Seroprevalence of human cysticercosis in Jataí, Goiás state, Brazil

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brazjinfectdis2018;22(2):146–149

w w w . e l s e v ie r . c o m / l o c a t e / b j i d

The

Brazilian

Journal

of

INFECTIOUS

DISEASES

Brief

communication

Seroprevalence

of

human

cysticercosis

in

Jataí,

Goiás

state,

Brazil

Kaliny

Xavier

da

Guarda

a

_,

_Julia

_Maria

_Costa-Cruz

b

_,

Ivanildes

Solange

da

Costa

Barcelos

a,∗

a_Universidade_Federal_de_Goiás,_Laboratório_de_Imunologia,_Jataí,_GO,_Brazil

b_Universidade_Federal_de_Uberlândia,_Instituto_de_Ciências_Biomédicas,_Laboratório_de_Diagnóstico_de_Parasitoses,_Uberlândia,_MG,_Brazil

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Articlehistory:

Received27September2017

Accepted18January2018

Availableonline10February2018

Keywords: Cysticercosis Taeniasolium Epidemiology Diagnosis Brazil

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TheTaeniasoliumcysticercosisaffectsmillionsofpeopleworldwideandisconsidereda

publichealthproblem,especiallyindevelopingcountries.Thediagnosisof

neurocysticer-cosisiscomplexandinvolvestheanalysisofepidemiological,clinical,neuroimaging,and

immunologicalhostdata.NeurocysticercosisisendemicinBrazil,andisrelated tothe

causeofdeathmainlyintheSoutheast,South,andCentral-Westregions.Theobjective

ofthisstudywastodeterminetheseroprevalenceofcysticercosisininhabitantsofthecity

ofJataí,Goiás,intheCentral-WestregionofBrazilfromApriltoAugust2012.Atotalof

529serumsampleswereanalyzedbyenzyme-linkedimmunosorbentassayfordetecting

IgGantibodiesagainstT.soliumlarvae,andWesternblottingwasusedforconﬁrmingthe

diagnosisthroughtherecognitionofatleasttwospeciﬁcpeptidesfromtheirserum

antibod-ies.The351/529(66.3%)reactivesampleswereanalyzedbyenzyme-linkedimmunosorbent

assayandWesternblottingconﬁrmedthediagnosisin73samplesthatrecognizedatleast

twoofthefollowingpeptidesspeciﬁcIgGantibodiesforcysticercosis:18,24,28–32,39–42,

47–52,64–68,and70kDa.Theseroprevalenceofcysticercosiswas13.8%(95%CI5.9–21.7),

demonstratingthatthestudiedareaisendemictothisdisease.

openaccessarticleundertheCCBY-NC-NDlicense(http://creativecommons.org/licenses/

by-nc-nd/4.0/).

The Taenia solium cysticercosis affects millions of people

worldwide and is considered a public health problem,

especiallyin developingcountries.1 _The _T._solium _life_cycle

involves both human and pig hosts. Cysticercosis occurs

afteringestionofeggsofT.soliumbyfecalcontaminationof

humanhost. Larvalinfectioncan bediagnosedindifferent

tissues,such asskeletal muscle,subcutaneous andcentral

∗ _{Corresponding}_author.

E-mailaddress:solbarcelos1@hotmail.com(I.S.C.Barcelos).

nervous systemtissue.2_Human_{neurocysticercosis} _(NCC)_is

the mostsevere form,because about ﬁve million casesof

epilepsy worldwideare related to this parasitosis.2 NCCis

aneglectedtropicaldiseaserecognizedbytheWorldHealth

Organizationsince2010;itisendemicinseveralcountriesin

LatinAmerica,sub-SaharanAfricaandAsia,includingIndia,

most of Southeast Asia, and China.3 _Agapejev4 _conducted

a critical analysis ofBrazilian literature (1915–2002) which

showedanNCCfrequencyof1.5%inautopsiesand3.0%in

clinicalstudies.Inseroepidemiologicalstudiesthepositivity

https://doi.org/10.1016/j.bjid.2018.01.002

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brazj infect dis.2018;22(2):146–149

