthy unvaccinated double-spurred francolins from a LBM suggests that they were subclinically infected with eitherfield or vaccine viruses and could thus serve as possible reservoirs of these viruses to domestic poultry Furthermore if they are tobe domesticated for intensive rearing a vaccination plan including ND and IBD should be developed and implemented

1 Introduction

Newcastle disease (ND) and infectious bursal disease (IBD)are the two most dreaded viral diseases of poultry in Nigeriaas they cause severe economic losses in domestic and wildbird populations resulting from illness reduced egg pro-duction immunosuppression and death following infectionwith pathogenic strains of their respective causative virusesDespite efforts to prevent and control them over the yearscirculation of the causative virus among free-roaming andwild birds has been reported as one of the factors responsiblefor the sporadic outbreaks of ND and IBD among free-roaming village chickens as well as commercial poultry flocks[1 2]

Newcastle disease (ND) is an acute highly contagiousrapidly spreading viral disease affecting birds of all ages [3]and is characterized in chickens by respiratory circulatorygastrointestinal and nervous signs [4]The clinical signs seen

in infected birds vary widely and are dependent on viralfactors like pathogenicity (which depends on virulence andtropism of the virus) host factors (species age and immunestatus) concurrent infections route of exposure durationand magnitude of the infection dose and external factorssuch as social and environmental stress [5] According toDocherty and Friend [6] it is capable of infecting over 230species from more than one-half of the 50 orders of birdsThese include domestic poultry [7 8] and wild birds such ashouse sparrows hawks crows double-breasted cormorantsand waterfowls [9ndash13] Wild birds constitute a natural reser-voir of low-virulence viruses while poultry are the mainreservoir of virulent strains Exchange of virus between thesereservoirs represents a risk for both bird populations [13]The most virulent form of ND virus (NDV) causes up to 100percent mortality in affected flocks [6]

On the other hand infectious bursal disease (IBD) isa highly contagious immunosuppressive viral infection of

Hindawi Publishing CorporationJournal of Veterinary MedicineVolume 2014 Article ID 106898 5 pageshttpdxdoiorg1011552014106898

2 Journal of Veterinary Medicine

chicks (3ndash6 weeks old) causing severe economic and produc-tion losses worldwide [14] Although turkeys ducks guineafowls and ostriches may be infected clinical disease occurssolely in chickens [15] However serological evidence of theinfection has been reported in free-living wild birds such ascordon bleu and village weaver [16] wild water birds [17]Antarctic penguins [18] cattle egrets [19] and wild turkeysand cranes [20] Moreover IBDV antibodies were detectablein the sera of sedentary and migratory wild bird species inJapan suggesting that they play a key role in the naturalhistory of IBD [21] while the virus was isolated from wildbirds in Korea [22]

Although several studies have been conducted on francol-ins in South Africa [23ndash25] sparse information exists on theWest African-based double-spurred francolin Francolinusbicalcaratus (synonym Pternistis bicalcaratus) which is agamebird in the pheasant family Phasianidae of the orderGalliformes It is a resident breeder in tropical West Africaand feeds on insects vegetable matter and seeds [26]According to Keith et al [27] seven Afrotropical francolinspecies including the double-spurred francolin have beenfound in Nigeria where they are widely consumed as bushmeat In a comparative biochemical study of meat qualityand digestive enzymes their meat was reported to be tastierjuicier more palatable and richer in protein than domesticchicken meat [28] Based on these desirable qualities offrancolinmeat it is needful to consider them as an alternativesource of affordable animal protein to the ever-increasingNigerian human population Moreover although they arefound mostly in the wild Mbinkar et al [29] noted that thespecies is considered a good candidate for future domes-tication due to the universal acceptability of its meat andits adaptability to anthropogenically altered environmentswhich may be occasioned by extensive bush burning andintensive grazing of grasslands Therefore since studies ontheir biology and ecology which can provide a basis for theireventual domestication have been conducted [28ndash31] thereis a need to also investigate the infections to which they aresusceptible Apart from few studies which showed that fran-colins are affected by diseases such as Marekrsquos disease [32]coccidiosis [33] toxoplasmosis [34] and bacterial sinusitis[35] there is sparse information on viral diseases such asND and IBD in francolinsThis study was therefore designedto investigate the presence of NDV and IBDV antibodies infree-living double-spurred francolins caught in the wild andsold at a popular live-bird market (LBM) located in ShashaIbadan southwest Nigeria

2 Materials and Methods

The study was conducted in Ibadan (latitude 7∘ 231015840 N andlongitude 3∘ 561015840 E) the capital city of Oyo State southwestNigeriaThis region is the core of the Nigerian poultry indus-try with Ibadan being a major city from where poultry (day-old chicks broilers and point-of-lay pullets) and poultryinputs (drugs vaccines and feed ingredients) are distributedto other parts of the countryThe city also has some LBMs ofwhich the one located at Shasha is popular as it is a sales point

for diverse bird species brought by peasant farmers fromadjoining rural communities and traders from the northernpart of the country that transport cattle sheep and goats tosouthwest Nigeria

Blood samples collected via the jugular vein betweenApril and August 2012 from 56 double-spurred francolinsat Shasha live-bird market were poured into sterile samplebottles without anticoagulant and allowed to clot at roomtemperature Separated sera were harvested and stored atminus20∘C Although larger sample population of spurred fran-colins existed in the market and prior consultations weremade with the traders on the importance of the project thenumber of birds available for bleeding was restricted to 56 inview of the refusal of traders to have their birds bledThe agesof the birds could not be determined

The 56 sera were screened for antibodies to NDV andIBDV using the haemagglutination inhibition (HI) test andagar gel precipitation test (AGPT) as described by Durojaiyeand Adene [36] and Hirai et al [37] respectively Positivecontrol ND and IBD sera were obtained from the NationalVeterinary Research Institute Vom Nigeria Antibodies toNDV and IBDVwere also detected and quantified in 42 fran-colin sera (the remaining 14 sera had been exhausted) usingcommercial ND and IBD enzyme-linked immunosorbentassay (ELISA) kits (ProFLOK Plus Synbiotics CorporationKansas City USA) respectively according to the manufac-turerrsquos instructions Positive and normal control ND and IBDsera were used to validate the tests As specified by the kitmanufacturer a serum dilution of 1 50 was used and opticaldensity (OD) values were read at 405 nm with an ELx800universal microplate reader (Bio-Tek Vermont USA) ValidNDV or IBDV ELISA results were obtained when the averageODvalue of the normal control serumwas less than 0250 andthe corrected positive control value range was between 0250and 0900

Data obtained were analysed with column statistics usingGraphPad prism version 50 (GraphPad software San DiegoCA USA) and 119875 values lt 005 were considered significant

3 Results and Discussion

Prevalence ofNDVantibodies in the tested francolin sera was250 (1456) and 357 (1542) using theHI andELISA testsrespectively (Table 1) The HI antibody titers ranged from1 2 to 1 32 while mean NDV antibody titer obtained withthe ELISA was 3206 (95 CI 1261ndash5152) There was a 595agreement between the HI test and the ELISA Only 3 (54)of the tested sera were positive for IBDV antibodies with theAGPT while 24 (571) were positive using ELISA (Table 2)Mean IBDV antibody titer obtained with the ELISA was 5735(95CI 2919ndash8550) and there was a 50 agreement betweenthe AGPT and ELISA Using the ELISA 286 (1242) of thetested sera had antibodies to both NDV and IBDV

This study investigated the presence of NDV and IBDVantibodies in free-living double-spurred francolins caught inthe wild and sold at Shasha LBM which is a trading centrewhere different avian species including indigenous chickenspigeons guinea fowls turkeys and francolins are sold in