147

ofspeciﬁcreactions was2.3%.In Brazil,between2000and

2011,a totalof1829deathsrelated toNCC were recorded,

which represented 0.015% of all deaths. High-risk clusters

ofNCC-relatedmortalitywere evidenced in endemicareas

in the Southeast, South and Central-West regions.5 _The

NorthandNortheastregionsusuallyhavenospeciﬁccontrol

programs; people from these areas have limited access to

healthservices,andthelivingconditionsarepoor.4,5

Thiszoonosis is directly related to risk factors such as

disorderedurbanization,precariousconditionsofbasicand

hygienicsanitation,closecontactwithpigs,andpoorhealth

surveillancepresentintheregionswherethehumaninfection

isendemic,aswellasculturalbehaviorandinternalmigration

ofpeoplefromruralareastourbancenters.5,6_Swine

cysticer-cosisisnotwellrecordedinBrazilduetoclandestineslaughter

ofpigs,alongwithlimitedinspectionandsanitarycontrol.7

NotiﬁcationofdiagnosedcasesofNCCisnotconsidered

com-pulsoryinBrazil,althoughitisrecommendedbytheMinistry

ofHealth.OnlytheSouthandSoutheastregionsofthecountry

usethisstrategytoinvestigateinfectioncasesforprevention

programswith increasedcoverage and accessto diagnosis

throughneuroimagingtests,alongwithclinicalandsurgical

treatment.4_Prevention_and_control_of_T._solium_transmission

shouldbeapriorityasinterventioncanreducethe

substan-tialhealthcareandeconomicburdensduetobothNCCand

taeniasis.8

Theobjective ofthis study was todetermine the

sero-prevalenceofhuman cysticercosis inserum samples from

inhabitants ofthe municipalityofJataí,Goiás state, inthe

Central-WestregionofBrazil.

ThestudywascarriedinthemunicipalityofJataí(17◦52

53S,51◦4252W),330kmfromthecapitalofGoiânia,located

inthesouthwestofthestateofGoiás,Brazil.The

municipal-ityarea of7174km2 _has_an_estimated_population_of_97,077

inhabitants.9_This_study_was_approved_by_the_Research_Ethics

CommitteeoftheFederalUniversityofGoiás(UFG),under

pro-tocolnumber230/2011.

A total of 529 individuals (301 females and 228 males)

participatedinthestudy.All individualsolder than18 and

representatives inthe caseofchildren(≥4years old), who

agreedtoparticipateinthestudy,signedtheinformed

con-sent.Theywereselectedvoluntarilyfromthewaitingroomsof

threeclinicalanalysislaboratories(twoprivateandonepublic)

inthecityunderstudy.Theparticipantsconsentedtodonate

2mLofserumremainingaftertheserologicaland/or

biochem-icalteststhatwererequestedbythedoctorswhotookcareof

them.TheserumsamplesweredonatedfromApriltoAugust

2012,transportedunderrefrigerationandstoredat−20◦_C_at

theImmunologyLaboratoryofUFGRegionalJataí.

Atotalsalineextractwith50metacestodesofT.soliumwas

preparedandtheobtainedextractwasdividedintoaliquots

of200␮L, identiﬁed and stored at−20◦_C _until _processing.

SerumsamplesweresubmittedtotheELISAtestinaccordance

withBarcelosetal.10_Some₂₀₀_␮g_of_the_total_saline_extract

proteinwasappliedby10×8cmgelandsubjectedto12%

poly-acrylamidegelelectrophoresis(SDS-PAGE)forseparationof

thepeptidespresentintheantigenicextractandsubsequent

transferto0.45␮mporesizenitrocellulosemembranes.After

transfer,the nitrocellulosemembraneswere cutinto 3mm

strips.Theantigenicpeptides recognizedbyIgG antibodies

Table1–FrequencyofpeptiderecognitionbyWestern

blottingtestthroughIgGantibodiesagainstthetotal

salineextractofTaeniasoliummetacestodesin314

serumsamplesfrominhabitantsofthemunicipalityof

Jataí,Goiás,fromApriltoAugust2012.

Typeand numberof peptides recognized Numberof reagent samples Recognition frequency(%) Only non-specific 85 27.1 Onespecific 156 49.7 Twospecific 62 19.7 Threespecific 5 1.6 Fourspecific 6 1.9 Total 314 100.0

Table2–Distributionofthe73individualreagentsinthe

ELISAandWesternblottingtestsinrelationtoagegroup

andsex.