Journal of Veterinary Medicine 3

Table 1 Correlation of HI and ELISA for NDV antibodies

Test Positive Negative TotalHI 10 32 42ELISA 15 27 42

Table 2 Correlation of AGPT and ELISA for IBDV antibodies

Test Positive Negative TotalAGPT 3 39 42ELISA 24 18 42

Ibadan Oyo State southwest Nigeria To our knowledgethere is no information available on viral diseases of double-spurred francolins which have been suggested as a goodcandidate for domestication in order to meet the animalprotein needs of the Nigerian populace [29] Therefore thisfirst report on the detection of NDV- and IBDV-specific IgYantibodies in the sera of free-living francolins in Nigeria isan indication of previous exposure of these birds to the twoviruses Since theywere not routinely vaccinated it is possiblethat the birds acquired their ND andor IBD seropositive sta-tus through exposure to other infected wild or domestic birdsthat were shedding the viruses The seropositivity detectedcould be due to circulating ND and IBD vaccine viruseswhich the birds might have contracted through interactionwith vaccinated free-range birds or even through operatingoccasionally around commercial poultry farms Karesh et al[38] noted that wild birds in the pet or exotic bird tradehave the potential to transmit parasites bacteria and viruseswhichmay ormay not be pathogenic in their normal host butpose threats when introduced to new geographic locationsand new host species In this study we observed that wildand domestic bird specieswere kept by the traders in the samecages and this is consistent with a previous report thatND forexample is spread by contact between birds and exacerbatedby birds being mixed together in rural markets [13] As thefrancolins were usually kept at the LBMs for about 3-4 weeksbefore being sold it is likely that they acquired the virusesfrom infected domestic or wild bird species with which theywere kept in the same cages

Moreover the francolins tested in this study were allapparently healthy except for one that showed signs of tor-ticollis at the time of sample collection (Figure 1) Thereforethe detection of NDV and IBDV antibodies in their serasuggests that they were subclinically infected and could serveas reservoirs shedding the viruses into the environmentPrevious studies have implicated wild birds as possiblereservoirsvectors of these viruses for domestic poultry [1621 39] It is also possible that the birds possess some hostfactors responsible for resistance Moreover Kim et al [40]suggested wild-type virus transmission between wild anddomestic birds as the origin of the similarity of NDV strainsfound in wild birds and domestic birds in LBMs

It is noteworthy that 286 (1242) of the sera testedby the ELISA technique had high titres of both NDV andIBDV antibodies which is an indication that the birds werecoinfected with the two viruses Additionally the detection

Figure 1 Francolin showing sign of torticollis at the time of bloodcollection

of a higher proportion of positive samples by the ELISAtechnique compared to the HI test and AGPT shows that itis more sensitive than the latter two tests for detecting NDV-and IBDV-specific antibodies respectively in francolin seraThis is consistent with the reports of Bell et al [41] andMarquardt et al [42] who also found that the ELISA wasmore sensitive than the HI test and AGPT for detection ofantibodies to NDV and IBDV respectively

4 Conclusions

This study has shown that free-living double-spurred francol-ins are susceptible to infection with NDV and IBDV and canserve as reservoirs of these viruses thus acting as a meansof transmission to domestic poultry Therefore if they areto be domesticated for intensive rearing as an alternativesource of animal protein a vaccination programme whichincludes ND and IBD vaccinations should be developedand implemented to protect them from clinical disease Inaddition the detection of NDV and IBDV antibodies infrancolins sold at LBMs where they have close interactionwith commercial and village chickens and other wild birdswarrants continuous surveillance for these diseases becauseof increased concerns that low-virulence wild bird virusescould become more virulent in domestic bird populationsFurther studies to isolate the two viruses from francolins anddetermine their level of pathogenicity should be conducted

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

References

[1] E N Okeke and A G Lamorde ldquoNewcastle disease and itscontrol in Nigeriardquo in Viral Disease of Animals in AfricaA O Williams and W N Masiga Eds CTAOAUSTRCPublication Lagos Nigeria 1988

[2] O J Ibu A Aba-Adulugba M A Adeleke and A Y TijjanildquoActivity of Newcastle disease and infectious bursal disease


viruses in ducks and guinea fowls in Jos area Nigeriardquo SokotoJournal of Veterinary Sciences vol 2 pp 45ndash46 2000

[3] P A Abdu ldquoEvolution and the pathogenicity of Newcastledisease virus and its implications for diagnosis and controlrdquoin Proceedings of the Workshop on Improved Disease DiagnosisHealth Nutrition and Risk Management Practice in PoultryAhmadu Bello University Zaria Nigeria November-December2005

[4] N J MacLachlan and E J Dubovi ldquoParamyxoviridaerdquo inFenners Veterinary Virology N J MacLachlan and E J DuboviEds pp 299ndash325 Academic Press London UK 4th edition2011

[5] J B McFerran and R M McCracken ldquoNewcastle diseaserdquo inNewcastle Disease D J Alexander Ed pp 161ndash183 KluwerAcademic Boston Mass USA 1988

[6] D E Docherty and M Friend ldquoNewcastle diseaserdquo in FieldManual of Wildlife Diseases General Field Procedures andDiseases of Birds M Friend and J C Franson Eds pp 175ndash180USGS-National Wildlife Health Center Madison Wis USA1999

[7] P H Joslashrgensen K J Handberg P Ahrens O RTherkildsen RJ Manvell and D J Alexander ldquoStrains of avian paramyxovirustype 1 of low pathogenicity for chickens isolated from poultryand wild birds in Denmarkrdquo Veterinary Record vol 154 no 16pp 497ndash500 2004

[8] B S Seal M G Wise J C Pedersen et al ldquoGenomicsequences of low-virulence avian paramyxovirus-1 (Newcastledisease virus) isolates obtained from live-bird markets in NorthAmerica not related to commonly utilized commercial vaccinestrainsrdquoVeterinaryMicrobiology vol 106 no 1-2 pp 7ndash16 2005

[9] H Takakuwa T Ito A Takada K Okazaki and H KidaldquoPotentially virulent Newcastle disease viruses are maintainedinmigratory waterfowl populationsrdquo Japanese Journal of Veteri-nary Research vol 45 no 4 pp 207ndash215 1998

[10] O J Ibu J O A Okoye E P Adulugba et al ldquoPrevalence ofnewcastle disease viruses in wild and captive birds in centralNigeriardquo International Journal of Poultry Science vol 8 no 6pp 574ndash578 2009

[11] W Zhu J Dong Z Xie Q Liu and M I Khan ldquoPhylogeneticandpathogenic analysis ofNewcastle disease virus isolated fromhouse sparrow (Passer domesticus) living around poultry farmin southernChinardquoVirus Genes vol 40 no 2 pp 231ndash235 2010

[12] D G Diel P J Miller P C Wolf et al ldquoCharacterization ofnewcastle disease viruses isolated from cormorant and gullspecies in the United States in 2010rdquo Avian Diseases vol 56 no1 pp 128ndash133 2012

[13] C J Snoeck M Marinelli E Charpentier et al ldquoCharacteri-zation of newcastle disease viruses in wild and domestic birdsin Luxembourg from 2006 to 2008rdquoApplied and EnvironmentalMicrobiology vol 79 no 2 pp 639ndash645 2013

[14] H Muller M R Islam and R Raue ldquoResearch on infectiousbursal diseasemdashthe past the present and the futurerdquo VeterinaryMicrobiology vol 97 no 1-2 pp 153ndash165 2003

[15] Office Internationale des Epizooties ldquoNewcastle diseaserdquo inOIEManual of Standards for Diagnostic Tests and Vaccines pp 221ndash232 OIE Paris France 2008

[16] D R Nawathe O Onunkwo and I M Smith ldquoSerologicalevidence of infection with the virus of infectious bursal diseasein wild and domestic birds in Nigeriardquo Veterinary Record vol102 no 20 article 444 1978

[17] G EWilcox R L P Flower W Baxendale and J S MackenzieldquoSerological survey of wild birds in Australia for the prevalence

of antibodies to egg drop syndrome 1976 (EDS-76) and infec-tious bursal disease virusesrdquo Avian Pathology vol 12 pp 135ndash139 1983

[18] H Gardner K Kerry M Riddle S Brouwer and L GleesonldquoPoultry virus infection in Antarctic penguinsrdquoNature vol 387no 6630 p 245 1997