Agegroup(years)a _Sex

Female Male 11–20 5 4 21–30 4 6 31–40 7 6 41–50 13a ₉ 51–60 6 2 61–70 3 3 71–80 0 2 81–90 0 3 Total 38 35 a _p₌_0.0182_by_two-way_ANOVA.

presentinthereagentserumsamplesintheELISAwere

ana-lyzedbyWesternblotting(WB).10_The_criterion_for_positivity_in

WBadoptedwastherecognitionofatleasttwoofthepeptides,

18,24,28–32,39–42,47–52,64–68,and70kDa,inWBspeciﬁc

forcysticercosis.10_The_results_obtained_in_the_ELISA_and_WB

wereanalyzedusingtwo-wayANOVAandFisher’sexacttests

withp≤0.05inthesoftwareGraphPadPrismversion5.0.The

total salineextractofT. soliummetacestodeswasprepared

foruseinbothtestsandpresentedaproteinconcentration

of3000␮g/mL.

All529serumsampleswereanalyzedbyELISAtodetectIgG

antibodiesagainstcysticercosisand351(66.3%)sampleswere

reactive.AllreagentserumsamplesanalyzedbyELISAwere

submittedtoWB,resultingin37samples(10.5%)withno

pep-tiderecognitionand314(89.5%)werereagents.Seventy-three

ofthe314sampleswithtwoormorepeptideswereconsidered

reagentaccordingtothediagnosticcriterionadoptedinWB

forcysticercosis10₍_Table₁_)._Therefore,_13.8%_(95%_CI_5.9–21.7)

ofthe 529 serum samplesanalyzed showed T.solium

anti-metacestodeantibodiesinELISAandrecognitionofspeciﬁc

peptidesforNCCinWB.Ofthe73participantsthatshowed

speciﬁc bandrecognitionin WB,38 (52%)were female and

35 (48%) male. Positivity amongfemales aged 41–50 years

wassigniﬁcantlyhigher(p=0.0182)(Table2).Thespeciﬁc

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braz j infect dis.2018;22(2):146–149 0 10 20 30 40 50 60 70 18 24 28-32 39-42 47-52 64-68 70 Fre qu e n cy ( % ) Peptides (kDa) ∗ ∗ ∗

Fig.1–FrequencyofWesternblottingrecognitionof

speciﬁcpeptides(kDa)forhumancysticercosisthroughIgG

antibodiespresentin73serumsamplesfrominhabitants

ofthemunicipalityofJataí,Goiás,Brazil(ApriltoAugust

2012).*_p_<_0.05,_Fisher’s_exact_test.

antibodies were 64–68 (45.2%), 47–52 (53.4%), and 70kDa

(65.7%)peptides(p=0.01)(Fig.1).