[19] O A Fagbohun D O Oluwayelu A A Owoade and F OOlayemi ldquoSurvey for antibodies to Newcastle disease virus incattle egrets pigeons and Nigerian laughing dovesrdquo AfricanJournal of Biomedical Research vol 3 no 3 pp 193ndash194 2000

[20] K L Candelora M G Spalding and H S Sellers ldquoSurveyfor antibodies to infectious bursal disease virus serotype 2 inwild Turkeys and Sandhill Cranes of Florida USArdquo Journal ofWildlife Diseases vol 46 no 3 pp 742ndash752 2010

[21] M Ogawa T Wakuda T Yamaguchi et al ldquoSeroprevalence ofinfectious bursal disease virus in free-livingwild birds in JapanrdquoJournal of Veterinary Medical Science vol 60 no 11 pp 1277ndash1279 1998

[22] W-J Jeon E-K Lee S-J Joh et al ldquoVery virulent infectiousbursal disease virus isolated from wild birds in Korea epidemi-ological implicationsrdquoVirus Research vol 137 no 1 pp 153ndash1562008

[23] J H van Niekerk ldquoSocial and breeding behaviour of thecrested francolin in the Rustenburg district South AfricardquoSouth African Journal of Wildlife Research vol 31 no 1-2 pp35ndash42 2001

[24] J H van Niekerk and D J Verwoerd ldquoAvian pox in Swainsonrsquosfrancolin in South Africardquo South African Journal of WildlifeResearch vol 32 no 1 pp 43ndash48 2002

[25] A C Uys and I G Horak ldquoTicks on crested francolinsFrancolinus sephaena and on the vegetation on a farm inLimpopo Province South Africardquo Onderstepoort Journal ofVeterinary Research vol 72 no 4 pp 339ndash343 2005

[26] Birdlife International ldquoFrancolinus bicalcaratusrdquo IUCN 2013IUCN Red List of Threatened Species Version 20132 2012httpwwwiucnredlistorg

[27] S Keith C H Fry and E K Urban The Birds of Africa vol 2Academic Press London UK 1993

[28] P C Onyenekwe Biochemical studies on the meat qualityand digestive enzymes of double-spurred francolin Francolinusbicalcaratus [MSc thesis] Ahmadu Bello University ZariaNigeria 1988

[29] D LMbinkar AU Ezealor and S J Oniye ldquoldquoFood habits of thedouble-spurred francolin Francolinus bicalcaratus (Linnaeus)in Zaria Nigeriardquo Journal of Biological Sciences vol 5 no 4 pp458ndash462 2005

[30] A U Ezealor Ecological profile of a Nigerian Sahelian wetlandtoward integrated vertebrate pest damage management [PhDthesis] Virginia Polytechnic Institute and State UniversityBlacksburg Va USA 1995

[31] J T C CodjiaM RM Ekue andGAMensah ldquoEcology of thedouble-spurred francolin Francolinus bicalcaratus in the south-east of BeninrdquoMalimbus vol 25 pp 77ndash84 2003

[32] J R Pettit P A Taylor and A W Gough ldquoMicroscopic lesionssuggestive of Marekrsquos disease in a Black Francolin (Francolinusf francolinus)rdquo Avian Diseases vol 20 no 2 pp 410ndash415 1976

[33] D F Adene and M Akande ldquoA diagnosis of coccidiosis incaptive bushfowl (Francolinus bicalcaratus) and identification ofthe causative coccidianrdquo African Journal of Ecology vol 16 no4 pp 227ndash230 1978


[34] T M Work J G Massey D S Lindsay and J P DubeyldquoToxoplasmosis in three species of native and introducedHawaiian birdsrdquo Journal of Parasitology vol 88 no 5 pp 1040ndash1042 2002

[35] W Yanhong Y Yuehui Z Shouchang and L Wenbo ldquoA reporton sinuitis of francolinrdquo Journal of Animal and VeterinaryAdvances vol 10 no 19 pp 2499ndash2500 2011

[36] O A Durojaiye and D F Adene ldquoNewcastle disease and eggdrop syndrome rsquo76 in guinea fowls (Numida meleagris galeataPallas)rdquo Journal of Veterinary Medicine Series B vol 35 no 2pp 152ndash154 1988

[37] K Hirai S Shimakura and M Hirose ldquoImmunodiffusionreaction to avian infectious bursal virusrdquo Avian Diseases vol16 no 4 pp 961ndash964 1972

[38] WBKaresh RACookMGilbert and JNewcomb ldquoImplica-tions of wildlife trade on the movement of avian influenza andother infectious diseasesrdquo Journal of Wildlife Diseases vol 43no 3 pp S55ndashS59 2007

[39] E Schelling BThur C Griot and L Audige ldquoEpidemiologicalstudy of Newcastle disease in backyard poultry and wild birdpopulations in Switzerlandrdquo Avian Pathology vol 28 no 3 pp263ndash272 1999

[40] B-Y Kim D-H Lee M-S Kim et al ldquoExchange of Newcastledisease viruses in Korea the relatedness of isolates betweenwild birds live bird markets poultry farms and neighboringcountriesrdquo Infection Genetics and Evolution vol 12 no 2 pp478ndash482 2012

[41] J G Bell D Ait Belarbi and A Amara ldquoA controlled vac-cination trial for Newcastle disease under village conditionsrdquoPreventive Veterinary Medicine vol 9 no 4 pp 295ndash300 1990

[42] W W Marquardt R B Johnson W F Odenwald and BA Schlotthober ldquoAn indirect enzyme-linked immunosorbentassay (ELISA) for measuring antibodies in chickens infectedwith infectious bursal disease virusrdquoAvian Diseases vol 24 no2 pp 375ndash385 1980

Submit your manuscripts athttpwwwhindawicom

Veterinary MedicineJournal of

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Veterinary Medicine International



International Journal of

Microbiology


AnimalsJournal of

EcologyInternational Journal of


PsycheHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Evolutionary BiologyInternational Journal of


Hindawi Publishing Corporationhttpwwwhindawicom

Applied ampEnvironmentalSoil Science

Volume 2014

Biotechnology Research International


Agronomy




Journal of Parasitology Research

Hindawi Publishing Corporation httpwwwhindawicom


Volume 2014

Zoology

GenomicsInternational Journal of


InsectsJournal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


VirusesJournal of

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Cell BiologyInternational Journal of



Case Reports in Veterinary Medicine


chicks (3ndash6 weeks old) causing severe economic and produc-tion losses worldwide [14] Although turkeys ducks guineafowls and ostriches may be infected clinical disease occurssolely in chickens [15] However serological evidence of theinfection has been reported in free-living wild birds such ascordon bleu and village weaver [16] wild water birds [17]Antarctic penguins [18] cattle egrets [19] and wild turkeysand cranes [20] Moreover IBDV antibodies were detectablein the sera of sedentary and migratory wild bird species inJapan suggesting that they play a key role in the naturalhistory of IBD [21] while the virus was isolated from wildbirds in Korea [22]

Although several studies have been conducted on francol-ins in South Africa [23ndash25] sparse information exists on theWest African-based double-spurred francolin Francolinusbicalcaratus (synonym Pternistis bicalcaratus) which is agamebird in the pheasant family Phasianidae of the orderGalliformes It is a resident breeder in tropical West Africaand feeds on insects vegetable matter and seeds [26]According to Keith et al [27] seven Afrotropical francolinspecies including the double-spurred francolin have beenfound in Nigeria where they are widely consumed as bushmeat In a comparative biochemical study of meat qualityand digestive enzymes their meat was reported to be tastierjuicier more palatable and richer in protein than domesticchicken meat [28] Based on these desirable qualities offrancolinmeat it is needful to consider them as an alternativesource of affordable animal protein to the ever-increasingNigerian human population Moreover although they arefound mostly in the wild Mbinkar et al [29] noted that thespecies is considered a good candidate for future domes-tication due to the universal acceptability of its meat andits adaptability to anthropogenically altered environmentswhich may be occasioned by extensive bush burning andintensive grazing of grasslands Therefore since studies ontheir biology and ecology which can provide a basis for theireventual domestication have been conducted [28ndash31] thereis a need to also investigate the infections to which they aresusceptible Apart from few studies which showed that fran-colins are affected by diseases such as Marekrsquos disease [32]coccidiosis [33] toxoplasmosis [34] and bacterial sinusitis[35] there is sparse information on viral diseases such asND and IBD in francolinsThis study was therefore designedto investigate the presence of NDV and IBDV antibodies infree-living double-spurred francolins caught in the wild andsold at a popular live-bird market (LBM) located in ShashaIbadan southwest Nigeria