Thisstudydemonstrated aseroprevalenceof13.8%(95%

CI 5.9–21.7) of human cysticercosis in the municipality of

Jataí,inthesouthwestofGoiásstate.Thisprevalence

char-acterizesthe study area asbeingendemic forcysticercosis

accordingtothePanAmericanHealthOrganization.11_There

wasahigherseroprevalenceamongfemalesaged41–50,but

itisknownthattheparasite’spredilectionisnotcorrelated

withsexperse, butoftenwith foodhabit orthe regionof

study,amongotherfactors.4,5_The_NCC-related_mortality_rates

inBrazil(2000–2011)showed1.59deaths/1,000,000inhabitants

forGoiás(Central-Westregion).5 _According_to_the _Brazilian

InstituteofGeographyand Statistics(IBGE),in2008,inthe

stateofGoiás,only33% ofthe householdswere servedby

ageneralsewagesystem.9_In_Goiânia,_out_of₉₂_swine_blood

samplessubmittedtotheindirectELISAtechnique,7.6%were

positiveforcysticercosis.12InthemunicipalityofJataíitiseasy

toﬁndhouseholdswherepigsareraisedintheperidomicileor

onfarms,whichisassociatedwithotherriskfactors,suchas

precarioussanitaryandlowsocioeconomicconditions,which

may explain the high seroprevalence of cysticercosis. Our

seroprevalenceofcysticercosisishigherthanthe11.3%found

inanotherstudyinthemunicipalityofCatalão,Goiás

includ-ing354serumsamplesfromthepopulationanalyzedbyELISA

and WB; the serologyreactivity was associated withareas

withoutasewagecollectionnetwork.13

Cysticercosis is endemic in Brazil according to several

studies.4,5 _In_the_northeastern_region,_a_cohort_study_carried

outinthemunicipalityofJoãodaCosta,inPiauístate,showed

aprevalenceof13.6% forcysticercosis byELISA intheﬁrst

stage ofthe study in169 individuals with a conﬁrmed or

suspectedhistoryofcysticercosisinfection.Subsequent

anal-ysisofthese92 reagentsamplesinthe ﬁrststagerevealed

that 24% presentedreagent samplesinthe ELISA testand

29%inWB.14 _In_the_city _of_Mulungu_do_Morro, _in_the _state

ofBahia,theprevalenceofhumancysticercosiswas1.6%in

694bloodsamplescollectedfromthegeneralpopulation;the

areaisconsideredanareaofcasualincidence,althoughithas

characteristicsthatindicatepoorhealthandsocioeconomic

proﬁles.15 _In _the _Southeast _region, _speciﬁc _IgG _antibodies

againstcysticercosisweredetectedin13.5%,5.0%,4.8%,and

4.7% ofserum samplesfrom blood donors inthe citiesof

Araguari,Tupaciguara,MonteAlegredeMinasandUberlândia,

respectively,inMinasGeraisstate.16_In_a_study_carried_out_in

1863inhabitantsoftheruralareaofthemunicipalityofCássia

dosCoqueiros,inSãoPaulostate,usingspeciﬁcELISAandWB

testsandconsideringtheuseofWBasconﬁrmatoryduetoits

highspeciﬁcity,theanti-cysticercusserumprevalenceinthis

populationwas2.1%.17_In_another_study,_conducted_in_a_rural

settlementinSãoPaulo,5.7%ofIgGantibodieswerefoundin

194serumsamplesbyELISAtestandWBconﬁrmedby18kDa

and 14kDa proteins puriﬁedfrom vesicularﬂuid

metaces-todesofT.solium.18_In_the_South_region,_a_study_demonstrated

thatcysticercosisseroprevalencewas3.4%inLagescity,Santa

Catarinastate,inwhich850bloodsampleswerecollectedon

ﬁlterpaperusing anantigenicextract ofvesicularﬂuidfor

testingbyWB.19

Thehighfrequencyofinfectionbyotherparasitesinthe

Brazilianpopulationmayinterferewiththeperformanceof

the ELISA testwherea totalparasite extractisused, asin

thepresentstudy,whoseresultsshowedapositivityof66.3%;

thetotalantigenicextractfromcysticercusofT.solium

corre-lateswithahighcross-reactivitytendencyinserumsamples

fromindividuals withtaeniasisintheELISAtest.10_The_use

ofWBasaconﬁrmatorytestfortheimmunological

diagno-sisofcysticercosisisjustiﬁedbecauseofthehighspeciﬁcity

ofthepeptidesrecognizedbyIgGantibodieswhenusingtotal

salineextractofT.soliummetacestodes.10,13,18_Elevated_levels

ofIgGpredominateinserumsamplesfromNCCpatientsat

differentevolutionarystagesofthediseaseandconstitutean

immunologicalmarkerofchronicexposure.2

ThedetectionofIgGantibodiesremainsareliablemethod

incysticercosisseroepidemiologicalstudiesasitis

noninva-sive and demonstrates previous exposureofindividuals to

theseparasites.Theresultssuggestthatthemunicipalityof

Jataíisendemicforcysticercosisandthereisaneedforhealth

managerstoadoptmeasurestocontrolandpreventthis

par-asitosis.

Funding

Fundac¸ãodeAmparoàPesquisadoEstadodeGoiás(FAPEG).

Conﬂicts

of

interest

Theauthorsdeclarenoconﬂictsofinterest.

Acknowledgements

Wegiveourdeepestthankstotheparticipantsofthestudy.

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