2 Materials and Methods

The study was conducted in Ibadan (latitude 7∘ 231015840 N andlongitude 3∘ 561015840 E) the capital city of Oyo State southwestNigeriaThis region is the core of the Nigerian poultry indus-try with Ibadan being a major city from where poultry (day-old chicks broilers and point-of-lay pullets) and poultryinputs (drugs vaccines and feed ingredients) are distributedto other parts of the countryThe city also has some LBMs ofwhich the one located at Shasha is popular as it is a sales point

for diverse bird species brought by peasant farmers fromadjoining rural communities and traders from the northernpart of the country that transport cattle sheep and goats tosouthwest Nigeria

Blood samples collected via the jugular vein betweenApril and August 2012 from 56 double-spurred francolinsat Shasha live-bird market were poured into sterile samplebottles without anticoagulant and allowed to clot at roomtemperature Separated sera were harvested and stored atminus20∘C Although larger sample population of spurred fran-colins existed in the market and prior consultations weremade with the traders on the importance of the project thenumber of birds available for bleeding was restricted to 56 inview of the refusal of traders to have their birds bledThe agesof the birds could not be determined

The 56 sera were screened for antibodies to NDV andIBDV using the haemagglutination inhibition (HI) test andagar gel precipitation test (AGPT) as described by Durojaiyeand Adene [36] and Hirai et al [37] respectively Positivecontrol ND and IBD sera were obtained from the NationalVeterinary Research Institute Vom Nigeria Antibodies toNDV and IBDVwere also detected and quantified in 42 fran-colin sera (the remaining 14 sera had been exhausted) usingcommercial ND and IBD enzyme-linked immunosorbentassay (ELISA) kits (ProFLOK Plus Synbiotics CorporationKansas City USA) respectively according to the manufac-turerrsquos instructions Positive and normal control ND and IBDsera were used to validate the tests As specified by the kitmanufacturer a serum dilution of 1 50 was used and opticaldensity (OD) values were read at 405 nm with an ELx800universal microplate reader (Bio-Tek Vermont USA) ValidNDV or IBDV ELISA results were obtained when the averageODvalue of the normal control serumwas less than 0250 andthe corrected positive control value range was between 0250and 0900

Data obtained were analysed with column statistics usingGraphPad prism version 50 (GraphPad software San DiegoCA USA) and 119875 values lt 005 were considered significant

3 Results and Discussion

Prevalence ofNDVantibodies in the tested francolin sera was250 (1456) and 357 (1542) using theHI andELISA testsrespectively (Table 1) The HI antibody titers ranged from1 2 to 1 32 while mean NDV antibody titer obtained withthe ELISA was 3206 (95 CI 1261ndash5152) There was a 595agreement between the HI test and the ELISA Only 3 (54)of the tested sera were positive for IBDV antibodies with theAGPT while 24 (571) were positive using ELISA (Table 2)Mean IBDV antibody titer obtained with the ELISA was 5735(95CI 2919ndash8550) and there was a 50 agreement betweenthe AGPT and ELISA Using the ELISA 286 (1242) of thetested sera had antibodies to both NDV and IBDV

This study investigated the presence of NDV and IBDVantibodies in free-living double-spurred francolins caught inthe wild and sold at Shasha LBM which is a trading centrewhere different avian species including indigenous chickenspigeons guinea fowls turkeys and francolins are sold in











4 Conclusions




References






















































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of
















4 Conclusions




References






















































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of

























































Microbiology


AnimalsJournal of








Volume 2014



Agronomy







Volume 2014

Zoology



InsectsJournal of




VirusesJournal of






research article occurrence of newcastle disease and...

Documents